78 results on '"Grubor-Bauk, Branka"'
Search Results
52. A HIV-Tat/C4-binding protein chimera encoded by a DNA vaccine is highly immunogenic and contains acute EcoHIV infection in mice
- Author
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Tomusange, Khamis, primary, Wijesundara, Danushka, additional, Gummow, Jason, additional, Garrod, Tamsin, additional, Li, Yanrui, additional, Gray, Lachlan, additional, Churchill, Melissa, additional, Grubor-Bauk, Branka, additional, and Gowans, Eric J., additional
- Published
- 2016
- Full Text
- View/download PDF
53. A Multiantigenic DNA Vaccine That Induces Broad Hepatitis C Virus-Specific T-Cell Responses in Mice
- Author
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Gummow, Jason, primary, Li, Yanrui, additional, Yu, Wenbo, additional, Garrod, Tamsin, additional, Wijesundara, Danushka, additional, Brennan, Amelia J., additional, Mullick, Ranajoy, additional, Voskoboinik, Ilia, additional, Grubor-Bauk, Branka, additional, and Gowans, Eric J., additional
- Published
- 2015
- Full Text
- View/download PDF
54. Immunocompetent mouse models to evaluate intrahepatic T cell responses to HCV vaccines
- Author
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Yu, Wenbo, primary, Grubor-Bauk, Branka, additional, Mullick, Ranajoy, additional, Das, Saumitra, additional, and Gowans, Eric J, additional
- Published
- 2014
- Full Text
- View/download PDF
55. Encoded novel forms of HSP70 or a cytolytic protein increase DNA vaccine potency
- Author
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Garrod, Tamsin, primary, Grubor-Bauk, Branka, additional, Yu, Stanley, additional, Gargett, Tessa, additional, and Gowans, Eric J, additional
- Published
- 2014
- Full Text
- View/download PDF
56. Increase in DNA vaccine efficacy by virosome delivery and co-expression of a cytolytic protein
- Author
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Gargett, Tessa, primary, Grubor-Bauk, Branka, additional, Miller, Darren, additional, Garrod, Tamsin, additional, Yu, Stanley, additional, Wesselingh, Steve, additional, Suhrbier, Andreas, additional, and Gowans, Eric J, additional
- Published
- 2014
- Full Text
- View/download PDF
57. DNA vaccines encoding membrane‐bound or secreted forms of heat shock protein 70 exhibit improved potency
- Author
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Garrod, Tamsin J., primary, Grubor‐Bauk, Branka, additional, Gargett, Tessa, additional, Li, Yanrui, additional, Miller, Darren S., additional, Yu, Wenbo, additional, Major, Lee, additional, Burrell, Christopher J., additional, Wesselingh, Steven, additional, Suhrbier, Andreas, additional, and Gowans, Eric J., additional
- Published
- 2014
- Full Text
- View/download PDF
58. Induction of antigen‐positive cell death by the expression of Perforin, but not DTa, from a DNA vaccine enhances the immune response
- Author
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Gargett, Tessa, primary, Grubor‐Bauk, Branka, additional, Garrod, Tamsin J, additional, Yu, Wenbo, additional, Miller, Darren, additional, Major, Lee, additional, Wesselingh, Steve, additional, Suhrbier, Andreas, additional, and Gowans, Eric J, additional
- Published
- 2013
- Full Text
- View/download PDF
59. Importance of NKT Cells in Resistance to Herpes Simplex Virus, Fate of Virus-Infected Neurons, and Level of Latency in Mice
- Author
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Grubor-Bauk, Branka, primary, Arthur, Jane Louise, additional, and Mayrhofer, Graham, additional
- Published
- 2008
- Full Text
- View/download PDF
60. Impaired Clearance of Herpes Simplex Virus Type 1 From Mice Lacking CD1d or NKT Cells Expressing the Semivariant Vα14-Jα281 TCR
- Author
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Grubor-Bauk, Branka, primary, Simmons, Anthony, additional, Mayrhofer, Graham, additional, and Speck, Peter G., additional
- Published
- 2003
- Full Text
- View/download PDF
61. Immunocompetent mouse models to evaluate intrahepatic T cell responses to HCV vaccines.
- Author
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Yu, Wenbo, Grubor-Bauk, Branka, Mullick, Ranajoy, Das, Saumitra, and Gowans, Eric J
- Published
- 2015
- Full Text
- View/download PDF
62. Long-term persistence of RBD+memory B cells encoding neutralizing antibodies in SARS-CoV-2 infection
- Author
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Abayasingam, Arunasingam, Balachandran, Harikrishnan, Agapiou, David, Hammoud, Mohamed, Rodrigo, Chaturaka, Keoshkerian, Elizabeth, Li, Hui, Brasher, Nicholas A., Christ, Daniel, Rouet, Romain, Burnet, Deborah, Grubor-Bauk, Branka, Rawlinson, William, Turville, Stuart, Aggarwal, Anupriya, Stella, Alberto Ospina, Fichter, Christina, Brilot, Fabienne, Mina, Michael, Post, Jeffrey J., Hudson, Bernard, Gilroy, Nicky, Dwyer, Dominic, Sasson, Sarah C., Tea, Fiona, Pilli, Deepti, Kelleher, Anthony, Tedla, Nicodemus, Lloyd, Andrew R., Martinello, Marianne, and Bull, Rowena A.
- Abstract
Considerable concerns relating to the duration of protective immunity against severe acute respiratory syndrome-coronavirus-2 (SARS-CoV-2) exist, with evidence of antibody titers declining rapidly after infection and reports of reinfection. Here, we monitor the antibody responses against SARS-CoV-2 receptor-binding domain (RBD) for up to 6 months after infection. While antibody titers are maintained, ∼13% of the cohort’s neutralizing responses return to background. However, encouragingly, in a selected subset of 13 participants, 12 have detectable RBD-specific memory B cells and these generally are increasing out to 6 months. Furthermore, we are able to generate monoclonal antibodies with SARS-CoV-2 neutralizing capacity from these memory B cells. Overall, our study suggests that the loss of neutralizing antibodies in plasma may be countered by the maintenance of neutralizing capacity in the memory B cell repertoire.
