Phalguni Gupta, PhD
Professor and Assistant Chair
426 Parran Hall
130 DeSoto Street
Pittsburgh, PA 15261
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.
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 immunostaining 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 to determine mechanism of sexual transmission of HIV-1, especially in elucidating viral and cellular factors that are involved in transmission through epithelial layer of cervical mucosa.
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 and a nonnucloside reverse transcriptase inhibitor 5-chloro-3-phenylsulfonylindole-2-carboxamide (CSIC) to block HIV transmission and be non-toxic to human. Our multidisciplinary approach to microbicide development targets two different sites of HIV-1 replication (entry and reverse transcription) through a controlled-release ring formulated product or a self dissolving film formulation, utilizes a novel ex vivo organ culture to test toxicity, inflammation and antiviral activity across cervicovaginal mucosa, and applies macaque models to study toxicity and efficacy. In designing the scope of 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 sexually transmitted infections (STI) in assessing antiviral activity of microbicides in the presence and absence of STI. Therefore, the proposed study will provide new tools and strategies for the prevention of HIV-1 transmission and their sequelae for women.
Origin and Dynamics of HIV-1 in Semen. We have demonstrated the 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 is further sum compartmentalized 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 seminal 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 antiretroviral 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.
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. In investigating the mechanism for this asymmetric distribution of HIV subtypes in India we found that subtype C is more replication fit and has higher transmission efficiency across cervical mucosa than subtype A. Recently we expanded our work in India by characterizing HIV-1 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-1 env gene revealed that regardless of the stage of disease, all the HIV-1 patient’s 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 sequences. We have isolated HIV-1 from Chinese subjects and characterized the genetic and biologic properties of these HIV isolates obtained at different stages of the disease. Virus isolates from asymptomatic virus grow slowly, are non-cytopathic and uses mostly CCR5 co-receptor. In contrast, isolates from patients with AIDS grow rapidly, highly cytopathic and uses both CCCR5 and CXCR4 co-receptors. Recently we are studying a group of Chinese subjects that developed AIDS within 2 years. Biologic and genetic approaches are being done to understand host and viral factors responsible for these rapid progressions in China.
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-1 and SIV at the cellular and molecular level, and to determine its role in HIV-1 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. Our recent data indicate that this HIV suppressive factor works by binding to target cell surface and transmitting an intracellular signal to repress transcription of HIV-1. Studies are underway to determine the mechanism of suppressive factor induced intracellular signaling pathway.
A novel cellular antiviral factor. Adenosine Deaminase acting on RNA 1(ADAR1) ADAR1 is a RNA editing enzyme which acts on complete or partially double-stranded RNA. Since HIV-1 RNA has such secondary structures, we have examined whether ADAR1 exhibits antiviral activity against HIV-1. Our results indicate that ADAR1 inhibits viral replication and infectious HIV-1 production in various cells including 293T, HeLa and Jurkat T cells, and is active against a number of X4 and R5 HIV-1 of different clades. Analysis of the level of intracellular HIV-1 RNA shows no change in levels of intracellular Gag and Env RNA in the presence of ADAR1 in spite of a significant inhibition of intracellular and virion associated HIV-1 protein production. Furthermore, our data indicate that ADAR1 introduces A-> G mutations in Rev Response Element (RRE) binding and nuclear export signal (NES) regions of HIV-1 Rev RNA and env RNA. Studies are being done to understand the mechanism of ADAR1 induced antiviral activity.
Cole, A.M., Patton, D.L., Rohan, L.C., Cole, A.L., Cosgrove-Sweeney, Y., Rogers, N.A., Ratner, D., Sassi, A.B., Lackman-Smith, C., Tarwater, P., Ramratnam, B., Ruchala, P., Lehrer, R.I., Waring, A.J., and Gupta, P. The formulated microbicide RC-101 was safe and antivirally active following intravaginal application in pigtailed macaques. PLoS One. 5, e15111, 2010.
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. 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. Hum. 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.
Chen, Y., Craigo, J.K., 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., and Gupta, P. Lack of differences in HIV-1 Nef functional domains in infected Chinese blood donors at different stages of diseases. AIDS Res. Hum. Retro. 23:1150-1154, 2007.
Cole, A.M., Herraimtschuck, A., Gupta, P., Waring, A.J., Lehrer, R.I., and 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 Res. Hum. Retro. 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. Retro. 24:661-6666, 2008.
Rodriguez, M.A., Ding, M., Ratner, D., Chen, Y., Paranjape, R.S., Kulkarni, S.S., Chatterjee, R.Tarwater, P.M., and Gupta, P. Higher replication fitness and transmission efficiency of HIV-1 subtype C than subtype A from India: implications for subtype C predominance. Virology 385:416-24, 2009.
Tumne, A., Shridhar, V., Chen, Y., Stolz, D.B., Ratner, D.M., Ding, M., Watkins, S.C., and Gupta, P. Noncytotoxic suppression of HIV-1 transcription by exosomes secreted from a CD8+ T cell line. J. Virol. 83:4354-4364, 2009.
Chen, Y., Shen, C., Wu, H., Caruso, L., Ratner, D., Rodriguez, M., Chen, X., and Gupta, P. Biological properties of HIV-1 subtype B' isolates from infected Chinese blood donors at different disease stages. Virology 384:161-168, 2009.
