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Faculty
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MS; University of Wisconsin; 1969
PhD; University of Wisconsin; 1972
Postdoctoral; Albert Einstein College; 1972-74
Postdoctoral; Hershey Medical Center; 1975-77
Cellular and molecular basis of HIV pathogenesis, Development of a novel vaccine candidates against HIV and SIV, Molecular mechanisms of sexual transmission of HIV, Development of microbicides against HIV, Non-lytic CD8+T cell mediated HIV suppression, International study on molecular epidemiology and pathogenesis of HIV in India and China.
Vaccine Development: A novel Clostridia perfringens based oral vaccine is being developed to induce mucosal
immunity against HIV and SIV. We have constructed a non-cytotoxic C. perfringens expressing large amount of
HIV p24 and SIV p27 proteins in the form of inclusion bodies. When they are orally administered into mouse and
these proteins were detected in the terminal ileum region of the small intestines. Furthermore, dendritic cells
beneath the mucosal surface in the Peyer’s Patches(PPs) captured both HIV and SIV antigens when
mouse PPs were exposed to recombinant expressing HIV and SIV antigens in the lumen. Mucosal immunization
with a combination of recombinant C. perfringens expressing HIV-1 protein and HIV-1 virus like particles
VLPs induced significant T cell immune responses in gut associated lymphoid tissues, such as lamina
propria and Peyer’s patches. compared to immunization with either recombinant clostridia or VLP alone.
In an ongoing study we are constructing C. perfringens expressing HIV and SIV reverse transcriptase (RT)
and envelop gp120 proteins and measure mocosal and systemic immune responses to HIV and SIV proteins
expressed in C. perfringens in mouse. The proposed vaccine strategy holds a great promise for developing
a practical vaccine against HIV due to its safety, low cost to produce and easy to administer.
Molecular Mechanism of Sexual Transmission of HIV : A novel in vitro cervical tissue based organ culture has been developed to study heterosexual transmission of HIV-1. This model system allows the maintenance of the natural architecture of the tissue in the organ culture as evidenced by histology and quantitative immnostaining of immune and non-immune cellular proteins. Infectious virus is found to be transmitted from cell-free as well as cell-associated HIV-1 across the mucosal barrier. Using simultaneous in situ hybridization and immunophenotyping techniques, HIV-1 expressing cells are detected to the basal layer of the epithelium and identified as being activated CD4+ T lymphocytes and CD14+ macrophages. The model is being used in elucidating viral and cellular factors that are involved in heterosexual transmission of HIV-1.
Studies are being done to investigate the origin and dynamics of HIV-1 in semen. This has demonstrated presence of high levels of HIV-1 in semen of infected subjects at all stages of the disease, but in most subjects HIV-1 present in semen is different than those present in blood. In addition, HIV-1 was further sumcompartmentalized between seminal cells and seminal fluid. By comparing sequences of HIV-1 in semen and male genital organ tissues, it seems that prostate is the origin of HIV-1 present in seminal fluid, Testis and rete testis are probably source of semnial cell associated HIV-1. Furthermore, studies are being conducted to determine the dynamics of HIV in semen and blood by measuring the viral decay in these two body compartments after antiretrovoiral therapy. In addition, studies are being conducted to elucidate the mechanism of sexual transmission of HIV-1 by comparing HIV-1 sequences in donor semen and blood in the female recipients and the effect of therapy on such transmission.
Development of anti-HIV Microbicides : An U19 Cooperative Agreement grant has been funded by NIH to perform studies of preclinical optimization of an antimicrobial peptide, retrocyclin in combination with cyanovirin-N-secreting lactobacilli (CV-N/LAB) that will have potent anti-HIV activity against cell-free and cell-associated virus, be non-toxic and non-inflammatory to the cervical and vaginal tissues and in monkeys. This study employs an array of in vitro and ex vivo laboratory tests of efficacy and toxicity, in vivo safety tests in monkey models, and studies to optimize the formulation of these two compounds. ). In designing the proposed topical microbicide program, we have focused on extensively evaluating safety and toxicity of candidate microbicides utilizing in vitro studies, tissue explants and animal models, as well as an expanded and innovative organ culture model to study the interaction of HIV-1 and common STI in evaluating antiviral activity of microbicides in the presence of STI. The development of a combination microbicide product that incorporates CV-N/LAB and retrocyclin would provide a novel strategy which is expected to have potent anti-HIV activity without inducing cellular toxicity.
