Associate Dean for Scientific Resources
Professor, Department of Pediatrics (Allergy & Immunology)
Professor, Department of Microbiology & Immunology
Director, Einstein/MMC Center for AIDS Research
Charles Michael Chair in Autoimmune Diseases
Generation of HIV-specific immunity using genetic engineering. Our laboratory is investigating the basis for generating an effective HIV-specific immune response and how we can use molecular engineering to reprogram T cells and B cells to increase the potency of their anti-HIV activity. For T cells, we demonstrated that functional HIV-specific CD8+ cytotoxic T lymphocytes (CTLs) can be generated by delivering genes encoding the alpha and beta chain of TCRs derived from potent HIV-specific CTLs into CD8+ lymphocytes (Joseph, et. al., J. Virology 2008;82:3078–3089). To this end we characterize the in vitro and in vivo function of human HIV-specific CTLs, clone out the TCR alpha and beta chain cDNA from the most potent HIV-specific CD8+ CTL clones and insert them into novel retroviral or lentiviral vectors. We are also performing structure-function analysis of the TCR structure to rationally design TCRs with increased affinity and avidity. We then determine the capacity of these vectors to introduce the cloned TCR genes into human hematopoietic precursor cells or CD8+ lymphocytes and program them to differentiate into HIV-specific CD8+ CTLs that display the in vitro and in vivo capacity to recognize and eliminate HIV-infected cells. Delivery of cDNA encoding TCR alpha and beta chains to CD34+ HSC or CD8+ lymphocytes to program them into protective CTLs provides a modality for generating fresh precursor cells to replace those CTLs undergoing replicative senescence and to redirect the immune response by creating CTLs that recognize immunoprotective epitopes not in the initial repertoire stimulated after infection.
While broadly neutralizing HIV-specific antibodies have the capacity to prevent or suppress HIV infection, they are rarely produced by infected individuals, due to the inherent immune evasion properties of the HIV envelope, This markedly compromises the ability of infected individuals to harness the humoral response to control HIV infection. We postulated that we could use genetic engineering to circumvent the restricted capacity of individuals to endogenously produce broadly neutralizing HIV-specific antibodies. We demonstrated that we could construct a single lentiviral vector that encoded the heavy and light chains of 2G12, a broadly neutralizing anti-HIV human antibody, and reprogram human hematopoietic stem cells to differentiate into 2G12-secreting B cells that protected humanized NOD/SCID/gcnull mice from in vivo HIV infection (Joseph et. al., J.Virology 2010;84:6645-53). These modalities have the capacity be new therapeutic approaches designed to augment the immune response of individuals infected with HIV-1 and other pathogens.
Examination of the pathogenesis of NeuroAIDS. We are also examining the pathogenesis of HIV-mediated CNS dysfunction using an HIV transgenic model. Many HIV-1-infected patients develop severe neurological complications including HIV-1-associated dementia (HAD). It has been postulated that HIV-1 infection initiates a neuroinflammatory cascade involving the production of cytokines, chemokines, neuromodulators and neurotoxic factors that leads to neural dysfunction and HAD. We have recently developed the JR-CSF/huCycT1 mouse line that is transgenic for two transgenes: HIV-1JR-CSF, a full-length, R5 HIV-1 provirus regulated by the endogenous HIV-1 LTR and human cyclin T1 controlled by a murine CD4 expression cassette that targets transgene expression to CD4 T lymphocytes and myeloid lineage cells including microglia (Sun et. al., J. Virology 2006;80:1850-62). Monocytes from these mice produced markedly higher levels of HIV-1 than monocytes from mice transgenic for the JR-CSF transgene alone. In addition, monocytes from the JR-CSF/huCycT1 mice produced significantly higher levels of MCP-1 after LPS stimulation when compared to monocytes from wild-type mice and JR-CSF mice. We are using these mice to generate a model for studying the mechanism by which HIV-1 induces neuroinflammation and induces neuronal dysfunction which ultimately could be used to test the efficacy of therapeutic interventions (Sun et. al., J. Virology 2008;82:5562-5572). We are also using these mice to investigate the effect of HIV-1 infection on the integrity of the blood brain barrier (BBB). We demonstrated that the JR-CSF mouse BBB was more susceptible to disruption by systemic LPS as compared to the control wild-type mouse BBB (Wang et. al., J. Virology 2008;82:7591-600). These results demonstrated that HIV-1 infection increased the ability of monocytes to enter the brain, and increased the sensitivity of the BBB to disruption by systemic LPS, which is elevated in HIV-1-infected individuals. We are using these mice using proteomics and microarray analysis as a new in vivo system to study the mechanism by which HIV-1-infected monocytes migrate into brain and to test therapies that reverse HIV-mediated compromise of the BBB.
Identification of the molecular basis of HIV-TB synergism. Our lab is also analyzing HIV-TB molecular interaction during in vitro and in vivo infection to investigate the co-pathogenesis of HIV and M. tuberculosis (with Dr. Bill Jacobs). Little is known about the effects of HIV infection on the initiation and spread of primary M. tuberculosis infection and the intracellular interactions between HIV and M. tuberculosis that promote reactivation of M. tuberculosis infection and increased M. tuberculosis and HIV replication. We are utilizing microarray analysis to identify cellular genes are induced by HIV infection of monocyte/macrophages that facilitate their productive infection with M. tuberculosis and the subsequent dissemination of M. tuberculosis. To evaluate the in vivo impact of HIV-1 replication on pulmonary infection with M. tuberculosis, we are using our unique in vivo model, JR-CSF/hu-cycT1 mice. Results of these studies should greatly increase our understanding of the effects of HIV and M. tuberculosis co-infection on macrophage function that could be applied to prevent dissemination of the looming HIV-XDR-TB outbreak that is nearly universally fatal (98% mortality) in HIV-infected individuals.
