109 results on '"Yasuhiro Yasutomi"'
Search Results
2. Highly susceptible SARS-CoV-2 model in CAG promoter–driven hACE2-transgenic mice
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Masamitsu N. Asaka, Daichi Utsumi, Haruhiko Kamada, Satoshi Nagata, Yutaka Nakachi, Tomokazu Yamaguchi, Yoshihiro Kawaoka, Keiji Kuba, and Yasuhiro Yasutomi
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COVID-19 ,Medicine - Abstract
COVID-19, caused by SARS-CoV-2, has spread worldwide with dire consequences. To urgently investigate the pathogenicity of COVID-19 and develop vaccines and therapeutics, animal models that are highly susceptible to SARS-CoV-2 infection are needed. In the present study, we established an animal model highly susceptible to SARS-CoV-2 via the intratracheal tract infection in CAG promoter–driven human angiotensin-converting enzyme 2–transgenic (CAG-hACE2) mice. The CAG-hACE2 mice showed several severe symptoms of SARS-CoV-2 infection, with definitive weight loss and subsequent death. Acute lung injury with elevated cytokine and chemokine levels was observed at an early stage of infection in CAG-hACE2 mice infected with SARS-CoV-2. Analysis of the hACE2 gene in CAG-hACE2 mice revealed that more than 15 copies of hACE2 genes were integrated in tandem into the mouse genome, supporting the high susceptibility to SARS-CoV-2. In the developed model, immunization with viral antigen or injection of plasma from immunized mice prevented body weight loss and lethality due to infection with SARS-CoV-2. These results indicate that a highly susceptible model of SARS-CoV-2 infection in CAG-hACE2 mice via the intratracheal tract is suitable for evaluating vaccines and therapeutic medicines.
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- 2021
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3. Diabetes mellitus accelerates Aβ pathology in brain accompanied by enhanced GAβ generation in nonhuman primates.
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Sachi Okabayashi, Nobuhiro Shimozawa, Yasuhiro Yasutomi, Katsuhiko Yanagisawa, and Nobuyuki Kimura
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Medicine ,Science - Abstract
Growing evidence suggests that diabetes mellitus (DM) is one of the strongest risk factors for developing Alzheimer's disease (AD). However, it remains unclear why DM accelerates AD pathology. In cynomolgus monkeys older than 25 years, senile plaques (SPs) are spontaneously and consistently observed in their brains, and neurofibrillary tangles are present at 32 years of age and older. In laboratory-housed monkeys, obesity is occasionally observed and frequently leads to development of type 2 DM. In the present study, we performed histopathological and biochemical analyses of brain tissue in cynomolgus monkeys with type 2 DM to clarify the relationship between DM and AD pathology. Here, we provide the evidence that DM accelerates Aβ pathology in vivo in nonhuman primates who had not undergone any genetic manipulation. In DM-affected monkey brains, SPs were observed in frontal and temporal lobe cortices, even in monkeys younger than 20 years. Biochemical analyses of brain revealed that the amount of GM1-ganglioside-bound Aβ (GAβ)--the endogenous seed for Aβ fibril formation in the brain--was clearly elevated in DM-affected monkeys. Furthermore, the level of Rab GTPases was also significantly increased in the brains of adult monkeys with DM, almost to the same levels as in aged monkeys. Intraneuronal accumulation of enlarged endosomes was also observed in DM-affected monkeys, suggesting that exacerbated endocytic disturbance may underlie the acceleration of Aβ pathology due to DM.
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- 2015
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4. Effects of mycobacteria major secretion protein, Ag85B, on allergic inflammation in the lung.
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Yusuke Tsujimura, Hiroyasu Inada, Misao Yoneda, Tomoyuki Fujita, Kazuhiro Matsuo, and Yasuhiro Yasutomi
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Medicine ,Science - Abstract
Many epidemiological studies have suggested that the recent increase in prevalence and severity of allergic diseases such as asthma is inversely correlated with Mycobacterium bovis bacillus Calmette Guerin (BCG) vaccination. However, the underlying mechanisms by which mycobacterial components suppress allergic diseases are not yet fully understood. Here we showed the inhibitory mechanisms for development of allergic airway inflammation by using highly purified recombinant Ag85B (rAg85B), which is one of the major protein antigens secreted from M. tuberculosis. Ag85B is thought to be a single immunogenic protein that can elicit a strong Th1-type immune response in hosts infected with mycobacteria, including individuals vaccinated with BCG. Administration of rAg85B showed a strong inhibitory effect on the development of allergic airway inflammation with induction of Th1-response and IL-17and IL-22 production. Both cytokines induced by rAg85B were involved in the induction of Th17-related cytokine-production innate immune cells in the lung. Administration of neutralizing antibodies to IL-17 or IL-22 in rAg85B-treated mice revealed that IL-17 induced the infiltration of neutrophils in BAL fluid and that allergen-induced bronchial eosinophilia was inhibited by IL-22. Furthermore, enhancement of the expression of genes associated with tissue homeostasis and wound healing was observed in bronchial tissues after rAg85B administration in a Th17-related cytokine dependent manner. The results of this study provide evidence for the potential usefulness of rAg85B as a novel approach for anti-allergic effect and tissue repair other than the role as a conventional TB vaccine.
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- 2014
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5. Intranasally administered antigen 85B gene vaccine in non-replicating human Parainfluenza type 2 virus vector ameliorates mouse atopic dermatitis.
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Hiroshi Kitagawa, Mitsuo Kawano, Keiichi Yamanaka, Masato Kakeda, Kenshiro Tsuda, Hiroyasu Inada, Misao Yoneda, Tadashi Sakaguchi, Akina Nigi, Koumei Nishimura, Hiroshi Komada, Masato Tsurudome, Yasuhiro Yasutomi, Tetsuya Nosaka, and Hitoshi Mizutani
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Medicine ,Science - Abstract
Atopic dermatitis (AD) is a refractory and recurrent inflammatory skin disease. Various factors including heredity, environmental agent, innate and acquired immunity, and skin barrier function participate in the pathogenesis of AD. T -helper (Th) 2-dominant immunological milieu has been suggested in the acute phase of AD. Antigen 85B (Ag85B) is a 30-kDa secretory protein well conserved in Mycobacterium species. Ag85B has strong Th1-type cytokine inducing activity, and is expected to ameliorate Th2 condition in allergic disease. To perform Ag85B function in vivo, effective and less invasive vaccination method is required. Recently, we have established a novel functional virus vector; recombinant human parainfluenza type 2 virus vector (rhPIV2): highly expressive, replication-deficient, and very low-pathogenic vector. In this study, we investigated the efficacy of rhPIV2 engineered to express Ag85B (rhPIV2/Ag85B) in a mouse AD model induced by repeated oxazolone (OX) challenge. Ear swelling, dermal cell infiltrations and serum IgE level were significantly suppressed in the rhPIV2/Ag85B treated mouse group accompanied with elevated IFN-γ and IL-10 mRNA expressions, and suppressed IL-4, TNF-α and MIP-2 mRNA expressions. The treated mice showed no clinical symptom of croup or systemic adverse reactions. The respiratory tract epithelium captured rhPIV2 effectively without remarkable cytotoxic effects. These results suggested that rhPIV2/Ag85B might be a potent therapeutic tool to control allergic disorders.
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- 2013
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6. Mycobacterium bovis Bacille Calmette-Guérin as a Vaccine Vector for Global Infectious Disease Control
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Kazuhiro Matsuo and Yasuhiro Yasutomi
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Medicine - Abstract
Mycobacterium bovis bacille Calmette-Guérin (BCG) is the only available vaccine for tuberculosis (TB). Although this vaccine is effective in controlling infantile TB, BCG-induced protective effects against pulmonary diseases in adults have not been clearly demonstrated. Recombinant BCG (rBCG) technology has been extensively applied to obtain more potent immunogenicity of this vaccine, and several candidate TB vaccines have currently reached human clinical trials. On the other hand, recent progress in the improvement of the BCG vector, such as the codon optimization strategy and combination with viral vector boost, allows us to utilize this bacterium in HIV vaccine development. In this paper, we review recent progress in rBCG-based vaccine studies that may have implications in the development of novel vaccines for controlling global infectious diseases in the near future.
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- 2011
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7. In vivo safety and persistence of endoribonuclease gene-transduced CD4+ T cells in cynomolgus macaques for HIV-1 gene therapy model.
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Hideto Chono, Naoki Saito, Hiroshi Tsuda, Hiroaki Shibata, Naohide Ageyama, Keiji Terao, Yasuhiro Yasutomi, Junichi Mineno, and Ikunoshin Kato
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Medicine ,Science - Abstract
BACKGROUND: MazF is an endoribonuclease encoded by Escherichia coli that specifically cleaves the ACA sequence of mRNA. In our previous report, conditional expression of MazF in the HIV-1 LTR rendered CD4+ T lymphocytes resistant to HIV-1 replication. In this study, we examined the in vivo safety and persistence of MazF-transduced cynomolgus macaque CD4+ T cells infused into autologous monkeys. METHODOLOGY/PRINCIPAL FINDINGS: The in vivo persistence of the gene-modified CD4+ T cells in the peripheral blood was monitored for more than half a year using quantitative real-time PCR and flow cytometry, followed by experimental autopsy in order to examine the safety and distribution pattern of the infused cells in several organs. Although the levels of the MazF-transduced CD4+ T cells gradually decreased in the peripheral blood, they were clearly detected throughout the experimental period. Moreover, the infused cells were detected in the distal lymphoid tissues, such as several lymph nodes and the spleen. Histopathological analyses of tissues revealed that there were no lesions related to the infused gene modified cells. Antibodies against MazF were not detected. These data suggest the safety and the low immunogenicity of MazF-transduced CD4+ T cells. Finally, gene modified cells harvested from the monkey more than half a year post-infusion suppressed the replication of SHIV 89.6P. CONCLUSIONS/SIGNIFICANCE: The long-term persistence, safety and continuous HIV replication resistance of the mazF gene-modified CD4+ T cells in the non-human primate model suggests that autologous transplantation of mazF gene-modified cells is an attractive strategy for HIV gene therapy.
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- 2011
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8. Ultrasound-guided, Transabdominal, Intrauterine Artificial Insemination for Cynomolgus Macaques (Macaca fascicularis) Based on Estimated Timing of Ovulation
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Shunya Nakayama, Naohide Ageyama, Hiroshi Koie, Nobuhiro Shimozawa, and Yasuhiro Yasutomi
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040301 veterinary sciences ,Offspring ,media_common.quotation_subject ,medicine.medical_treatment ,Uterus ,Physiology ,Insemination ,Macaque ,0403 veterinary science ,03 medical and health sciences ,biology.animal ,medicine ,Ovulation ,Cervical canal ,030304 developmental biology ,media_common ,0303 health sciences ,Pregnancy ,biology ,business.industry ,Artificial insemination ,04 agricultural and veterinary sciences ,medicine.disease ,medicine.anatomical_structure ,Animal Science and Zoology ,business - Abstract
Intrauterine sperm injection for artificial insemination is difficult in cynomolgus macaques (Macaca fascicularis) and rhesus macaques (M. mulatta) due to the complex structure of the cervical canal, which differs from that of humans. Despite the availability of several artificial insemination methods for macaques, pregnancy rates are inconsistent, and details regarding ovulation are unclear, thus warranting more effective methods. Therefore, we developed an effective, ultrasound-guided, transabdominal intrauterine artificial insemination method for cynomolgus macaques that involves timing sperm injection to coincide with the periovulation phase estimated according to rapid hormone measurement. We performed our intrauterine artificial insemination on 6 female macaques; 4 of the 5 animals that were predicted to have ovulated soon after insemination became pregnant, whereas the 1 macaque that was predicted not to have ovulated did not. Furthermore, we saw no evidence of injury, such as a conspicuous needle hole or bleeding on the surface of or inside the uterus, nor did our method result in any abnormalities in the mothers or their offspring. Thus, our ultrasound-guided, transabdominal, intrauterine artificial insemination method is rapid, safe, and effective in cynomolgus macaques.
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- 2021
9. Usefulness of cardiac hormones for evaluating valvular disease in cynomolgus monkeys (Macaca fascicularis)
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Yasuyo Ito-Fujishiro, Kiichi Kanayama, Yoshiko Munesue, Naohide Ageyama, Hiroshi Koie, Yasuhiro Yasutomi, Tadashi Sankai, Shunya Nakayama, and Chungyu Pai
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medicine.medical_specialty ,Heart disease ,040301 veterinary sciences ,Disease ,Cardiac hormones ,0403 veterinary science ,03 medical and health sciences ,Atrial natriuretic peptide ,Valvular disease ,Internal medicine ,medicine ,cardiovascular diseases ,030304 developmental biology ,0303 health sciences ,General Veterinary ,medicine.diagnostic_test ,business.industry ,04 agricultural and veterinary sciences ,medicine.disease ,Brain natriuretic peptide ,Nonhuman primate ,cardiovascular system ,Cardiology ,business ,Electrocardiography ,hormones, hormone substitutes, and hormone antagonists ,circulatory and respiratory physiology - Abstract
Nonhuman primates are commonly used as experimental animals due to their biological resemblance to humans. In patients with cardiac disease, the levels of atrial natriuretic peptide (ANP) and brain natriuretic peptide (BNP) tend to increase in response to cardiac damage, and they are thus used as indicators for the diagnosis of human heart failure. However, no reference values for ANP and BNP have been reported for heart disease in nonhuman primates. In this study, we recorded the age, sex, and body weight of 202 cynomolgus monkeys, and performed evaluations to assess the ANP and BNP levels, electrocardiography and echocardiography, and accordingly divided the monkeys into two groups: healthy monkeys and those with spontaneous cardiac disease. Statistical analysis was performed to determine the relationship of ANP and BNP with the factors of age, sex, and body weight. No significant relationship was found between the levels of ANP and BNP and the factors of age, sex, and body weight. However, both the ANP and BNP levels were significantly different between the healthy monkeys and monkeys with valvular disease. Similar to humans, the ANP and BNP levels tended to increase with the progression of cardiac disease in monkeys. Based on these results, we concluded that ANP and BNP are indicators of cardiac disease in nonhuman primates, and that this nonhuman primate cardiac disease model is applicable for cardiology research in humans.
