90 results on '"Retroviruses, Simian genetics"'
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2. High Prevalences and a Wide Genetic Diversity of Simian Retroviruses in Non-human Primate Bushmeat in Rural Areas of the Democratic Republic of Congo.
- Author
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Steve AM, Ahidjo A, Placide MK, Caroline F, Mukulumanya M, Simon-Pierre NK, Octavie LM, Valentin MA, Jean-Jacques MT, Eric D, and Martine P
- Subjects
- Animals, Antibodies, Viral blood, Deltaretrovirus Infections diagnosis, Deltaretrovirus Infections transmission, Democratic Republic of the Congo, Genetic Variation, Humans, Meat, Phylogeny, Prevalence, Retroviruses, Simian classification, Simian Acquired Immunodeficiency Syndrome diagnosis, Simian Acquired Immunodeficiency Syndrome transmission, Zoonoses transmission, Zoonoses virology, Primates virology, Retroviruses, Simian genetics, Retroviruses, Simian isolation & purification
- Abstract
Like the majority of emerging infectious diseases, HIV and HTLV are of zoonotic origin. Here we assess the risk of cross-species transmissions of their simian counterparts, SIV and STLV, from non-human primates (NHP) to humans in the Democratic Republic of Congo (DRC). A total of 331 samples, derived from NHP bushmeat, were collected as dried blood spots (DBS, n = 283) or as tissue samples (n = 36) at remote forest sites mainly in northern and eastern DRC. SIV antibody prevalences in DBS were estimated with a novel high throughput immunoassay with antigens representing the actual known diversity of HIV/SIV lineages. Antibody-positive samples were confirmed by PCR and sequence analysis. Screening for STLV infection was done with universal primers in tax, and new strains were further characterized in LTR. SIV and STLV infection in tissue samples was done by PCR only. Overall, 5 and 15.4% of NHP bushmeat was infected with SIV and STLV, respectively. A new SIV lineage was identified in Allen's swamp monkeys (Allenopithecus nigroviridis). Three new STLV-1 subtypes were identified in Allen's swamp monkeys (Allenopithecus nigroviridis), blue monkeys (Cercopithecus mitis), red-tailed guenons (Cercopithecus ascanius schmidti) and agile mangabeys (Cercocebus agilis). SIV and STLV prevalences varied according to species and geographic region. Our study illustrates clearly, even on a small sample size from a limited number of geographic areas, that our knowledge on the genetic diversity and geographic distribution of simian retroviruses is still limited and that humans continue to be exposed to relative high proportions on infected NHP bushmeat.
- Published
- 2017
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3. A novel simian retrovirus subtype discovered in cynomolgus monkeys (Macaca fascicularis).
- Author
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Zao CL, Tomanek L, Cooke A, Berger R, Yang L, Xie C, Chen S, Shi C, and Rong R
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- Animals, Macaca fascicularis virology, Open Reading Frames, Phylogeny, Retroviridae Infections virology, Retroviruses, Simian classification, Retroviruses, Simian genetics, Viral Proteins genetics, Monkey Diseases virology, Retroviridae Infections veterinary, Retroviruses, Simian isolation & purification
- Abstract
A new simian retrovirus (SRV) subtype was discovered in China and the USA from Cambodian-origin cynomolgus monkeys. Histopathological examination from necropsied animals showed multifocal lymphoplasmacystic and histocytic inflammation. The complete genome sequences demonstrated that the US virus isolates were nearly identical (99.91-99.93 %) and differed only slightly (99.13-99.16 % identical) from the China isolate. Phylogenetic analysis showed that the new virus isolates formed a distinct branch of SRV-1 through -7, and therefore were named this subtype, SRV-8. This SRV-8 variant was also phylogenetically and serologically more closely related to SRV-4 than any other SRV subtype.
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- 2016
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4. Experimental evaluation of the zoonotic infection potency of simian retrovirus type 4 using humanized mouse model.
- Author
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Sato K, Kobayashi T, Misawa N, Yoshikawa R, Takeuchi JS, Miura T, Okamoto M, Yasunaga J, Matsuoka M, Ito M, Miyazawa T, and Koyanagi Y
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- APOBEC Deaminases, Animals, Cell Line, Cytidine Deaminase, Cytokines metabolism, Cytosine Deaminase metabolism, Female, HEK293 Cells, Hematopoietic Stem Cell Transplantation, Hematopoietic Stem Cells cytology, Hematopoietic Stem Cells metabolism, Hematopoietic Stem Cells virology, Humans, Japan, Male, Mice, Mice, Inbred NOD, Mice, Transgenic, Models, Animal, Polymerase Chain Reaction, RNA, Viral analysis, Retroviridae Infections pathology, Retroviridae Infections virology, Retroviruses, Simian genetics, Retroviruses, Simian isolation & purification, Transplantation, Heterologous, Zoonoses virology, Retroviridae Infections transmission, Retroviruses, Simian pathogenicity, Zoonoses transmission
- Abstract
During 2001-2002 and 2008-2011, two epidemic outbreaks of infectious hemorrhagic disease have been found in Japanese macaques (Macaca fuscata) in Kyoto University Primate Research Institute, Japan. Following investigations revealed that the causative agent was simian retrovirus type 4 (SRV-4). SRV-4 was isolated by using human cell lines, which indicates that human cells are potently susceptible to SRV-4 infection. These raise a possibility of zoonotic infection of pathogenic SRV-4 from Japanese macaques into humans. To explore the possibility of zoonotic infection of SRV-4 to humans, here we use a human hematopoietic stem cell-transplanted humanized mouse model. Eight out of the twelve SRV-4-inoculated humanized mice were infected with SRV-4. Importantly, 3 out of the 8 infected mice exhibited anemia and hemophagocytosis, and an infected mouse died. To address the possibility that SRV-4 adapts humanized mouse and acquires higher pathogenicity, the virus was isolated from an infected mice exhibited severe anemia was further inoculated into another 6 humanized mice. However, no infected mice exhibited any illness. Taken together, our findings demonstrate that the zoonotic SRV-4 infection from Japanese macaques to humans is technically possible under experimental condition. However, such zoonotic infection may not occur in the real society.
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- 2015
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5. Emergence of infectious malignant thrombocytopenia in Japanese macaques (Macaca fuscata) by SRV-4 after transmission to a novel host.
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Okamoto M, Miyazawa T, Morikawa S, Ono F, Nakamura S, Sato E, Yoshida T, Yoshikawa R, Sakai K, Mizutani T, Nagata N, Takano J, Okabayashi S, Hamano M, Fujimoto K, Nakaya T, Iida T, Horii T, Miyabe-Nishiwaki T, Watanabe A, Kaneko A, Saito A, Matsui A, Hayakawa T, Suzuki J, Akari H, Matsuzawa T, and Hirai H
- Subjects
- Animals, Communicable Diseases, Emerging diagnosis, Communicable Diseases, Emerging transmission, Female, Genome, Viral, Macaca, Metagenomics methods, Phylogeny, RNA, Viral, Retroviridae Infections diagnosis, Retroviridae Infections transmission, Retroviruses, Simian isolation & purification, Retroviruses, Simian ultrastructure, Thrombocytopenia diagnosis, Communicable Diseases, Emerging complications, Communicable Diseases, Emerging virology, Retroviridae Infections complications, Retroviridae Infections virology, Retroviruses, Simian classification, Retroviruses, Simian genetics, Thrombocytopenia etiology
- Abstract
We discovered a lethal hemorrhagic syndrome arising from severe thrombocytopenia in Japanese macaques kept at the Primate Research Institute, Kyoto University. Extensive investigation identified that simian retrovirus type 4 (SRV-4) was the causative agent of the disease. SRV-4 had previously been isolated only from cynomolgus macaques in which it is usually asymptomatic. We consider that the SRV-4 crossed the so-called species barrier between cynomolgus and Japanese macaques, leading to extremely severe acute symptoms in the latter. Infectious agents that cross the species barrier occasionally amplify in virulence, which is not observed in the original hosts. In such cases, the new hosts are usually distantly related to the original hosts. However, Japanese macaques are closely related to cynomolgus macaques, and can even hybridize when given the opportunity. This lethal outbreak of a novel pathogen in Japanese macaques highlights the need to modify our expectations about virulence with regards crossing species barriers.
- Published
- 2015
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6. The genome landscape of the african green monkey kidney-derived vero cell line.
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Osada N, Kohara A, Yamaji T, Hirayama N, Kasai F, Sekizuka T, Kuroda M, and Hanada K
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- Animals, Chlorocebus aethiops, Female, Humans, Vero Cells, Chromosome Aberrations, Genome, Multigene Family, Proviruses genetics, Retroviruses, Simian genetics
- Abstract
Continuous cell lines that originate from mammalian tissues serve as not only invaluable tools for life sciences, but also important animal cell substrates for the production of various types of biological pharmaceuticals. Vero cells are susceptible to various types of microbes and toxins and have widely contributed to not only microbiology, but also the production of vaccines for human use. We here showed the genome landscape of a Vero cell line, in which 25,877 putative protein-coding genes were identified in the 2.97-Gb genome sequence. A homozygous ∼9-Mb deletion on chromosome 12 caused the loss of the type I interferon gene cluster and cyclin-dependent kinase inhibitor genes in Vero cells. In addition, an ∼59-Mb loss of heterozygosity around this deleted region suggested that the homozygosity of the deletion was established by a large-scale conversion. Moreover, a genomic analysis of Vero cells revealed a female Chlorocebus sabaeus origin and proviral variations of the endogenous simian type D retrovirus. These results revealed the genomic basis for the non-tumourigenic permanent Vero cell lineage susceptible to various pathogens and will be useful for generating new sub-lines and developing new tools in the quality control of Vero cells., (© The Author 2014. Published by Oxford University Press on behalf of Kazusa DNA Research Institute.)
- Published
- 2014
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7. miR-190b is markedly upregulated in the intestine in response to simian immunodeficiency virus replication and partly regulates myotubularin-related protein-6 expression.