International studies in India and China: Our laboratory is also involved in International study of HIV infection in India and China. Our previous genetic studies of HIV-1 circulating at different parts of India indicate that the subtype C predominates in India with a small proportion of infection caused by subtype A or B. Studies are being conducted to investigate the mechanism for this asymmetric distribution of HIV subtypes in India. Three mechanisms are being investigated: 1) enhanced replication fitness of type C subtypes, 2) higher transmission efficiency of type C subtypes and 3) Founder effect: unique characteristics of founder virus resulting in disproportionate spread of subtype C viruses. Sequence analysis, organ culture and replication fitness methods are being used to investigate these possibilities. Recently we expanded our work on the characterization of HIV in semen in India. We are investigating whether HIV in semen is the same as that in blood and effect of therapy on viral load in semen and blood in HIV-infected subjects from India.
During the last 3 years we have established a collaborative study with Investigators at You An Hospital at Beijing, China to characterize HIV circulating among Chinese blood donors in central China for more than 10 years. Sequence analysis of HIV env gene revealed that regardless of the stage of disease, all the patient’s HIV-1 belonged to Clade B’subtype. However, the env sequences from AIDS patients clustered differently in the phylogenetic tree than those present in asymptomatic patients and showed a significantly higher divergence and lower diversity compared to those from asymptomatic patients. We have also analyzed nef and vpr sequnces Recently we have isolated HIV from Chinese subjects who are asymptomatic or developed AIDS. Studies are underway to characterize the genetic and biologic properties of these HIV isolates obtained at different stages of the disease.
Nonlytic CD8+ mediated HIV suppression: A number of studies are being conducted to investigate the mechanism of nonlytic CD8+ T cell mediated suppression of HIV and SIV at the cellular and molecular level, and to determine its role in HIV and SIV pathogenesis. To date, the precise cellular and molecular determinants mediating this CD8+ T cell effector function remain unsolved. Recently using a CD8+ T cell line displaying potent noncytotoxic HIV-1 suppression activity, we have identified a membrane-localized HIV-1 suppressing activity that is concomitantly secreted as 30-100nm sized endosome-derived tetraspanin-rich vesicles known as exosomes. Purified exosomes from CD8+ T cell culture supernatant noncytotoxically suppressed CCR5-tropic (R5) and CXCR4-tropic (X4) replication of HIV-1 in vitro through a protein moiety. The antiviral exosomes specifically inhibited HIV-1 transcription in both acute and chronic models of infection. Our results, for the first time, indicate the existence of an antiviral membrane-bound protein factor consistent with the hallmarks defining noncytotoxic CD8+ T cell suppression of HIV-1. Localization of this antiviral activity to secreted exosomes suggests a possible role for tetraspanin microdomains in regulating this antiviral activity.
Collins, K.B*., Patterson, B.K., Naus, G.J., Landers, D.V., and Gupta, P. Development of an in vitro organ culture model to study transmission of HIV-1 in the female genital tract. Nature Med. 6:475-479, 2000.
Gupta, P., Leroux, C., Patterson, B.K., Kingsley, L., Rinaldo, C., Ding, M., Chen, Y*., Kulka, K., Buchanan, W., McKeon, B., and Montelaro, R.. Human immunodeficiency virus type 1 shedding pattern in semen correlates with the compartmentalization of viral quasi species between blood and semen. J. Infect. Dis. 182:79-87, 2000.
Shankarappa, R., Chatterjee, R., Learn, G.H., Neogi, D., Ding, M., Roy, P., Ghosh, A., Kingsley, L., Harrrison, L., Mullins, J.I., and Gupta, P. (2001). Human immunodeficiency virus type 1 Env sequences from Calcutta in
Eastern India; identification of features that distinguish subtype C sequences in India from other subtype C sequences. J. Virol. 75:10479-10487, 2001.
Gupta, P., Collins, K.B*., Ratner, D., Watkins, S., Naus, G.J., Landers, D.V., and Patterson, B.K. Memory CD4+ T cells are the earliest detectable HIV-1 infected cells in the female genital mucosal tissue during HIV-1 transmission in an organ culture system. J. Virol. 76, 9868-9876, 2002.
Paranjpe, S*., Craigo, J., Patterson, B., Ding, M., Barroso, P., Harrison, L., Montelaro, R., and Gupta, P. Subcampartmentalization of HIV-1 quasispecies between seminal cells and seminal plasma indicates their origin in distinct genital tissues. AIDS. Res. Hum.Retro, 17, 1271-1280, 2002.
Gupta, P., Kingsley, L., Sheppard, H.W., Harrison, L.H., Chatterjee, R., Ghosh, A., Roy, P., and Neogi, D.K. High incidence and prevalence of HIV-1 infection in high risk population in Calcutta, India. Intl. J. STD & AIDS 14:463-468, 2003.
