EXCEED THE SPACE PROVIDED. This is a competing renewal of an R01 (AI47062) initially submitted in response to the RFA: 'Laboratory Methods to Assess Responses to HIV Vaccine Candidates.' Since 2/1/00, substantial progress has been made on the first two Specific Aims of the original proposal, focused on development, optimization and utilization of a manual cytokine flow cytometry (CFC) assay for the detection of antigen-induced IFNy responses by CD4+ and CD8+ human T cells specific for the human immunodeficiency virus, type 1 (HIV-1). In studies that are currently ongoing, we anticipate that additional progress will be made in the use of this 'Gag-IFN'l, CFC assay' to determine whether various HIV-1 vaccines may induce specific CD4+ and/or CD8+ T cell immunity against HIV-1 in vivo. This work has generated 7 papers published in peer-reviewed journals and 5 abstracts presented at international meetings. Our objective now is to assess laboratory correlates of immune protection by automating new flow cytometric assays of antigen-specific immune responses, to identify candidate laboratory correlates of immune protection by comparing the results of these assays with results from other assays of immune phenotype and function in long-term nonprogressors (LTNP), untreated subjects with progressive HIV-1 disease, and recipients of candidate HIV-1 vaccines, and to examine HIV-1-specific immune responses in HIV-l-infected individuals who appear to exhibit significant immune protection from HIV-1 disease.
Specific Aim 1 To develop, optimize, and standardize the methods, hardware, and software for automated stimulus-response flow cytometric assays of immune function. The manual Gag-IFN_, CFC assay will be configured into an automated assay platform, amenable for use with additional stimulants and for the measurement of additional intracellular and cell surface markers.
Specific Aim 2 To cross-sectionally compare Gag-IFN_ CFC results to those obtained from other assays of immune phenotype and function in a) long-term nonprogressors, b) untreated subjects with progressive HIV-1 disease, and c) recipients of candidate HIV-1 vaccine(s). The automated Gag-IFNy CFC assay will be compared to CFC assays which measure additional intracellular and cell surface markers; and to more traditional measures of CD4+ and CD8+ T cell function (e.g., proliferation and cytolysis). Such comparisons will be made by cross-sectional analysis of two distinct groups of HIV-l-infected patients (long-term nonprogressors and untreated subjects with progressive HIV-1 disease) and two distinct groups of HIV- vaccinees ('responders' and 'nonresponders' as tested by lymphoproliferation, 5_Cr release, and ELISPOT).
Specific Aim 3 To prospectively characterize antigen-specific immunity in HIV-l-infected individuals who appear to exhibit significant immune protection from HIV-1 disease. Using a cohort of patients with partial virologic control, a prospective analysis will be carried out to define the temporal relationship between loss of immune function, as measured in CFC assay(s) developed in the first two Aims, and disease progression. PERFORMANCE SITE ========================================Section End===========================================

Agency
National Institute of Health (NIH)
Institute
National Institute of Allergy and Infectious Diseases (NIAID)
Type
Research Project (R01)
Project #
5R01AI047062-06
Application #
6838132
Study Section
Special Emphasis Panel (ZRG1-AARR-2 (02))
Program Officer
D'Souza, Patricia D
Project Start
2000-02-01
Project End
2007-12-31
Budget Start
2005-01-01
Budget End
2005-12-31
Support Year
6
Fiscal Year
2005
Total Cost
$451,832
Indirect Cost
Name
J. David Gladstone Institutes
Department
Type
DUNS #
099992430
City
San Francisco
State
CA
Country
United States
Zip Code
94158
Emu, Brinda; Moretto, Walter J; Hoh, Rebecca et al. (2014) Composition and function of T cell subpopulations are slow to change despite effective antiretroviral treatment of HIV disease. PLoS One 9:e85613
Stoddart, Cheryl A; Keir, Mary E; McCune, Joseph M (2010) IFN-alpha-induced upregulation of CCR5 leads to expanded HIV tropism in vivo. PLoS Pathog 6:e1000766
Contreras, Xavier; Schweneker, Marc; Chen, Ching-Shih et al. (2009) Suberoylanilide hydroxamic acid reactivates HIV from latently infected cells. J Biol Chem 284:6782-9
Barbour, Jason D; Ndhlovu, Lishomwa C; Xuan Tan, Qi et al. (2009) High CD8+ T cell activation marks a less differentiated HIV-1 specific CD8+ T cell response that is not altered by suppression of viral replication. PLoS One 4:e4408
Maecker, Holden T (2009) Multiparameter flow cytometry monitoring of T cell responses. Methods Mol Biol 485:375-91
Schweneker, Marc; Favre, David; Martin, Jeffrey N et al. (2008) HIV-induced changes in T cell signaling pathways. J Immunol 180:6490-500
Napolitano, Laura A; Schmidt, Diane; Gotway, Michael B et al. (2008) Growth hormone enhances thymic function in HIV-1-infected adults. J Clin Invest 118:1085-98
Jacobson, Mark A; Tan, Qi Xuan; Girling, Valerie et al. (2008) Poor predictive value of cytomegalovirus (CMV)-specific T cell assays for the development of CMV retinitis in patients with AIDS. Clin Infect Dis 46:458-66
Emu, Brinda; Sinclair, Elizabeth; Hatano, Hiroyu et al. (2008) HLA class I-restricted T-cell responses may contribute to the control of human immunodeficiency virus infection, but such responses are not always necessary for long-term virus control. J Virol 82:5398-407
Owen, Rachel E; Sinclair, Elizabeth; Emu, Brinda et al. (2007) Loss of T cell responses following long-term cryopreservation. J Immunol Methods 326:93-115

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