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Mohan M, Chandra LC, Torben W, Aye PP, Alvarez X, and Lackner AA
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- Animals, CD4-Positive T-Lymphocytes immunology, CD4-Positive T-Lymphocytes virology, Colon immunology, Colon metabolism, Colon virology, Down-Regulation genetics, Down-Regulation immunology, Genes, Reporter, Intestinal Mucosa immunology, Intestinal Mucosa virology, Jejunum immunology, Jejunum metabolism, Jejunum virology, Luciferases genetics, Macaca mulatta, MicroRNAs biosynthesis, Protein Tyrosine Phosphatases, Non-Receptor biosynthesis, RNA, Viral genetics, RNA, Viral immunology, Retroviruses, Simian immunology, Simian Acquired Immunodeficiency Syndrome immunology, Simian Immunodeficiency Virus immunology, Up-Regulation immunology, Virus Replication immunology, Intestinal Mucosa metabolism, MicroRNAs genetics, Protein Tyrosine Phosphatases, Non-Receptor genetics, Retroviruses, Simian genetics, Simian Acquired Immunodeficiency Syndrome genetics, Simian Immunodeficiency Virus genetics, Up-Regulation genetics, Virus Replication genetics
- Abstract
HIV replication and the cellular micro-RNA (miRNA) machinery interconnect at several posttranscriptional levels. To understand their regulatory role in the intestine, a major site of HIV/SIV replication, dissemination, and CD4(+) T cell depletion, we profiled miRNA expression in colon following SIV infection (10 acute SIV, 5 uninfected). Nine (four up and five down) miRNAs showed statistically significant differential expression. Most notably, miR-190b expression showed high statistical significance (adjusted p = 0.0032), the greatest fold change, and was markedly elevated in colon and jejunum throughout SIV infection. In addition, miR-190b upregulation was detected before peak viral replication and the nadir of CD4(+) T cell depletion predominantly in lamina propria leukocytes. Interestingly non-SIV-infected macaques with diarrhea and colitis failed to upregulate miR-190b, suggesting that its upregulation was neither inflammation nor immune-activation driven. SIV infection of in vitro-cultured CD4(+) T cells and primary intestinal macrophages conclusively identified miR-190b upregulation to be driven in response to viral replication. Further miR-190b expression levels in colon and jejunum positively correlated with tissue viral loads. In contrast, mRNA expression of myotubularin-related protein 6 (MTMR6), a negative regulator of CD4(+) T cell activation/proliferation, significantly decreased in SIV-infected macrophages. Luciferase reporter assays confirmed MTMR6 as a direct miR-190b target. To our knowledge, this is the first report, which describes dysregulated miRNA expression in the intestine, that identifies a potentially significant role for miR-190b in HIV/SIV pathogenesis. More importantly, miR-190b-mediated MTMR6 downregulation suggests an important mechanism that could keep infected cells in an activated state, thereby promoting viral replication. In the future, the mechanisms driving miR-190b upregulation including other cellular processes it regulates in SIV-infected cells need determination., (Copyright © 2014 by The American Association of Immunologists, Inc.)
- Published
- 2014
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8. Nonhuman primate retroviruses from Cambodia: high simian foamy virus prevalence, identification of divergent STLV-1 strains and no evidence of SIV infection.
- Author
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Ayouba A, Duval L, Liégeois F, Ngin S, Ahuka-Mundeke S, Switzer WM, Delaporte E, Ariey F, Peeters M, and Nerrienet E
- Subjects
- Animals, Antibodies, Viral blood, Cambodia, DNA, Viral blood, Lorisidae virology, Molecular Sequence Data, Phylogeny, Prevalence, Retroviridae Infections blood, Retroviridae Infections virology, Retroviruses, Simian genetics, Retroviruses, Simian isolation & purification, Simian foamy virus genetics, Simian foamy virus isolation & purification, Tumor Virus Infections blood, Tumor Virus Infections virology, Catarrhini virology, Primate Diseases virology, Retroviridae Infections veterinary, Retroviruses, Simian classification, Simian foamy virus classification, Tumor Virus Infections veterinary
- Abstract
Nonhuman primates (NHPs) carry retroviruses such as simian immunodeficiency viruses (SIV), simian T-cell lymphotropic viruses (STLV) and simian foamy viruses (SFV). Here, we revisited NHPs from Cambodia to assess the prevalence and diversity of these retroviruses using updated viral detection tools. We screened blood from 118 NHPs consisting of six species (Macaca fascicularis (n=91), Macaca leonine (n=8), Presbytis cristata (n=3), Nycticebus coucang (n=1), Hylobates pileatus (n=14), and Pongo pygmaeus) (n=1) by using a Luminex-based multiplex serology assay that allows the detection of all known SIV/HIV and SFV lineages. We also used highly sensitive PCR assays to detect each simian retrovirus group. Positive PCR products were sequenced and phylogenetically analyzed to infer evolutionary histories. Fifty-three of 118 (44.9%) NHPs tested positive for SFV by serology and 8/52 (15.4%), all from M. fascicularis, were PCR-confirmed. The 8 novel SFV sequences formed a highly supported distinct lineage within a clade composed of other macaque SFV. We observed no serological or molecular evidence of SIV infection among the 118 NHP samples tested. Four of 118 (3.3%) NHPs were PCR-positive for STLV, including one M. fascicularis, one P. cristata, and two H. pileatus. Phylogenetic analyses revealed that the four novel STLV belonged to the PTLV-1 lineage, outside the African radiation of PTLV-1, like all Asian PTLV identified so far. Sequence analysis of the whole STLV-1 genome from a H. pileatus (C578_Hp) revealed a genetic structure characteristic of PTLV. Similarity analysis comparing the STLV-1 (C578_Hp) sequence with prototype PTLVs showed that C578_Hp is closer to PTLV-1s than to all other types across the entire genome. In conclusion, we showed a high frequency of SFV infection but found no evidence of SIV infection in NHPs from Cambodia. We identified for the first time STLV-1 in a P. cristata and in two H. pileatus., (Copyright © 2013 Elsevier B.V. All rights reserved.)
- Published
- 2013
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9. Isolation and DNA characterization of a simian retrovirus 5 from a Japanese monkey (Macaca fuscata).
- Author
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Takano JI, Leon A, Kato M, Abe Y, and Fujimoto K
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- Amino Acid Sequence, Animals, Base Sequence, DNA Primers genetics, DNA, Viral genetics, Genes, env genetics, Genes, gag genetics, Genes, pol genetics, Japan, Molecular Sequence Data, Phylogeny, Retroviruses, Simian classification, Retroviruses, Simian genetics, Sequence Analysis, DNA, Genes, Viral genetics, Genome, Viral genetics, Macaca, Monkey Diseases virology, Retroviridae Infections virology, Retroviruses, Simian isolation & purification
- Abstract
An SRV-like virus was isolated from a colony-born Japanese monkey. To identify this SRV-like virus, we designed universal primers at regions that were conserved among the reported SRV sequences in the 5'-LTR and the short ORF and we obtained plasmid clones containing the complete gag, prt, pol and env genes. The full-length sequences of the isolate were determined from the plasmids and by direct sequencing. Sequence comparisons and phylogenetic analyses indicated that this SRV-like virus had a sequence identical to the reported 626 bp of SRV-5. In this study, we isolated SRV5/JPN/2005/V1 from a Japanese monkey and characterized the full-length SRV-5 sequence.
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- 2013
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10. Simian retroviruses in African apes.
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Peeters M and Delaporte E
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- Animals, Disease Reservoirs, Disease Transmission, Infectious, Gorilla gorilla, Humans, Molecular Epidemiology, Pan paniscus, Pan troglodytes, Primate Diseases transmission, Retroviridae Infections epidemiology, Retroviridae Infections transmission, Retroviridae Infections virology, Retroviruses, Simian classification, Retroviruses, Simian genetics, Zoonoses epidemiology, Zoonoses transmission, Primate Diseases epidemiology, Primate Diseases virology, Retroviridae Infections veterinary, Retroviruses, Simian isolation & purification
- Abstract
It is now well established that simian immunodeficiency viruses (SIVs) from chimpanzees (SIVcpz) and gorillas (SIVgor) from west Central Africa are at the origin of HIV-1/AIDS. Apes are also infected with other retroviruses, notably simian T-cell lymphotropic viruses (STLVs) and simian foamy viruses (SFVs), that can be transmitted to humans. We discuss the actual knowledge on SIV, STLV and SFV infections in chimpanzees, gorillas, and bonobos. We especially elaborate on how the recent development of non-invasive methods has allowed us to identify the reservoirs of the HIV-1 ancestors in chimpanzees and gorillas, and increased our knowledge of the natural history of SIV infections in chimpanzees. Multiple cross-species events with retroviruses from apes to humans have occurred, but only one transmission of SIVcpz from chimpanzees in south-eastern Cameroon spread worldwide, and is responsible for the actual HIV pandemic. Frequent SFV transmissions have been recently reported, but no human-to-human transmission has been documented yet. Because humans are still in contact with apes, identification of pathogens in wild ape populations can signal which pathogens may be cause risk for humans, and allow the development of serological and molecular assays with which to detect transmissions to humans. Finally, non-invasive sampling also allows the study of the impact of retroviruses and other pathogens on the health and survival of endangered species such as chimpanzees, gorillas, and bonobos., (© 2012 The Authors. Clinical Microbiology and Infection © 2012 European Society of Clinical Microbiology and Infectious Diseases.)
- Published
- 2012
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11. [Epidemiological survey of a captive Chinese rhesus macaque breeding colony in Yunnan for SRV, STLV and BV].
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Zhu L, Han JB, Zhang XH, Ma JP, Lv LB, Zhang GH, and Zheng YT
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- Animals, Breeding, China epidemiology, Female, Herpesviridae Infections epidemiology, Herpesviridae Infections virology, Herpesvirus 1, Cercopithecine genetics, Humans, Macaca mulatta genetics, Male, Primate Diseases epidemiology, Retroviridae Infections epidemiology, Retroviridae Infections virology, Retroviruses, Simian genetics, Simian T-lymphotropic virus 1 genetics, Herpesviridae Infections veterinary, Herpesvirus 1, Cercopithecine isolation & purification, Macaca mulatta virology, Primate Diseases virology, Retroviridae Infections veterinary, Retroviruses, Simian isolation & purification, Simian T-lymphotropic virus 1 isolation & purification
- Abstract
Nonhuman primates are critical resources for biomedical research. Rhesus macaque is a popularly used laboratory nonhuman primate that share many characteristics with humans. However, rhesus macaques are the natural host of two exogenous retroviruses, SRV (simian type D retrovirus) and STLV (simian T lymphotropic virus). SRV and STLV may introduce potentially significant confounding factors into the study of AIDS model. Moreover, B virus (ceropithecine herpesvirus 1) is likely to harm not only rhesus macaque but also humans in experiments involving rhesus macaque. Yunnan province has large-scale breeding colonies of Chinese rhesus macaque. Therefore there is an urgent need for SPF Chinese rhesus macaque colonies. Here we investigated SRV, STLV and BV infections in 411 Chinese rhesus macaque by PCR technique. The results showed that the prevalence of SRV, STLV and BV among Chinese rhesus macaque breeding colony was 19.71% (81/411), 13.38% (55/411) and 23.11% (95/411), respectively. Comparison of viruses infection in different age-groups and male/female of Chinese rhesus macaque was also analyzed. This study will contribute to establishment of SPF Chinese rhesus macaque breeding colony.
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- 2012
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12. Hemophagocytic syndrome in a pancytopenic simian retrovirus-infected male rhesus macaque (Macaca mulatta).