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- 2021
10. Echocardiographic evaluation of cardiac function in cynomolgus monkeys over a wide age range
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Shunya Nakayama, Yasuhiro Yasutomi, Tadashi Sankai, Kiichi Kanayama, Naohide Ageyama, Hiroshi Koie, Yasuyo Ito-Fujishiro, and Chungyu Pai
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Male ,0301 basic medicine ,Cardiac function curve ,Aging ,medicine.medical_specialty ,Cardiac output ,Original ,elderly ,General Biochemistry, Genetics and Molecular Biology ,03 medical and health sciences ,0302 clinical medicine ,cardiovascular disease ,Internal medicine ,medicine ,Animals ,Ejection fraction ,General Veterinary ,medicine.diagnostic_test ,business.industry ,Ketamine hydrochloride ,Heart ,Organ Size ,General Medicine ,Fractional shortening ,Disease Models, Animal ,Macaca fascicularis ,030104 developmental biology ,Blood pressure ,medicine.anatomical_structure ,Echocardiography ,Ventricle ,Cardiology ,Female ,cynomolgus monkey ,Animal Science and Zoology ,cardiac function ,business ,Electrocardiography ,030217 neurology & neurosurgery - Abstract
Various cardiovascular diseases can be detected and diagnosed using echocardiography. The demand for cardiovascular system research using nonhuman primates is increasing, but echocardiographic references for nonhuman primates are limited. This report describes the first comparison of echocardiographic reference values in 247 normal cynomolgus monkeys (135 females, 112 males) over a wide age range. Echocardiography, electrocardiography, blood pressure and chest X-ray images were acquired under immobilization with intramuscular ketamine hydrochloride, then cardiac structure, function, and flow velocity were assessed. Cardiac hormone levels were also tested. We found that cardiac structures positively correlated with weight, that the size of these structures stabilized after reaching maturity and that cardiac output increased according to heart size. In contrast, fractional shortening of the left ventricle, ejection fraction and flow velocity showed no significant correlations with weight or age, and age and E wave correlated negatively. These findings appear sufficiently similar to those in humans to suggest that cynomolgus monkeys can serve as a suitable model of human cardiac disease. Our data should also prove useful for surveying cardiac dysfunction in monkeys.
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- 2020
11. ACE2-like carboxypeptidase B38-CAP protects from SARS-CoV-2-induced lung injury
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Daichi Utsumi, Akari Saku, Yoshihiro Kawaoka, Kwok-Yung Yuen, Chris Chung-Sing Chan, Satoru Motoyama, Saori Takahashi, Jasper Fuk-Woo Chan, Jianbo An, Takafumi Minato, Satoshi Nagata, Vincent Kwok-Man Poon, Masaki Imai, Keiji Kuba, Maki Kiso, Midori Hoshizaki, Atsuhiro Yasuhara, Josef M. Penninger, Tomokazu Yamaguchi, Ken Maeda, Yasuhiro Yasutomi, Yumiko Imai, Haruhiko Kamada, Mayumi Niiyama, Akihiko Uda, Ryota Nukiwa, Masamitsu N Asaka, Satoru Nirasawa, Wataru Kamitani, and Yuji Fujino
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Male ,viruses ,General Physics and Astronomy ,Endogeny ,Carboxypeptidases ,Pharmacology ,Mice ,Cricetinae ,Chlorocebus aethiops ,skin and connective tissue diseases ,Receptor ,Lung ,Multidisciplinary ,biology ,Angiotensin II ,Lung Injury ,respiratory system ,medicine.anatomical_structure ,Spike Glycoprotein, Coronavirus ,Female ,Angiotensin-Converting Enzyme 2 ,hormones, hormone substitutes, and hormone antagonists ,Genetically modified mouse ,Respiratory distress syndrome ,Science ,Acute Lung Injury ,Mice, Transgenic ,Pulmonary Edema ,Lung injury ,Article ,General Biochemistry, Genetics and Molecular Biology ,In vivo ,medicine ,Animals ,Humans ,Vero Cells ,SARS-CoV-2 ,business.industry ,fungi ,COVID-19 ,General Chemistry ,Virus Internalization ,Carboxypeptidase ,COVID-19 Drug Treatment ,respiratory tract diseases ,Disease Models, Animal ,biology.protein ,Peptides ,business - Abstract
Angiotensin-converting enzyme 2 (ACE2) is a receptor for cell entry of SARS-CoV-2, and recombinant soluble ACE2 protein inhibits SARS-CoV-2 infection as a decoy. ACE2 is a carboxypeptidase that degrades angiotensin II, thereby improving the pathologies of cardiovascular disease or acute lung injury. Here we show that B38-CAP, an ACE2-like enzyme, is protective against SARS-CoV-2-induced lung injury. Endogenous ACE2 expression is downregulated in the lungs of SARS-CoV-2-infected hamsters, leading to elevation of angiotensin II levels. Recombinant Spike also downregulates ACE2 expression and worsens the symptoms of acid-induced lung injury. B38-CAP does not neutralize cell entry of SARS-CoV-2. However, B38-CAP treatment improves the pathologies of Spike-augmented acid-induced lung injury. In SARS-CoV-2-infected hamsters or human ACE2 transgenic mice, B38-CAP significantly improves lung edema and pathologies of lung injury. These results provide the first in vivo evidence that increasing ACE2-like enzymatic activity is a potential therapeutic strategy to alleviate lung pathologies in COVID-19 patients., Endogenous ACE2 is a receptor for SARS-CoV-2 and a recombinant soluble ACE2 protein can inhibit SARS-CoV-2 infection acting as a decoy. Here the authors show that B38-CAP, an ACE2-like enzyme but not a decoy for the virus, is protective against SARS-CoV-2-induced lung injury in animal models.
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- 2021
12. COVID-19 cynomolgus macaque model reflecting human COVID-19 pathological conditions
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Satoshi Nagata, Wataru Kamitani, Tomotaka Okamura, Shiori Ueno, Mahoko Higuchi, Yoshiharu Matsuura, Yasuhiro Yasutomi, Yoshihiro Kawaoka, Haruhiko Kamada, Emiko Urano, Chikako Ono, and Mugi Furukawa
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Male ,Chemokine ,nonhuman primate ,Antibodies, Viral ,Microbiology ,elderly ,Immunoglobulin G ,Animals ,Humans ,Medicine ,underlying disease ,Viral shedding ,Lung ,Pathological ,Multidisciplinary ,biology ,SARS-CoV-2 ,business.industry ,Primate Diseases ,COVID-19 ,Biological Sciences ,medicine.disease ,Pathophysiology ,Virus Shedding ,Disease Models, Animal ,Macaca fascicularis ,medicine.anatomical_structure ,Immunology ,biology.protein ,Female ,Tomography, X-Ray Computed ,business ,Pneumonia (non-human) - Abstract
Significance COVID-19 has shown severe pathogenicity in people with underlying diseases and in elderly people. It is necessary to elucidate the pathological conditions in patients with underlying diseases and in elderly patients. The use of appropriate animal models of COVID-19 is required for the development of pharmaceutical products; however, usually healthy young animals are used as experimental animals. Cynomolgus macaques with various clinical conditions and ages were infected with severe acute respiratory syndrome coronavirus 2 and there was a difference in pathological condition between young individuals and old-aged individuals with underlying diseases; therefore, the COVID-19 cynomolgus monkey model reflecting the pathophysiology of humans would be useful for elucidating the pathophysiology and developing therapeutic and prophylactic agents., The pandemic of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is a global threat to human health and life. A useful pathological animal model accurately reflecting human pathology is needed to overcome the COVID-19 crisis. In the present study, COVID-19 cynomolgus monkey models including monkeys with underlying diseases causing severe pathogenicity such as metabolic disease and elderly monkeys were examined. Cynomolgus macaques with various clinical conditions were intranasally and/or intratracheally inoculated with SARS-CoV-2. Infection with SARS-CoV-2 was found in mucosal swab samples, and a higher level and longer period of viral RNA was detected in elderly monkeys than in young monkeys. Pneumonia was confirmed in all of the monkeys by computed tomography images. When monkeys were readministrated SARS-CoV-2 at 56 d or later after initial infection all of the animals showed inflammatory responses without virus detection in swab samples. Surprisingly, in elderly monkeys reinfection showed transient severe pneumonia with increased levels of various serum cytokines and chemokines compared with those in primary infection. The results of this study indicated that the COVID-19 cynomolgus monkey model reflects the pathophysiology of humans and would be useful for elucidating the pathophysiology and developing therapeutic agents and vaccines.
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- 2021
13. Highly susceptible SARS-CoV-2 model in CAG promoter-driven hACE2-transgenic mice
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Yutaka Nakachi, Satoshi Nagata, Keiji Kuba, Masamitsu N Asaka, Haruhiko Kamada, Yasuhiro Yasutomi, Tomokazu Yamaguchi, Yoshihiro Kawaoka, and Daichi Utsumi
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Genetically modified mouse ,Male ,Chemokine ,medicine.medical_treatment ,viruses ,Mice, Transgenic ,Lung injury ,Biology ,Mouse models ,Genome ,Weight loss ,medicine ,Animals ,skin and connective tissue diseases ,Promoter Regions, Genetic ,Gene ,SARS-CoV-2 ,fungi ,COVID-19 ,General Medicine ,Virology ,respiratory tract diseases ,body regions ,Mice, Inbred C57BL ,Disease Models, Animal ,Cytokine ,Immunization ,Resource and Technical Advance ,biology.protein ,Female ,Angiotensin-Converting Enzyme 2 ,Disease Susceptibility ,medicine.symptom - Abstract
COVID-19, caused by SARS-CoV-2, has spread worldwide with dire consequences. To urgently investigate the pathogenicity of COVID-19 and develop vaccines and therapeutics, animal models that are highly susceptible to SARS-CoV-2 infection are needed. In the present study, we established an animal model highly susceptible to SARS-CoV-2 via the intratracheal tract infection in CAG promoter-driven human angiotensin-converting enzyme 2-transgenic (CAG-hACE2) mice. The CAG-hACE2 mice showed several severe symptoms of SARS-CoV-2 infection, with definitive weight loss and subsequent death. Acute lung injury with elevated cytokine and chemokine levels was observed at an early stage of infection in CAG-hACE2 mice infected with SARS-CoV-2. Analysis of the hACE2 gene in CAG-hACE2 mice revealed that more than 15 copies of hACE2 genes were integrated in tandem into the mouse genome, supporting the high susceptibility to SARS-CoV-2. In the developed model, immunization with viral antigen or injection of plasma from immunized mice prevented body weight loss and lethality due to infection with SARS-CoV-2. These results indicate that a highly susceptible model of SARS-CoV-2 infection in CAG-hACE2 mice via the intratracheal tract is suitable for evaluating vaccines and therapeutic medicines.
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- 2021
14. Development and Characterization of a Highly Sensitive NanoLuciferase-Based Immunoprecipitation System for the Detection of Anti-Influenza Virus HA Antibodies
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Fumihiko Yasui, Kenzaburo Yamaji, Yasushi Itoh, Daisuke Yamane, Michinori Kohara, Tomoko Honda, Tsubasa Munakata, Sumiko Gomi, Kazumasa Ogasawara, Takuya Yamamoto, Takahiro Sanada, Kyoko Tsukiyama-Kohara, and Yasuhiro Yasutomi
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0301 basic medicine ,medicine.drug_class ,030106 microbiology ,Hemagglutinin (influenza) ,Hemagglutinin Glycoproteins, Influenza Virus ,Monoclonal antibody ,Antibodies, Viral ,Sensitivity and Specificity ,Microbiology ,Epitope ,Virus ,03 medical and health sciences ,Epitopes ,Mice ,Antigen ,Orthomyxoviridae Infections ,broad dynamic range ,medicine ,Animals ,Immunoprecipitation ,luciferase immunoprecipitation system (LIPS) ,influenza virus hemagglutinin (HA) protein ,Luciferases ,Molecular Biology ,NanoLuciferase (NLuc) ,mouse ,Mice, Inbred BALB C ,Hemagglutination assay ,biology ,Chemistry ,Tupaiidae ,Antibodies, Monoclonal ,Hemagglutination Inhibition Tests ,Virology ,Fusion protein ,Antibodies, Neutralizing ,QR1-502 ,Macaca fascicularis ,030104 developmental biology ,trimeric complex ,Influenza Vaccines ,biology.protein ,Female ,Antibody ,cynomolgus macaque ,tree shrew ,Research Article - Abstract
Influenza virus HA-specific antibodies can be detected via the hemagglutination inhibition (HI) assay, the neutralization (NT) assay, and the enzyme-linked immunosorbent assay (ELISA). However, these assays have some drawbacks, including narrow dynamic range and the requirement for large amounts of sera., Antibody detection is crucial for monitoring host immune responses to specific pathogen antigens (Ags) and evaluating vaccine efficacies. The luciferase immunoprecipitation system (LIPS) was developed for sensitive detection of Ag-specific antibodies in sera from various species. In this study, we describe NanoLIPS, an improved LIPS assay based on NanoLuciferase (NLuc), and employ the assay for monitoring antibody responses following influenza virus infection or vaccination. We generated recombinant influenza virus hemagglutinin (HA) proteins tagged with N-terminal (N-NLuc-HA) or C-terminal (C-NLuc-HA) NLuc reporters. NLuc-HA yielded an at least 20-fold higher signal-to-noise ratio than did a LIPS assay employing a recombinant HA-Gaussia princeps luciferase (GLuc) fusion protein. NanoLIPS-based detection of anti-HA antibodies yielded highly reproducible results with a broad dynamic range. The levels of antibodies against C-NLuc-HA generated by mice vaccinated with recombinant vaccinia virus DIs strain expressing an influenza virus HA protein (rDIs-HA) was significantly correlated with the protective effect elicited by the rDIs-HA vaccine. C-NLuc-HA underwent glycosylation with native conformations and assembly to form a trimeric structure and was detected by monoclonal antibodies that detect conformational epitopes present on the globular head or stalk domain of HA. Therefore, NanoLIPS is applicable for evaluating vaccine efficacy. We also showed that C-NLuc-HA is applicable for detection of HA-specific antibodies in sera from various experimental species, including mouse, cynomolgus macaque, and tree shrew. Thus, NanoLIPS-based detection of HA offers a simple and high-sensitivity method that detects native conformational epitopes and can be used in various experimental animal models. IMPORTANCE Influenza virus HA-specific antibodies can be detected via the hemagglutination inhibition (HI) assay, the neutralization (NT) assay, and the enzyme-linked immunosorbent assay (ELISA). However, these assays have some drawbacks, including narrow dynamic range and the requirement for large amounts of sera. As an alternative to an ELISA-based method, luciferase immunoprecipitation system (LIPS) was developed. We focused on NanoLuciferase (NLuc), which has a small size, higher intensity, and longer stability. In this study, we developed a technically feasible and highly sensitive method for detecting influenza virus-specific antibodies using a NLuc-tagged recombinant HA protein produced in mammalian cells. HA with a C-terminal NLuc extension (C-NLuc-HA) was glycosylated and formed trimeric complexes when expressed in mammalian cells. Furthermore, C-NLuc-HA was recognized not only by monoclonal antibodies that bind to the globular head domain but also by those that bind to the stalk domain. We also demonstrated that the data obtained by this assay correlate with the protection of an experimental vaccine in animal models.
