TY - JOUR KW - CD4-Positive T-Lymphocytes KW - CD8-Positive T-Lymphocytes KW - Cytokines KW - Cytoplasm KW - Flow Cytometry KW - Humans KW - Interferon-gamma KW - Interleukin-10 KW - Interleukin-4 KW - leprosy KW - Mycobacterium leprae KW - Tuberculin KW - Tuberculosis, Pulmonary KW - Tumor Necrosis Factor-alpha AU - Antas P R Z AU - Sales J S AU - Pereira K C AU - Oliveira E B AU - Cunha K S AU - Sarno E N AU - Sampaio E P AB -
Using a short-term bulk culture protocol designed for an intracellular-staining method based on a flow cytometry approach to the frequencies of cytokine-producing cells from tuberculosis and leprosy patients, we found distinct patterns of T cell subset expression. The method also reveals the profile of peak cytokine production and can provide simultaneous information about the phenotype of cytokine-producing cells, providing a reliable assay for monitoring the immunity of these patients. The immune response of Mycobacterium leprae and purified protein derivative (PPD) in vitro to a panel of mycobacteria-infected patients from an endemic area was assessed in primary mononuclear cell cultures. The kinetics and source of the cytokine pattern were measured at the single-cell level. IFN-gamma-, TNF-alpha-, IL-4- and IL-10-secreting T cells were intracytoplasmic evaluated in an attempt to identify M. leprae- and PPD-specific cells directly from the peripheral blood. The analysis by this approach indicated that TNF-alpha was the first (8 h) to be produced, followed by IFN-gamma (16 h), IL-10 (20 h) and IL-4 (24 h), and double-staining experiments confirmed that CD4+ were a greater source of TNF-alpha than of CD8+ T cells (P < 0.05). Both T cell subsets secreted similar amounts of IFN-gamma. We conclude that the protocol permits rapid evaluation of cytokine production by different T cell populations. The method can also be used to define immune status in non-infected and contact individuals.
BT - Brazilian journal of medical and biological research = Revista brasileira de pesquisas medicas e biologicas C1 - http://www.ncbi.nlm.nih.gov/pubmed/15273814?dopt=Abstract CN - ANTAS2004 DA - 2004 Aug DO - 10.1590/s0100-879x2004000800003 IS - 8 J2 - Braz. J. Med. Biol. Res. LA - eng N2 -Using a short-term bulk culture protocol designed for an intracellular-staining method based on a flow cytometry approach to the frequencies of cytokine-producing cells from tuberculosis and leprosy patients, we found distinct patterns of T cell subset expression. The method also reveals the profile of peak cytokine production and can provide simultaneous information about the phenotype of cytokine-producing cells, providing a reliable assay for monitoring the immunity of these patients. The immune response of Mycobacterium leprae and purified protein derivative (PPD) in vitro to a panel of mycobacteria-infected patients from an endemic area was assessed in primary mononuclear cell cultures. The kinetics and source of the cytokine pattern were measured at the single-cell level. IFN-gamma-, TNF-alpha-, IL-4- and IL-10-secreting T cells were intracytoplasmic evaluated in an attempt to identify M. leprae- and PPD-specific cells directly from the peripheral blood. The analysis by this approach indicated that TNF-alpha was the first (8 h) to be produced, followed by IFN-gamma (16 h), IL-10 (20 h) and IL-4 (24 h), and double-staining experiments confirmed that CD4+ were a greater source of TNF-alpha than of CD8+ T cells (P < 0.05). Both T cell subsets secreted similar amounts of IFN-gamma. We conclude that the protocol permits rapid evaluation of cytokine production by different T cell populations. The method can also be used to define immune status in non-infected and contact individuals.
PY - 2004 SP - 1119 EP - 29 T2 - Brazilian journal of medical and biological research = Revista brasileira de pesquisas medicas e biologicas TI - Patterns of intracellular cytokines in CD4 and CD8 T cells from patients with mycobacterial infections. VL - 37 SN - 0100-879X ER -