- Author
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Cotroneo TM, Colby LA, and Bergin IL
- Subjects
- Animals, Autopsy veterinary, Bone Marrow pathology, Bone Marrow virology, Euthanasia, Animal, Humans, Immunohistochemistry, Lymphohistiocytosis, Hemophagocytic complications, Lymphohistiocytosis, Hemophagocytic pathology, Lymphohistiocytosis, Hemophagocytic virology, Macrophages virology, Male, Monkey Diseases pathology, Pancytopenia complications, Pancytopenia pathology, Pancytopenia virology, Polymerase Chain Reaction, Retroviridae Infections complications, Retroviridae Infections pathology, Retroviruses, Simian genetics, Spleen pathology, Spleen virology, Tumor Virus Infections complications, Tumor Virus Infections pathology, Tumor Virus Infections veterinary, Lymphohistiocytosis, Hemophagocytic veterinary, Macaca mulatta virology, Monkey Diseases virology, Pancytopenia veterinary, Retroviridae Infections veterinary, Retroviruses, Simian isolation & purification
- Abstract
Hemophagocytic syndrome (HPS) is a macrophage hyperactivation disorder triggered by disrupted T-cell macrophage cytokine interaction. HPS has been reported in humans, dogs, cats, and cattle, and it is infrequent and poorly characterized in animals. A 16-year-old male rhesus macaque was euthanized because of severe pancytopenia, including nonregenerative anemia (hematocrit = 5.5%), neutropenia (0.29 K/μl), and thrombocytopenia (21 K/μl). Bone marrow was hypocellular with normal maturation, myeloid hypoplasia, and few megakaryocytes. There were numerous morphologically normal macrophages (12% of nucleated cells), with 6% of nucleated cells being hemophagocytic macrophages in the bone marrow. Serology was negative, but polymerase chain reaction and immunohistochemistry were positive for simian retrovirus type 2. Blood and bone marrow findings were consistent with HPS. Cytopenias are common in simian retrovirus-infected macaques, but HPS has not been reported. An association between simian retrovirus infection and HPS is undetermined, but retrovirus-associated HPS has been observed in humans.
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- 2011
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13. Virological and serological characterization of SRV-4 infection in cynomolgus macaques.
- Author
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Zao CL, Ward JA, Tomanek L, Cooke A, Berger R, and Armstrong K
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- Animals, Macaca fascicularis, Molecular Sequence Data, Retroviridae Infections immunology, Retroviridae Infections virology, Retroviruses, Simian genetics, Retroviruses, Simian isolation & purification, Retroviruses, Simian physiology, Antibodies, Viral immunology, Monkey Diseases immunology, Monkey Diseases virology, Retroviridae Infections veterinary, Retroviruses, Simian immunology
- Abstract
The nature of SRV-4 infection in cynomolgus macaques remains unclear to date. Here, we report the monitoring of 24 cynomolgus monkeys that were naturally infected with SRV-4 for virus isolation, proviral load and antibody. The results indicated that the SRV-4 antibody status was statistically correlated to environmental temperature.
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- 2011
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14. The complete genome and genetic characteristics of SRV-4 isolated from cynomolgus monkeys (Macaca fascicularis).
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Zao CL, Armstrong K, Tomanek L, Cooke A, Berger R, Estep JS, Marx PA, Trask JS, Smith DG, Yee JL, and Lerche NW
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- Animals, California, Japan, Molecular Sequence Data, Retroviridae Infections virology, Retroviruses, Simian classification, Retroviruses, Simian isolation & purification, Texas, Tumor Virus Infections virology, Viral Proteins genetics, Genome, Viral, Macaca fascicularis virology, Monkey Diseases virology, Retroviridae Infections veterinary, Retroviruses, Simian genetics, Sequence Analysis, DNA, Tumor Virus Infections veterinary
- Abstract
At least 5 serotypes of exogenous simian retrovirus type D (SRV/D) have been found in nonhuman primates, but only SRV-1, 2 and 3 have been completely sequenced. SRV-4 was recovered once from cynomolgus macaques in California in 1984, but its genome sequences are unknown. Here we report the second identification of SRV-4 and its complete genome from infected cynomolgus macaques with Indochinese and Indonesian/Indochinese mixed ancestry. Phylogenetic analysis demonstrated that SRV-4 was distantly related to SRV-1, 2, 3, 5, 6 and 7. SRV/D-T, a new SRV/D recovered in 2005 from cynomolgus monkeys at Tsukuba Primate Center in Japan, clustered with the SRV-4 isolates from California and Texas and was shown to be another occurrence of SRV-4 infection. The repeated occurrence of SRV-4 in cynomolgus monkeys in different areas of the world and across 25years suggests that this species is the natural host of SRV-4., (Copyright 2010 Elsevier Inc. All rights reserved.)
- Published
- 2010
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15. Detection of EBV, HBV, HCV, HIV-1, HTLV-I and -II, and SMRV in human and other primate cell lines.
- Author
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Uphoff CC, Denkmann SA, Steube KG, and Drexler HG
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- Animals, Blotting, Southern, Cell Line, DNA, Circular analysis, HIV-1 genetics, HIV-1 isolation & purification, Hepacivirus genetics, Hepacivirus isolation & purification, Hepatitis B virus genetics, Hepatitis B virus isolation & purification, Herpesvirus 4, Human genetics, Herpesvirus 4, Human isolation & purification, Human T-lymphotropic virus 1 genetics, Human T-lymphotropic virus 1 isolation & purification, Human T-lymphotropic virus 2 genetics, Human T-lymphotropic virus 2 isolation & purification, Humans, In Situ Hybridization, Fluorescence, Polymerase Chain Reaction, Retroviruses, Simian genetics, Retroviruses, Simian isolation & purification, Saimiri virology, Viral Proteins analysis, Primates virology, Viruses genetics, Viruses isolation & purification
- Abstract
The high prevalence of contaminated cell cultures suggests that viral contaminations might be distributed among cultures. We investigated more than 460 primate cell lines for Epstein-Barr (EBV), hepatitis B (HBV), hepatitis C (HCV), human immunodeficiency virus type 1 (HIV-1), human T-cell leukemia/lymphoma virus I and II (HTLV-I/-II), and squirrel monkey retrovirus (SMRV) infections for risk assessment. None of the cell lines were infected with HCV, HIV-1, or HTLV-I/-II. However, one cell line displayed reverse transcriptase activity. Thirty-nine cell lines harbored EBV DNA sequences. Studies on the lytic phase of EBV revealed that five cell lines produce EBV particles and six further cell lines produced EBV upon stimulation. One cell line contained an integrated HBV genome fragment but showed no virus production. Six cell lines were SMRV-infected. Newly established cell lines should be tested for EBV infections to detect B-lymphoblastoid cell lines (B-LCL). B-LCLs established with EBV from cell line B95-8 should be tested for SMRV infections.
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- 2010
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16. [Alternative methods for the study of simian retroviral genetic diversity].
- Author
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Calvignac S
- Subjects
- Animals, Chlorocebus aethiops, Haplorhini, Macaca mulatta, Genetic Variation, Retroviruses, Simian genetics, Simian Immunodeficiency Virus genetics
- Published
- 2008
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17. Rev proteins of human and simian immunodeficiency virus enhance RNA encapsidation.
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Brandt S, Blissenbach M, Grewe B, Konietzny R, Grunwald T, and Uberla K
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- Animals, Biological Transport, Cell Nucleus virology, Cytoplasm virology, Gene Expression Regulation, Viral, Gene Products, gag genetics, Gene Products, gag metabolism, Gene Products, rev genetics, Humans, Molecular Sequence Data, Mutation genetics, Nuclear Proteins metabolism, Plasmids genetics, Response Elements genetics, rev Gene Products, Human Immunodeficiency Virus, Gene Products, rev metabolism, HIV-1 genetics, RNA Splice Sites genetics, RNA, Messenger metabolism, RNA, Viral metabolism, Retroviruses, Simian genetics, Virus Assembly genetics
- Abstract
The main function attributed to the Rev proteins of immunodeficiency viruses is the shuttling of viral RNAs containing the Rev responsive element (RRE) via the CRM-1 export pathway from the nucleus to the cytoplasm. This restricts expression of structural proteins to the late phase of the lentiviral replication cycle. Using Rev-independent gag-pol expression plasmids of HIV-1 and simian immunodeficiency virus and lentiviral vector constructs, we have observed that HIV-1 and simian immunodeficiency virus Rev enhanced RNA encapsidation 20- to 70-fold, correlating well with the effect of Rev on vector titers. In contrast, cytoplasmic vector RNA levels were only marginally affected by Rev. Binding of Rev to the RRE or to a heterologous RNA element was required for Rev-mediated enhancement of RNA encapsidation. In addition to specific interactions of nucleocapsid with the packaging signal at the 5' end of the genome, the Rev/RRE system provides a second mechanism contributing to preferential encapsidation of genomic lentiviral RNA.
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- 2007
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18. New simian beta retroviruses from rhesus monkeys (Macaca mulatta) and langurs (Semnopithecus entellus) from Rajasthan, India.
- Author
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Nandi JS, Van Dooren S, Chhangani AK, and Mohnot SM
- Subjects
- Amino Acid Sequence, Animals, Base Sequence, Cell Line, Tumor, Cells, Cultured, Humans, India, Leukocytes, Mononuclear, Molecular Sequence Data, Retroviruses, Simian classification, Macaca mulatta virology, Retroviruses, Simian genetics
- Abstract
Natural infection of feral Indian rhesus monkeys (Macaca mulatta) by a new simian beta retrovirus, provisionally called simian retrovirus-7 (SRV-7) is described. The virus is capable of in vitro replication in primary human peripheral blood lymphocytes (PBL) and B and T cell lines. We have earlier reported a novel SRV, SRV-6 from Indian langurs (Semnopithecus entellus). Additional sequence analyses from gp20 transmembrane (TM) env genes of SRV-6 and SRV-7 place them in a separate cluster, related to but distinct from known exogenous SRVs and also close to the simian endogenous beta retrovirus, (SERV) from African baboon. Phylogenetic analyses of pol gene of SRV-7 place it closer to SERV when the stop codons of the SERV genes are removed. On the other hand, additional sequence data from gp70, surface glycoprotein (SU) region of the env gene of SRV-6 suggest it is more closely related to known exogenous SRVs, (SRV-1 to 3). It is also related to the endogenous langur virus, Po-1-Lu. We hypothesize that SRV-6 and SRV-7 probably originated from a progenitor exogenous SRV which recombined with an endogenous SERV in the TM env and pol genes during evolution, based on the phylogenetic analyses.
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- 2006
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19. Intestinal stromal tumors in a simian immunodeficiency virus-infected, simian retrovirus-2 negative rhesus macaque (Macaca mulatta).
- Author
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Bielefeldt-Ohmann H, Barouch DH, Bakke AM, Bruce AG, Durning M, Grant R, Letvin NL, Ryan JT, Schmidt A, Thouless ME, and Rose TM
- Subjects
- Animals, Antigens, Nuclear metabolism, DNA Primers, Gastrointestinal Stromal Tumors complications, Gastrointestinal Stromal Tumors pathology, Herpesvirus 8, Human metabolism, Immunohistochemistry veterinary, Polymerase Chain Reaction veterinary, Retroviruses, Simian genetics, Reverse Transcriptase Polymerase Chain Reaction veterinary, Vimentin metabolism, Gastrointestinal Stromal Tumors veterinary, Macaca mulatta, Monkey Diseases pathology, Monkey Diseases virology, Simian Acquired Immunodeficiency Syndrome complications, Simian Immunodeficiency Virus
- Abstract
Multifocal submucosal stromal tumors were diagnosed in a 5.5-year-old rhesus macaque (Macaca mulatta) experimentally infected with simian immunodeficiency virus, strain SIVsmE660, and CD4+ T cell depleted. The animal was negative for simian retroviruses, SRV-1, -2, and -5. Polymerase chain reaction analysis of DNA from tumor and spleen tissue revealed abundant, preferential presence of retroperitoneal fibromatosis herpesvirus, the macaque homologue of the Kaposi sarcoma-associated herpesvirus (human herpesvirus-8), in the tumors. This was corroborated by demonstration of viral latent nuclear antigen-1 in the nuclei of a majority of the spindeloid tumor cells. Low levels of an additional macaque herpesvirus, rhesus rhadinovirus, were also detected in the spleen and tumor tissues. The spindeloid cells labeled positively for vimentin and CD117 but were negative for CD31, CD68, desmin, and smooth muscle cell actin. Collectively, these findings suggest a relation to but not absolute identity with simian mesenchymoproliferative disorders (MPD) or typical gastrointestinal stromal tumors (GISTs).