Ding, M., Tarwater, P., Rodriguez, M., Chatterjee, R., Ratner, D., Yamamura, Y., Roy, P., Mellors, J., Neogi, D., Chen, Y., and Gupta, P. Estimation of the predictive role of plasma viral load on CD4 decline in HIV-1-subtype C infected subjects in India. J. AIDS 50:119-125, 2009.
Helmus, R.A., Poonam, P., Chen, Y., Caruso, L., Kolls, J.K., and Gupta, P. Induction of SIV p27-specific multifunctional T cells in the gut following prime-boost immunization with Clostridium perfringens and adenovirus vaccines expressing SIV p27. Curr. HIV Res. 8:101-112, 2010
Shen, C., Ding, M., Craigo, J.,Tarwater, P.,Chatterjee ,R., Roy P., Guha, S. K, Saha, B, Modak, D, Neogi, D, Chen, Y., and Gupta, P. Genetic characterization of HIV-1 from semen and blood from clade C-infected subjects from India and effect of therapy in these body compartments. Virology 401:190-196, 2010.
Sassi, A.B., Cost, M.R., Cole, A.L., Cole, A.M., Patton, D.L., Gupta, P., and Rohan, L.C. Formulation development of Retrocyclin-1 analog RC-101 as an anti-HIV vaginal microbicide product. Antimicrob Agents Chemother. 55:2282-2289, 2011.
Ding, M., Bullotta, A., Caruso, L., Gupta, P., Rinaldo, C.R., and Chen, Y. An optimized sensitive method for quantitation of DNA/RNA viruses in heparinized and cyropreserved plasma. J. Virol. Methods. 176:1-8, 2011.
Sassi, A.B., Bunge, K.E., Hood, B.L., Conrads, T.P., Cole, A.M., Gupta, P., and Rohan, L.C. Preformulation and stability in biological fluids of the retrocyclin RC-101, a potential anti-HIV topical microbicide. AIDS Res Ther. 8:27, 2011.
Pegu, P., Helmus, R., Gupta, P., Tarwater, P., Caruso, L., Shen, C., Ross, T., and Chen, Y. Induction of strong anti-HIV cellular immunity by a combination of Clostridium Perfringens expressing HIV gag and virus like particles. Current HIV Res. 9:613-622, 2011.
Shen, C., Craigo, J., Ding, M., Chen, Y., and Gupta, P. Origin and dynamics of HIV-1 subtype C infection in India. PLoS ONE 6:e25956, 2011.
Biswas, N., Wang, T., Ding, M., Tumne, A., Chen, Y., Wang, Q., and Gupta, P. ADAR1 is a novel multi targeted anti-HIV-1 cellular protein. Virology 422:265-277, 2012.
Shen, C., Ding, M., Ratner, D., Montelaro, R.C., Chen, Y., and Gupta, P. Evaluation of cervical mucosa in transmission bottleneck during acute HIV-1 infection using a cervical tissue based organ culture. PLoS One, accepted for publication, 2012. e32539, 2012
Li, M., Patton, D.L., Cosgrove-Sweeney, Y., Ratner, D., Rohan, L.C., Cole, A.M., Tarwater, P.M., Gupta, P., and Ramratnam, B. Incorporation of the HIV-1 microbicide cyanovirin-N in a food product. J. AIDS.58, 379-84, 2011
Gupta, P., Ratner, D., Ding,M1.,Patterson,B., Rohan, LC,. Reinhart, T.R.,Ayyavoo, V,Huang,X., Patton, D. L., Ramratnam, BL, and Cole, A.C. J. AIDS. In Press, 2012
Soto-Rivera, J., Patterson, B.K, Tumne, A., Shen, C., Ratner, D., Ding, M., Chen, Y. and Gupta,P. Study of HIV-1 transmission across cervical mucosa and migration to lymph nodes using an organ culture. Am.J.Reproductive. Immunol. In Press. 2012.
Gupta, P., Lackman-Smith, C., Snyder,B , Ratner,B., Rohan, L.C., Patton, D ., Ramratnam, B and Cole, A.M. Antiviral Activity of Retrocyclin RC-101, a Candidate Microbicide Against Cell-Associated HIV-1. Submitted. AIDS Res. Hum Retro, 2012.
Shridhar, V., Chen, Y., Tumne, A., Caruso, L. and Gupta, P.
Identification of the region on the HIV-1 Long Terminal Repeat responsible for transcriptional suppression by the cd8 antiviral factor. Revision, Virology. 2012.
Shridhar, V., Tumne, A., Chen, Y., Caruso, L and Gupta, P. STAT1-independent suppression of HIV-1 transcription by primary CD8+ T cell derived antiviral factor. Submitted to Immunology, 2012.
Dr. Gupta's Lab
- June 2012
Lori Caruso; Lab Technician
422 Parran Hall; 412-383-9709; email@example.com
Yue Chen; Research Assistant Professor
421 Parran Hall; 412-624-5398; firstname.lastname@example.org
Ming Ding; PCR Lab Technician
423 Parran Hall; 412-383-9710; email@example.com
Deena Ratner; Lab Technician
438 Parran Hall; 412-624-2057; firstname.lastname@example.org
Mary White; Lab Technician
440 Parran Hall; 412-624-6929
440 Parran Hall; 412-624-6929; email@example.com
440 Parran Hall; 412-624-6929; firstname.lastname@example.org
Visiting Assistant Professor
425 Parran Hall; 412-383-8694; email@example.com