Patterns of intracellular cytokines in CD4 and CD8 T cells from patients with mycobacterial infections

Brazilian Journal of Medical and Biological Research

P.r.z. Antas²  J.s. Sales²  K.c. Pereira²  E.b. Oliveira²  K.s. Cunha¹  E.n. Sarno²  E.p. Sampaio² 

[1] ,FIOCRUZ Instituto Oswaldo Cruz Departamento de Medicina TropicalRio de Janeiro RJ ,Brasil

关键词: Tuberculosis;    Leprosy;    IFN-gamma;    TNF-alpha;    Anti-hCD28 mAb;   

DOI  :  10.1590/S0100-879X2004000800003

来源: SciELO

【 摘 要 】

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.

【 授权许可】

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