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- 2005
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20. Survey of captive cynomolgus macaque colonies for SRV/D infection using polymerase chain reaction assays.
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Hara M, Kikuchi T, Ono F, Takano J, Ageyama N, Fujimoto K, Terao K, Baba T, and Mukai R
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- Animals, DNA, Viral analysis, Female, Genes, gag, Health Surveys, Japan epidemiology, Male, Monkey Diseases epidemiology, Monkey Diseases pathology, Retroviridae Infections epidemiology, Retroviridae Infections pathology, Retroviruses, Simian genetics, Specific Pathogen-Free Organisms, Tumor Virus Infections epidemiology, Tumor Virus Infections pathology, Macaca fascicularis virology, Monkey Diseases virology, Polymerase Chain Reaction veterinary, Retroviridae Infections veterinary, Retroviruses, Simian isolation & purification, Tumor Virus Infections veterinary
- Abstract
The exogenous simian type D retroviruses (SRV/Ds) are prevalent in macaque monkeys and sometimes cause immunodeficiency with anemia, weight loss, and persistent unresponsive diarrhea. SRV/D isolates are classified as subtypes 1 to 6, and the entire sequences of the gag region of SRV/D-1, -2, and -3 and SRV/D-Tsukuba (SRV/D-T) have been determined. We designed specific primers in the gag region of SRV/D-T that enabled us to directly detect by polymerase chain reaction (PCR) SRV/D-T proviral DNA sequences in DNA extracted from whole blood. Using this assay and another PCR assay that detects multiple SRV/D subtypes, we performed a survey for SRV/D infection in our specific pathogen-free (SPF) and conventional colonies at Tsukuba Primate Center (TPC). In the SPF colony, no SRV/D signal was detected in any animal. On the other hand, SRV/D-T was detected in 11 of 49 animals (22.5%) in the conventional colony. SRV/D-T was the only SRV/D subtype detected. Consequently, SRV/D-T is the major SRV/D subtype present in cynomolgus monkeys at TPC.
- Published
- 2005
21. Isolation and characterization of a new simian retrovirus type D subtype from monkeys at the Tsukuba Primate Center, Japan.
- Author
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Hara M, Sata T, Kikuchi T, Nakajima N, Uda A, Fujimoto K, Baba T, and Mukai R
- Subjects
- Amino Acid Sequence, Animals, Betaretrovirus classification, Betaretrovirus genetics, Female, Genes, gag, Japan, Molecular Sequence Data, Monkey Diseases pathology, Phylogeny, Retroviridae Infections pathology, Retroviruses, Simian classification, Retroviruses, Simian genetics, Sequence Alignment, Sequence Homology, Nucleic Acid, Tumor Virus Infections pathology, Betaretrovirus isolation & purification, Macaca fascicularis virology, Monkey Diseases virology, Retroviridae Infections veterinary, Retroviruses, Simian isolation & purification, Tumor Virus Infections veterinary
- Abstract
Exogenous type D simian retroviruses (SRV/D) are prevalent in captive and feral populations of various macaque monkeys. Thus far, five subtypes of SRV/Ds have been reported, three of which (SRV-1, -2 and -3) have been molecularly characterized. Two SRV/D strains (N27 and T150) were isolated from seropositive cynomolgus macaques at the Tsukuba Primate Center (TPC) in Japan, showing clinical signs of SRV/D infection, including anemia and persistent unresponsive diarrhea. Electron microscopy demonstrated that both SRV/D isolates have a virion morphology typical of type D retrovirus. The SRV/D N27 and T150 isolates were essentially the same based on sequence analysis. From homology analysis of the entire gag sequence, the N27 isolate is closely related to the other known SRV/Ds but is distinct from the three molecularly characterized SRV/Ds. Thus, we have tentatively designated the N27 and T150 viruses isolated from TPC cynomolgus macaques as SRV/D-Tsukuba (SRV/D-T).
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- 2005
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22. Exchange of genetic sequences of long terminal repeat and the env gene by a promiscuous primate type D retrovirus.
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Chiu CN, Mitra R, and Chiu IM
- Subjects
- Amino Acid Sequence, Animals, DNA, Viral, Endogenous Retroviruses classification, Evolution, Molecular, Molecular Sequence Data, Recombination, Genetic, Retroviruses, Simian classification, Sequence Homology, Amino Acid, Endogenous Retroviruses genetics, Gene Products, env genetics, Genome, Viral, Papio virology, Retroviruses, Simian genetics, Terminal Repeat Sequences
- Abstract
Squirrel monkey retrovirus (SMRV) is a New World primate type D retrovirus. It was shown that SMRV-related sequences could be detected in another New World species, the skunk. It was further suggested that SMRV and an Old World primate type C retrovirus, baboon endogenous virus (BaEV), may have exchanged their env gene sequences. In this study, we sought to understand which sequences were exchanged between the genomic DNAs of SMRV and skunk. We also sought to determine the sequences exchanged between SMRV and BaEV. Here, we demonstrate that the long terminal repeat of SMRV is present in the skunk genome. We also show, by nucleotide sequence analysis, that the env gene that encodes the p15E glycoprotein of BaEV was most likely transduced from the corresponding gene of a primate type D retrovirus. Our results demonstrate that SMRV is a promiscuous virus with its pol gene homologous to the pol genes of type A, type B and avian type C viruses and a portion of its env gene homologous to the env genes of primate type C retroviruses. However, the primer binding sequence is unique to type D retroviruses. These kinds of recombination are likely to occur more than once in the evolution of retroviruses. The promiscuous nature of retroviruses and the recent incidence of unintended retroviral integration into a gene therapy patient underscore the importance of understanding how retroviral sequences are recombined among themselves and how they are integrated into the mammalian genome.
- Published
- 2003
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23. Natural infection by simian retrovirus-6 (SRV-6) in Hanuman langurs (Semnopithecus entellus) from two different geographical regions of India.
- Author
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Nandi JS, Tikute SA, Chhangani AK, Potdar VA, Tiwari-Mishra M, Ashtekar RA, Kumari J, Walimbe A, and Mohnot SM
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- Animals, Antibodies, Viral blood, Cercopithecidae blood, Cross Reactions, Cytopathogenic Effect, Viral, Gene Products, env genetics, Genome, Viral, Glycoproteins genetics, Humans, India, Leukocytes, Mononuclear virology, Molecular Sequence Data, Monkey Diseases virology, Phylogeny, Retroviridae Infections epidemiology, Retroviruses, Simian genetics, Seroepidemiologic Studies, Tumor Virus Infections epidemiology, Viral Envelope Proteins genetics, Viral Proteins genetics, Cercopithecidae virology, Monkey Diseases epidemiology, Retroviridae Infections veterinary, Retroviruses, Simian isolation & purification, Tumor Virus Infections veterinary
- Abstract
We have previously reported natural infection of Hanuman langurs (Semnopithecus entellus) from Lucknow, India by a novel simian retrovirus, SRV-6, a beta-retrovirus (type D retrovirus). Here we describe infection by a closely related SRV-6 in an isolated feral population of Hanuman langurs from Jodhpur in the Northwestern desert region of India. Serological analyses, using in-house ELISA and WB, genomic amplification, and sequencing of env region (gp70 and gp20) of the viral genome were carried out. SRV-6-infected langurs from the two regions were serologically cross-reactive. The env gene was used for phylogenetic analyses, being the most variable part of a retroviral genome. The surface glycoproteins (gp70) were almost identical between the two SRV-6 isolates and related to but distinct from equivalent regions from other exogenous SRVs. We could sequence the transmembrane glycoprotein gp20 from SRV-6 infecting the Jodhpur langurs, which was again shown to be related to but unique compared to the other known SRVs. The study suggests that natural infection by related strains of SRV-6 occurs in wild langurs from different parts of India.
- Published
- 2003
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24. Development of simian retroviral vectors for gene delivery.
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Li B and Machida CA
- Subjects
- Animals, Cell Line, Gene Expression Regulation, Viral, Gene Targeting, Genetic Therapy, Humans, Polymerase Chain Reaction methods, Retroviridae Proteins genetics, Retroviruses, Simian physiology, Terminal Repeat Sequences, Transduction, Genetic, Transfection, Gene Transfer Techniques, Genetic Vectors, Retroviruses, Simian genetics
- Published
- 2003
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25. Solution structure of the pseudoknot of SRV-1 RNA, involved in ribosomal frameshifting.
- Author
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Michiels PJ, Versleijen AA, Verlaan PW, Pleij CW, Hilbers CW, and Heus HA
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- Adenine metabolism, Base Pairing, Base Sequence, Genes, Viral genetics, Models, Genetic, Models, Molecular, Molecular Sequence Data, Mutation genetics, Nuclear Magnetic Resonance, Biomolecular, RNA Stability genetics, RNA, Viral genetics, Thermodynamics, Frameshifting, Ribosomal genetics, Gene Expression Regulation, Viral, Nucleic Acid Conformation, RNA, Viral chemistry, RNA, Viral metabolism, Retroviruses, Simian genetics
- Abstract
RNA pseudoknots play important roles in many biological processes. In the simian retrovirus type-1 (SRV-1) a pseudoknot together with a heptanucleotide slippery sequence are responsible for programmed ribosomal frameshifting, a translational recoding mechanism used to control expression of the Gag-Pol polyprotein from overlapping gag and pol open reading frames. Here we present the three-dimensional structure of the SRV-1 pseudoknot determined by NMR. The structure has a classical H-type fold and forms a triple helix by interactions between loop 2 and the minor groove of stem 1 involving base-base and base-sugar interactions and a ribose zipper motif, not identified in pseudoknots so far. Further stabilization is provided by a stack of five adenine bases and a uracil in loop 2, enforcing a cytidine to bulge. The two stems of the pseudoknot stack upon each other, demonstrating that a pseudoknot without an intercalated base at the junction can induce efficient frameshifting. Results of mutagenesis data are explained in context with the present three-dimensional structure. The two base-pairs at the junction of stem 1 and 2 have a helical twist of approximately 49 degrees, allowing proper alignment and close approach of the three different strands at the junction. In addition to the overwound junction the structure is somewhat kinked between stem 1 and 2, assisting the single adenosine in spanning the major groove of stem 2. Geometrical models are presented that reveal the importance of the magnitude of the helical twist at the junction in determining the overall architecture of classical pseudoknots, in particular related to the opening of the minor groove of stem 1 and the orientation of stem 2, which determines the number of loop 1 nucleotides that span its major groove.
