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

Abstract

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.

More information

Type
Journal Article
Author
Antas P R Z
Sales J S
Pereira K C
Oliveira E B
Cunha K S
Sarno E N
Sampaio E P
Year of Publication
2004
Journal
Brazilian journal of medical and biological research = Revista brasileira de pesquisas medicas e biologicas
Volume
37
Issue
8
Number of Pages
1119-29
Date Published
2004 Aug
Language
eng
ISSN Number
0100-879X
Call Number
ANTAS2004
DOI
10.1590/s0100-879x2004000800003
Alternate Journal
Braz. J. Med. Biol. Res.
Publication Language
eng

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