Chen Y., Helmus,R**., McClane B., Hoffman,R., Watkins,S., Wehrli,T and Gupta, P. Use of a Clostridium Perfringens vector to express high levels of SIV p27 protein for the development of an oral SIV vaccine. Virology. 329, 226-233, 2004.
Craigo, J.K., Patterson, B.K., Paranjpe, S*., Kulka, S., Ding, M., Mellors., Montelaro, R.C., and Gupta, P. Persistent viral Infection and sequence evolution in semen and blood compartments in HIV-infected patients following long term potent antiretroviral therapy. AIDS Res Human. Retro. 20, 1196-1204, 2004.
Kalia, V., Sarkar, S., Gupta, P., and Montelaro, R. Antibody neutralization escape mediated by point mutations in the intracytoplasmic tail of human immunodeficiency virus type 1 gp41. J. Virol. 79, 2097-2107, 2005.
Chen, Y., McClean, B., Fisher, D., Rood, J., and Gupta, P. Construction of an alpha toxin gene inactivated mutant of clostridia perfringens type A using a moblile II group II intron. J Appl Environmental Microbiol. 71:7542-7547, 2005.
Rodriguez, M *., Chen,Y., Craigo, J., Chatterjee, R., Roy, P., Neogi, D., and Gupta, P. Construction and characterization of a full length infectious molecular clone of subtype A HIV-1 from India. Virology, 345, 328-336, 2006.
Rappocciolo, G., Piazza, P., Fuller, C.L., Reinhart, T.A., Watkins, S.C., Rowe, D.T., Jais, M., Jaffe, R., Gupta, P., and Rinaldo, C.R., Jr. DC-SIGN on B lymphocytes is required for transmission of HIV-1 to T lymphocytes. In press, PLOS, 2006.
Chen, Y., Craigo, JK., Ding, M., Guo,Y., Shen,C., Zhou,Y., Wu,H and Gupta, P. Characterization of gp120 of circulating HIV type 1 in a group of infected Chinese blood/plasma donors with asymptomatic and symptomatic stages of the disease. AIDS Res. Hum. Retro. 22, 1161-1170, 2006.
Chen, Y., Caruso, L., Shen, S.L., Wu,H., Zhou,Y., Phalguni Gupta. Lack of differences in HIV-1 Nef functional domains in infected Chinese blood donors at different stages of diseases. AIDS Research and Human Retroviruses.23, 1150-1154, 2007.
Cole, A.M., Herraimtschuck, A., Gupta, P., Waring, A.J., Lehrer, R.I., Cole, A.M. The retrocyclin analog RC101 prevents human immunodeficiency virus type 1 infection of a model human cervicovaginal tissue construct. Immunology, 121, 140-145, 2007
Rodriguez, M.A., Cheng, S.L., Ratner, D., Paranjape, R.S., Kulkarni, R.S., Chatterjee, R., and Gupta, P. Genetic and Functional Characterization of the LTR of HIV-1 Subtypes A and C Circulating in India. AIDS Research and Human Retroviruses, 23, 1428-1433, 2007 .
Shen ,C., Gupta, P., Wu, H., Chen, X., Huang,X., Zhou, Y and Chen Y. Molecular characterization of the HIV type 1 vpr gene in infected Chinese former blood/plasma donors at different stages of diseases.AIDS Res Hum Retroviruses. 24:661-6666, 2008.
*Postdoctoral fellows and **graduate students of Dr. Gupta.
Caruso, Lori; Lab Technician
615 Parran Hall; 412-624-2057; lcaruso@pitt.edu
Chen, Yue; Research Assistant Professor
434 Parran Hall; 412-624-5772; cheny@pitt.edu
Ding, Ming; PCR Lab Technician
525A Parran Hall; 412-624-0776; mding@pitt.edu
Kulka, Kathy; Lab Supervisor
A419 Crabtree Hall; 412-624-0867; kulka@pitt.edu
Ratner, Deena; Lab Technician
525A Parran Hall; 412-624-0776; dampf@pitt.edu
White, Mary; Lab Technician
A419 Crabtree Hall; 412-624-0867
Poonam, Poonam
404 Parran Hall; 412-624-6929; pop2@pitt.edu
Jackeline Soto
431 Parran Hall; 412-624-5772; jas180@pitt.edu
Varsha Sridhar
431 Parran Hall; 412-624-5772; vas30@pitt.edu
Tumne, Ashwin
433 Parran Hall; 412-624-5772; astst9@pitt.edu
Chengli Shen
425 Parran Hall, 412-383-8926: chs97@pitt.edu
Last Updated: June 2, 2008
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