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- 2021
15. Avian H5N1 influenza virus infection causes severe pneumonia in the Northern tree shrew (Tupaia belangeri)
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Kyoko Tsukiyama-Kohara, Takahiro Sanada, Yasuhiro Yasutomi, Michinori Kohara, Mohammad Enamul Hoque Kayesh, Fumihiko Yasui, Yumiko Shiogama, Jun-ichiro Takano, and Tomoko Honda
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viruses ,Tupaia ,Pneumonia, Viral ,Influenza A Virus, H7N9 Subtype ,medicine.disease_cause ,Virus ,Proinflammatory cytokine ,Tree shrew ,03 medical and health sciences ,Tupaia belangeri ,Orthomyxoviridae Infections ,Virology ,medicine ,Animals ,Lung ,Infectious virus ,030304 developmental biology ,0303 health sciences ,Influenza A Virus, H5N1 Subtype ,biology ,030302 biochemistry & molecular biology ,virus diseases ,biology.organism_classification ,medicine.disease ,Influenza A virus subtype H5N1 ,Pneumonia (non-human) - Abstract
Avian-origin influenza viruses like H5N1 and H7N9 often cause severe symptoms with high mortality in humans. Animal models are useful for clarification of the mechanisms of pathogenicity of these infections. In this study, to expand the potential utility of the Northern tree shrew (Tupaia belangeri) for influenza virus infection, we assessed the pathogenicity of H5N1 and H7N9 avian influenza viruses in tupaia. Infectious virus was detected continuously from nasal, oral, tracheal, and conjunctival swab samples in the animals infected with these viruses. H5N1 influenza virus infection of tupaia caused severe diffuse pneumonia with fever and weight loss. In contrast, H7N9 influenza virus infection caused focal pneumonia. The severity of pneumonia was correlated with proinflammatory cytokine transcript levels. These results indicated that tupaia can be another suitable animal model for avian influenza virus research.
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- 2019
16. Elevated Membrane Cholesterol Disrupts Lysosomal Degradation to Induce β-Amyloid Accumulation
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Nobuyuki Kimura, Nobuhiro Shimozawa, Naoya Ueda, Yasuhiro Yasutomi, Keiko Suzuki, and Shingo Takeuchi
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0301 basic medicine ,Pathology ,medicine.medical_specialty ,endocrine system diseases ,Membrane cholesterol ,Chemistry ,Cholesterol ,Endocytic cycle ,nutritional and metabolic diseases ,Type 2 Diabetes Mellitus ,Metabolism ,medicine.disease ,Pathology and Forensic Medicine ,Pathogenesis ,03 medical and health sciences ,chemistry.chemical_compound ,030104 developmental biology ,0302 clinical medicine ,β amyloid ,medicine ,Alzheimer's disease ,030217 neurology & neurosurgery - Abstract
The endocytic membrane trafficking system is altered in the brains of early-stage Alzheimer disease (AD) patients, and endocytic disturbance affects the metabolism of β-amyloid (Aβ) protein, a key molecule in AD pathogenesis. It is widely accepted that type 2 diabetes mellitus (T2DM) is one of the strongest risk factors for development of AD. Supporting this link, experimentally induced T2DM enhances AD pathology in various animal models. Spontaneous T2DM also enhances Aβ pathology with severe endocytic pathology, even in nonhuman primate brains. However, it remains unclear how T2DM accelerates Aβ pathology. Herein, we demonstrate that cholesterol metabolism–related protein levels are increased and that membrane cholesterol level is elevated in spontaneous T2DM-affected cynomolgus monkey brains. Moreover, in vitro studies that manipulate cellular cholesterol reveal that elevated membrane cholesterol disrupts lysosomal degradation and enhances chemical-induced endocytic disturbance, resulting in great accumulation of Aβ in Neuro2a cells. These findings suggest that an alteration of cerebral cholesterol metabolism may be responsible for augmentation of Aβ pathology in T2DM-affected brains, which, in turn, may increase the risk for developing AD.
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- 2019
17. Successful Inflammation Imaging of Non-Human Primate Hearts Using an Antibody Specific for Tenascin-C
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Hiroyuki Kurosawa, Ousuke Fujimoto, Tomoya Uehara, Yasushi Arano, Yasuhiro Yasutomi, Michiaki Hiroe, Toshimichi Yoshida, Kyoko Imanaka-Yoshida, and Naohide Ageyama
- Subjects
Primates ,Pathology ,medicine.medical_specialty ,Myocardial Infarction ,Inflammation ,030204 cardiovascular system & hematology ,Indium ,03 medical and health sciences ,Pericarditis ,0302 clinical medicine ,Animals ,Medicine ,030212 general & internal medicine ,Myocardial infarction ,Ventricular remodeling ,Ligation ,Tomography, Emission-Computed, Single-Photon ,Environmental Biomarkers ,Ventricular Remodeling ,biology ,business.industry ,Tenascin C ,Heart ,Tenascin ,General Medicine ,musculoskeletal system ,medicine.disease ,Coronary Vessels ,Extracellular Matrix ,Molecular Imaging ,Rats ,Macaca fascicularis ,medicine.anatomical_structure ,Ventricle ,Heart failure ,biology.protein ,medicine.symptom ,Cardiology and Cardiovascular Medicine ,business ,Immunostaining - Abstract
Inflammation after myocardial infarction (MI) may be a major factor influencing ventricular remodeling, leading to congestive heart failure and arrhythmia. Therefore, inflammation in the heart needs to be monitored. Tenascin-C (TNC) is an extracellular matrix molecule not normally expressed, but it is strongly upregulated when associated with active inflammation. Based on this characteristic, we successfully imaged in vivo inflammatory lesions in rat models using 111Indium (111In)-labeled anti-TNC antibodies. The aim of the present study was to further assess the applicability of this molecular imaging probe to detect inflammatory activity in primate hearts.We generated an MI model of cynomolgus monkeys (Macaca fascicularis) by coronary artery ligation and performed dual-isotope single-photon emission computed tomography (SPECT) imaging with an 111In-labeled anti-TNC antibody Fab' fragment (111In-TNC Fab') and 99mtechnetium methoxy-isobutyl isonitrile (99mTc-MIBI). Dual autoradiography was used to compare the uptake of 111In-TNC Fab' with histology and immunostaining for TNC. Dual-isotope SPECT showed the regional myocardial uptake of 111In-TNC Fab' complementary to a defect in the perfusion image by 99mTc-MIBI. The high radioactivity of 111In-TNC Fab' by autoradiography corresponded to immunostaining for TNC, which was observed in inflammatory lesions at the border zone between the infarcted and non-infarcted areas of the left ventricle and at the epi/pericarditis lesions of the right ventricle. These results demonstrate the potential of 111In-TNC-Fab' imaging to monitor myocardial injury and inflammation and suggest the feasibility of the non-invasive detection of cardiac inflammation following acute MI in a preclinical stage before testing in humans.
- Published
- 2019
18. A unique nanoparticulate TLR9 agonist enables a HA split vaccine to confer FcγR-mediated protection against heterologous lethal influenza virus infection
- Author
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Takuya Yamamoto, Yasuhiro Yasutomi, Masatoshi Momota, Barney S. Graham, Tomohiro Kanuma, Ken Ishii, Shoukichi Takahama, Masaru Kanekiyo, Yoshimasa Takahashi, Takashi Saito, Eiko Moriishi, and Yuji Masuta
- Subjects
0301 basic medicine ,Influenza vaccine ,medicine.medical_treatment ,Immunology ,Orthomyxoviridae ,Hemagglutinins, Viral ,Hemagglutinin (influenza) ,Heterologous ,Receptors, Fc ,Virus ,Mice ,03 medical and health sciences ,0302 clinical medicine ,Orthomyxoviridae Infections ,Influenza, Human ,medicine ,Animals ,Humans ,Immunology and Allergy ,biology ,General Medicine ,Vaccine efficacy ,biology.organism_classification ,Virology ,Mice, Inbred C57BL ,Vaccination ,030104 developmental biology ,Influenza Vaccines ,Toll-Like Receptor 9 ,biology.protein ,Nanoparticles ,Featured Article of the Month ,Female ,Adjuvant ,030215 immunology - Abstract
The development of a universal influenza vaccine that can provide a robust and long-lasting protection against a broader range of influenza virus strains is a global public health priority. One approach to improve vaccine efficacy is to use an adjuvant to boost immune responses to the target antigens; nevertheless, the role of adjuvants in the context of influenza vaccines is not fully understood. We have previously developed the K3-schizophyllan (SPG) adjuvant, which is composed of nanoparticulated oligodeoxynucleotides K3, a TLR9 agonist, with SPG, a non-agonistic β-glucan ligand of Dectin-1. In this study, K3-SPG given with conventional influenza hemagglutinin (HA) split vaccine (K3-SPG HA) conferred protection against antigenically mismatched heterologous virus challenge. While K3-SPG HA elicited robust cross-reactive HA-specific IgG2c and CD8 T-cell responses, CD8 T-cell depletion had no impact on this cross-protection. In contrast, K3-SPG HA was not able to confer protection against heterologous virus challenge in FcRγ-deficient mice. Our results indicated that FcγR-mediated antibody responses induced by the HA antigen and K3-SPG adjuvant were important for potent protection against antigenically mismatched influenza virus infection. Thus, we demonstrated that the K3-SPG-adjuvanted vaccine strategy broadens protective immunity against influenza and provides a basis for the development of next-generation influenza vaccines.
- Published
- 2018
19. CD8 T Cells Show Protection against Highly Pathogenic Simian Immunodeficiency Virus (SIV) after Vaccination with SIV Gene-Expressing BCG Prime and Vaccinia Virus/Sendai Virus Vector Boosts
- Author
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Seiichi Kato, Taeko K Naruse, Yasuhiro Yasutomi, Tomotaka Okamura, Tomoyuki Miura, Makoto Inoue, Tsugumine Shu, Xianfeng Zhang, Akinori Kimura, Tetsu Mukai, Hisatoshi Shida, and Kazuhiro Matsuo
- Subjects
viruses ,recombinant BCG ,Simian Acquired Immunodeficiency Syndrome ,CD8-Positive T-Lymphocytes ,medicine.disease_cause ,Sendai virus ,Mice ,chemistry.chemical_compound ,0302 clinical medicine ,Cricetinae ,recombinant vaccinia ,Cytotoxic T cell ,030212 general & internal medicine ,AIDS Vaccines ,0303 health sciences ,human immunodeficiency virus ,biology ,Immunogenicity ,Vaccination ,SAIDS Vaccines ,Provirus ,SIV ,BCG Vaccine ,Rabbits ,simian immunodeficiency virus ,Genetic Vectors ,Immunology ,Vaccinia virus ,Microbiology ,Virus ,Cell Line ,03 medical and health sciences ,Virology ,Vaccines and Antiviral Agents ,medicine ,Animals ,Humans ,Tuberculosis ,030304 developmental biology ,Acquired Immunodeficiency Syndrome ,HIV ,Simian immunodeficiency virus ,biology.organism_classification ,Macaca mulatta ,Mice, Inbred C57BL ,Disease Models, Animal ,chemistry ,Insect Science ,CTL ,HIV-1 ,Vaccinia - Abstract
Because both AIDS and tuberculosis are serious health threats in middle/low-income countries, development of a dual vaccine against them would be highly beneficial. To approach the goal, here we first assessed a urease-deficient bacillus Calmette-Guérin (BCG) for improvement of immunogenicity against both Mycobacterium tuberculosis and SIV. Second, we demonstrated the usefulness of Asian-origin cynomolgus monkeys for development of a preclinical AIDS vaccine by direct comparison with Indian rhesus macaques as the only validated hosts that identically mirror the outcomes of clinical trials, since the availability of Indian rhesus macaques is limited in countries other than the United States. Finally, we report the protective effect of a vaccination regimen comprising BCG, the highly attenuated vaccinia virus LC16m8Δ strain, and nontransmissible Sendai virus as safe vectors expressing SIV genes using repeated mucosal challenge with highly pathogenic SIVmac251. Identification of CD8+ T cells as a protective immunity suggests a future direction of AIDS vaccine development., Toward development of a dual vaccine for human immunodeficiency virus type 1 (HIV-1) and tuberculosis infections, we developed a urease-deficient bacillus Calmette-Guérin (BCG) strain Tokyo172 (BCGΔurease) to enhance its immunogenicity. BCGΔurease expressing a simian immunodeficiency virus (SIV) Gag induced BCG antigen-specific CD4+ and CD8+ T cells more efficiently and more Gag-specific CD8+ T cells. We evaluated its protective efficacy against SIV infection in cynomolgus monkeys of Asian origin, shown to be as susceptible to infection with SIVmac251 as Indian rhesus macaques. Priming with recombinant BCG (rBCG) expressing SIV genes was followed by a boost with SIV gene-expressing LC16m8Δ vaccinia virus and a second boost with SIV Env-expressing Sendai virus. Eight weeks after the second boost, monkeys were repeatedly challenged with a low dose of SIVmac251 intrarectally. Two animals out of 6 vaccinees were protected, whereas all 7 control animals were infected without any early viral controls. In one vaccinated animal, which had the most potent CD8+ T cells in an in vitro suppression activity (ISA) assay of SIVmac239 replication, plasma viremia was undetectable throughout the follow-up period. Protection was confirmed by the lack of anamnestic antibody responses and detectable cell-associated provirus in various organs. Another monkey with a high ISA acquired a small amount of SIV, but it later became suppressed below the detection limit. Moreover, the ISA score correlated with SIV acquisition. On the other hand, any parameter relating anti-Env antibody was not correlated with the protection. IMPORTANCE Because both AIDS and tuberculosis are serious health threats in middle/low-income countries, development of a dual vaccine against them would be highly beneficial. To approach the goal, here we first assessed a urease-deficient bacillus Calmette-Guérin (BCG) for improvement of immunogenicity against both Mycobacterium tuberculosis and SIV. Second, we demonstrated the usefulness of Asian-origin cynomolgus monkeys for development of a preclinical AIDS vaccine by direct comparison with Indian rhesus macaques as the only validated hosts that identically mirror the outcomes of clinical trials, since the availability of Indian rhesus macaques is limited in countries other than the United States. Finally, we report the protective effect of a vaccination regimen comprising BCG, the highly attenuated vaccinia virus LC16m8Δ strain, and nontransmissible Sendai virus as safe vectors expressing SIV genes using repeated mucosal challenge with highly pathogenic SIVmac251. Identification of CD8+ T cells as a protective immunity suggests a future direction of AIDS vaccine development.