- Published
- 2021
- Full Text
- View/download PDF
63. Immunological Memory in Imiquimod-Induced Murine Model of Psoriasiform Dermatitis
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Danushka K. Wijesundara, Kevin A. Fenix, Allison J. Cowin, Branka Grubor-Bauk, Zlatko Kopecki, Fenix, Kevin, Wijesundara, Danushka K, Cowin, Allison J, Grubor-Bauk, Branka, and Kopecki, Zlatko
- Subjects
tissue-resident memory T cells ,T-Lymphocytes ,T cell ,Integrin alpha1 ,Imiquimod ,Granzymes ,Article ,Catalysis ,GZMB ,Inorganic Chemistry ,lcsh:Chemistry ,Mice ,Psoriasis ,Animals ,Humans ,Medicine ,Cell Lineage ,Physical and Theoretical Chemistry ,Molecular Biology ,Psoriasiform Dermatitis ,lcsh:QH301-705.5 ,Spectroscopy ,resolved lesions ,business.industry ,Organic Chemistry ,General Medicine ,psoriasis ,medicine.disease ,Computer Science Applications ,Granzyme B ,Immunosurveillance ,Disease Models, Animal ,medicine.anatomical_structure ,Gene Expression Regulation ,lcsh:Biology (General) ,lcsh:QD1-999 ,Immunology ,Epidermis ,business ,Immunologic Memory ,CD8 ,medicine.drug - Abstract
Psoriasis is a common chronic inflammatory skin condition manifested by T cell responses and characterized by preferential recurrence at previously inflamed sites upon withdrawal of treatment. The site-specific disease memory in psoriasis has been linked to CD8+CD103+ tissue-resident memory T cells (Trm) in the epidermis which were previously thought to only provide &ldquo, frontline&rdquo, protection against pathogens and immunosurveillance during cancer development. In this study, we correlated the presence of a subset of the Trm cells which are also CD49a+ with disease severity in human psoriatic lesions with acute and chronic disease. Using an imiquimod (IMQ)-induced murine model of psoriasiform dermatitis, we also investigated the level of CD49a+ Trm cells in acute, chronic and resolved psoriatic lesions. Investigation of clinical human samples showed that patient disease severity highly correlated with the numbers of epidermal CD49a+ Trm cells. Additionally, this subset of Trm cells was shown to persist in resolved lesions of murine psoriasiform dermatitis once clinical disease features had subsided. Importantly, these CD49a+ Trm cells showed significantly higher levels of granzyme B (GzmB) production compared to acute disease, suggesting a potential role of CD49a+ Trm cells for psoriatic re-occurrence in resolved patients. Better understanding of epidermal CD49a+ Trm cell activity is necessary for development of advanced treatment strategies for psoriasis to permit long-term, continuous disease control.
- Published
- 2020
64. Safety Profile of a Multi-Antigenic DNA Vaccine Against Hepatitis C Virus
- Author
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Zelalem A Mekonnen, Branka Grubor-Bauk, Ilia Voskoboinik, Ashish C. Shrestha, Makutiro Ghislain Masavuli, Eric J. Gowans, Jason Gummow, Danushka K. Wijesundara, Yanrui Li, Gummow, Jason, Masavuli, Makutiro G, Mekonnen, Zelalem A, Li, Yanrui, Wijesundara, Danushka K, Shrestha, Ashish C, Voskoboinik, Ilia, Gowans, Eric J, and Grubor-Bauk, Branka
- Subjects
hepatitis C virus ,0301 basic medicine ,DNA vaccine ,immune breadth ,Hepatitis C Virus ,pre-clinical ,Hepatitis C virus ,Immunology ,lcsh:Medicine ,medicine.disease_cause ,Article ,DNA vaccination ,03 medical and health sciences ,chemistry.chemical_compound ,0302 clinical medicine ,Immunity ,Drug Discovery ,medicine ,Pharmacology (medical) ,NS5B ,perforin ,Pharmacology ,Reactogenicity ,biology ,business.industry ,ELISPOT ,lcsh:R ,Hepatitis C ,medicine.disease ,Virology ,digestive system diseases ,030104 developmental biology ,Infectious Diseases ,cell death ,chemistry ,Perforin ,Pre-clinical ,biology.protein ,030211 gastroenterology & hepatology ,pathology ,business ,toxicology - Abstract
Despite direct acting antivirals (DAAs) curing >, 95% of individuals infected with hepatitis C (HCV), in order to achieve the World Health Organization HCV Global Elimination Goals by 2030 there are still major challenges that need to be overcome. DAAs alone are unlikely to eliminate HCV in the absence of a vaccine that can limit viral transmission. Consequently, a prophylactic HCV vaccine is necessary to relieve the worldwide burden of HCV disease. DNA vaccines are a promising vaccine platform due to their commercial viability and ability to elicit robust T-cell-mediated immunity (CMI). We have developed a novel cytolytic DNA vaccine that encodes non-structural HCV proteins and a truncated mouse perforin (PRF), which is more immunogenic than the respective canonical DNA vaccine lacking PRF. Initially we assessed the ability of the HCV pNS3-PRF and pNS4/5-PRF DNA vaccines to elicit robust long-term CMI without any adverse side-effects in mice. Interferon-&gamma, (IFN-&gamma, ) enzyme-linked immunosorbent spot (ELISpot) assay was used to evaluate CMI against NS3, NS4 and NS5B in a dose-dependent manner. This analysis showed a dose-dependent bell-curve of HCV-specific responses in vaccinated animals. We then thoroughly examined the effects associated with reactogenicity of cytolytic DNA vaccination with the multi-antigenic HCV DNA vaccine (pNS3/4/5B). Hematological, biochemical and histological studies were performed in male Sprague Dawley rats with a relative vaccine dose 10&ndash, 20-fold higher than the proposed dose in Phase I clinical studies. The vaccine was well tolerated, and no toxicity was observed. Thus, the cytolytic multi-antigenic DNA vaccine is safe and elicits broad memory CMI.