- Published
- 2001
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26. Simian retrovirus vectors for gene transfer in nonhuman primate cells.
- Author
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Li B, Nguyen S, Li X, and Machida CA
- Subjects
- 5' Untranslated Regions genetics, Animals, COS Cells, Cell Line, Clone Cells, DNA, Recombinant genetics, DNA, Viral genetics, Gene Products, gag genetics, Genes, Reporter genetics, Genome, Viral, Humans, Lac Operon genetics, Rats, Transduction, Genetic, Transfection, Tumor Cells, Cultured, Virion genetics, Virion metabolism, Virus Assembly genetics, beta-Galactosidase genetics, Gene Transfer Techniques, Genetic Vectors genetics, Retroviruses, Simian genetics
- Abstract
We have recently identified and sequenced a molecular clone of the serogroup 2 simian retrovirus (SRV), D2/RHE/OR/V1, that retains an enhanced ability to infect specific T cell lines. In this report, using deletion mutagenesis, we localized the psi packaging signal, necessary for packaging of D2/RHE/OR/V1 particles, to the genomic region 345-650, which comprises the 5' intergenic region (IR) and the extreme 5' portion of the gag gene. To build an SRV-based gene transfer system and to reduce the possibility of recombination and regeneration of replication-competent viruses, we constructed split-genome D2/RHE/OR/V1 plasmid recombinants containing distinct and non-overlapping retroviral gene regions and several replacement components. For the retrovirus gene transfer vehicle, we deleted the D2/RHE/OR/V1 structural genes and substituted a cassette including the psi-packaging region, the beta-galactosidase reporter gene, and the 3' IR. Both packaging cell recombinants were used to generate stable monkey packaging cell lines; the gene transfer vehicle was subsequently transfected into the packaging cell lines, and replication-defective viruses were recovered for subsequent infection into fresh monkey cells. Successful infection by the recovered viruses verifies the potential efficacy of the SRV-based system as a research tool for gene transfer of heterologous genes into nonhuman primate cells.
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- 2001
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27. Evidence of infection with simian type D retrovirus in persons occupationally exposed to nonhuman primates.
- Author
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Lerche NW, Switzer WM, Yee JL, Shanmugam V, Rosenthal AN, Chapman LE, Folks TM, and Heneine W
- Subjects
- Animals, Antibodies, Viral blood, DNA, Viral blood, Humans, Macaca mulatta, Monkey Diseases virology, Neutralization Tests, Polymerase Chain Reaction, Retroviridae Infections virology, Retroviruses, Simian genetics, Retroviruses, Simian immunology, Tumor Virus Infections virology, Monkey Diseases transmission, Occupational Exposure, Retroviridae Infections transmission, Retroviruses, Simian isolation & purification, Tumor Virus Infections transmission, Zoonoses
- Abstract
Simian type D retrovirus (SRV) is enzootic in many populations of Asian monkeys of the genus Macaca and is associated with immunodeficiency diseases. However, the zoonotic potential of this agent has not been well defined. Screening for antibodies to SRV was performed as part of an ongoing study looking for evidence of infection with simian retroviruses among persons occupationally exposed to nonhuman primates (NHPs). Of 231 persons tested, 2 (0.9%) were found to be strongly seropositive, showing reactivity against multiple SRV antigens representing gag, pol, and env gene products by Western immunoblotting. Persistent long-standing seropositivity, as well as neutralizing antibody specific to SRV type 2, was documented in one individual (subject 1), while waning antibody with eventual seroreversion was observed in a second (subject 2). Repeated attempts to detect SRV by isolation in tissue culture and by using sensitive PCR assays for amplification of two SRV gene regions (gag and pol) were negative. Both individuals remain apparently healthy. We were also unable to transmit this seropositivity to an SRV-negative macaque by using inoculation of whole blood from subject 1. The results of this study provide evidence that occupational exposure to NHPs may increase the risk of infection with SRV and underscore the importance of both occupational safety practices and efforts to eliminate this virus from established macaque colonies.
- Published
- 2001
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28. Intrabodies as antiviral agents.
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Marasco WA
- Subjects
- Acquired Immunodeficiency Syndrome therapy, Animals, CD4-Positive T-Lymphocytes immunology, Cytoplasm immunology, Cytoplasm virology, Disease Models, Animal, Genetic Therapy methods, Genetic Vectors, HIV Antibodies therapeutic use, HIV Infections therapy, HIV-1 genetics, HIV-1 immunology, Humans, Protein Engineering, Receptors, CCR5 genetics, Recombination, Genetic, Retroviruses, Simian genetics, Transfection, Virus Replication, Antibodies, Viral therapeutic use, Antiviral Agents therapeutic use
- Published
- 2001
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29. A novel type D simian retrovirus naturally infecting the Indian Hanuman langur (Semnopithecus entellus).
- Author
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Nandi JS, Bhavalkar-Potdar V, Tikute S, and Raut CG
- Subjects
- Amino Acid Sequence, Animals, Evolution, Molecular, Gene Products, env chemistry, Genes, env, India, Malaysia, Molecular Sequence Data, Retroviridae Infections virology, Retroviruses, Simian genetics, Retroviruses, Simian isolation & purification, Sequence Alignment, Sequence Homology, Amino Acid, Serotyping, Tumor Virus Infections virology, Cercopithecidae virology, Gene Products, env genetics, Phylogeny, Primate Diseases virology, Retroviridae Infections veterinary, Retroviruses, Simian classification, Tumor Virus Infections veterinary
- Abstract
As a simian species, the langurs are not known to harbor simian retroviruses, except for one report on a simian Type D endogenous retrovirus from the spectacled langur (Trachypithecus obscurus) from Malaysia. The present report describes for the first time natural infection of the common Hanuman langur (Semnopithecus entellus) from India by a novel simian retrovirus (SRV). The new SRV is phylogenetically related to but distinct from the three molecularly characterized serotypes, SRV 1-3, of the five known serotypes of SRVs, based on sequence analyses from the 3'orf and env regions of the viral genome. The novel SRV isolated from the Indian Hanuman langur is provisionally named SRV-6., (Copyright 2000 Academic Press.)
- Published
- 2000
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30. Simian retrovirus serogroup 5: partial gag-prt sequence and viral RNA distribution in an infected rhesus macaque.
- Author
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Li B, Axthelm MK, and Machida CA
- Subjects
- Amino Acid Sequence, Animals, Base Sequence, Cell Line, DNA, Viral, Humans, Macaca mulatta, Male, Molecular Sequence Data, Retroviridae Infections pathology, Sequence Homology, Amino Acid, Tissue Distribution, Tumor Virus Infections pathology, Gene Products, gag genetics, RNA, Viral metabolism, Retroviridae Infections virology, Retroviruses, Simian genetics, Tumor Virus Infections virology
- Abstract
The simian type D retroviruses (SRVs) are one of the causative agents of simian acquired immunodeficiency syndrome (SAIDS) in Asian macaques. In this report, we describe the infection of a rhesus macaque with the SRV serogroup 5 isolate, D5/RHE/OR. Using reverse transcriptase-polymerase chain reaction (RT-PCR) and dot blot hybridization analyses, we have determined the tissue distribution of D5/RHE/OR in this infected rhesus macaque, and have demonstrated viral mRNA in the majority of the surveyed tissues, including robust loads in the bone marrow, seminal vesicle, submaxillary salivary gland, prostate, and skeletal muscle. Microscopic examination of necropsy tissues revealed generalized lymphoid hyperplasia that was most severe in the salivary gland, bone marrow, kidney, and spleen. We also describe the first sequence analyses of portions of the D5/RHE/OR gag-prt region, obtained as a RT-PCR amplification product from infected rhesus macaque tissue, and report the first confirmation using Northern blot analyses that the SRV serogroups, including D5/RHE/OR, express similarly-sized genomic and subgenomic env mRNAs.
- Published
- 2000
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31. Improved titers of HIV-based lentiviral vectors using the SRV-1 constitutive transport element.
- Author
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Mautino MR, Keiser N, and Morgan RA
- Subjects
- Gene Products, rev, Humans, Mutagenesis, Site-Directed, rev Gene Products, Human Immunodeficiency Virus, Acquired Immunodeficiency Syndrome therapy, Genetic Therapy methods, Genetic Vectors genetics, Retroviruses, Simian genetics
- Abstract
The development of lentiviral vectors that use Rev-independent mechanisms of nuclear export for their genomic RNA could facilitate the construction of novel anti-HIV vectors. We have improved the titers of Rev-independent lentiviral vectors having the SRV-1 CTE by mutating the major splice donor and acceptor sites present in the vector and by relocalization of the CTE sequences adjacent to the HIV-1 3'LTR. These two modifications have additive beneficial effects on vector titers and packaging efficiency. Packaging these CTE+ vectors expressing marker genes with a Rev-dependent HIV-1 helper vector yields higher titers than are obtained using a Rev-dependent lentiviral vector.
- Published
- 2000
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32. Eradication of simian retrovirus type D from a colony of cynomolgus, rhesus, and stump-tailed macaques by using serial testing and removal.
- Author
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Schroder MA, Fisk SK, and Lerche NW
- Subjects
- Animal Husbandry, Animals, DNA, Viral analysis, Disease Transmission, Infectious prevention & control, Disease Transmission, Infectious veterinary, Female, Male, Polymerase Chain Reaction, Retroviruses, Simian genetics, Retroviruses, Simian pathogenicity, Serologic Tests, Simian Acquired Immunodeficiency Syndrome transmission, Animals, Laboratory, Infection Control, Macaca fascicularis virology, Macaca mulatta virology, Retroviruses, Simian isolation & purification, Simian Acquired Immunodeficiency Syndrome prevention & control
- Abstract
The markedly compromised health of animals in a macaque colony and the problematic interpretation of data from two drug safety assessment studies prompted a review of the effect of simian retrovirus type D on the drug-development process at a Midwest pharmaceutical company. After reviewing relevant literature and consulting with an expert in simian retroviruses, we initiated a program of eradication. During a 16-month period, all cynomolgus (Macaca fascicularis), rhesus (Macaca mulatta), and stump-tailed (Macaca arctoides) macaques housed in the facility were evaluated as many as eight times for the presence of simian retrovirus type D by using serology, virus isolation, and/or polymerase chain reaction tests. All animals with positive test results were removed from the colony immediately. No test results indicative of simian retrovirus type D infection have occurred during the subsequent 2.5 years. We attribute the successful eradication and prevention of re-introduction of the virus to regular testing, purchasing animals from sources free of simian retrovirus type D, and assiduous application of procedures designed to prevent transmission between animals.
- Published
- 2000
33. Human spontaneous laryngeal carcinoma HEp-2 cells are chronically infected with SRV-1 virus, a variant of simian type D retrovirus.