- Published
- 2021
20. Long-term sterile immunity induced by an adjuvant-containing live-attenuated AIDS virus
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Masamitsu N Asaka, Tomotaka Okamura, Yusuke Tsujimura, Takuya Yamamoto, Kazuhiro Matsuo, Yasuhiro Yasutomi, Tomohiro Kanuma, and Yuya Shimizu
- Subjects
viruses ,animal diseases ,medicine.medical_treatment ,T cell ,virus diseases ,Biology ,Virology ,Virus ,Vaccination ,Immune system ,medicine.anatomical_structure ,Antigen ,Immunity ,medicine ,HIV vaccine ,Adjuvant - Abstract
Antigen 85B (Ag85B) is one of the most dominant proteins secreted from most mycobacterial species, and it induces Th1-type immune responses as an adjuvant. We genetically constructed a live attenuated simian human immunodeficiency virus to express the adjuvant molecule Ag85B (SHIV-Ag85B). SHIV-Ag85B could not be detected 4 weeks after injection in cynomolgus macaques, and strong SHIV-specific T cell responses were induced in these macaques. When these macaques in which SHIV-Ag85B had become undetectable were challenged with pathogenic SHIV89.6P at 37 weeks after SHIV-Ag85B became undetectable, SHIV89.6P could not be detected after the challenge. Eradication of SHIV89.6P was confirmed by adoptive transfer experiments and CD8-depletion studies. The SHIV-Ag85B-inoculated macaques showed enhancement of Gag-specific monofunctional and polyfunctional CD8+T cells in the acute phase of pathogenic SHIV challenge. The results suggest that SHIV-Ag85B elicited strong sterile immune responses against pathogenic SHIV and that it may lead to the development of a vaccine for AIDS virus infection.ImportanceDevelopment of an effective HIV vaccine has been a major priority to control the worldwide AIDS epidemic. The moderately attenuated prototypic vaccine strain SIVmac239Δnef has been used in various studies; however, it does not provide sufficient effects to prevent infection. The use of adjuvant in vaccination is thought to be useful for enhancing the immune responses to various pathogens. In the present study, we constructed a live attenuated SHIV virus expressing adjuvant molecule Ag85B and assessed vaccine effects in cynomolgus macaques. The present study shows that live-attenuated SHIV expressing Ag85B elicits viral antigen-specific polyfunctional CD8+T cell responses against pathogenic SHIV and provide the possibility of eradicating a pathogenic lentivirus from infected animals.
- Published
- 2020
21. Development of a Mucosal TB Vaccine Using Human Parainfluenza Type 2 Virus
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Yusuke Tsujimura and Yasuhiro Yasutomi
- Subjects
Tuberculosis ,biology ,business.industry ,Mucous membrane ,medicine.disease ,biology.organism_classification ,Virus ,Mycobacterium tuberculosis ,Vaccination ,medicine.anatomical_structure ,Immune system ,Immunopathology ,Immunology ,medicine ,Vector (molecular biology) ,business - Abstract
Tuberculosis (TB) is a major chronic infectious disease, and the majority of TB patients show latent infection with a risk of reactivation for their entire lifetime. The phase of latent TB infection involves an ongoing balance between anti-mycobacterial activity and regulatory mechanisms to minimize immunopathology on the pulmonary mucous membrane. Therefore, future study should target the development of vaccines that could induce a mucosal immune system response against Mycobacterium tuberculosis (Mtb) in any period, including dormancy. Human parainfluenza type 2 virus (HPIV2) is a human respiratory pathogen without visible pathogenicity. Recently, by reverse genetics technology, a recombinant HPIV2 has been constructed that has not only replication-incompetent behavior, but also expression systems of multiple TB antigens that are selectable at any stage of TB infection, such as latency, resuscitation, and activity in the respiratory tract. In this review, we show recent findings related to current and future vaccination strategies against TB, and we provide further evidences for the potential usefulness of HPIV2 as a novel nasal vaccine vector that could elicit Mtb-specific systemic and mucosal immunity.
- Published
- 2020
22. Gene expression dataset for whole cochlea of Macaca fascicularis
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Fuyuki Miya, Tatsuhiko Tsunoda, Hideki Mutai, Yasuhiro Yasutomi, Tatsuo Matsunaga, and Hiroaki Shibata
- Subjects
0301 basic medicine ,Hearing loss ,lcsh:Medicine ,Computational biology ,Biology ,Macaque ,Article ,03 medical and health sciences ,0302 clinical medicine ,biology.animal ,medicine ,otorhinolaryngologic diseases ,Auditory system ,Animals ,Humans ,lcsh:Science ,Gene ,Cochlea ,Multidisciplinary ,Microarray analysis techniques ,Gene Expression Profiling ,lcsh:R ,Microarray Analysis ,Ear morphogenesis ,Gene expression profiling ,Macaca fascicularis ,030104 developmental biology ,medicine.anatomical_structure ,lcsh:Q ,sense organs ,medicine.symptom ,030217 neurology & neurosurgery - Abstract
Macaca fascicularis is a highly advantageous model in which to study human cochlea with regard to both evolutionary proximity and physiological similarity of the auditory system. To better understand the properties of primate cochlear function, we analyzed the genes predominantly expressed in M. fascicularis cochlea. We compared the cochlear transcripts obtained from an adult male M. fascicularis by macaque and human GeneChip microarrays with those in multiple macaque and human tissues or cells and identified 344 genes with expression levels more than 2-fold greater than in the other tissues. These “cochlear signature genes” included 35 genes responsible for syndromic or nonsyndromic hereditary hearing loss. Gene set enrichment analysis revealed groups of genes categorized as “ear development” and “ear morphogenesis” in the top 20 gene ontology categories in the macaque and human arrays, respectively. This dataset will facilitate both the study of genes that contribute to primate cochlear function and provide insight to discover novel genes associated with hereditary hearing loss that have yet to be established using animal models.
- Published
- 2018
23. Superparamagnetic Iron Oxide-enhanced Magnetic Resonance Imaging of Spontaneous Hepatic Neoplasia in a Cynomolgus Macaque (Macaca fascicularis)
- Author
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Kiichi Kanayama, Shunya Nakayama, Hiroaki Shibata, Naohide Ageyama, Yasuhiro Yasutomi, Sachi Okabayashi, Yasuyo Ito Fujishiro, Hiroshi Koie, and Yuko Katakai
- Subjects
Pathology ,medicine.medical_specialty ,General Veterinary ,medicine.diagnostic_test ,040301 veterinary sciences ,business.industry ,Radiography ,Magnetic resonance imaging ,04 agricultural and veterinary sciences ,Anorexia ,medicine.disease ,Cynomolgus macaque ,humanities ,General Biochemistry, Genetics and Molecular Biology ,030218 nuclear medicine & medical imaging ,Serology ,0403 veterinary science ,03 medical and health sciences ,0302 clinical medicine ,Hepatocellular carcinoma ,Hepatic neoplasms ,medicine ,medicine.symptom ,business ,Superparamagnetic iron oxide - Abstract
Although the number of reports describing tumors in aged NHP has increased, spontaneous neoplasias in NHP are extremely rare, with the notable exception of prosimians, in which spontaneous hepatic neoplasms arise. In addition to radiography and ultrasonography, superparamagnetic iron oxide (SPIO)-enhanced MRI tends to be applied in human practice to non-invasively locate, identify, and size liver tumors and to define the border between neoplastic and normal tissues. Here we report a 13-y-old female cynomolgus monkey with anorexia and serologically normal liver enzymes. After fluid therapy, the condition remained in remission for several months. Later, however, a palpable mass was assessed by using ultrasonography, radiology, and SPIO-MRI; T2-weighted images revealed a clear border between a hepatocellular carcinoma and normal liver tissue. Findings at necropsy supported the imaging data. Serologic assessment after euthanasia revealed a positive reaction to an abnormal form of prothrombin (PIVKA-II). We recommend SPIO-MRI as a practical and useful for diagnosing hepatocellular neoplasias in NHP. This study is the first to demonstrate the applicability of SPIO-MRI for the identification of hepatocellular carcinoma in NHP.
- Published
- 2018
24. An Antigen-Free, Plasmacytoid Dendritic Cell–Targeting Immunotherapy To Bolster Memory CD8+ T Cells in Nonhuman Primates
- Author
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Ken Ishii, Kenji Mizuguchi, Yasuhiro Yasutomi, Yayoi Natsume-Kitatani, Takuya Yamamoto, Eiko Moriishi, Yuji Masuta, Kouji Kobiyama, and Tomohiro Kanuma
- Subjects
0301 basic medicine ,Chemistry ,medicine.medical_treatment ,Immunology ,TLR9 ,Plasmacytoid dendritic cell ,Immunotherapy ,Proinflammatory cytokine ,03 medical and health sciences ,CTL ,030104 developmental biology ,0302 clinical medicine ,Antigen ,030220 oncology & carcinogenesis ,Cancer research ,medicine ,Immunology and Allergy ,Cytotoxic T cell ,CD8 - Abstract
The priming, boosting, and restoration of memory cytotoxic CD8+ T lymphocytes by vaccination or immunotherapy in vivo is an area of active research. Particularly, nucleic acid–based compounds have attracted attention due to their ability to elicit strong Ag-specific CTL responses as a vaccine adjuvant. Nucleic acid–based compounds have been shown to act as anticancer monotherapeutic agents even without coadministration of cancer Ag(s); however, so far they have lacked efficacy in clinical trials. We recently developed a second-generation TLR9 agonist, a humanized CpG DNA (K3) complexed with schizophyllan (SPG), K3-SPG, a nonagonistic Dectin-1 ligand. K3-SPG was previously shown to act as a potent monoimmunotherapeutic agent against established tumors in mice in vivo. In this study we extend the monoimmunotherapeutic potential of K3-SPG to a nonhuman primate model. K3-SPG activated monkey plasmacytoid dendritic cells to produce both IFN-α and IL-12/23 p40 in vitro and in vivo. A single injection s.c. or i.v. with K3-SPG significantly increased the frequencies of activated memory CD8+ T cells in circulation, including Ag-specific memory CTLs, in cynomolgus macaques. This increase did not occur in macaques injected with free CpG K3 or polyinosinic-polycytidylic acid. Injection of 2 mg K3-SPG induced mild systemic inflammation, however, levels of proinflammatory serum cytokines and circulating neutrophil influx were lower than those induced by the same dose of polyinosinic-polycytidylic acid. Therefore, even in the absence of specific Ags, we show that K3-SPG has potent Ag-specific memory CTL response–boosting capabilities, highlighting its potential as a monoimmunotherapeutic agent for chronic infectious diseases and cancer.
- Published
- 2018
25. Relationship between menarche and fertility in long-tailed macaques (Macaca fascicularis)
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Maiko Kobayashi, Takamasa Koyama, Yasuhiro Yasutomi, and Tadashi Sankai
- Subjects
0301 basic medicine ,Pregnancy ,biology ,media_common.quotation_subject ,Physiology ,Fertility ,030204 cardiovascular system & hematology ,medicine.disease ,Fecundity ,Menstruation ,03 medical and health sciences ,030104 developmental biology ,0302 clinical medicine ,Animal model ,biology.animal ,medicine ,Menarche ,Animal Science and Zoology ,Primate ,Reproduction ,media_common - Abstract
We attempted to elucidate female reproduction in long-tailed macaques (Macaca fascicularis). These monkeys have a non-seasonal menstruation cycle, which makes them suitable subjects for studies in a variety fields including medical science and regenerative medicine. We analyzed individual breeding data including time of menarche, start of regular menstruation, and first pregnancy. These three events are related to the maturation of female long-tailed macaques. All research subjects were female long-tailed macaques bred at the Tsukuba Primate Research Center. The study comprised 45 females; we included time of menstruation, male-female cohabitation, and first pregnancy in their growth records. We extracted age and weight data relating to menarche, start of regular menstruation, and first pregnancy from these records. In the two years typically required from menarche to first pregnancy, the body weight increased by approximately 500 g (21% of the weight at menarche); it is clear that there is a significant physical change after menarche. Our findings suggest that female monkeys are not necessarily mature enough for pregnancy at menarche. Therefore, the use of the word "maturity" in terms of fecundity may be more accurate after the start of regular menstruation. This is what we term "adolescence" in the developmental process. Therefore, M. fascicularis monkeys are candidates for an animal model of human adolescence.
- Published
- 2018
26. Utility of arterial blood gas, CBC, biochemistry and cardiac hormones as evaluation parameters of cardiovascular disease in nonhuman primates
- Author
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Yasuyo Ito-Fujishiro, Shunya Nakayama, Naohide Ageyama, Kiichi Kanayama, Hiroshi Koie, Tadashi Sankai, Yasuhiro Yasutomi, and Yuko Katakai
- Subjects
Male ,Primates ,complete blood count ,040301 veterinary sciences ,Cardiomyopathy ,nonhuman primate ,Inflammation ,Disease ,030204 cardiovascular system & hematology ,Cardiac hormones ,0403 veterinary science ,03 medical and health sciences ,0302 clinical medicine ,Laboratory Animal Science ,cardiovascular disease ,medicine ,Animals ,Acidosis ,Full Paper ,General Veterinary ,medicine.diagnostic_test ,business.industry ,Complete blood count ,Red blood cell distribution width ,arterial blood gas ,04 agricultural and veterinary sciences ,cardiac hormone ,medicine.disease ,Blood Cell Count ,Biochemistry ,Cardiovascular Diseases ,Quality of Life ,Arterial blood ,Female ,Blood Gas Analysis ,medicine.symptom ,business - Abstract
Cardiovascular disease (CVD) has a tremendous impact on the quality of life of humans. While experimental animals are valuable to medical research as models of human diseases, cardiac systems differ widely across various animal species. Thus, we examined a CVD model in cynomolgus monkeys. Laboratory primates are precious resources, making it imperative that symptoms of diseases and disorders are detected as early as possible. Thus, in this study we comprehensively examined important indicators of CVD in cynomolgus monkeys, including arterial blood gas, complete blood count (CBC), biochemistry and cardiac hormones. The control group included 20 healthy macaques showing non-abnormal findings in screening tests, whereas the CVD group included 20 macaques with valvular disease and cardiomyopathy. An increase of red blood cell distribution width was observed in the CBC, indicating chronic inflammation related to CVD. An increase of HCO3 was attributed to the correction of acidosis. Furthermore, development of the CVD model was supported by significant increases in natriuretic peptides. It is suggested that these results indicated a correlation between human CVD and the model in monkeys. Moreover, blood tests including arterial blood gas are non-invasive and can be performed more easily than other technical tests. CVD affected animals easily change their condition by anesthesia and surgical invasion. Pay attention to arterial blood gas and proper respond to their condition are important for research. This data may facilitate human research and aid in the management and veterinary care of nonhuman primates.