- Published
- 2020
- Full Text
- View/download PDF
65. Enhanced T Cell Responses Induced by a Necrotic Dendritic Cell Vaccine, Expressing HCV NS3
- Author
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Zelalem A. Mekonnen, Makutiro G. Masavuli, Wenbo Yu, Jason Gummow, Dawn M. Whelan, Zahraa Al-Delfi, Joseph Torresi, Eric J. Gowans, Branka Grubor-Bauk, Mekonnen, Zelalem A, Masavuli, Makutiro G, Yu, Wenbo, Gummow, Jason, Whelan, Dawn M, Al-Delfi, Zahraa, Torresi, Joseph, Gowans, Eric J, and Grubor-Bauk, Branka
- Subjects
and promotion of well-being ,viruses ,lcsh:QR1-502 ,lcsh:Microbiology ,necrosis ,Hepatitis ,0302 clinical medicine ,vaccine ,Original Research ,0303 health sciences ,Immunogenicity ,Liver Disease ,Cross-presentation ,virus diseases ,Vaccination ,medicine.anatomical_structure ,Infectious Diseases ,3.4 Vaccines ,030220 oncology & carcinogenesis ,HCV ,HIV/AIDS ,Infection ,Biotechnology ,Microbiology (medical) ,Environmental Science and Management ,T cell ,Chronic Liver Disease and Cirrhosis ,Biology ,hepatitis (C) virus ,Microbiology ,DNA vaccination ,Vaccine Related ,03 medical and health sciences ,Immune system ,Antigen ,cell mediated immunity ,Hepatitis - C ,medicine ,030304 developmental biology ,cross presentation ,Prevention ,Inflammatory and immune system ,biochemical phenomena, metabolism, and nutrition ,Prevention of disease and conditions ,Virology ,digestive system diseases ,Emerging Infectious Diseases ,Good Health and Well Being ,Soil Sciences ,Immunization ,Digestive Diseases ,CD8 - Abstract
A vaccine that induces potent, broad and sustained cell-mediated immunity, resulting in effective memory has the potential to restrict hepatitis C (HCV) virus infection. Early, multi-functional CD4+ and CD8+ T cell responses against non-structural protein 3 (NS3) have been associated with HCV clearance. Necrotic cells generate strong immune responses and represent a major antigenic source used by dendritic cells (DC) for processing and presentation, but there is conflicting evidence as to their immunogenicity in vaccination. Immunization with DC loaded with viral antigens has been done in the past, but to date the immunogenicity of live vs. necrotic DC vaccines has not been investigated. We developed a DC2.4 cell line stably expressing HCV NS3, and compared the NS3-specific responses of live vs. necrotic NS3 DC. Vaccination of mice with necrotic NS3 DC increased the breadth of T-cell responses and enhanced the production of IL-2, TNF-α, and IFN-γ by effector memory CD4+ and CD8+T cells, compared to mice vaccinated with live NS3 DC. A single dose of necrotic NS3 DC vaccine induced a greater influx and activation of cross-presenting CD11c+ CD8α+ DC and necrosis-sensing Clec9A+ DC in the draining lymph nodes. Furthermore, using a hydrodynamic challenge model necrotic NS3 DC vaccination resulted in enhanced clearance of NS3-positive hepatocytes from the livers of vaccinated mice. Taken together, the data demonstrate that necrotic DC represent a novel and exciting vaccination strategy capable of inducing broad and multifunctional T cell memory.
- Published
- 2020
66. Induction of Genotype Cross-Reactive, Hepatitis C Virus-Specific, Cell-Mediated Immunity in DNA-Vaccinated Mice
- Author
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Charani Ranasinghe, Wenbo Yu, Benjamin J Quah, Joseph Torresi, Danushka K. Wijesundara, Eric J. Gowans, Branka Grubor-Bauk, Jason Gummow, Yanrui Li, Wijesundara, Danushka K, Gummow, Jason, Li, Yanrui, Yu, Wenbo, Quah, Benjamin J, Ranasinghe, Charani, Torresi, Joseph, Gowans, Eric J, and Grubor-Bauk, Branka
- Subjects
Viral Hepatitis Vaccines ,0301 basic medicine ,Genotype ,NS5 ,T-Lymphocytes ,viruses ,Hepatitis C virus ,Immunology ,ELISpot ,Hepacivirus ,Cross Reactions ,Viral Nonstructural Proteins ,Biology ,medicine.disease_cause ,Microbiology ,Virus ,DNA vaccines ,DNA vaccination ,Mice ,03 medical and health sciences ,chemistry.chemical_compound ,Immunity ,vaccine ,Virology ,Vaccines and Antiviral Agents ,Vaccines, DNA ,medicine ,Animals ,Humans ,hepatitis ,NS5B ,Immunity, Cellular ,Mice, Inbred BALB C ,NS3 ,Immunogenicity ,multiantigenic ,virus diseases ,Hepatitis C ,digestive system diseases ,Vaccination ,multigenotypic ,HEK293 Cells ,030104 developmental biology ,chemistry ,consensus ,Insect Science ,HCV ,Female - Abstract
A universal hepatitis C virus (HCV) vaccine should elicit multiantigenic, multigenotypic responses, which are more likely to protect against challenge with the range of genotypes and subtypes circulating in the community. A vaccine cocktail and vaccines encoding consensus HCV sequences are attractive approaches to achieve this goal. Consequently, in a series of mouse vaccination studies, we compared the immunogenicity of a DNA vaccine encoding a consensus HCV nonstructural 5B (NS5B) protein to that of a cocktail of DNA plasmids encoding the genotype 1b (Gt1b) and Gt3a NS5B proteins. To complement this study, we assessed responses to a multiantigenic cocktail regimen by comparing a DNA vaccine cocktail encoding Gt1b and Gt3a NS3, NS4, and NS5B proteins to a single-genotype NS3/4/5B DNA vaccine. To thoroughly evaluate in vivo cytotoxic T lymphocyte (CTL) and T helper (Th) cell responses against Gt1b and Gt3a HCV peptide-pulsed target cells, we exploited a novel fluorescent-target array (FTA). FTA and enzyme-linked immunosorbent spot (ELISpot) analyses collectively indicated that the cocktail regimens elicited higher responses to Gt1b and Gt3a NS5B proteins than those with the consensus vaccine, while the multiantigenic DNA cocktail significantly increased the responses to NS3 and NS5B compared to those elicited by the single-genotype vaccines. Thus, a DNA cocktail vaccination regimen is more effective than a consensus vaccine or a monovalent vaccine at increasing the breadth of multigenotypic T cell responses, which has implications for the development of vaccines for communities where multiple HCV genotypes circulate. IMPORTANCE Despite the development of highly effective direct-acting antivirals (DAA), infections with hepatitis C virus (HCV) continue, particularly in countries where the supply of DAA is limited. Furthermore, patients who eliminate the virus as a result of DAA therapy can still be reinfected. Thus, a vaccine for HCV is urgently required, but the heterogeneity of HCV strains makes the development of a universal vaccine difficult. To address this, we developed a novel cytolytic DNA vaccine which elicits robust cell-mediated immunity (CMI) to the nonstructural (NS) proteins in vaccinated animals. We compared the immune responses against genotypes 1 and 3 that were elicited by a consensus DNA vaccine or a DNA vaccine cocktail and showed that the cocktail induced higher levels of CMI to the NS proteins of both genotypes. This study suggests that a universal HCV vaccine can most readily be achieved by use of a DNA vaccine cocktail.
