title: Detection of human immunodeficiency virus type 1-specific memory cytotoxic T lymphocytes in freshly donated and frozen-thawed peripheral blood mononuclear cells.
abstract: Loss of anti-human immunodeficiency virus type 1 (HIV-1) memory cytotoxic T-lymphocyte (CTLm) responses is associated with disease progression in HIV-1 infection. In this study, nonspecific stimulation of peripheral blood mononuclear cells (PBMC) from HIV-1-infected homosexual men with anti-CD3 monoclonal antibody (MAb) was compared with antigen-specific stimulation with inactivated, autologous B lymphoblastoid cells (B-LCL) infected with a vaccinia virus vector encoding HIV-1 IIIb Gag, Pol, and Env (VV-GPE) for activation of HIV-1-specific CTLm responses in a bulk lysis assay and by precursor frequency analysis. The results show that VV-GPE-infected B-LCL stimulated on average 10-fold greater anti-HIV-1 CTLm activity, as detected in the bulk lysis assay, and 55-fold-greater CTLm precursor frequencies specific for the three HIV-1 structural proteins than did stimulation with anti-CD3 MAb. This effect was noted with both freshly donated and frozen-thawed PBMC. The lysis was mediated by CD8+ T cells and was restricted by the major histocompatibility class I complex. These data indicate that antigen-specific stimulation with VV-GPE-infected B-LCL is a highly efficient method for detection of anti-HIV-1 CTLm responses that is applicable to noncurrent prospective studies with frozen PBMC.
authors.affiliation: Department of Infectious Diseases and Microbiology, University of Pittsburgh Graduate School of Public Health, Pennsylvania 15261, USA.
authors.forename: X L
authors.initials: XL
authors.lastname: Huang
authors.forename: Z
authors.initials: Z
authors.lastname: Fan
authors.forename: J
authors.initials: J
authors.lastname: Liebmann
authors.forename: C
authors.initials: C
authors.lastname: Rinaldo
mesh_terms: D000163:Acquired Immunodeficiency Syndrome / Q000097:blood / Q000821:virology; D000911:Antibodies, Monoclonal; D000917:Antibody Formation / Q000276:immunology; D000956:Antigens, Viral / Q000276:immunology; D001402:B-Lymphocytes / Q000276:immunology / Q000821:virology; D001800:Blood Specimen Collection; D018414:CD8-Positive T-Lymphocytes / Q000276:immunology; D002470:Cell Survival / Q000276:immunology; D005615:Freezing; D015686:Gene Products, env / Q000276:immunology; D015683:Gene Products, gag / Q000276:immunology; D015684:Gene Products, pol / Q000276:immunology; D015497:HIV-1 / Q000302:isolation & purification*; D015395:Histocompatibility Antigens Class I / Q000276:immunology; D006801:Humans; D007156:Immunologic Memory; D016130:Immunophenotyping; D007963:Leukocytes, Mononuclear / Q000166:cytology*; D008297:Male; D013602:T-Lymphocytes, Cytotoxic / Q000821:virology*; D014616:Vaccinia virus / Q000276:immunology
chemical_list: D000911:Antibodies, Monoclonal; D000956:Antigens, Viral; D015686:Gene Products, env; D015683:Gene Products, gag; D015684:Gene Products, pol; D015395:Histocompatibility Antigens Class I