Open AccessFlow cytometric analysis of the stimulatory response of T cell subsets from normal and HIV‐1 + individuals to various mitogenic stimuli in vitro
Author(s) -
MEDINA E.,
BORTHWICK N.,
JOHNSON M. A.,
MILLER S.,
BOFILL M.
Publication year - 1994
Publication title -
clinical & experimental immunology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.329
H-Index - 135
eISSN - 1365-2249
pISSN - 0009-9104
DOI - 10.1111/j.1365-2249.1994.tb06079.x
Subject(s) - phytohaemagglutinin , pokeweed mitogen , t cell , immunology , cd8 , t lymphocyte , cd3 , biology , lymphoblast , lymphocyte , population , stimulation , microbiology and biotechnology , endocrinology , in vitro , immune system , medicine , cell culture , peripheral blood mononuclear cell , biochemistry , genetics , environmental health
SUMMARY 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‐P and HIV‐1 + patients using a new five‐parameter flow cytometric method. We found that normal T ceils responded faster to PHA than lo 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 HlV‐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 (50‐40%). and that AALD affected different lymphocyte subsets to different extents. AALD occurred more frequently in total CD4 + and CD45RO + T ceils compared with CD4 + and CD45RA + T cells, but memory CD4 + T cells were the population most severely affected in samples from HIV‐I + donors.