M. S. Bennett, A. Joseph, H. L. Ng, H. Goldstein, O. O. Yang. Fine Tuning of T Cell Receptor Avidity to Increase HIV Epitope Variant Recognition by Cytotoxic T Lymphocytes. AIDS 2010;24:2619-28..
K. Sango, A. Joseph, M. Patel, K. Osiecki, M. Dutta, H. Goldstein. Highly Active Antiretroviral Therapy Potently Suppresses HIV Infection in Humanized Rag2-/-gc-/- Mice. AIDS Res. Hum. Retro. 2010;26:735-46.
K. L. Schaubert, D. A. Price, J. R. Salkowitz, A. K. Sewell, J. Sidney, T. E. Asher, S. E. Blondelle, S. Adams, F. M. Marincola, A. Joseph, A. Sette, D. C. Douek, V. Ayyavoo, W. Storkus, L. M. Yeung, H. L. Ng, O. O. Yang, H. Goldstein, D. B. Wilson, J. Kan-Mitchell. Generation of robust CD8(+) T-cell responses against subdominant epitopes in conserved regions of HIV-1 by repertoire mining with mimotopes. Eur J Immunol. 2010;40:1950-1962.
A. Joseph, J. H. Zheng, K. Chen, M. Dutta, C. Chen, G. Stiegler, R. Kunert, A.Follenzi, and H. Goldstein. Inhibition of in vivo HIV Infection in humanized mice by gene therapy of human hematopoietic stem cells with a lentiviral vector encoding a broadly neutralizing anti-HIV antibody. J.Virology 2010;84:6645-53. PMCID: 2903239
S. S. Toussi, A. Joseph, J. H. Zheng, M. Dutta, L. Santambrogio and H. Goldstein. Methamphetamine treatment increases in vitro and in vivo HIV replication. AIDS Res. Hum. Retro. 2009;25:1117-21. PMCID: 2828189
S.G. Kitchen, M. Bennett, Z. Galiæ, J. Kim, Q. Xu, A. Young, A. Lieberman, A. Joseph, H. Goldstein, H. Ng, O. Yang, J.A. Zack. Engineering antigen-specific T cells from genetically modified human hematopoietic stem cells in immunodeficient mice. PLoS One 2009 7:4:e8208. PMCID: 2785433
T. Kojaoghlanian, A. Joseph, A. Follenzi, J. H. Zheng, M. Leiser, N. Fleischer, M. S. Horwitz, T. P. DiLorenzo, and H. Goldstein. Lentivectors encoding immunosuppressive proteins genetically engineer pancreatic β cells to correct diabetes in allogeneic mice. Gene Therapy 2009:16(3):340-348. PMCID: 2901156.
A. Joseph, K. Sango, and H. Goldstein. Novel mouse models for understanding HIV-1 pathogenesis. Methods Mol Biol. 2009;485:311-27.
H. Wang, J. Sun and H. Goldstein. HIV-1 infection increases the in vivo capacity of peripheral monocytes to cross the blood brain barrier into the brain and the in vivo sensitivity of the blood brain barrier to disruption by LPS. J.Virology 2008;82:7591-7600. PMCID: 2493310
J. Sun, J. H. Zheng, M. Zhao, Sunhee Lee and H. Goldstein. Increased in vivo activation of microglia and astrocytes in the brains of mice transgenic for an infectious R5 HIV-1 provirus and for CD4-specific expression of human cyclin T1 in response to LPS stimulation. J.Virology, 2008;82:5562–5572. PMCID:2395169
A. Joseph, J. Zheng, A. Follenzi, T. DiLorenzo, K. Sango, J. Hyman, K. Chen ,E. Hooijberg, D. A. Vignali,, B. D. Walker and H. Goldstein. Lentiviral vectors encoding human immunodeficiency virus type 1 (HIV-1)-specific T-cell receptor genes efficiently convert peripheral blood CD8 T lymphocytes into cytotoxic T lymphocytes with potent in vitro and in vivo HIV-1-specific inhibitory activity.. J. Virology 2008;82: 3078–3089. PMCID: 2258988
H. Goldstein, NIH/NIAID New Humanized Rodent Models 2007 Workshop, AIDS Research and Therapy, 2008 ;5:3. PMCID: 2276217
J. Sun, K. Osiecki, J. Zheng, L. Falkin, L. Santambrogio, D. R. Littman, and H. Goldstein. CD4-specific transgenic expression of human cyclin T1 markedly increases HIV-1 production by CD4+ T lymphocytes and myeloid committed cells in mice transgenic for a provirus encoding a monocyte-tropic HIV-1 isolate. J. Virology 2006;80:1850-62. PMCID: 1367149
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Science Daily and BBC report on a technique developed by Einstein researchers that could influence the way type 1 diabetes is treated in the future, using transplanted insulin-producing pancreatic cells.