- Published
- 2018
67. Loss of long term protection with the inclusion of HIV pol to a DNA vaccine encoding gag
- Author
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Branka Grubor-Bauk, Lee Major, Tamsin J. Garrod, Wenbo Yu, Tessa Gargett, Eric J. Gowans, Steven Lodewyk Wesselingh, Andreas Suhrbier, Christopher J. Burrell, Garrod, Tamsin J, Gargett, Tessa, Yu, Wenbo, Major, Lee, Burrell, Christopher J, Wesselingh, Steven, Suhrbier, Andreas, Grubor-Bauk, Branka, and Gowans, Eric J
- Subjects
DNA vaccine ,Cancer Research ,HIV Antigens ,T cell ,HIV Infections ,CD8-Positive T-Lymphocytes ,Biology ,gag Gene Products, Human Immunodeficiency Virus ,DNA vaccination ,Immune system ,Antigen ,Immunity ,Virology ,medicine ,Animals ,HIV vaccine ,AIDS Vaccines ,Vaccines ,HIV ,Mice, Inbred C57BL ,Vaccination ,Disease Models, Animal ,Infectious Diseases ,medicine.anatomical_structure ,pol Gene Products, Human Immunodeficiency Virus ,Immunology ,Female ,EcoHIV challenge ,Spleen ,CD8 - Abstract
Traditional vaccine strategies that induce antibody responses have failed to protect against HIV infection in clinical trials, and thus cell-mediated immunity is now an additional criterion. Recent clinical trials that aimed to induce strong T cell responses failed to do so. Therefore, to enhance induction of protective T cell responses, it is crucial that the optimum antigen combination is chosen. Limited research has been performed into the number of antigens selected for an HIV vaccine. This study aimed to compare DNA vaccines encoding either a single HIV antigen or a combination of two antigens, using intradermal vaccination of C57BL/6 mice. Immune assays were performed on splenocytes, and in vivo protection was examined by challenge with a chimeric virus, EcoHIV, able to infect mouse but not human leukocytes, at 10 days (short term) and 60 days (long term) post final vaccination. At 60 days there was significantly lower frequency of induced antigen-specific CD8+ T cells in the spleens of pCMVgag-pol-vaccinated mice compared with mice which received pCMVgag only. Most importantly, short term viral control of EcoHIV was similar for pCMVgag and pCMVgag-pol-vaccinated mice at day 10, but only the pCMVgag-vaccinated significantly controlled EcoHIV at day 60 compared with pCMV-vaccinated mice, showing that control was reduced with the inclusion of the HIV pol gene. Refereed/Peer-reviewed
- Published
- 2014
68. Cytolytic DNA vaccine encoding lytic perforin augments the maturation of- and antigen presentation by- dendritic cells in a time-dependent manner
- Author
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Wenbo Yu, John D. Hayball, Kerrilyn R. Diener, Ilia Voskoboinik, Eric J. Gowans, Branka Grubor-Bauk, Danushka K. Wijesundara, Preethi Eldi, Ben J. C. Quah, Wijesundara, Danushka K, Yu, Wenbo, Quah, Ben JC, Eldi, Preethi, Hayball, John D, Diener, Kerrilyn R, Voskoboinik, Ilia, Gowans, Eric J, and Grubor-Bauk, Branka
- Subjects
0301 basic medicine ,Time Factors ,Science ,T cell ,Antigen presentation ,T cells ,Priming (immunology) ,chemical and pharmacologic phenomena ,Article ,DNA vaccination ,03 medical and health sciences ,plasmid DNA ,0302 clinical medicine ,Vaccines, DNA ,medicine ,Animals ,Humans ,dendritic cells ,Antigens ,cytomegalovirus ,self-antigens ,Cell Proliferation ,CD86 ,Antigen Presentation ,Multidisciplinary ,biology ,Perforin ,Dendritic Cells ,Molecular biology ,Multidisciplinary Sciences ,Mice, Inbred C57BL ,cytolytic DNA immunisation ,HEK293 Cells ,030104 developmental biology ,medicine.anatomical_structure ,Lytic cycle ,biology.protein ,Medicine ,CD80 ,B lymphocytes ,T-Lymphocytes, Cytotoxic ,030215 immunology - Abstract
The use of cost-effective vaccines capable of inducing robust CD8+ T cell immunity will contribute significantly towards the elimination of persistent viral infections and cancers worldwide. We have previously reported that a cytolytic DNA vaccine encoding an immunogen and a truncated mouse perforin (PRF) protein significantly augments anti-viral T cell (including CD8+ T cell) immunity. Thus, the current study investigated whether this vaccine enhances activation of dendritic cells (DCs) resulting in greater priming of CD8+ T cell immunity. In vitro data showed that transfection of HEK293T cells with the cytolytic DNA resulted in the release of lactate dehydrogenase, indicative of necrotic/lytic cell death. In vitro exposure of this lytic cell debris to purified DCs from naïve C57BL/6 mice resulted in maturation of DCs as determined by up-regulation of CD80/CD86. Using activation/proliferation of adoptively transferred OT-I CD8+ T cells to measure antigen presentation by DCs in vivo, it was determined that cytolytic DNA immunisation resulted in a time-dependent increase in the proliferation of OT-I CD8+ T cells compared to canonical DNA immunisation. Overall, the data suggest that the cytolytic DNA vaccine increases the activity of DCs which has important implications for the design of DNA vaccines to improve their translational prospects.