- Author
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Imamova LR, Kzhyshkowska YG, Ostashkin AS, Itkes AV, and Il'in KV
- Subjects
- Animals, Base Sequence, Electrophoresis, Female, Humans, Laryngeal Neoplasms genetics, Molecular Sequence Data, Polymerase Chain Reaction, Retroviruses, Simian classification, Retroviruses, Simian genetics, Tumor Cells, Cultured, Laryngeal Neoplasms virology, Retroviruses, Simian isolation & purification
- Abstract
A type D retrovirus chronically persisting in HEp-2 cells from human laryngeal carcinoma was analyzed by PCR and sequenced. This virus is most similar to SRV-1 and probably represents one of its subtypes.
- Published
- 2000
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34. Two distinct lineages of macaque gamma herpesviruses related to the Kaposi's sarcoma associated herpesvirus.
- Author
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Strand K, Harper E, Thormahlen S, Thouless ME, Tsai C, Rose T, and Bosch ML
- Subjects
- Amino Acid Sequence, Animals, Base Sequence, Coculture Techniques, DNA-Directed DNA Polymerase genetics, Gammaherpesvirinae classification, Herpesviridae Infections virology, Humans, Leukocytes virology, Lymphoid Tissue cytology, Lymphoid Tissue virology, Molecular Sequence Data, Polymerase Chain Reaction, Retroperitoneal Fibrosis veterinary, Retroperitoneal Fibrosis virology, Retroviruses, Simian genetics, Species Specificity, Viral Proteins chemistry, Viral Proteins genetics, Gammaherpesvirinae genetics, Herpesviridae Infections veterinary, Herpesvirus 8, Human genetics, Macaca virology, Monkey Diseases virology, Phylogeny
- Abstract
Background: KSHV, Kaposi's sarcoma-associated herpesvirus, is a necessary cofactor for the development of Kaposi's sarcoma (KS). We have previously reported KSHV-related DNA sequences in retroperitoneal fibromatosis (RF) tissue from two species of macaque. The putative herpesvirus was called RFHV for RF-associated herpesvirus. These data suggested that KSHV is a human representative of a larger family of primate herpesviruses., Objective: To identify and characterize other members of a putative family of KSHV-related herpesviruses in macaques in order to obtain information on the evolutionary history of KSHV infection in humans., Study Design: Lymphoid tissue cells and blood leukocytes from rhesus-, cynomolgus- and pigtailed-macaques were tested for the presence of unknown herpesviruses using degenerate primer-driven PCR amplification. The sequences obtained were compared against known herpesvirus sequences., Results: We have identified new herpesvirus DNA sequences in each of the three macaque species. Sequence comparisons indicate that these new viruses are most related to each other and form a separate phylogenetic lineage within the gamma herpesviruses. Screening of PBMC from Indonesian-origin quarantine animals suggests that these viruses (MGV, macaque gamma virus) are species-specific, and highly prevalent in the wild. They are readily cultured in vivo, and share a common tissue tropism with the previously identified RFHV., Conclusions: MGV and RFHV represent two independent introductions of an ancestral gamma herpesvirus into macaque precursors.
- Published
- 2000
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35. Structure, stability and function of RNA pseudoknots involved in stimulating ribosomal frameshifting.
- Author
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Giedroc DP, Theimer CA, and Nixon PL
- Subjects
- Base Sequence, Cations metabolism, Cations pharmacology, Infectious bronchitis virus genetics, Luteovirus genetics, Mammary Tumor Virus, Mouse genetics, Models, Genetic, RNA, Messenger genetics, RNA, Viral chemistry, RNA, Viral genetics, RNA, Viral metabolism, Retroviruses, Simian genetics, Frameshifting, Ribosomal genetics, Nucleic Acid Conformation drug effects, RNA Stability drug effects, RNA, Messenger chemistry, RNA, Messenger metabolism
- Abstract
Programmed -1 ribosomal frameshifting has become the subject of increasing interest over the last several years, due in part to the ubiquitous nature of this translational recoding mechanism in pathogenic animal and plant viruses. All cis-acting frameshift signals encoded in mRNAs are minimally composed of two functional elements: a heptanucleotide "slippery sequence" conforming to the general form X XXY YYZ, followed by an RNA structural element, usually an H-type RNA pseudoknot, positioned an optimal number of nucleotides (5 to 9) downstream. The slippery sequence itself promotes a low level ( approximately 1 %) of frameshifting; however, downstream pseudoknots stimulate this process significantly, in some cases up to 30 to 50 %. Although the precise molecular mechanism of stimulation of frameshifting remains poorly understood, significant advances have been made in our knowledge of the three-dimensional structures, thermodynamics of folding, and functional determinants of stimulatory RNA pseudoknots derived from the study of several well-characterized frameshift signals. These studies are summarized here and provide new insights into the structural requirements and mechanism of programmed -1 ribosomal frameshifting., (Copyright 2000 Academic Press.)
- Published
- 2000
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36. Modified human immunodeficiency virus-based lentiviral vectors display decreased sensitivity to trans-dominant Rev.
- Author
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Mautino MR, Ramsey WJ, Reiser J, and Morgan RA
- Subjects
- Base Sequence, Cell Line, Gene Products, rev chemistry, Gene Products, rev metabolism, Genes, Dominant, Genetic Vectors biosynthesis, Humans, Mutation, Purines chemistry, RNA Splicing, Retroviruses, Simian genetics, Transgenes, rev Gene Products, Human Immunodeficiency Virus, Gene Products, rev genetics, Genetic Vectors genetics, HIV genetics
- Abstract
As a first step toward the development of HIV-based conditionally replicating defective interfering particles expressing trans-dominant Rev (TdRev), we studied whether mutation of the splicing signals and replacement of the RRE by the SRV-1 CTE would render these vectors less sensitive to TdRev. Vectors with mutations in the splicing signals (SD-/RRE+) yielded high titers (5 X 10(6) CFU/ml) and showed higher levels of cytoplasmic unspliced mRNA than the corresponding SD+/RRE+ vectors either in the absence of Rev, in the presence of TdRev, or in the presence of both TdRev and Rev. Proviral copies of SD-/RRE+ vectors were rescued more efficiently than SD+/RRE+ vectors when TdRev was expressed. Vectors with the SRV-1 CTE (SD+/CTE+ and SD-/CTE+) expressed high levels of cytoplasmic unspliced mRNA in the absence of Rev expression. Titers obtained with the SD-/CTE+ vectors (10(6) CFU/ml) were higher than the titers obtained with SD+/CTE+ vectors. We also tested the effect of other structural modifications such as the orientation of the expression cassette and the presence of the central polypurine tract (cPPT/CTS). We show that an expression cassette cloned in the reverse orientation with respect to the LTRs or elimination of the cPPT/CTS element severely affected vector titers. We also demonstrated that these vectors can be efficiently mobilized from their proviral state by HIV trans-complementing functions, and transduced into secondary target cells without suffering any genomic rearrangement.
- Published
- 2000
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37. Virus load and sequence variation in simian retrovirus type 2 infection.
- Author
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Rosenblum LL, Weiss RA, and McClure MO
- Subjects
- Animals, Antibodies, Viral blood, DNA, Viral blood, Genetic Variation genetics, Macaca fascicularis, Molecular Sequence Data, Neutralization Tests, RNA, Viral blood, Retroviridae Infections virology, Retroviruses, Simian immunology, Retroviruses, Simian isolation & purification, Sequence Analysis, DNA, Tumor Virus Infections virology, Viral Load, Monkey Diseases virology, Retroviridae Infections veterinary, Retroviruses, Simian genetics, Retroviruses, Simian physiology, Tumor Virus Infections veterinary
- Abstract
The natural history of type D simian retrovirus (SRV) infection is poorly characterized in terms of viral load, antibody status, and sequence variation. To investigate this, blood samples were taken from a small cohort of mostly asymptomatic cynomolgus macaques (Macaca fascicularis), naturally infected with SRV type 2 (SRV-2), some of which were followed over an 8-month period with blood taken every 2 months. Provirus and RNA virus loads were obtained, the samples were screened for presence of antibodies to SRV-2 and neutralizing antibody titers to SRV-2 were assayed. env sequences were aligned to determine intra- and intermonkey variation over time. Virus loads varied greatly among cohort individuals but, conversely, remained steady for each macaque over the 8-month period, regardless of their initial levels. No significant sequence variation was found within an individual over time. No clear picture emerged from these results, which indicate that the variables of SRV-2 infection are complex, differ from those for lentivirus infection, and are not distinctly related to disease outcome.
- Published
- 2000
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38. Nucleocytoplasmic export of type D simian retrovirus genomic RNA: identification of important genetic subregions and interacting cellular proteins.
- Author
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Li B, Wyman TE, Moudgil T, Marracci GH, Ju CF, and Machida CA
- Subjects
- Animals, Base Sequence, Cell Line, Cell Nucleus virology, Chlorocebus aethiops, Cytoplasm virology, DNA, Ribosomal chemistry, DNA, Ribosomal genetics, Genome, Viral, Humans, Introns, Models, Molecular, Molecular Sequence Data, Mutagenesis, Site-Directed, Nucleic Acid Conformation, Tumor Cells, Cultured, RNA Helicases metabolism, RNA, Viral genetics, Retroviruses, Simian chemistry, Retroviruses, Simian genetics
- Abstract
The simian retrovirus (SRV) genome contains a constitutive transport element (CTE) within its 3' intergenic region (IR) that mediates the nuclear export of unspliced SRV RNA. The serogroup 2 SRV CTE is predicted to form a stable stem-loop structure containing two major internal loops exhibiting 180 degrees inverse symmetry, with loop face sequences A, A', B, and B' and additional minor internal and terminal loops. To begin the identification of potential CTE-interacting proteins and to assess structural requirements for protein interaction, we conducted RNA mobility shift assays using IR fragments that obliterated this region's known stable stem-loop structure. Using immunoblotting assays, we have determined that RNA helicase A, implicated in the nuclear export of unspliced SRV genomic RNA, does not appear to interact directly with either the complete serogroup 2 SRV 3' IR or the subregion RNAs and that formation of RNA-protein complexes is conferred by interaction with other novel proteins. UV crosslinking of RNA-protein complexes, coupled with RNase T1/A digestion, indicates that a novel protein of 120 kDa molecular weight interacts with the complete CTE or with individual subregion RNAs. Transfection analyses indicate that SRV recombinants containing A, A', B, or B' sequences forming the faces for two open loops undergo RNA export; only the complete sense CTE recombinant or a second recombinant containing two subregions in sense orientation that reconstitute the 3' two-thirds of the 3' IR, and contain only A' and B that form the faces for two terminal loops, are capable of SRV RNA export. These experiments indicate that secondary structural determinants of the 3' IR and multiple protein interactions may be important factors in the nuclear export of unspliced SRV RNA., (Copyright 1999 Academic Press.)
