title: Flow cytometric analysis of the stimulatory response of T cell subsets from normal and HIV-1+ individuals to various mitogenic stimuli in vitro.
abstract: A novel technique is described which allows the study of the responses of T cell subpopulations stimulated in bulk cultures without interfering with cell-cell interactions. The number and phenotype of lymphoblasts developing following stimulation with phytohaemagglutinin (PHA), anti-CD3, staphylococcal protein A (SPA) and pokeweed mitogen (PWM) was determined in HIV-1- and HIV-1+ patients using a new five-parameter flow cytometric method. We found that normal T cells responded faster to PHA than to any of the other mitogens tested. The peak of the PHA response occurred on day 3, followed by anti-CD3 and SPA on day 4 and PWM mitogen on day 5. Although PHA and anti-CD3 stimulated up to 95% and 80% of lymphocytes, respectively, SPA and PWM stimulated only 40% and 30% of cells, respectively. A defective T cell response was observed in lymphocytes cultured from asymptomatic HIV-1+ patients compared with negative controls. This loss of response was related to a selective mortality of T cells following mitogenic stimulation, referred to as activation-associated lymphocyte death (AALD). The results showed that stronger mitogens (PHA and anti-CD3) induced AALD in a larger proportion (50-60%) of T cells than weaker mitogens such as SPA and PWM (30-40%), and that AALD affected different lymphocyte subsets to different extents. AALD occurred more frequently in total CD8+ and CD45RO+ T cells compared with CD4+ and CD45RA+ T cells, but memory CD4+ T cells were the population most severely affected in samples from HIV-1+ donors.
authors.affiliation: Department of Clinical Immunology, Royal Free Hospital and School of Medicine, London, UK.
authors.forename: E
authors.initials: E
authors.lastname: Medina
authors.forename: N
authors.initials: N
authors.lastname: Borthwick
authors.forename: M A
authors.initials: MA
authors.lastname: Johnson
authors.forename: S
authors.initials: S
authors.lastname: Miller
authors.forename: M
authors.initials: M
authors.lastname: Bofill
mesh_terms: D000163:Acquired Immunodeficiency Syndrome / Q000276:immunology; D017252:CD3 Complex / Q000276:immunology; D015496:CD4-Positive T-Lymphocytes / Q000276:immunology; D016827:CD8 Antigens / Q000276:immunology; D005434:Flow Cytometry; D015658:HIV Infections / Q000276:immunology*; D006679:HIV Seropositivity / Q000276:immunology; D015497:HIV-1; D006801:Humans; D016130:Immunophenotyping; D008213:Lymphocyte Activation / Q000502:physiology*; D008934:Mitogens / Q000276:immunology*; D010835:Phytohemagglutinins / Q000276:immunology; D011043:Pokeweed Mitogens / Q000276:immunology; D013205:Staphylococcal Protein A / Q000276:immunology; D016176:T-Lymphocyte Subsets / Q000276:immunology*
chemical_list: D017252:CD3 Complex; D016827:CD8 Antigens; D008934:Mitogens; D010835:Phytohemagglutinins; D011043:Pokeweed Mitogens; D013205:Staphylococcal Protein A