- Published
- 2017
69. Encoded novel forms of HSP70 or a cytolytic protein increase DNA vaccine potency
- Author
-
Branka Grubor-Bauk, Eric J. Gowans, Stanley Yu, Tamsin J. Garrod, Tessa Gargett, Garrod, Tamsin, Grubor-Bauk, Branka, Yu, Stanley, Gargett, Tessa, and Gowans, Eric J
- Subjects
DNA vaccine ,Immunogen ,medicine.medical_treatment ,Immunology ,heat shock protein ,chemical and pharmacologic phenomena ,Biology ,DNA vaccination ,protective immunity ,Immune system ,adjuvant ,Adjuvants, Immunologic ,Heat shock protein ,Vaccines, DNA ,medicine ,Animals ,Immunology and Allergy ,HSP70 Heat-Shock Proteins ,AIDS Vaccines ,Pharmacology ,Innate immune system ,Perforin ,Immunogenicity ,cytolytic ,Pattern recognition receptor ,Virology ,Biotechnology & Applied Microbiology ,Commentary ,Adjuvant - Abstract
In humans, DNA vaccines have failed to demonstrate the equivalent levels of immunogenicity that were shown in smaller animals. Previous studies have encoded adjuvants, predominantly cytokines, within these vaccines in an attempt to increase antigen-specific immune responses. However, these strategies have lacked breadth of innate immune activation and have led to disappointing results in clinical trials. Damage associated molecular patterns (DAMPs) have been identified as pattern recognition receptor (PRR) agonists. DAMPs can bind to a wide range of PRRs on dendritic cells (DCs) and thus our studies have aimed to utilize this characteristic to act as an adjuvant in a DNA vaccine approach. Specifically, HSP70 has been identified as a DAMP, but has been limited by its lack of accessibility to PRRs in and on DCs. Here, we discuss the promising results achieved with the inclusion of membrane-bound or secreted HSP70 into a DNA vaccine encoding HIV gag as the model immunogen. Refereed/Peer-reviewed
- Published
- 2014
70. A novel challenge model to evaluate the efficacy of hepatitis C virus vaccines in mice
- Author
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Tamsin J. Garrod, Wenbo Yu, Tessa Gargett, Eric J. Gowans, Branka Grubor-Bauk, Yu, Wenbo, Grubor-Bauk, Branka, Gargett, Tessa, Garrod, Tamsin, and Gowans, Eric J
- Subjects
hepatitis C virus ,NS3 ,General Veterinary ,General Immunology and Microbiology ,Hepatitis C virus ,Immunology ,Public Health, Environmental and Occupational Health ,Transfection ,Research & Experimental Medicine ,Biology ,medicine.disease_cause ,Virology ,Molecular biology ,animal models ,Infectious Diseases ,Plasmid ,Medicine, Research & Experimental ,Antigen ,hydrodynamic injection ,vaccine ,Gene expression ,medicine ,Molecular Medicine ,Luciferase ,CD8 - Abstract
An effective hepatitis C virus (HCV) vaccine should elicit robust humoral and cell mediated immunity (CMI). A small animal challenge model is required to assess the efficacy of vaccines which elicit CMI. In this study, HCV proteins were expressed in hepatocytes of immunocompetent mice after hydrodynamic injection of a plasmid encoding the HCV NS3/4A protein. This vector, constructed as the "challenge", was optimized for long term, specific gene expression in hepatocytes. To monitor HCV antigen expression in transfected hepatocytes, the plasmid also encoded secreted alkaline phosphatase (SEAP), which was detected in the mouse serum. The design of this novel challenge plasmid was based on studies using luciferase and SEAP as reporter molecules to examine the kinetics of the proteins expressed in hepatocytes and secreted into blood. We designed two constructs to control SEAP expression. In one construct, SEAP expression was controlled by the EMCV IRES, while in the other, a SEAP and luciferase polyprotein was cleaved by the FMDV2A proteinase. We found that SEAP expressed after FMDV2A self cleavage was more sensitive and showed a higher correlation with luciferase expressed in liver. The NSIMA challenge model using the FMDV2A design provided a window period of 50 days to monitor changes in SEAP expression after hydrodynamic injection of DNA. In a challenge experiment, mice which received an adenovirus-based HCV vaccine showed accelerated clearance of SEAP and thus, of NS3/4A positive hepatocytes compared with a mock vaccinated group, that coincided with an increased number of CD8(+) lymphocytes in the liver. Refereed/Peer-reviewed
- Published
- 2014
71. A HIV-Tat/C4-binding protein chimera encoded by a DNA vaccine is highly immunogenic and contains acute EcoHIV infection in mice
- Author
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Danushka K. Wijesundara, Branka Grubor-Bauk, Khamis Tomusange, Eric J. Gowans, Jason Gummow, Tamsin J. Garrod, Yanrui Li, Lachlan Robert Gray, Melissa J Churchill, Tomusange, Khamis, Wijesundara, Danushka, Gummow, Jason, Garrod, Tamsin, Li, Yanrui, Gray, Lachlan, Churchill, Melissa, Grubor-Bauk, Branka, and Gowans, Eric J
- Subjects
0301 basic medicine ,Recombinant Fusion Proteins ,Survivin ,HIV Infections ,HIV Antibodies ,Biology ,Article ,DNA vaccines ,DNA vaccination ,Pathogenesis ,Mice ,03 medical and health sciences ,Chimera (genetics) ,0302 clinical medicine ,Immune system ,In vivo ,Immunity ,Vaccines, DNA ,Animals ,Humans ,030212 general & internal medicine ,Immunity, Cellular ,Multidisciplinary ,C4b-binding protein ,Immunogenicity ,Virology ,HIV-Tat ,030104 developmental biology ,Immunology ,HIV-1 ,tat Gene Products, Human Immunodeficiency Virus ,Apoptosis Regulatory Proteins - Abstract
DNA vaccines are cost-effective to manufacture on a global scale and Tat-based DNA vaccines have yielded protective outcomes in preclinical and clinical models of human immunodeficiency virus (HIV), highlighting the potential of such vaccines. However, Tat-based DNA vaccines have been poorly immunogenic and despite the administration of multiple doses and/or the addition of adjuvants, these vaccines are not in general use. In this study, we improved Tat immunogenicity by fusing it with the oligomerisation domain of a chimeric C4-binding protein (C4b-p), termed IMX313, resulting in Tat heptamerisation and linked Tat to the leader sequence of tissue plasminogen activator (TPA) to ensure that the bulk of heptamerised Tat is secreted. Mice vaccinated with secreted Tat fused to IMX313 (pVAX-sTat-IMX313) developed higher titres of Tat-specific serum IgG, mucosal sIgA and cell-mediated immune (CMI) responses and showed superior control of EcoHIV infection, a surrogate murine HIV challenge model, compared with animals vaccinated with other test vaccines. Given the crucial contribution of Tat to HIV-1 pathogenesis and the precedent of Tat-based DNA vaccines in conferring some level of protection in animal models, we believe that the virologic control demonstrated with this novel multimerised Tat vaccine highlights the promise of this vaccine candidate for humans.