- Published
- 1999
- Full Text
- View/download PDF
39. Squirrel monkey retrovirus (SMRV) sequence from an SMRV-negative cell line?
- Author
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Miyazawa M, Yanai Y, and Kurimoto M
- Subjects
- Animals, Genes, pol, Humans, Polymerase Chain Reaction, Retroviruses, Simian genetics, DNA, Viral analysis, Drug Contamination, Interferons, Retroviruses, Simian isolation & purification, Saimiri virology
- Published
- 1999
- Full Text
- View/download PDF
40. Many human endogenous retrovirus K (HERV-K) proviruses are unique to humans.
- Author
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Barbulescu M, Turner G, Seaman MI, Deinard AS, Kidd KK, and Lenz J
- Subjects
- Animals, Base Sequence, Gorilla gorilla virology, Humans, Male, Molecular Sequence Data, Pan troglodytes virology, Polymerase Chain Reaction, Pongo pygmaeus virology, Retroviruses, Simian genetics, Sequence Alignment, Sequence Homology, Species Specificity, Terminal Repeat Sequences genetics, Genes, Viral, Proviruses genetics, Retroviridae genetics
- Abstract
Background: Endogenous retroviruses contribute to the evolution of the host genome and can be associated with disease. Human endogenous retrovirus K (HERV-K) is related to the mouse mammary tumor virus and is present in the genomes of humans, apes and cercopithecoids (Old World monkeys). It is unknown how long ago in primate evolution the full-length HERV-K proviruses that are in the human genome today were formed., Results: Ten full-length HERV-K proviruses were cloned from the human genome. Using provirus-specific probes, eight of the ten were found to be present in a genetically diverse set of humans but not in other extant hominoids. Intact preintegration sites for each of these eight proviruses were present in the apes. A ninth provirus was detected in the human, chimpanzee, bonobo and gorilla genomes, but not in the orang-utan genome. The tenth was found only in humans, chimpanzees and bonobos. Complete sequencing of six of the human-specific proviruses showed that full-length open reading frames for the retroviral protein precursors Gag-Pro-Pol or Env were each present in multiple proviruses., Conclusions: At least eight full-length HERV-K genomes that are in the human germline today integrated after humans diverged from chimpanzees. All of the viral open reading frames and cis-acting sequences necessary for HERV-K replication must have been intact during the recent time when these proviruses formed. Multiple full-length open reading frames for all HERV-K proteins are present in the human genome today.
- Published
- 1999
- Full Text
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41. Molecular cloning and cell-specific growth characterization of polymorphic variants of type D serogroup 2 simian retroviruses.
- Author
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Marracci GH, Avery NA, Shiigi SM, Couch G, Palmer H, Pilcher KY, Nichols H, Hallick LM, Axthelm MK, and Machida CA
- Subjects
- Amino Acid Sequence, Animals, Cells, Cultured, DNA, Recombinant, Endopeptidases genetics, Genes, env genetics, Genes, gag genetics, Genes, pol genetics, Genetic Variation, Macaca, Molecular Sequence Data, Monkey Diseases virology, Proviruses genetics, Retroviruses, Simian classification, Sequence Analysis, DNA, T-Lymphocytes virology, Terminal Repeat Sequences genetics, Cloning, Molecular, Genes, Viral, Polymorphism, Genetic, Retroviruses, Simian genetics, Retroviruses, Simian growth & development, Simian Acquired Immunodeficiency Syndrome virology
- Abstract
Simian retroviruses (SRVs), the etiological agent of a spontaneous Simian acquired immunodeficiency syndrome, endemically infects large percentages of Asian macaques housed in biomedical research colonies and severely compromises the effective use of these species as a viable research animal. We recently described the molecular cloning of a serogroup 2 SRV, D2/RHE/OR, which causes mild immunosuppression in rhesus macaques. A restriction site variant, D2/RHE/OR/V1, has also been recovered from severely ill animals endemically infected with D2/RHE/OR. We now report the complete nucleotide sequences of D2/RHE/OR and D2/RHE/OR/V1. Both infectious molecular clones retain the genetic structure typical of type D SRVs (5' LTR-gag-prt-pol-env-3'LTR) and encode identically sized 8105-bp proviruses. D2/RHE/OR and D2/RHE/OR/V1 are 99.3% similar at the amino acid level, exhibiting only 17 residue differences, of which 10 are located in the envelope glycoproteins. The molecular clones and reciprocal chimeric viruses were used to assess the contribution of different genetic domains to virus infectivity in a T cell infection assay. These experiments indicate that D2/RHE/OR has a reduced ability to infect specific T cell lines, especially Hut-78 and MT-4 cells, and that the envelope gene is not the sole determinant of in vitro tropism., (Copyright 1999 Academic Press.)
- Published
- 1999
- Full Text
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42. One-step fluorescent probe product-enhanced reverse transcriptase assay.
- Author
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Arnold BA, Hepler RW, and Keller PM
- Subjects
- Animals, Avian Myeloblastosis Virus enzymology, Avian Myeloblastosis Virus genetics, Cell Line, DNA Probes chemistry, DNA Probes genetics, Fluorescent Dyes, HIV-1 enzymology, HIV-1 genetics, Humans, Leukemia Virus, Murine enzymology, Leukemia Virus, Murine genetics, Polymerase Chain Reaction, Retroviruses, Simian enzymology, Retroviruses, Simian genetics, Sensitivity and Specificity, Tumor Cells, Cultured cytology, Tumor Cells, Cultured enzymology, Tumor Cells, Cultured virology, Clinical Enzyme Tests methods, RNA-Directed DNA Polymerase metabolism, Retroviridae Infections diagnosis
- Abstract
Recently developed PCR-based reverse transcriptase (RT) assays are useful in the detection of retroviruses since they are approximately a millionfold more sensitive than conventional RT assays. However, these assays are both labor- and time-intensive. The previously described product-enhanced reverse transcriptase (PERT) assay involves a two-step RT-PCR followed by detection and quantitation of PCR products by either Southern blot or enzyme-linked immunosorbent assay (ELISA). We have modified the PERT assay to be a one-step, fluorescent probe, PCR-based RT assay that can be completed from sample dilution to final quantitative assay results in approximately 5 h without loss of assay sensitivity or specificity. The assay has a dynamic range of 6 logs, and therefore, extensive sample dilution is not necessary for quantitation. This newly enhanced fluorescent PERT assay can play an important role in the high-throughput detection of retroviral infection and characterization of RT activity.
- Published
- 1998
- Full Text
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43. Amplification of simian retroviral sequences from human recipients of baboon liver transplants.
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Allan JS, Broussard SR, Michaels MG, Starzl TE, Leighton KL, Whitehead EM, Comuzzie AG, Lanford RE, Leland MM, Switzer WM, and Heneine W
- Subjects
- Adult, Animals, Base Sequence, DNA, Viral, Gene Amplification, Humans, Male, Middle Aged, Molecular Sequence Data, Papio, Phylogeny, Retroviridae Infections virology, Retroviruses, Simian classification, Tumor Virus Infections virology, Liver Transplantation adverse effects, Retroviridae Infections transmission, Retroviruses, Simian genetics, Spumavirus genetics, Transplantation, Heterologous adverse effects, Tumor Virus Infections transmission
- Abstract
Investigations into the use of baboons as organ donors for human transplant recipients, a procedure called xenotransplantation, have raised the specter of transmitting baboon viruses to humans and possibly establishing new human infectious diseases. Retrospective analysis of tissues from two human transplant recipients with end-stage hepatic disease who died 70 and 27 days after the transplantation of baboon livers revealed the presence of two simian retroviruses of baboon origin, simian foamy virus (SFV) and baboon endogenous virus (BaEV), in multiple tissue compartments. The presence of baboon mitochondrial DNA was also detected in these same tissues, suggesting that xenogeneic "passenger leukocytes" harboring latent or active viral infections had migrated from the xenografts to distant sites within the human recipients. The persistence of SFV and BaEV in human recipients throughout the posttransplant period underscores the potential infectious risks associated with xenotransplantation.
- Published
- 1998
- Full Text
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44. Mutational analysis of the RNA pseudoknot involved in efficient ribosomal frameshifting in simian retrovirus-1.
- Author
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Sung D and Kang H
- Subjects
- Animals, Base Composition, Base Sequence, Cats, Immunodeficiency Virus, Feline genetics, In Vitro Techniques, Mammary Tumor Virus, Mouse genetics, Mice, Mutagenesis, Site-Directed, Nucleic Acid Conformation, Rabbits, Reticulocytes metabolism, Species Specificity, Frameshifting, Ribosomal, Mutation, RNA, Viral chemistry, RNA, Viral genetics, Retroviruses, Simian genetics
- Abstract
Mutational effects on frameshifting efficiency of the RNA pseudoknot involved in ribosomal frameshifting in simian retrovirus-1 (SRV-1) have been investigated. The primary sequence and the proposed secondary structure of the SRV-1 pseudoknot are similar to those of other efficient frameshifting pseudoknots in mouse mammary tumor virus (MMTV) and feline immunodeficiency virus (FIV), where an unpaired adenine nucleotide intercalates between stem 1 and stem 2. In SRV-1 pseudoknot, the adenine nucleotide in between stem 1 and stem 2 has a potential to form an A*U base pair with the last uridine nucleotide in the loop 2, resulting in a continuous A-form helix with coaxially stacked stem 1 and stem 2. To test whether this A*U base pairing and coaxial stacking of stem 1 and stem 2 is absolutely required for efficient frameshifting in SRV-1, a series of mutants changing this potential A.U base pair to either G.C base pair or A.A, A.G, A.C, G.A, G.G mismatch is generated, and their frameshifting efficiencies are investigated in vitro using rabbit reticulocyte lysate translation assay. The frameshifting abilities of these mutant pseudoknots are similar to that of the wild-type pseudoknot, suggesting that the A*U base pair in between stem 1 and stem 2 is not necessary to promote efficient frameshifting in SRV-1. These results reveal that coaxial stacking of stem 1 and stem 2 with a Watson-Crick A.U base pair in between two stems is not a required structural feature of the pseudoknot for promoting efficient frameshifting in SRV-1. Our mutational data suggest that SRV-1 pseudoknot adopts similar structural features common to other efficient frameshifting pseudoknots as observed in MMTV and FIV.
- Published
- 1998
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- View/download PDF
45. Detection of squirrel monkey retroviral sequences in interferon samples.
- Author
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Pienkowska M and Seth A
- Subjects
- Animals, Base Sequence, Cell Line virology, DNA genetics, DNA, Viral analysis, DNA, Viral genetics, Genome, Humans, Interferons isolation & purification, Polymerase Chain Reaction methods, Interferons genetics, Retroviruses, Simian genetics, Saimiri virology
- Abstract
Background/aims: Interferons have been used therapeutically in viral infections, and as immunomodulants in the treatment of different types of cancers. Interferons have been prepared from human lymphoid cell-lines, such as Namalwa, that contain integrated copies of squirrel monkey retrovirus proviral DNA. Squirrel monkey retrovirus is related to simian type D retroviruses, such as Mason-Pfizer monkey virus. Thus it is important to determine if these retroviral sequences are present in interferon preparations purified from human cell lines., Methods: DNA samples were prepared from 75 commercial interferon preparations and analyzed for squirrel monkey retrovirus sequences by polymerase chain reaction and DNA sequencing. Since single polymerase chain reaction is not as sensitive, a nested polymerase chain reaction strategy was devised in order to detect squirrel monkey retrovirus-pol sequences. Amplification of beta-actin (human) sequences was used to confirm that samples contained human genomic DNA. To determine the authenticity of squirrel monkey retrovirus sequences, we analyzed amplified products by Southern blot hybridization and direct DNA sequencing., Results/conclusions: Thirty-nine samples were positive for squirrel monkey retrovirus-pol sequences by nested polymerase chain reaction. It is noteworthy that 29 samples were either weakly or very weakly positive by single polymerase chain reaction, thus stressing the importance of our sensitive polymerase chain reaction assay. However, it remains to be determined whether the residual DNA sequences detected by our sensitive nested polymerase chain reaction assay have biological consequences.