- Published
- 2018
27. Establishment of reference values for complete blood count and blood gases in cynomolgus monkeys (Macaca fascicularis)
- Author
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Yuko Katakai, Tadashi Sankai, Yasuyo Ito-Fujishiro, Yasuhiro Yasutomi, Kiichi Kanayama, Shunya Nakayama, Hiroshi Koie, and Naohide Ageyama
- Subjects
0301 basic medicine ,General Veterinary ,medicine.diagnostic_test ,business.industry ,Physiology ,Complete blood count ,030204 cardiovascular system & hematology ,pCO2 ,03 medical and health sciences ,030104 developmental biology ,0302 clinical medicine ,Animal management ,Reference values ,Circulatory system ,medicine ,Arterial blood ,Respiratory system ,business ,Hormone - Abstract
Cynomolgus monkeys are closely related to humans phylogenetically, and this has resulted in their widespread use as a preclinical model. Hematological data with regard to these monkeys are thus important. Although reference values for blood components and sex hormones have been established for cynomolgus monkeys, those for arterial blood gases have not. The arterial blood gases quickly reflect respiratory and circulatory dynamics, and are thus useful for animal management and safe general anesthesia and surgical operations. Furthermore, since O2 is transported by RBC, CBC and blood gases are closely related. The present study aimed to establish reference values for arterial blood gases and CBC in cynomolgus monkeys over a wide age range. Blood gases and CBC of arterial blood, collected from 41 female and 21 male anesthetized monkeys, were measured. Age correlated with RBC, HGB and HCT in the CBC. Values differed significantly between males and females in pCO2, CO2 concentration, MCV and MCH. The pH of blood was equivalent to that of humans and pCO2 was more stable, whereas MCV and MCH were lower than those in humans. Erythrocytes were smaller and less pigmented than in other Macaca species. Several relationships between gender and age, and blood gases and CBC were identified in cynomolgus monkeys. In conclusion, these reference values will be useful as markers for veterinary applications and in the care and maintenance of these animals.
- Published
- 2017
28. Simian immunodeficiency virus SIVmac239 infection and simian human immunodeficiency virus SHIV89.6P infection result in progression to AIDS in cynomolgus macaques of Asian origin
- Author
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Tomotaka Okamura, Ichiro Takahashi, Shogo Soma, Yusuke Tsujimura, Yasuhiro Yasutomi, and Kazuhiro Matsuo
- Subjects
CD4-Positive T-Lymphocytes ,Male ,0301 basic medicine ,Asia ,animal diseases ,viruses ,T cell ,Biology ,medicine.disease_cause ,Pathogenesis ,03 medical and health sciences ,Acquired immunodeficiency syndrome (AIDS) ,Virology ,medicine ,Animals ,Humans ,Infectivity ,Acquired Immunodeficiency Syndrome ,Simian human immunodeficiency virus ,HIV ,virus diseases ,Viral Load ,Simian immunodeficiency virus ,medicine.disease ,Vaccination ,Disease Models, Animal ,Macaca fascicularis ,030104 developmental biology ,medicine.anatomical_structure ,Immunology ,Disease Progression ,Simian Immunodeficiency Virus ,Viral load - Abstract
Simian immunodeficiency virus (SIV) infection models in cynomolgus macaques are important for analysis of the pathogenesis of immunodeficiency virus and for studies on the efficacy of new vaccine candidates. However, very little is known about the pathogenesis of SIV or simian human immunodeficiency virus (SHIV) in cynomolgus macaques from different Asian countries. In the present study, we analysed the infectivity and pathogenicity of CCR5-tropic SIVmac and those of dual-tropic SHIV89.6P inoculated into cynomolgus macaques in Indonesian, Malaysian or Philippine origin. The plasma viral loads in macaques infected with either SIVmac239 or SHIV89.6P were maintained at high levels. CD4+ T cell levels in macaques infected with SIVmac239 gradually decreased. All of the macaques infected with SHIV89.6P showed greatly reduced CD4+ T-cell numbers within 6 weeks of infection. Eight of the 11 macaques infected with SIVmac239 were killed due to AIDS symptoms after 2-4.5 years, while four of the five macaques infected with SHIV89.6P were killed due to AIDS symptoms after 1-3.5 years. We also analysed cynomolgus macaques infected intrarectally with repeated low, medium or high doses of SIVmac239, SIVmac251 or SHIV89.6P. Infection was confirmed by quantitative RT-PCR at more than 5000, 300 and 500 TCID50 for SIVmac239, SIVmac251 and SHIV89.6P, respectively. The present study indicates that cynomolgus macaques of Asian origin are highly susceptible to SIVmac and SHIV infection by both intravenous and mucosal routes. These models will be useful for studies on virus pathogenesis, vaccination and therapeutics against human immunodeficiency virus/AIDS.
- Published
- 2016
29. Leucine-rich alpha 2 glycoprotein is a new marker for active disease of tuberculosis
- Author
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Yusuke Tsujimura, Tetsuji Naka, Minoru Fujimoto, Tomoshige Matsumoto, Yasuhiro Yasutomi, Satoshi Serada, and Shoji Hashimoto
- Subjects
0301 basic medicine ,Tuberculosis ,lcsh:Medicine ,Blood Sedimentation ,Article ,Mycobacterium tuberculosis ,03 medical and health sciences ,0302 clinical medicine ,medicine ,Animals ,Humans ,lcsh:Science ,Lung ,Glycoproteins ,Multidisciplinary ,biology ,business.industry ,Interleukin-6 ,lcsh:R ,Diagnostic markers ,biology.organism_classification ,medicine.disease ,Vaccination ,Disease Models, Animal ,Macaca fascicularis ,030104 developmental biology ,medicine.anatomical_structure ,C-Reactive Protein ,Giant cell ,030220 oncology & carcinogenesis ,Case-Control Studies ,Immunology ,BCG Vaccine ,Biomarker (medicine) ,Immunohistochemistry ,lcsh:Q ,business ,Epithelioid cell ,Biomarkers - Abstract
Tuberculosis (TB) caused by Mycobacterium tuberculosis (Mtb) is a global health problem. At present, prior exposure to Mtb can be determined by blood-based interferon-gamma release assay (IGRA), but active TB is not always detectable by blood tests such as CRP and ESR. This study was undertaken to investigate whether leucine-rich alpha-2 glycoprotein (LRG), a new inflammatory biomarker, could be used to assess active disease of TB. Cynomolgus macaques pretreated with or without Bacille Calmette-Guerin (BCG) vaccination were inoculated with Mtb to induce active TB. Blood was collected over time from these animals and levels of LRG as well as CRP and ESR were quantified. In the macaques without BCG vaccination, Mtb inoculation caused extensive TB and significantly increased plasma CRP and LRG levels, but not ESR. In the macaques with BCG vaccination, whereas Mtb challenge caused pulmonary TB, only LRG levels were significantly elevated. By immunohistochemical analysis of the lung, LRG was visualized in epithelioid cells and giant cells of the granulation tissue. In humans, serum LRG levels in TB patients were significantly higher than those in healthy controls and declined one month after anti-tubercular therapy. These findings suggest that LRG is a promising biomarker when performed following IGRA for the detection of active TB.
- Published
- 2019
30. Intranasal vaccination with HBs and HBc protein combined with carboxyl vinyl polymer induces strong neutralizing antibody, anti-HBs IgA, and IFNG response
- Author
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Miyazaki Takashi, Goh Yasumasa, Osamu Yoshida, Hideki Hasegawa, Naoki Yamamoto, Yoichi Hiasa, Yumiko Shiogama, Michinori Kohara, Takahiro Sanada, Mohammad Enamul Hoque Kayesh, Kyoko Tsukiyama-Kohara, Yasuhiro Yasutomi, and Jun-ichiro Takano
- Subjects
0301 basic medicine ,Hepatitis B virus ,Immunogen ,Genotype ,Polymers ,Biophysics ,medicine.disease_cause ,Antibodies, Viral ,Biochemistry ,03 medical and health sciences ,Interferon-gamma ,Mice ,0302 clinical medicine ,Antigen ,Interferon ,Neutralization Tests ,medicine ,Animals ,Humans ,Neutralizing antibody ,Molecular Biology ,Administration, Intranasal ,Hepatitis B Surface Antigens ,biology ,business.industry ,Tupaiidae ,virus diseases ,Cell Biology ,Hep G2 Cells ,Hepatitis B ,medicine.disease ,Virology ,Antibodies, Neutralizing ,Hepatitis B Core Antigens ,digestive system diseases ,Immunoglobulin A ,Vaccination ,030104 developmental biology ,Immunization ,Liver ,030220 oncology & carcinogenesis ,Immunoglobulin G ,biology.protein ,business ,medicine.drug - Abstract
Hepatitis B virus (HBV) infection causes acute and chronic hepatitis, which is a major public health concern worldwide. Immunization methods incorporating hepatitis B surface-small (HBs-S) antigen and hepatitis B core antigen (HBc) have been proposed as candidate therapeutic vaccines, but the elimination of existing HBV infection remains a challenge. To enhance the efficacy of HBs and HBc vaccination, we investigated HBs-large (HBs-L) as an immunogen, and carboxyl vinyl polymer (CVP) as an excipient. HBs-S or HBs-L, in combination with HBc antigen, was administered subcutaneously (without CVP) or intranasally (with or without CVP) for the evaluation of immune response in the tree shrew, which is considered to be a suitable small animal model of HBV infection. Immunization with HBs-L antigen by either route induced a rapid IgG response. Intranasal immunization with HBs-S or HBs-L and HBc formulated with CVP strongly induced neutralizing antibody activity, IgA response, and HBc-specific expression of the interferon gamma-encoding gene. These data indicated the potential of HBs-L and HBc intranasal immunization with CVP, not only as a therapeutic vaccine, but also as a prophylactic vaccine candidate.
- Published
- 2019
31. Establishment of a new formula for QT interval correction using a large colony of cynomolgus monkeys
- Author
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Yasuyo Ito-Fujishiro, Tadashi Sankai, Shunya Nakayama, Yasuhiro Yasutomi, Kiichi Kanayama, Chungyu Pai, Hiroshi Koie, Naohide Ageyama, and Miyoko Kato-Tateishi
- Subjects
0301 basic medicine ,Male ,QT interval ,medicine.medical_specialty ,congenital, hereditary, and neonatal diseases and abnormalities ,Correction method ,Original ,the corrected QT interval ,electrocardiogram ,General Biochemistry, Genetics and Molecular Biology ,03 medical and health sciences ,Electrocardiography ,0302 clinical medicine ,Heart Rate ,cardiovascular disease ,Internal medicine ,Heart rate ,medicine ,Repolarization ,Animals ,cardiovascular diseases ,General Veterinary ,business.industry ,Mean value ,Prolongation ,Corrected qt ,General Medicine ,Long QT Syndrome ,Macaca fascicularis ,030104 developmental biology ,Underlying disease ,Cardiology ,cardiovascular system ,Animal Science and Zoology ,Female ,cynomolgus monkey ,business ,030217 neurology & neurosurgery ,circulatory and respiratory physiology - Abstract
The demand for monkeys for medical research is increasing, because their ionic mechanism of repolarization is similar to that of humans. The QT interval is the distance between the Q wave and T wave, but this interval is affected by heart rate. Therefore, QT correction methods are commonly used in clinical settings. However, an accurate correction formula for the QT interval in cynomolgus monkeys has not been reported. We assessed snapshot electrocardiograms (ECGs) of 353 ketamine-immobilized monkeys, including aged animals, and contrived a new formula for the corrected QT interval (QTc) as a marker of QT interval prolongation in cynomolgus monkeys. Values for QTc were calculated using the formula [QTc] = [QT] / [RR]n, along with several other formulas commonly used to calculate QTc. We found that the optimal exponent of the QT interval corrected for heart rate, n, was 0.576. The mean value of QTc in healthy monkeys determined using the new formula was 373 ± 31 mm, and there were no significant differences between the sexes. Other ECG parameters were not significantly different between the sexes and there were no age-related effects on QTc. Prolongation of QTc to over 405 ms, as calculated by the new formula, was observed in 50 monkeys with underlying diseases. Additionally, all monkeys with QTc above 440 ms by the new formula had some underlying disease. The results resemble those in humans, suggesting that the new QTc formula could be useful for diagnosis of QT interval prolongation in cynomolgus monkeys.
- Published
- 2019
32. STING agonists activate latently infected cells and enhance SIV-specific responses ex vivo in naturally SIV controlled cynomolgus macaques
- Author
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Takuya Yamamoto, Kazutaka Terahara, Ken Ishii, Yasuhiro Yasutomi, Eiko Moriishi, Tomotaka Okamura, Shokichi Takahama, and Tomohiro Kanuma
- Subjects
0301 basic medicine ,Male ,medicine.medical_treatment ,Simian Acquired Immunodeficiency Syndrome ,lcsh:Medicine ,Biology ,CD8-Positive T-Lymphocytes ,Peripheral blood mononuclear cell ,Article ,03 medical and health sciences ,0302 clinical medicine ,Immune system ,medicine ,Cytotoxic T cell ,Animals ,lcsh:Science ,Multidisciplinary ,lcsh:R ,Toll-Like Receptors ,Imidazoles ,virus diseases ,Membrane Proteins ,Viral Load ,Virology ,Virus Latency ,Sting ,CTL ,Macaca fascicularis ,030104 developmental biology ,Cytokine ,Leukocytes, Mononuclear ,lcsh:Q ,Simian Immunodeficiency Virus ,Virus Activation ,Nucleotides, Cyclic ,030217 neurology & neurosurgery ,CD8 ,Ex vivo ,Dinucleoside Phosphates ,T-Lymphocytes, Cytotoxic - Abstract
To achieve a functional cure for HIV, treatment regimens that eradicate latently HIV-infected cells must be established. For this, many groups have attempted to reactivate latently-infected cells to induce cytopathic effects and/or elicit cytotoxic T lymphocyte (CTL)/NK cell-mediated immune responses to kill these cells. We believe that not only the reactivation of latently-infected cells, but also the induction of strong CTL responses, would be required for this. Here, we used typical immune activators that target pattern recognition receptors (PRRs). For our experimental model, we identified eight SIV-infected cynomolgus monkeys that became natural controllers of viremia. Although plasma viral loads were undetectable, we could measure SIV-DNA by qPCR in peripheral blood mononuclear cells (PBMCs). Using these PBMCs, we screened 10 distinct PRR ligands to measure IFN-α and IFN-γ production. Among these, STING ligands, cGAMP and c-di-AMP, and the TLR7/8 agonist R848 markedly increased cytokine levels. Both R848 and STING ligands could reactivate latently-infected cells in both cynomolgus monkeys and human PBMCs in vitro. Furthermore, c-di-AMP increased the frequency of SIV Gag-specific CD8+ T cells including polyfunctional CD8+ T cells, as compared to that in untreated control or R848-treated cells. Together, STING ligands might be candidates for HIV treatment.