- Published
- 2016
72. Engineering human rhinovirus serotype-A1 as a vaccine vector
- Author
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Danushka K. Wijesundara, Andreas Suhrbier, Wenbo Yu, Branka Grubor-Bauk, Khamis Tomusange, Eric J. Gowans, Tomusange, Khamis, Yu, Wenbo, Suhrbier, Andreas, Wijesundara, Danushka, Grubor-Bauk, Branka, and Gowans, Eric J
- Subjects
Cancer Research ,vaccine vector ,Rhinovirus ,HIV Tat ,medicine.medical_treatment ,Genetic Vectors ,Human immunodeficiency virus (HIV) ,Biology ,Cleavage (embryo) ,medicine.disease_cause ,gag Gene Products, Human Immunodeficiency Virus ,law.invention ,Rhinovirus serotype ,law ,Virology ,medicine ,Humans ,Mucosal immunity ,Gene ,AIDS Vaccines ,Recombination, Genetic ,Genetics ,Drug Carriers ,Vaccines, Synthetic ,Protease ,HIV Gag ,Group-specific antigen ,Infectious Diseases ,Recombinant DNA ,tat Gene Products, Human Immunodeficiency Virus ,human rhinovirus A1 ,Genetic Engineering - Abstract
Herein we describe the construction of recombinant human rhinoviruses (rHRVs) encoding HIV Gag or Tat by inserting the full length tat gene or regions of the gag gene flanked by sequences encoding the HRV 2A protease cleavage site into the junction between HRV genes encoding structural (P1) and nonstructural (P2) proteins. Most recombinants were unstable, but this was corrected by mutation of the flanking cleavage sites. Thereafter, all rHRV constructs retained the inserts throughout six passages. Such constructs may find utility as vaccine vectors to generate mucosal immunity. Refereed/Peer-reviewed
- Published
- 2015
73. A Multiantigenic DNA Vaccine That Induces Broad Hepatitis C Virus-Specific T-Cell Responses in Mice
- Author
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Danushka K. Wijesundara, Branka Grubor-Bauk, Ranajoy Mullick, Tamsin J. Garrod, Amelia J. Brennan, Jason Gummow, Eric J. Gowans, Wenbo Yu, Ilia Voskoboinik, Yanrui Li, Gummow, Jason, Yanrui, Li, Wenbo, Yu, Garrod, Tamsin, Wijesundara, Danushka, Brennan, Amelia J, Mullick, Ranajoy, Voskoboinik, Ilia, Grubor-Bauk, Branka, and Gowansa, Eric J
- Subjects
Male ,Viral Hepatitis Vaccines ,T-Lymphocytes ,viruses ,Hepatitis C virus ,Immunology ,Hepacivirus ,Viral Nonstructural Proteins ,Biology ,Antibodies, Viral ,medicine.disease_cause ,in-vivo ,Microbiology ,Virus ,DNA vaccination ,Mice ,antigen ,Virology ,Vaccines and Antiviral Agents ,innate ,Vaccines, DNA ,medicine ,Animals ,Humans ,dendritic cells ,cleavage ,cross-presentation ,Microbiology & Cell Biology ,Immunity, Cellular ,NS3 ,Attenuated vaccine ,Liver infection ,Immunogenicity ,adaptive immunity ,Hepatitis C ,infection ,digestive system diseases ,Mice, Inbred C57BL ,Vaccination ,Insect Science ,Female ,Immunization ,protein ,complex - Abstract
There are 3 to 4 million new hepatitis C virus (HCV) infections annually around the world, but no vaccine is available. Robust T-cell mediated responses are necessary for effective clearance of the virus, and DNA vaccines result in a cell-mediated bias. Adjuvants are often required for effective vaccination, but during natural lytic viral infections damage-associated molecular patterns (DAMPs) are released, which act as natural adjuvants. Hence, a vaccine that induces cell necrosis and releases DAMPs will result in cell-mediated immunity (CMI), similar to that resulting from natural lytic viral infection. We have generated a DNA vaccine with the ability to elicit strong CMI against the HCV nonstructural (NS) proteins (3, 4A, 4B, and 5B) by encoding a cytolytic protein, perforin (PRF), and the antigens on a single plasmid. We examined the efficacy of the vaccines in C57BL/6 mice, as determined by gamma interferon enzyme-linked immunosorbent spot assay, cell proliferation studies, and intracellular cytokine production. Initially, we showed that encoding the NS4A protein in a vaccine which encoded only NS3 reduced the immunogenicity of NS3, whereas including PRF increased NS3 immunogenicity. In contrast, the inclusion of NS4A increased the immunogenicity of the NS3, NS4B, andNS5B proteins, when encoded in a DNA vaccine that also encoded PRF. Finally, vaccines that also encoded PRF elicited similar levels of CMI against each protein after vaccination with DNA encoding NS3, NS4A, NS4B, and NS5B compared to mice vaccinated with DNA encoding only NS3 or NS4B/5B. Thus, we have developed a promising “multiantigen” vaccine that elicits robust CMI. IMPORTANCE Since their development, vaccines have reduced the global burden of disease. One strategy for vaccine development is to use commercially viable DNA technology, which has the potential to generate robust immune responses. Hepatitis C virus causes chronic liver infection and is a leading cause of liver cancer. To date, no vaccine is currently available, and treatment is costly and often results in side effects, limiting the number of patients who are treated. Despite recent advances in treatment, prevention remains the key to efficient control and elimination of this virus. Here, we describe a novel DNA vaccine against hepatitis C virus that is capable of inducing robust cell-mediated immune responses in mice and is a promising vaccine candidate for humans.