- Published
- 1998
- Full Text
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46. Development and characterization of conditionally immortalized osteoblast precursor cell lines from human bone marrow stroma.
- Author
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Hicok KC, Thomas T, Gori F, Rickard DJ, Spelsberg TC, and Riggs BL
- Subjects
- Adipocytes metabolism, Antigens, Viral, Tumor genetics, Ascorbic Acid pharmacology, Calcitriol pharmacology, Cell Differentiation, Cell Division, Cell Line, Dexamethasone pharmacology, Genetic Vectors, Glycerophosphates pharmacology, Humans, Immunohistochemistry, Mutation, Osteoblasts metabolism, Osteogenesis drug effects, RNA, Messenger analysis, Retroviruses, Simian genetics, Stromal Cells cytology, Transfection, Adipocytes cytology, Bone Marrow Cells cytology, Osteoblasts cytology
- Abstract
Although the differentiation of mature osteoblasts has been well studied, there is still a need for a convenient way to study preosteoblast differentiation. Our laboratory has recently described a method for isolating small numbers of authentic osteoblast precursor cells from human bone marrow (Rickard et al., J Bone Miner Res 11:312-324, 1996). Here we describe the conditional immortalization of these cells by retroviral transfection with the amphotrophic vector, pZipSV40tsa58, which encodes for a temperature-sensitive mutant form of the simian virus large T-antigen. At the permissive temperature of 34 degrees C, the cell lines proliferated, but differentiation was arrested, whereas at the restrictive temperature of 39.5 degrees C, proliferation was decreased and differentiation was induced. As assessed by semiquantitative reverse transcriptase PCR after 4 days of culture at 39.5 degrees C, the six cell lines expressed similar mRNA levels both constitutively and in response to dexamethasone (Dex) and 1alpha,25-dihydroxyvitamin D3 (1,25(OH2)D3) for osteoblast (alkaline phosphatase [ALP], type I collagen [Col I], osteocalcin [OC], and parathyroid hormone receptor [PTH-R] and adipocyte (lipoprotein lipase [LPL]) genes. In the presence of 10(-8) M Dex, gene expression for ALP, PTH-R, and LPL increased, but that for OC decreased. Stimulation with 10(-8) M 1,25(OH2)D3 increased gene expression for ALP, OC, and Col I. Changes in protein production for ALP, OC, and type I procollagen in response to Dex and 1,25(OH2)D3 were similar to changes in mRNA levels. When cultured at 39.5 degrees C with ascorbate and beta1-glycerolphosphate for 21 days, mineralization of matrix occurred, whereas culture with Dex plus 1,25(OH2)D3, or rabbit serum led to enhanced formation of cytoplasmic lipid droplets within 6 days. Thus, these cell lines are capable of bipotential differentiation and should serve as an excellent tool to study the molecular mechanisms that regulate and select for osteoblast and adipocyte differentiation in humans.
- Published
- 1998
- Full Text
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47. Assignment of the human endogenous retrovirus S71 (SSAV1) to chromosome 18 band q12.3 by radiation hybrid mapping.
- Author
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Kim HS and Crow TJ
- Subjects
- Animals, Chromosome Banding, Chromosome Mapping, Humans, Hybrid Cells, Proviruses genetics, Sarcoma Virus, Woolly Monkey drug effects, Chromosomes, Human, Pair 18 genetics, Endogenous Retroviruses genetics, Retroviruses, Simian genetics
- Published
- 1998
- Full Text
- View/download PDF
48. DNA immunization of mice against SIVmac239 Gag and Env using Rev-independent expression plasmids.
- Author
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Indraccolo S, Feroli F, Minuzzo S, Mion M, Rosato A, Zamarchi R, Titti F, Verani P, Amadori A, and Chieco-Bianchi L
- Subjects
- Animals, Antibodies, Antinuclear immunology, Gene Products, gag genetics, Gene Products, gag immunology, Genes, env genetics, Genes, env immunology, Genetic Vectors genetics, Genetic Vectors immunology, Mice, Mice, Inbred BALB C, Retroviruses, Simian genetics, Simian Immunodeficiency Virus genetics, Immunization methods, SAIDS Vaccines immunology, Vaccines, DNA immunology
- Abstract
Simian immunodeficiency virus (SIV) structural gene expression, including gag and env, strictly depends on the interaction of the viral posttranscriptional regulator Rev with its target RNA, the Rev-responsive element (RRE). A small RNA element, termed the constitutive transport element (CTE), located in the 3' portion of simian retrovirus 1 (SRV-1) mRNA, can efficiently substitute for the human immunodeficiency virus (HIV) Rev-RRE interaction, and thus render HIV expression and replication Rev independent. We tested the ability of the SRV-1 CTE to drive the expression of SIVmac239 env and gag from subgenomic constructs designed for possible use in vaccine trials. In vitro expression studies showed that when the SRV-1 sequence is coupled to the SIV gag and env mRNAs, it functions in an orientation-dependent fashion, and leads to strong expression of SIV Gag and Env in human and monkey cell lines; levels of CTE-mediated protein expression were similar to those obtained with a functional Rev-RRE system. On the other hand, in murine fibroblast-like cells, SIV Gag and Env were expressed from constructs at relatively high levels even in the absence of Rev-RRE; nevertheless, their expression was increased by the presence of the SRV-1 CTE. As reported previously for HIV, the murine cell lines appeared to be defective for Rev-RRE activity, and required overexpression of Rev to induce a Rev response. Intramuscular injection of the gag-CTE and env-CTE constructs in BALB/c mice resulted in the expression of the corresponding mRNAs, and the production of anti-Gag and anti-Env antibodies, thus suggesting that these vectors might be used for genetic immunization approaches.
- Published
- 1998
- Full Text
- View/download PDF
49. The simian retrovirus-1 constitutive transport element, unlike the HIV-1 RRE, uses factors required for cellular mRNA export.
- Author
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Saavedra C, Felber B, and Izaurralde E
- Subjects
- Animals, Biological Transport, Cell Nucleus virology, GTP Phosphohydrolases metabolism, GTP-Binding Proteins metabolism, Gene Products, rev genetics, Heterogeneous Nuclear Ribonucleoprotein A1, Heterogeneous-Nuclear Ribonucleoproteins, Humans, Introns, Nuclear Proteins metabolism, Oocytes metabolism, RNA Splicing, Ribonucleoproteins metabolism, Serum Albumin, Bovine metabolism, Xenopus, ran GTP-Binding Protein, rev Gene Products, Human Immunodeficiency Virus, Gene Products, rev metabolism, HIV-1 genetics, Heterogeneous-Nuclear Ribonucleoprotein Group A-B, RNA, Messenger metabolism, RNA, Viral metabolism, Retroviruses, Simian genetics
- Abstract
Background: A hallmark of retroviral gene expression is that unspliced retroviral genomic RNA is exported to the cytoplasm, whereas endogenous intron-containing cellular RNAs are usually retained in the nucleus. In complex retroviruses, such as human immunodeficiency virus-1 (HIV-1), nuclear export is accomplished by the interaction of a virally encoded protein, Rev, with a cis-acting RNA element, the Rev-responsive element (RRE). In type D retroviruses, such as the simian retrovirus type 1 (SRV-1), however, genomic RNA is exported by cellular factor(s) that interact with a conserved cis-acting RNA element, the constitutive transport element (CTE)., Results: We found that the CTE was exported in a specific and saturable fashion from Xenopus oocyte nuclei. When inserted into the intron of an adenovirus-derived pre-mRNA, the CTE did not affect splicing efficiency but promoted the nuclear export of the excised intron lariat that is normally retained within the nucleus. Export of CTE-containing RNAs to the cytoplasm was not affected by the heterogeneous nuclear ribonucleoprotein A1 or an excess of peptides corresponding to the Rev nuclear export signal. Microinjection of saturating amounts of CTE RNA did not affect tRNA export or Rev-mediated export but did inhibit mRNA export. CTE-mediated export was found to be dependent on Ran-mediated GTP hydrolysis., Conclusion: The Rev-RRE system and the CTE direct intron-containing RNAs to distinct export pathways. Although previous data have suggested that Rev uses the same export pathway as uracil-rich small nuclear RNAs and 5S ribosomal RNA, the CTE seems to interact with evolutionarily conserved factors that are essential for cellular mRNA export.
- Published
- 1997
- Full Text
- View/download PDF
50. Screening for simian type-D retrovirus infection in macaques, using nested polymerase chain reaction.
- Author
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Lerche NW, Cotterman RF, Dobson MD, Yee JL, Rosenthal AN, and Heneine WM
- Subjects
- Animals, Blotting, Western, Macaca fascicularis, Macaca mulatta, Macaca nemestrina, Mass Screening veterinary, Polymerase Chain Reaction methods, Retroviridae Infections diagnosis, Retroviridae Infections virology, Retroviruses, Simian chemistry, Retroviruses, Simian genetics, Sensitivity and Specificity, Tumor Virus Infections diagnosis, Tumor Virus Infections virology, Polymerase Chain Reaction veterinary, Retroviridae Infections veterinary, Retroviruses, Simian isolation & purification, Tumor Virus Infections veterinary
- Abstract
A nested polymerase chain reaction (PCR) assay was developed to detect proviral DNA in peripheral blood mononuclear cells (PBMC) of macaques infected with simian type-D retrovirus (SRV/D). Primers were designed to amplify gag gene sequences of SRV/D serotype 1, 2, and 3 viral genomes and were used in a single assay for simultaneous detection of infection with SRV/D-1, SRV/D-2, or SRV/D-3. Results of plasmid dilution studies indicate sensitivity of nested PCR in the range of 1 to 10 genomic copies. The PBMC samples from 395 macaques of unknown SRV/D status, obtained from several primate facilities, were tested in parallel by Western blot (immunoblot) analysis, virus isolation, and nested PCR. Infection was detected in 60 (15.2%) animals by nested PCR, in 40 (10.1%) animals by virus isolation, and in 28 (7.1%) animals by immunoblot. All 40 culture-positive samples were positive by nested PCR. In addition, 11 of 23 immunoblot-positive/virus isolation-negative samples, 2 of 20 immunoblot-indeterminate/virus isolation-negative samples, and 7 of 312 immunoblot-negative/virus isolation-negative samples were identified as positive by nested PCR. Nested PCR is a sensitive and specific assay for simultaneous screening for infection with serotypes 1, 2, and 3 of simian type D retrovirus, and is a powerful tool for rapid screening and surveillance in macaque colonies.
- Published
- 1997
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