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We have developed a single-cell enzyme immunoassay (enzyme-linked immunospot assay) specific for gamma interferon (IFN-\u03b3) production stimulated by either autologous B-lymphoblastoid cell lines (B-LCL) infected with vaccinia virus vectors expressing HIV-1 proteins or synthetic peptides representing known HIV-1 CD8<jats:sup>+<\/jats:sup>cytotoxic T-lymphocyte (CTL) epitopes. Single-cell IFN-\u03b3 production stimulated by HIV-1 Gag-, Pol-, and Env-expressing B-LCL was a reliable measure of HIV-1-specific T-cell immunity in peripheral blood CD8<jats:sup>+<\/jats:sup>T cells from HIV-1 infected individuals. This method was more sensitive than stimulation of IFN-\u03b3 by direct infection of the cultures with HIV-1\u2013vaccinia virus vectors. Comparable results were found for IFN-\u03b3 production in CD8<jats:sup>+<\/jats:sup>T cells from HIV-1-negative, cytomegalovirus (CMV)-seropositive, healthy donors stimulated with B-LCL expressing the CMV pp65 lower matrix protein. HIV-1 peptides were immunodominant for both CD8<jats:sup>+<\/jats:sup>single-cell IFN-\u03b3 production and CTL precursor frequencies. The number of cells producing IFN-\u03b3 decreased in individuals with late-stage HIV-1 infection and was temporally enhanced during combination antiretroviral therapy with two reverse transcriptase nucleoside inhibitors and a protease inhibitor.<\/jats:p>","DOI":"10.1128\/cdli.7.2.279-287.2000","type":"journal-article","created":{"date-parts":[[2002,7,27]],"date-time":"2002-07-27T09:55:30Z","timestamp":1027763730000},"page":"279-287","update-policy":"http:\/\/dx.doi.org\/10.1128\/asmj-crossmark-policy-page","source":"Crossref","is-referenced-by-count":19,"title":["CD8<sup>+<\/sup>T-Cell Gamma Interferon Production Specific for Human Immunodeficiency Virus Type 1 (HIV-1) in HIV-1-Infected Subjects"],"prefix":"10.1128","volume":"7","author":[{"given":"Xiao-Li","family":"Huang","sequence":"first","affiliation":[{"name":"<!--label omitted: 1-->University of Pittsburgh Graduate School of Public Health1 and"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Zheng","family":"Fan","sequence":"additional","affiliation":[{"name":"<!--label omitted: 1-->University of Pittsburgh Graduate School of Public Health1 and"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Christine","family":"Kalinyak","sequence":"additional","affiliation":[{"name":"<!--label omitted: 1-->University of Pittsburgh Graduate School of Public Health1 and"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"John W.","family":"Mellors","sequence":"additional","affiliation":[{"name":"<!--label omitted: 1-->University of Pittsburgh Graduate School of Public Health1 and"},{"name":"<!--label omitted: 2-->School of Medicine2 and"},{"name":"<!--label omitted: 3-->Veterans Affairs Medical Center,3 Pittsburgh, Pennsylvania 15261"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"suffix":"Jr.","given":"Charles R.","family":"Rinaldo","sequence":"additional","affiliation":[{"name":"<!--label omitted: 1-->University of Pittsburgh Graduate School of Public Health1 and"},{"name":"<!--label omitted: 2-->School of Medicine2 and"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"235","reference":[{"key":"e_1_3_2_2_2","doi-asserted-by":"publisher","DOI":"10.1111\/j.1365-2249.1995.tb02285.x"},{"key":"e_1_3_2_3_2","doi-asserted-by":"publisher","DOI":"10.1016\/S0952-7915(99)80076-4"},{"key":"e_1_3_2_4_2","doi-asserted-by":"publisher","DOI":"10.1016\/S1074-7613(00)80469-0"},{"key":"e_1_3_2_5_2","doi-asserted-by":"publisher","DOI":"10.1016\/0022-1759(88)90079-8"},{"key":"e_1_3_2_6_2","doi-asserted-by":"crossref","first-page":"3971","DOI":"10.4049\/jimmunol.151.8.3971","article-title":"Alkali hydrolysis of recombinant proteins allows for the rapid identification of class I MHC-restricted CTL epitopes.","volume":"151","author":"Gavin M. 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