- Published
- 2015
74. Immunocompetent mouse models to evaluate intrahepatic T cell responses to HCV vaccines
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Ranajoy Mullick, Eric J. Gowans, Branka Grubor-Bauk, Saumitra Das, Wenbo Yu, Yu, Wenbo, Grubor-Bauk, Branka, Mullick, Ranajoy, Das, Saumitra, and Gowans, Eric J
- Subjects
Viral Hepatitis Vaccines ,NOVEL VACCINES/Commentaries ,T-Lymphocytes ,T cell ,Immunology ,mouse challenge models ,hepatitis HCV vaccine ,Hepacivirus ,Viral Nonstructural Proteins ,Biology ,Transfection ,T cell response ,Cell Line ,Antigen ,Small animal ,medicine ,Immunology and Allergy ,Animals ,Immunocompetent mouse ,Pharmacology ,Immunity, Cellular ,Surrogate endpoint ,C virus ,Alkaline Phosphatase ,Virology ,Mice, Inbred C57BL ,Vaccination ,Disease Models, Animal ,medicine.anatomical_structure ,Biotechnology & Applied Microbiology ,Liver ,Female ,SCID - Severe combined immunodeficiency - Abstract
An effective hepatitis C virus (HCV) vaccine should elicit robust humoral and cell mediated immunity (CMI). A small animal challenge model is required to assess the efficacy of vaccines which elicit CMI. In this study, HCV proteins were expressed in hepatocytes of immunocompetent mice after hydrodynamic injection of a plasmid encoding the HCV NS3/4A protein. This vector, constructed as the "challenge", was optimized for long term, specific gene expression in hepatocytes. To monitor HCV antigen expression in transfected hepatocytes, the plasmid also encoded secreted alkaline phosphatase (SEAP), which was detected in the mouse serum. The design of this novel challenge plasmid was based on studies using luciferase and SEAP as reporter molecules to examine the kinetics of the proteins expressed in hepatocytes and secreted into blood. We designed two constructs to control SEAP expression. In one construct, SEAP expression was controlled by the EMCV IRES, while in the other, a SEAP and luciferase polyprotein was cleaved by the FMDV2A proteinase. We found that SEAP expressed after FMDV2A self cleavage was more sensitive and showed a higher correlation with luciferase expressed in liver. The NS3/4A challenge model using the FMDV2A design provided a window period of 50 days to monitor changes in SEAP expression after hydrodynamic injection of DNA. In a challenge experiment, mice which received an adenovirus-based HCV vaccine showed accelerated clearance of SEAP and thus, of NS3/4A positive hepatocytes compared with a mock vaccinated group, that coincided with an increased number of CD8(+) lymphocytes in the liver.
- Published
- 2014
75. Increase in DNA vaccine efficacy by virosome delivery and co-expression of a cytolytic protein
- Author
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Andreas Suhrbier, Branka Grubor-Bauk, Steven Lodewyk Wesselingh, Tessa Gargett, Darren S. Miller, Stanley Yu, Tamsin J. Garrod, Eric J. Gowans, Gargett, Tessa, Grubor-Bauk, Branka, Miller, Darren, Garrod, Tamsin, Yu, Stanley, Wesselingh, Steven, Suhrbier, Andreas, and Gowans, Eric J
- Subjects
DNA vaccine ,Immunogen ,biology ,cell-mediated immune response ,NSP4 ,Immunology ,Antigen presentation ,virosome ,biochemical phenomena, metabolism, and nutrition ,luciferase ,Virology ,targeted IN delivery ,DNA vaccination ,Virosome ,Immune system ,cytolytic protein ,Perforin ,Antigen ,Immunity ,biology.protein ,Immunology and Allergy ,Original Article ,General Nursing - Abstract
The potential of DNA vaccines has not been realised due to suboptimal delivery, poor antigen expression and the lack of localised inflammation, essential for antigen presentation and an effective immune response to the immunogen. Initially, we examined the delivery of a DNA vaccine encoding a model antigen, luciferase (LUC), to the respiratory tract of mice by encapsulation in a virosome. Virosomes that incorporated influenza virus haemagglutinin effectively delivered DNA to cells in the mouse respiratory tract and resulted in antigen expression and systemic and mucosal immune responses to the immunogen after an intranasal (IN) prime/intradermal (ID) boost regimen, whereas a multidose ID regimen only generated systemic immunity. We also examined systemic immune responses to LUC after ID vaccination with a DNA vaccine, which also encoded one of the several cytolytic or toxic proteins. Although the herpes simplex virus thymidine kinase, in the presence of the prodrug, ganciclovir, resulted in cell death, this failed to increase the humoral or cell-mediated immune responses. In contrast, the co-expression of LUC with the rotavirus non-structural protein 4 (NSP4) protein or a mutant form of mouse perforin, proteins which are directly cytolytic, resulted in increased LUC-specific humoral and cell-mediated immunity. On the other hand, co-expression of LUC with diphtheria toxin subunit A or overexpression of perforin or NSP4 resulted in a lower level of immunity. In summary, the efficacy of DNA vaccines can be improved by targeted IN delivery of DNA or by the induction of cell death in vaccine-targeted cells after ID delivery. Refereed/Peer-reviewed
- Published
- 2014
76. Specific and off-target immune responses following COVID-19 vaccination with ChAdOx1-S and BNT162b2 vaccines-an exploratory sub-study of the BRACE trial.
