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

• Main Authors: P.R.Z. Antas, J.S. Sales, K.C. Pereira, E.B. Oliveira, K.S. Cunha, E.N. Sarno, E.P. Sampaio
• Format: Article
• Language: English
• Published: Associação Brasileira de Divulgação Científica 2004-08-01
• Series: Brazilian Journal of Medical and Biological Research
• Subjects: Tuberculosis; Leprosy; IFN-gamma; TNF-alpha; Anti-hCD28 mAb
• Online Access: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S0100-879X2004000800003
• ISSN: 0100-879X; 1414-431X

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.