- Published
- 2019
33. Immunization with Components of the Viral Fusion Apparatus Elicits Antibodies That Neutralize Epstein-Barr Virus in B Cells and Epithelial Cells
- Author
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Sandeep Narpala, Yusuke Tsujimura, Masaru Kanekiyo, Adrian B. McDermott, Dalton V. Banh, Rebecca A. Gillespie, Sarah F. Andrews, M. Gordon Joyce, Moh Lan Yap, Fiona Aguilar, Wei Bu, Zeshan Tariq, Yasuhiro Yasutomi, Gary J. Nabel, Jeffrey I. Cohen, Michael G. Rossmann, Yaroslav Tsybovsky, and Hanh Nguyen
- Subjects
0301 basic medicine ,Male ,Epstein-Barr Virus Infections ,Herpesvirus 4, Human ,Immunology ,Virus Attachment ,CHO Cells ,medicine.disease_cause ,Antibodies, Viral ,Virus ,Epitope ,Article ,Cell Fusion ,03 medical and health sciences ,Mice ,0302 clinical medicine ,Cricetulus ,Viral Envelope Proteins ,hemic and lymphatic diseases ,Cell Line, Tumor ,medicine ,Immunology and Allergy ,Animals ,Humans ,Vaccines, Virus-Like Particle ,B cell ,B-Lymphocytes ,Mice, Inbred BALB C ,Cell fusion ,Membrane Glycoproteins ,biology ,Viral Vaccine ,Immune Sera ,Epithelial Cells ,Viral Vaccines ,Epstein–Barr virus ,Virology ,Antibodies, Neutralizing ,Macaca fascicularis ,030104 developmental biology ,Infectious Diseases ,medicine.anatomical_structure ,HEK293 Cells ,Immunization ,030220 oncology & carcinogenesis ,biology.protein ,Female ,Antibody ,HeLa Cells - Abstract
Summary Epstein-Barr virus (EBV) causes infectious mononucleosis and is associated with epithelial-cell cancers and B cell lymphomas. An effective EBV vaccine is not available. We found that antibodies to the EBV glycoprotein gH/gL complex were the principal components in human plasma that neutralized infection of epithelial cells and that antibodies to gH/gL and gp42 contributed to B cell neutralization. Immunization of mice and nonhuman primates with nanoparticle vaccines that displayed components of the viral-fusion machinery EBV gH/gL or gH/gL/gp42 elicited antibodies that potently neutralized both epithelial-cell and B cell infection. Immune serum from nonhuman primates inhibited EBV-glycoprotein-mediated fusion of epithelial cells and B cells and targeted an epitope critical for virus-cell fusion. Therefore, unlike the leading EBV gp350 vaccine candidate, which only protects B cells from infection, these EBV nanoparticle vaccines elicit antibodies that inhibit the virus-fusion apparatus and provide cell-type-independent protection from virus infection.
- Published
- 2018
34. A New Mouse Model of Chronic Myocarditis Induced by Recombinant Bacille Calmette–Guèrin Expressing a T-Cell Epitope of Cardiac Myosin Heavy Chain-α
- Author
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Kyoko Imanaka-Yoshida, Michiaki Hiroe, Yasuhiro Yasutomi, Masaki Ieda, Kazuhiro Matsuo, Yusuke Tsujimura, Kazutaka Aonuma, and Kazuko Tajiri
- Subjects
Male ,0301 basic medicine ,Adoptive cell transfer ,recombinant BCG ,Epitopes, T-Lymphocyte ,Lymphocyte Activation ,medicine.disease_cause ,Epitope ,Autoimmunity ,lcsh:Chemistry ,0302 clinical medicine ,BCG ,lcsh:QH301-705.5 ,Spectroscopy ,Mice, Inbred BALB C ,Interleukin-17 ,autoimmunity ,Dilated cardiomyopathy ,General Medicine ,Recombinant Proteins ,Computer Science Applications ,medicine.anatomical_structure ,Echocardiography ,030220 oncology & carcinogenesis ,BCG Vaccine ,cardiovascular system ,Cytokines ,inflammatory dilated cardiomyopathy ,Cardiomyopathy, Dilated ,Cardiac function curve ,Myocarditis ,T cell ,Article ,Catalysis ,Proinflammatory cytokine ,Ventricular Myosins ,Inorganic Chemistry ,03 medical and health sciences ,medicine ,Animals ,Humans ,Physical and Theoretical Chemistry ,Molecular Biology ,business.industry ,Organic Chemistry ,Th1 Cells ,medicine.disease ,autoimmune myocarditis ,Disease Models, Animal ,030104 developmental biology ,lcsh:Biology (General) ,lcsh:QD1-999 ,Chronic Disease ,Immunology ,Th17 Cells ,myocarditis ,business - Abstract
Dilated cardiomyopathy (DCM) is a potentially lethal disorder characterized by progressive impairment of cardiac function. Chronic myocarditis has long been hypothesized to be one of the causes of DCM. However, owing to the lack of suitable animal models of chronic myocarditis, its pathophysiology remains unclear. Here, we report a novel mouse model of chronic myocarditis induced by recombinant bacille Calmette-Gué, rin (rBCG) expressing a CD4+ T-cell epitope of cardiac myosin heavy chain-&alpha, (rBCG-MyHC&alpha, ). Mice immunized with rBCG-MyHC&alpha, developed chronic myocarditis, and echocardiography revealed dilation and impaired contraction of ventricles, similar to those observed in human DCM. In the heart, CD62L&minus, CD4+ T cells were increased and produced significant amounts of IFN-&gamma, and IL-17 in response to cardiac myosin. Adoptive transfer of CD62L&minus, CD4+ T cells induced myocarditis in the recipient mice, which indicated that CD62L&minus, CD4+ T cells were the effector cells in this model. rBCG-MyHC&alpha, infected dendritic cells produced proinflammatory cytokines and induced MyHC&alpha, specific T-cell proliferation and Th1 and Th17 polarization. This novel chronic myocarditis mouse model may allow the identification of the central pathophysiological and immunological processes involved in the progression to DCM.
- Published
- 2021
35. Suppression of SARS-CoV-2-induced lung injury by ACE2-like carboxypeptidase B38-CAP in COVID-19 mouse model
- Author
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Midori Hoshizaki, Yumiko Imai, Satoru Nirasawa, Masaki Imai, Yoshihiro Kawaoka, Yasuhiro Yasutomi, Takafumi Minato, Saori Takahashi, Satoshi Nagata, Daichi Utsumi, Jianbo An, Haruhiko Kamada, Tomokazu Yamaguchi, Masamitsu N Asaka, Mayumi Niiyama, and Keiji Kuba
- Subjects
Lung ,medicine.anatomical_structure ,biology ,Chemistry ,Applied Mathematics ,General Mathematics ,medicine ,biology.protein ,Angiotensin-converting enzyme ,Virology ,Virus - Published
- 2021
36. Persistent replication of a hepatitis C virus genotype 1b-based chimeric clone carrying E1, E2 and p6 regions from GB virus B in a New World monkey
- Author
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Noboru Maki, Atsunori Higashino, Kenichi Mori, Hirofumi Akari, Yasuhiro Yasutomi, Yuki Iwasaki, and Saori Suzuki
- Subjects
0301 basic medicine ,Hepatitis B virus DNA polymerase ,Hepatitis C virus ,Immunology ,virus diseases ,Viral transformation ,Biology ,medicine.disease_cause ,Microbiology ,Virology ,digestive system diseases ,Hepatitis B virus PRE beta ,Virus ,03 medical and health sciences ,Chimera (genetics) ,030104 developmental biology ,0302 clinical medicine ,Chimeric RNA ,medicine ,030211 gastroenterology & hepatology ,Oncovirus - Abstract
The development of effective hepatitis C virus (HCV) vaccines is essential for the prevention of further HCV dissemination, especially in developing countries. Therefore the aim of this study is to establish a feasible and immunocompetent surrogate animal model of HCV infection that will help in evaluation of the protective efficacy of newly developing HCV vaccine candidates. To circumvent the narrow host range of HCV, an HCV genotype 1b-based chimeric clone carrying E1, E2 and p6 regions from GB virus B (GBV-B), which is closely related to HCV, was generated. The chimera between HCV and GBV-B, named HCV/G, replicated more efficiently as compared with the HCV clone in primary marmoset hepatocytes. Furthermore, it was found that the chimera persistently replicated in a tamarin for more than 2 years after intrahepatic inoculation of the chimeric RNA. Although relatively low (
- Published
- 2016
37. Tracking cells implanted into cynomolgus monkeys (Macaca fascicularis) using MRI
- Author
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Sachi Okabayashi, Kiichi Kanayama, Hiroshi Koie, Yuko Katakai, Yasuhiro Yasutomi, Chieko Ohno, Hiroaki Shibata, Naohide Ageyama, and Yasuyo Ito-Fujishiro
- Subjects
0301 basic medicine ,Pathology ,medicine.medical_specialty ,General Veterinary ,medicine.diagnostic_test ,business.industry ,Magnetic resonance imaging ,General Medicine ,Blood flow ,030204 cardiovascular system & hematology ,medicine.disease ,Peripheral blood mononuclear cell ,Regenerative medicine ,General Biochemistry, Genetics and Molecular Biology ,Transplantation ,03 medical and health sciences ,030104 developmental biology ,0302 clinical medicine ,Triceps surae muscle ,Heart failure ,medicine ,Animal Science and Zoology ,Stem cell ,business - Abstract
Regenerative therapy with stem cell transplantation is used to treat various diseases such as coronary syndrome and Buerger's disease. For instance, stem-cell transplantation into the infarcted myocardium is an innovative and promising strategy for treating heart failure due to ischemic heart disease. Basic studies using small animals have shown that transplanted cells improve blood flow in the infarcted region. Magnetic resonance imaging (MRI) can noninvasively identify and track transplanted cells labeled with superparamagnetic iron oxide (SPIO). Although clinical regenerative therapies have been clinically applied to patients, the fate of implanted cells remains unknown. In addition, follow-up studies have shown that some adverse events can occur after recovery. Therefore, the present study evaluated the ability of MRI using a 3T scanner to track implanted peripheral blood mononuclear cells labeled with SPIO on days 0 and 7 after intramuscular (i.m.) and intravenous (i.v.) injection into a cynomolgus monkey. Labeled cells were visualized at the liver and triceps surae muscle on MR images using T1- and T2-weighted sequences and histologically localized by Prussian blue staining. The transplanted cells were tracked without abnormal clinical manifestations throughout this study. Hence, MRI of cynomolgus monkey transplanted SPIO-labeled cells is a safe and efficient method of tracking labeled cells that could help to determine the mechanisms involved in regenerative therapy.
- Published
- 2016
38. Adaptation of the Plasmodium falciparum FCB strain for in vitro and in vivo analysis in squirrel monkeys (Saimiri sciureus)
- Author
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Sawako Itagaki, Takahiro Tougan, Yuko Katakai, Yasuhiro Yasutomi, Nobuko Arisue, and Toshihiro Horii
- Subjects
0301 basic medicine ,Plasmodium falciparum ,Biology ,Parasitemia ,03 medical and health sciences ,In vivo ,parasitic diseases ,medicine ,Parasite hosting ,Animals ,Saimiri ,Squirrel monkey ,Saimiri sciureus ,medicine.disease ,biology.organism_classification ,Virology ,Adaptation, Physiological ,Red blood cell ,Disease Models, Animal ,030104 developmental biology ,Infectious Diseases ,medicine.anatomical_structure ,Parasitology ,Laboratories ,Genome, Protozoan ,Malaria ,Spleen - Abstract
The asexual blood stages of the Plasmodium falciparum parasite are responsible for inducing the clinical symptoms and the most severe presentations of malaria infection that causes frequent mortality and morbidity in tropical and subtropical areas of the world, making the blood stages of infection a key target of new malaria treatment and prevention strategies. Progress towards the development of more effective treatment and prevention strategies has been hindered by the limited availability of infection models that permit the sequential analysis of blood stage parasites in vitro followed by in vivo analysis to confirm therapeutic benefits. To advance a model for in vitro and in vivo analysis of blood stage parasites, we examined nine laboratory strains of P. falciparum to determine which strains could adapt to growth in vivo in splenectomized squirrel monkeys (Saimiri sciureus). Only one of the nine laboratory strains tested, the FCB strain, adapted to in vivo growth. Morphological analysis show that the adapted ring-stage parasites have a different morphology from original parasites cultured in vitro, and more often they were found to localize at the edge of the infected red blood cell. No remarkable differences were observed for both trophozoites and schizonts. The adapted strain can be cultured back in vitro similar to the original parasite, indicating that the adapted parasite can develop both in vitro and in vivo. This squirrel monkey-adapted P. falciparum parasite is expected to be suitable and is advantageous for the research and development of vaccines and antimalarial drugs.
- Published
- 2018
39. Immunological association of inducible bronchus-associated lymphoid tissue organogenesis in Ag85B-rHPIV2 vaccine-induced anti-tuberculosis mucosal immune responses in mice
- Author
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Ken Ishii, Sakiko Morimoto, Kentaro Ogami, Yusuke Tsujimura, Jun Kunisawa, Yasuko Wada, Yasuhiro Yasutomi, Takahiro Nagatake, Etsushi Kuroda, Ayaka Nasu, Soichiro Hirata, Norifumi Iijima, Tetsuya Nosaka, Michiko Shimojou, Koji Hosomi, Naomi Matsumoto, Mitsuo Kawano, and Hidehiko Suzuki
- Subjects
0301 basic medicine ,Immunoglobulin A ,Lymphoid Tissue ,Organogenesis ,Immunology ,Inflammation ,03 medical and health sciences ,Mice ,Immune system ,Antigen ,medicine ,Immunology and Allergy ,Animals ,Tuberculosis Vaccines ,Antigens, Bacterial ,biology ,General Medicine ,Mycobacterium tuberculosis ,Eosinophil ,Parainfluenza Virus 2, Human ,Mice, Inbred C57BL ,030104 developmental biology ,Lymphatic system ,medicine.anatomical_structure ,biology.protein ,Antibody ,medicine.symptom ,Tuberculosis vaccines ,Acyltransferases - Abstract
We previously reported that Ag85B-expressing human parainfluenza type 2 virus (Ag85B-rHPIV2) was effective as a nasal vaccine against tuberculosis in mice; however, the mechanism by which it induces an immune response remains to be investigated. In the present study, we found that organogenesis of inducible bronchus-associated lymphoid tissue (iBALT) played a role in the induction of antigen-specific T cells and IgA antibody responses in the lung of mice intra-nasally administered Ag85B-rHPIV2. We found that expression of Ag85B was dispensable for the development of iBALT, suggesting that HPIV2 acted as an iBALT-inducing vector. When iBALT organogenesis was disrupted in Ag85B-rHPIV2-immunized mice, either by neutralization of the lymphotoxin pathway or depletion of CD11b+ cells, Ag85B-specific immune responses (i.e. IFN γ-producing T cells and IgA antibody) were diminished in the lung. Furthermore, we found that immunization with Ag85B-rHPIV2 induced neutrophil and eosinophil infiltration temporally after the immunization in the lung. Thus, our results show that iBALT organogenesis contributes to the induction of antigen-specific immune responses by Ag85B-rHPIV2 and that Ag85B-rHPIV2 provokes its immune responses without inducing long-lasting inflammation.