- Author
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Messina NL, Germano S, McElroy R, Bonnici R, Grubor-Bauk B, Lynn DJ, McDonald E, Nicholson S, Perrett KP, Pittet LF, Rudraraju R, Stevens NE, Subbarao K, and Curtis N
- Subjects
- Humans, Female, Male, Adult, Middle Aged, Vaccination, COVID-19 Vaccines immunology, COVID-19 Vaccines administration & dosage, Prospective Studies, Health Personnel, Spike Glycoprotein, Coronavirus immunology, BNT162 Vaccine immunology, BNT162 Vaccine administration & dosage, COVID-19 prevention & control, COVID-19 immunology, SARS-CoV-2 immunology, Cytokines metabolism, Antibodies, Viral immunology, Antibodies, Viral blood, ChAdOx1 nCoV-19 immunology
- Abstract
Background: The COVID-19 pandemic led to the rapid development and deployment of several highly effective vaccines against SARS-CoV-2. Recent studies suggest that these vaccines may also have off-target effects on the immune system. We sought to determine and compare the off-target effects of the adenovirus vector ChAdOx1-S (Oxford-AstraZeneca) and modified mRNA BNT162b2 (Pfizer-BioNTech) vaccines on immune responses to unrelated pathogens., Methods: Prospective sub-study within the BRACE trial. Blood samples were collected from 284 healthcare workers before and 28 days after ChAdOx1-S or BNT162b2 vaccination. SARS-CoV-2-specific antibodies were measured using ELISA, and whole blood cytokine responses to specific (SARS-CoV-2) and unrelated pathogen stimulation were measured by multiplex bead array., Findings: Both vaccines induced robust SARS-CoV-2 specific antibody and cytokine responses. ChAdOx1-S vaccination increased cytokine responses to heat-killed (HK) Candida albicans and HK Staphylococcus aureus and decreased cytokine responses to HK Escherichia coli and BCG. BNT162b2 vaccination decreased cytokine response to HK E. coli and had variable effects on cytokine responses to BCG and resiquimod (R848). After the second vaccine dose, BNT162b2 recipients had greater specific and off-target cytokine responses than ChAdOx1-S recipients., Interpretation: ChAdOx1-S and BNT162b2 vaccines alter cytokine responses to unrelated pathogens, indicative of potential off-target effects. The specific and off-target effects of these vaccines differ in their magnitude and breadth. The clinical relevance of these findings is uncertain and needs further study., Funding: Bill & Melinda Gates Foundation, National Health and Medical Research Council, Swiss National Science Foundation and the Melbourne Children's. BRACE trial funding is detailed in acknowledgements., Competing Interests: Declaration of interests All authors declare no conflicts of interest. The BRACE trial consortium funding is detailed in the acknowledgements., (Copyright © 2024 The Author(s). Published by Elsevier B.V. All rights reserved.)
- Published
- 2024
- Full Text
- View/download PDF
77. Human Rhinovirus-A1 as an Expression Vector.
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Tomusange K, Wijesundara D, Gowans EJ, and Grubor-Bauk B
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- Enterovirus genetics, Genetic Vectors, HeLa Cells, Humans, Virus Replication, gag Gene Products, Human Immunodeficiency Virus immunology, tat Gene Products, Human Immunodeficiency Virus immunology, Enterovirus physiology, gag Gene Products, Human Immunodeficiency Virus genetics, tat Gene Products, Human Immunodeficiency Virus genetics
- Abstract
Expression vectors that are based on live human rhinoviruses (HRVs) are attractive, yet often overlooked in vaccine development due to their limited capacity for foreign gene inserts and poor genetic stability. This chapter describes a novel methodology to engineer a replication-competent genetically stable recombinant HRV (rHRV) without affecting viral replication capability. We have previously used these methods to generate live, genetically stable recombinant HRVs encoding HIV Gag and Tat proteins (rHRV-Gag-Tat), a potential mucosally targeted HIV vaccine.
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- 2017
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78. Impaired clearance of herpes simplex virus type 1 from mice lacking CD1d or NKT cells expressing the semivariant V alpha 14-J alpha 281 TCR.
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Grubor-Bauk B, Simmons A, Mayrhofer G, and Speck PG
- Subjects
- Animals, Antigens, CD1d, Antigens, Viral metabolism, Ganglia, Sensory immunology, Ganglia, Sensory virology, Genetic Variation immunology, Herpes Simplex genetics, Herpesvirus 1, Human growth & development, Killer Cells, Natural metabolism, Killer Cells, Natural pathology, Mice, Mice, Inbred C57BL, Mice, Knockout, Protein Isoforms genetics, Receptors, Antigen, T-Cell, alpha-beta genetics, Severity of Illness Index, T-Lymphocyte Subsets metabolism, T-Lymphocyte Subsets pathology, Time Factors, Viral Load, Virus Latency genetics, Virus Latency immunology, Antigens, CD1 genetics, Herpes Simplex immunology, Herpes Simplex virology, Herpesvirus 1, Human immunology, Killer Cells, Natural immunology, Receptors, Antigen, T-Cell, alpha-beta biosynthesis, Receptors, Antigen, T-Cell, alpha-beta deficiency, T-Lymphocyte Subsets immunology
- Abstract
Ag-presenting molecule CD1 and CD1-restricted NKT cells are known to contribute to defense against a range of infectious pathogens, including some viruses. CD1-restricted NKT cells, a distinct subpopulation of T cells, have striking and rapid effector functions that contribute to host defense, including rapid production of IFN-gamma and IL-4, and activation of NK cells. Consideration of the important contributions of innate and adaptive immunity to clearance of HSV prompted us to investigate the role of CD1 and of NKT cells expressing the V alpha 14-J alpha 281 TCR in the pathogenesis of HSV infection. To address this issue, we compared infection in wild-type mice with that in CD1 gene knockout (GKO) and J alpha 281 GKO mice. In this study, we report impaired clearance of virus and viral Ags, and more florid acute infection in mice lacking CD1 (and by inference, CD1-restricted T cells), in comparison with parental C57BL6 mice. In J alpha 281 GKO mice there was also impairment of virus clearance, resembling that seen in CD1 GKO mice. These results imply roles for the V alpha 14-J alpha 281 subset of NKT cells and for CD1d in control of HSV infection.
- Published
- 2003
- Full Text
- View/download PDF
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