- Published
- 2018
40. Nanogel-based pneumococcal surface protein A nasal vaccine induces microRNA-associated Th17 cell responses with neutralizing antibodies against Streptococcus pneumoniae in macaques
- Author
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David E. Briles, Kazunari Akiyoshi, Yoshikazu Yuki, Haruko Takahashi, Hideo Tsukada, Shiho Kurokawa, Norihiro Harada, Shigeo Takeda, Hiroshi Kiyono, Mio Mejima, Yasuhiro Yasutomi, H Shibata, Shin-ichi Sawada, Kohtaro Fujihashi, Yuko Katakai, Sunyi Joo, Yoshiko Fukuyama, and Eun Jeong Park
- Subjects
Male ,Nasal cavity ,Immunology ,medicine.disease_cause ,Article ,Microbiology ,Th2 Cells ,Immune system ,Bacterial Proteins ,Streptococcus pneumoniae ,otorhinolaryngologic diseases ,medicine ,Animals ,Humans ,Immunology and Allergy ,Glucans ,Administration, Intranasal ,Drug Carriers ,biology ,Pneumonia, Pneumococcal ,biology.organism_classification ,Antibodies, Bacterial ,Antibodies, Neutralizing ,Vaccination ,Macaca fascicularis ,MicroRNAs ,Rhesus macaque ,medicine.anatomical_structure ,biology.protein ,Nanoparticles ,Female ,Nasal administration ,Antibody ,Gels ,Respiratory tract - Abstract
We previously established a nanosized nasal vaccine delivery system by using a cationic cholesteryl group-bearing pullulan nanogel (cCHP nanogel), which is a universal protein-based antigen-delivery vehicle for adjuvant-free nasal vaccination. In the present study, we examined the central nervous system safety and efficacy of nasal vaccination with our developed cCHP nanogel containing pneumococcal surface protein A (PspA-nanogel) against pneumococcal infection in nonhuman primates. When [(18)F]-labeled PspA-nanogel was nasally administered to a rhesus macaque (Macaca mulatta), longer-term retention of PspA was noted in the nasal cavity when compared with administration of PspA alone. Of importance, no deposition of [(18)F]-PspA was seen in the olfactory bulbs or brain. Nasal PspA-nanogel vaccination effectively induced PspA-specific serum IgG with protective activity and mucosal secretory IgA (SIgA) Ab responses in cynomolgus macaques (Macaca fascicularis). Nasal PspA-nanogel-induced immune responses were mediated through T-helper (Th) 2 and Th17 cytokine responses concomitantly with marked increases in the levels of miR-181a and miR-326 in the serum and respiratory tract tissues, respectively, of the macaques. These results demonstrate that nasal PspA-nanogel vaccination is a safe and effective strategy for the development of a nasal vaccine for the prevention of pneumonia in humans.
- Published
- 2015
41. Contrasting infection susceptibility of the Japanese macaques and cynomolgus macaques to closely related malaria parasites, Plasmodium vivax and Plasmodium cynomolgi
- Author
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Yasuhiro Yasutomi, Toshihiro Horii, Yuko Katakai, Hideo Takahashi, Toru Nakade, Kazuyuki Tanabe, Shin-Ichiro Tachibana, and Satoru Kawai
- Subjects
Serotype ,Molecular Sequence Data ,Plasmodium vivax ,Protozoan Proteins ,Antigens, Protozoan ,Receptors, Cell Surface ,Old World monkey ,Macaque ,Species Specificity ,Antigen ,biology.animal ,parasitic diseases ,Malaria, Vivax ,medicine ,Animals ,Humans ,Amino Acid Sequence ,Serotyping ,Infectivity ,biology ,Monkey Diseases ,biology.organism_classification ,medicine.disease ,Macaca mulatta ,Virology ,Malaria ,Macaca fascicularis ,Japanese macaque ,Infectious Diseases ,Macaca ,Parasitology ,Disease Susceptibility ,Plasmodium cynomolgi - Abstract
Although the human malaria parasite Plasmodium vivax is closely related to Asian Old World monkey malaria parasites, there are no reports of P. vivax infections in macaques. In this study, we compared the infectivity of P. vivax and Plasmodium cynomolgi in Japanese macaques (Macaca fuscata) and in cynomolgus macaques (Macaca fascicularis). The Japanese macaques were highly susceptible to P. cynomolgi but not to P. vivax, whereas cynomolgus macaques showed mild/limited P. cynomolgi infection and were, also, not susceptible to P. vivax. Serotyping and amino acid sequence comparison of erythrocyte surface Duffy antigen/receptor for chemokines (DARC) indicate that the Japanese macaque DARC sequence is nearly identical to that of rhesus (Macaca mulatta) and cynomolgus macaques. This suggests that the macaques share a common mechanism for preventing P. vivax infection. Comparison of amino acid sequences of the Duffy-binding-like (DBL) domain from several different Plasmodium species suggests that P. vivax DBLs will not bind to macaque DARCs, which can explain the lack of P. vivax infectivity. The DBL sequence analyses also suggest that P. cynomolgi DBLs may target Japanese macaque erythrocytes through a DARC-independent interaction.
- Published
- 2015
42. Interferon-β response is impaired by hepatitis B virus infection in Tupaia belangeri
- Author
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Naoki Yamamoto, Mohammad Enamul Hoque Kayesh, Yasuhito Tanaka, Takahiro Sanada, Sayeh Ezzikouri, Chimène Nze Nkogue, Jun-ichiro Takano, Yasuhiro Yasutomi, Hitoshi Hatai, Haiying Chi, Michinori Kohara, Yumiko Shiogama, Takumi Haraguchi, Kyoko Tsukiyama-Kohara, Bouchra Kitab, Shuko Murakami, Noriaki Miyoshi, and Rika Matsuyama
- Subjects
0301 basic medicine ,Cancer Research ,HBsAg ,Hepatitis B virus ,Tupaia ,medicine.disease_cause ,Virus Replication ,03 medical and health sciences ,Tupaia belangeri ,Hepatitis B, Chronic ,Interferon ,Virology ,medicine ,Animals ,Immune Evasion ,Hepatitis B Surface Antigens ,biology ,Liver cell ,Gene Expression Profiling ,Toll-Like Receptors ,virus diseases ,TLR9 ,Interferon-beta ,biology.organism_classification ,digestive system diseases ,Immunity, Innate ,Chronic infection ,Disease Models, Animal ,030104 developmental biology ,Infectious Diseases ,Immunology ,Host-Pathogen Interactions ,Hepatocytes ,medicine.drug - Abstract
To date, the chimpanzee has been used as the natural infection model for hepatitis B virus (HBV). However, as this model is very costly and difficult to use because of ethical and animal welfare issues, we aimed to establish the tupaia (Tupaia belangeri) as a new model for HBV infection and characterized its intrahepatic innate immune response upon HBV infection. First, we compared the propagation of HBV genotypes A2 and C in vivo in tupaia hepatocytes. At 8-10days post infection (dpi), the level of HBV-A2 propagation in the tupaia liver was found to be higher than that of HBV-C. Abnormal architecture of liver cell cords and mitotic figures were also observed at 8 dpi with HBV-A2. Moreover, we found that HBV-A2 established chronic infection in some tupaias. We then aimed to characterize the intrahepatic innate immune response in this model. First, we infected six tupaias with HBV-A2 (strains JP1 and JP4). At 28 dpi, intrahepatic HBV-DNA and serum hepatitis B surface antigens (HBsAg) were detected in all tupaias. The levels of interferon (IFN)-β were found to be significantly suppressed in the three tupaias infected with HBV A2_JP4, while no significant change was observed in the three infected with HBV A2_JP1. Expression of toll-like receptor (TLR) 1 was suppressed, while that of TLR3 and TLR9 were induced, in HBV A2_JP1-infected tupaias. Expression of TLR8 was induced in all tupaias. Next, we infected nine tupaias with HBV-A2 (JP1, JP2, and JP4), and characterized the infected animals after 31 weeks. Serum HBsAg levels were detected at 31 weeks post-infection (wpi) and IFN-β was found to be significantly suppressed in all tupaias. TLR3 was not induced, except in tupaia #93 and #96. Suppression of TLR9 was observed in all tupaias, except tupaia #93. Also, we investigated the expression levels of cyclic GMP-AMP synthase, which was found to be induced in all tupaias at 28 dpi and in four tupaias at 31 wpi. Additionally, we evaluated the expression levels of sodium-taurocholate cotransporting polypeptide, which was found to be suppressed during chronic HBV infection. Thus, the tupaia infection model of HBV clearly indicated the suppression of IFN-β at 31 wpi, which might have contributed to the establishment of chronic HBV infection.
- Published
- 2016
43. Tau pathology in aged cynomolgus monkeys is progressive supranuclear palsy/corticobasal degeneration- but not Alzheimer disease-like -Ultrastructural mapping of tau by EDX
- Author
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Nobuhiro Shimozawa, Hiromi Kondo, Yasuhiro Yasutomi, Eijiro Adachi, Sachi Okabayashi, Kentaro Endo, Nobuyuki Kimura, and Toshiki Uchihara
- Subjects
0301 basic medicine ,Male ,Pathology ,medicine.medical_specialty ,Aging ,Amyloid ,Hippocampus ,tau Proteins ,Biology ,Pathology and Forensic Medicine ,Progressive supranuclear palsy ,03 medical and health sciences ,Cellular and Molecular Neuroscience ,0302 clinical medicine ,Alzheimer Disease ,Basal ganglia ,mental disorders ,medicine ,Corticobasal degeneration ,Immuno electron microscopy ,Animals ,Microscopy, Immunoelectron ,Neurons ,Neocortex ,Research ,Aged monkey ,Brain ,Spectrometry, X-Ray Emission ,Human brain ,medicine.disease ,Immunohistochemistry ,Four-repeat tau ,Macaca fascicularis ,Oligodendroglia ,030104 developmental biology ,medicine.anatomical_structure ,Astrocytes ,Female ,Neurology (clinical) ,Energy-dispersive X-ray analysis ,Supranuclear Palsy, Progressive ,Alzheimer's disease ,Neuroscience ,030217 neurology & neurosurgery - Abstract
Concomitant deposition of amyloid -beta protein (Aβ) and neuronal tau as neurofibrillary tangles in the human brain is a hallmark of Alzheimer disease (AD). Because these deposits increase during normal aging, it has been proposed that aging brains may also undergo AD-like changes. To investigate the neuropathological changes that occur in the aging primate brain, we examined 21 brains of cynomolgus monkeys (7–36 years old) for Aβ- and tau-positive lesions. We found, 1) extensive deposition of Aβ in brains of cynomolgus monkeys over 25 years of age, 2) selective deposition of 4-repeat tau as pretangles in neurons, and as coiled body-like structures in oligodendroglia-like cells and astrocytes, 3) preferential distribution of tau in the basal ganglia and neocortex rather than the hippocampus, and 4) age-associated increases in 30–34 kDa AT8- and RD4-positive tau fragments in sarkosyl-insoluble fractions. We further labeled tau-positive structures using diaminobezidine enhanced with nickel, and visualized nickel-labeled structures by energy-dispersive X-ray (EDX) analysis of ultrathin sections. This allowed us to distinguish between nickel-labeled tau and background electron-dense structures, and we found that tau localized to 20–25 nm straight filaments in oligodendroglia-like cells and neurons. Our results indicate that the cytopathology and distribution of tau deposits in aged cynomolgus brains resemble those of progressive supranuclear palsy (PSP) and corticobasal degeneration (CBD) rather than AD. Thus, even in the presence of Aβ, age-associated deposition of tau in non-human primates likely does not occur through AD-associated mechanisms. Electronic supplementary material The online version of this article (doi:10.1186/s40478-016-0385-5) contains supplementary material, which is available to authorized users.
- Published
- 2016
44. Pitavastatin Regulates Helper T-Cell Differentiation and Ameliorates Autoimmune Myocarditis in Mice
- Author
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Michiaki Hiroe, Kyoko Imanaka-Yoshida, Kazutaka Aonuma, Yasuhiro Yasutomi, Tomoko Machino-Ohtsuka, Kazuko Tajiri, Nobutake Shimojo, Yusuke Tsujimura, Satoshi Sakai, Akira Sato, and Taizo Kimura
- Subjects
Male ,Autoimmune myocarditis ,Statin ,medicine.drug_class ,Cardiomyopathy ,Inflammation ,Mice, SCID ,medicine.disease_cause ,Autoimmune Diseases ,Autoimmunity ,Interferon-gamma ,Mice ,medicine ,Animals ,Pharmacology (medical) ,Pitavastatin ,Cell Proliferation ,Pharmacology ,Helper T cell differentiation ,Mice, Inbred BALB C ,business.industry ,Interleukin-17 ,Cell Differentiation ,General Medicine ,Th1 Cells ,medicine.disease ,Myocarditis ,Immunology ,Quinolines ,Th17 Cells ,Hydroxymethylglutaryl-CoA Reductase Inhibitors ,medicine.symptom ,Cardiology and Cardiovascular Medicine ,business ,Th17 cytokines ,medicine.drug - Abstract
Experimental autoimmune myocarditis (EAM) is a mouse model of inflammatory cardiomyopathy, and the involvement of T helper (Th) 1 and Th17 cytokines has been demonstrated. Accumulated evidence has shown that statins have anti-inflammatory and immunomodulatory effects; however, the mechanism has not been fully elucidated. This study was designed to test the hypothesis that pitavastatin affects T cell-mediated autoimmunity through inhibiting Th1 and Th17 responses and reduces the severity of EAM in mice.The EAM model was established in BALB/c mice by immunization with murine α-myosin heavy chain. Mice were fed pitavastatin (5 mg/kg) or vehicle once daily for 3 weeks from day 0 to day 21 after immunization.Pitavastatin reduced the pathophysiological severity of the myocarditis. Pitavastatin treatment inhibited the phosphorylation of signal transducer and activator of transcription (STAT)3 and STAT4, which have key roles in the Th1 and Th17 lineage commitment, respectively, in the heart, and suppressed production of Th1 cytokine interferon-γ and Th17 cytokine interleukin-17 from autoreactive CD4(+) T cells. In in vitro T-cell differentiation experiments, pitavastatin-treated T cells failed to differentiate into Th1 and Th17 cells through inhibiting the transcription of T-box expressed in T-cells (T-bet) and RAR-related orphan receptor γt (RORγT) which have critical roles in the development of Th1 and Th17 cells, respectively, and this failure was rescued by adding mevalonate.Pitavastatin inhibits Th1 and Th17 responses and ameliorates EAM. These results suggest that statins may be a promising novel therapeutic strategy for the clinical treatment of myocarditis and inflammatory cardiomyopathy.
- Published
- 2013
45. Dynamics of cellular immune responses in the acute phase of dengue virus infection
- Author
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Hirofumi Akari, Tomohiko Takasaki, Tomoyuki Yoshida, Atsunori Higashino, Yuko Katakai, Akatsuki Saito, Terue Kurosawa, Yasuhiro Yasutomi, Yuki Iwasaki, Shinichiro Nakamura, Tsutomu Omatsu, Ichiro Kurane, and Masataka Hamano
- Subjects
CD4-Positive T-Lymphocytes ,Male ,medicine.medical_specialty ,T-Lymphocytes ,viruses ,CD8-Positive T-Lymphocytes ,Dengue virus ,Virus Replication ,medicine.disease_cause ,Dengue fever ,Dengue ,Medical microbiology ,Immune system ,Virology ,medicine ,Animals ,Severe Dengue ,Neutralizing antibody ,Immunity, Cellular ,biology ,virus diseases ,Callithrix ,General Medicine ,Dengue Virus ,biochemical phenomena, metabolism, and nutrition ,medicine.disease ,Natural killer T cell ,Antibodies, Neutralizing ,Immunity, Humoral ,Viral replication ,Immunology ,biology.protein ,Natural Killer T-Cells ,Immunologic Memory ,CD8 - Abstract
In this study, we examined the dynamics of cellular immune responses in the acute phase of dengue virus (DENV) infection in a marmoset model. Here, we found that DENV infection in marmosets greatly induced responses of CD4/CD8 central memory T and NKT cells. Interestingly, the strength of the immune response was greater in animals infected with a dengue fever strain than in those infected with a dengue hemorrhagic fever strain of DENV. In contrast, when animals were re-challenged with the same DENV strain used for primary infection, the neutralizing antibody induced appeared to play a critical role in sterilizing inhibition against viral replication, resulting in strong but delayed responses of CD4/CD8 central memory T and NKT cells. The results in this study may help to better understand the dynamics of cellular and humoral immune responses in the control of DENV infection.
- Published
- 2013
46. Gag-CA Q110D mutation elicits TRIM5-independent enhancement of HIV-1mt replication in macaque cells
- Author
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Akatsuki Saito, Ken Kono, Masako Nomaguchi, Hirofumi Akari, Emi E. Nakayama, Hironori Sato, Yasuhiro Yasutomi, Masaru Yokoyama, Tatsuo Shioda, Akio Adachi, and Tetsuro Matano
- Subjects
Immunology ,Mutagenesis (molecular biology technique) ,HIV Infections ,Biology ,Virus Replication ,medicine.disease_cause ,gag Gene Products, Human Immunodeficiency Virus ,Microbiology ,Macaque ,Cell Line ,Adaptive mutation ,biology.animal ,medicine ,Animals ,Humans ,Infectivity ,Mutation ,Virology ,Infectious Diseases ,Viral replication ,Capsid ,Cell culture ,Host-Pathogen Interactions ,Cats ,HIV-1 ,Mutagenesis, Site-Directed ,Macaca ,Capsid Proteins ,Carrier Proteins - Abstract
HIV-1 is strictly adapted to humans, and cause disease-inducing persistent infection only in humans. We have generated a series of macaque-tropic HIV-1 (HIV-1mt) to establish non-human primate models for basic and clinical studies. HIV-1mt clones available to date grow poorly in macaque cells relative to SIVmac239. In this study, viral adaptive mutation in macaque cells, G114E in capsid (CA) helix 6 of HIV-1mt, that enhances viral replication was identified. Computer-assisted structural analysis predicted that another Q110D mutation in CA helix 6 would also increase viral growth potential. A new proviral construct MN4Rh-3 carrying CA-Q110D exhibited exquisitely enhanced growth property specifically in macaque cells. Susceptibility of MN4Rh-3 to macaque TRIM5α/TRIMCyp proteins was examined by their expression systems. HIV-1mt clones so far constructed already completely evaded TRIMCyp restriction, and further enhancement of TRIMCyp resistance by Q110D was not observed. In addition, Q110D did not contribute to evasion from TRIM5α restriction. However, the single-cycle infectivity of MN4Rh-3 in macaque cells was enhanced relative to the other HIV-1mt clones. Our results here indicate that CA-Q110D accelerates viral growth in macaque cells irrelevant to TRIM5 proteins restriction.
- Published
- 2013
47. Single systemic administration of Ag85B of mycobacteria DNA inhibits allergic airway inflammation in a mouse model of asthma
- Author
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Yasuhiro Yasutomi, Hiroyasu Inada, Yumiko Shiogama, Mitsuo Kawano, Katsuo Karamatsu, Kazuhiro Matsuo, Akihiro Matsubara, and Yusuke Tsujimura
- Subjects
Pulmonary and Respiratory Medicine ,lcsh:Immunologic diseases. Allergy ,Allergy ,mycobacteria ,medicine.medical_treatment ,Ag85B ,Immunoglobulin E ,Immune system ,Antigen ,Journal of Asthma and Allergy ,medicine ,Immunology and Allergy ,Original Research ,biology ,business.industry ,Immunotherapy ,Eosinophil ,respiratory system ,asthma ,medicine.disease ,allergy ,Ovalbumin ,medicine.anatomical_structure ,Immunology ,Systemic administration ,biology.protein ,immunotherapy ,business ,lcsh:RC581-607 - Abstract
Katsuo Karamatsu,1,2 Kazuhiro Matsuo,3 Hiroyasu Inada,4 Yusuke Tsujimura,1 Yumiko Shiogama,1,2 Akihiro Matsubara,1,2 Mitsuo Kawano,5 Yasuhiro Yasutomi1,21Laboratory of Immunoregulation and Vaccine Research, Tsukuba Primate Research Center, National Institute of Biomedical Innovation, Tsukuba, 2Division of Immunoregulation, Department of Molecular and Experimental Medicine, Mie University Graduate School of Medicine, Tsu, 3Department of Research and Development, Japan BCG Laboratory, Tokyo, 4Department of Pathology, Suzuka University of Medical Science, Suzuka, 5Department of Microbiology and Molecular Genetics, Mie University Graduate School of Medicine, Tsu, JapanAbstract: The immune responses of T-helper (Th) and T-regulatory cells are thought to play a crucial role in the pathogenesis of allergic airway inflammation observed in asthma. The correction of immune response by these cells should be considered in the prevention and treatment of asthma. Native antigen 85B (Ag85B) of mycobacteria, which cross-reacts among mycobacteria species, may play an important biological role in host–pathogen interaction since it elicits various immune responses by activation of Th cells. The current study investigated the antiallergic inflammatory effects of DNA administration of Ag85B from Mycobacterium kansasii in a mouse model of asthma. Immunization of BALB/c mice with alum-adsorbed ovalbumin followed by aspiration with aerosolized ovalbumin resulted in the development of allergic airway inflammation. Administration of Ag85B DNA before the aerosolized ovalbumin challenge protected the mice from subsequent induction of allergic airway inflammation. Serum and bronchoalveolar lavage immunoglobulin E levels, extent of eosinophil infiltration, and levels of Th2-type cytokines in Ag85B DNA-administered mice were significantly lower than those in control plasmid-immunized mice, and levels of Th1- and T-regulatory-type cytokines were enhanced by Ag85B administration. The results of this study provide evidence for the potential utility of Ag85B DNA inoculation as a novel approach for the treatment of asthma.Keywords: immunotherapy, asthma, Ag85B, mycobacteria, allergy
- Published
- 2012
48. Allergy Vaccines Using a Mycobacterium-Secreted Antigen, Ag85B, and an IL-4 Antagonist
- Author
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Yusuke Tsujimura and Yasuhiro Yasutomi
- Subjects
0301 basic medicine ,Allergy ,Atopic dermatitis ,Biology ,medicine.disease ,Acquired immune system ,030207 dermatology & venereal diseases ,03 medical and health sciences ,030104 developmental biology ,0302 clinical medicine ,Immune system ,Antigen ,Immunology ,medicine ,Hay fever ,Interleukin 4 ,Asthma - Abstract
In recent decades, the prevalence of allergic diseases, including bronchial asthma, airway hypersensitivity, hay fever, and atopic dermatitis, has been increasing in the industrialized world, and effective treatments probably require manipulating the inflammatory response to pathogenic allergens. T helper (Th) 2 cells are thought to play a crucial role in the initiation, progression, and persistence of allergic responses in association with production of interleukin (IL)-4, IL-5, and IL-13. Therefore, a strategy of a shift from Th2- to Th1-type immune response may be valuable in the prophylaxis and management of allergic diseases. It is also necessary to develop prophylactic and therapeutic treatment that induces homeostatic functions in the multifaceted allergic environment, because various factors including innate and adaptive immunity, mucosal immune response, and functional and structural maintenance of local tissue might be involved in the pathogenesis of allergic disorders. We review herein recent findings related to the curative effect for mouse models of asthma and atopic dermatitis using DNA-, virus-, and protein-based vaccines of a Mycobacterium secretion antigen, Ag85B, and a plasmid encoding cDNA of antagonistic IL-4 mutant.
- Published
- 2016
49. Suppressor of Cytokine Signaling 1 DNA Administration Inhibits Inflammatory and Pathogenic Responses in Autoimmune Myocarditis
- Author
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Satoshi Sakai, Yasuhiro Yasutomi, Nobutake Shimojo, Kazuko Tajiri, Tetsuji Naka, Yusuke Tsujimura, Michiaki Hiroe, Akihiro Matsubara, Kyoko Imanaka-Yoshida, and Kazutaka Aonuma
- Subjects
Male ,Myocarditis ,T-Lymphocytes ,medicine.medical_treatment ,Immunology ,Cardiomyopathy ,Suppressor of Cytokine Signaling Proteins ,Mice, SCID ,Real-Time Polymerase Chain Reaction ,Transfection ,medicine.disease_cause ,Autoimmune Diseases ,Autoimmunity ,Mice ,Suppressor of Cytokine Signaling 1 Protein ,medicine ,Animals ,Immunology and Allergy ,Inflammation ,Mice, Inbred BALB C ,Innate immune system ,Reverse Transcriptase Polymerase Chain Reaction ,business.industry ,Suppressor of cytokine signaling 1 ,Gene Transfer Techniques ,Dilated cardiomyopathy ,DNA ,Flow Cytometry ,medicine.disease ,Acquired immune system ,Adoptive Transfer ,Cytokine ,Models, Animal ,business - Abstract
Myocarditis and subsequent dilated cardiomyopathy are major causes of heart failure in young adults. Myocarditis in humans is highly heterogeneous in etiology. Recent studies have indicated that a subgroup of myocarditis patients may benefit from immune-targeted therapies, because autoimmunity plays an important role in myocarditis as well as contributing to the progression to cardiomyopathy and heart failure. Suppressor of cytokine signaling (SOCS) 1 plays a key role in the negative regulation of both TLR- and cytokine receptor-mediated signaling, which is involved in innate immunity and subsequent adaptive immunity. In this study, we investigated the therapeutic effect of SOCS1 DNA administration on experimental autoimmune myocarditis (EAM) in mice. EAM was induced by s.c. immunization with cardiac-specific peptides derived from α myosin H chain in BALB/c mice. In contrast to control myocarditis mice, SOCS1 DNA-injected mice were protected from development of EAM and heart failure. SOCS1 DNA administration was effective for reducing the activation of autoreactive CD4+ T cells by inhibition of the function of Ag-presenting dendritic cells. Our findings suggest that SOCS1 DNA administration has considerable therapeutic potential in individuals with autoimmune myocarditis and dilated cardiomyopathy.
- Published
- 2012
50. Discovery of a Cynomolgus Monkey Family With Retinitis Pigmentosa
- Author
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Toshio Hisatomi, Yasuhiro Ikeda, Shigeo Yoshida, Toru Nakazawa, Koh Hei Sonoda, Kohta Fujiwara, Tatsuhiko Tsunoda, Nobuhiro Shimozawa, Jun Funatsu, Fuyuki Miya, Shunji Nakatake, Koji M. Nishiguchi, Yasuhiro Yasutomi, Takashi Tachibana, Yusuke Murakami, and Tatsuro Ishibashi
- Subjects
Male ,0301 basic medicine ,Retinal degeneration ,medicine.medical_specialty ,genetic structures ,Polymerase Chain Reaction ,Macular Edema ,Retina ,Ophthalmoscopy ,03 medical and health sciences ,biology.animal ,Ophthalmology ,Exome Sequencing ,Retinitis pigmentosa ,Electroretinography ,medicine ,Animals ,Primate ,Fluorescein Angiography ,biology ,medicine.diagnostic_test ,business.industry ,Fundus photography ,medicine.disease ,Fluorescein angiography ,eye diseases ,Pedigree ,Disease Models, Animal ,Macaca fascicularis ,030104 developmental biology ,medicine.anatomical_structure ,Female ,sense organs ,business ,Retinitis Pigmentosa ,Tomography, Optical Coherence - Abstract
Purpose To accelerate the development of new therapies, an inherited retinal degeneration model in a nonhuman primate would be useful to confirm the efficacy in preclinical studies. In this study, we describe the discovery of retinitis pigmentosa in a cynomolgus monkey (Macaca fascicularis) pedigree. Methods First, screening with fundus photography was performed on 1443 monkeys at the Tsukuba Primate Research Center. Ophthalmic examinations, such as indirect ophthalmoscopy, ERGs using RETeval, and optic coherent tomography (OCT) measurement, were then performed to confirm diagnosis. Results Retinal degeneration with cystoid macular edema was observed in both eyes of one 14-year-old female monkey. In her examinations, the full-field ERGs were nonrecordable and the outer layer of the retina in the parafoveal area was not visible on OCT imaging. Moreover, less frequent pigmentary retinal anomalies also were observed in her 3-year-old nephew. His full-field ERGs were almost nonrecordable and the outer layer was not visible in the peripheral retina. His father was her cousin (the son of her mother's older brother) and his mother was her younger half-sibling sister with a different father. Conclusions The hereditary nature is highly probable (autosomal recessive inheritance suspected). However, whole-exome analysis performed identified no pathogenic mutations in these monkeys.
- Published
- 2018
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