title: Cytokine production at the site of disease in human tuberculosis.
abstract: Clinical and immunologic evidence suggests that tuberculous pleuritis provides a model to understand protective immune mechanisms against Mycobacterium tuberculosis. We therefore evaluated the pattern of cytokine mRNA expression and cytokine production in pleural fluid and blood of patients with tuberculous pleuritis. RNA was extracted from mononuclear cells, reverse transcribed to cDNA, and amplified by polymerase chain reaction (PCR). After normalization for T-cell cDNA, cDNA from pleural fluid cells and peripheral blood mononuclear cells (PBMC) was amplified with cytokine-specific primers. PCR product was quantified by Southern blot. For the Th1 cytokines gamma interferon (IFN-gamma) and interleukin-2 (IL-2), PCR product was greater in pleural fluid than in blood, whereas PCR product for the Th2 cytokine IL-4 was decreased in pleural fluid compared with blood. Concentrations of IFN-gamma were elevated in pleural fluid compared with serum, but IL-2, IL-4, and IL-5 were not detectable. Mean concentrations of IFN-gamma and IL-2 in supernatants of M. tuberculosis-stimulated pleural fluid cells were significantly greater than corresponding concentrations in supernatants of stimulated PBMC. In situ hybridization showed that increased IFN-gamma production by pleural fluid cells was associated with a 20- to 60-fold increase in the frequency of antigen-reactive IFN-gamma-mRNA-expressing cells. Because IL-10 can be produced by T cells and macrophages, pleural fluid cells and PBMC were normalized for beta-actin cDNA content and then amplified by PCR with IL-10-specific primers. IL-10 mRNA was greater in pleural fluid cells than in PBMC and was expressed predominantly by macrophages. IL-10 concentrations were elevated in pleural fluid versus serum. These data provide strong evidence for compartmentalization of Th1 cytokines and IL-10 at the site of disease in humans with a resistant immune response to mycobacterial infection.
authors.affiliation: Department of Medicine, University of Southern California School of Medicine, Los Angeles 90033.
authors.forename: P F
authors.initials: PF
authors.lastname: Barnes
authors.forename: S
authors.initials: S
authors.lastname: Lu
authors.forename: J S
authors.initials: JS
authors.lastname: Abrams
authors.forename: E
authors.initials: E
authors.lastname: Wang
authors.forename: M
authors.initials: M
authors.lastname: Yamamura
authors.forename: R L
authors.initials: RL
authors.lastname: Modlin
mesh_terms: D016207:Cytokines / Q000096:biosynthesis* / Q000235:genetics; D004247:DNA / Q000235:genetics; D006801:Humans; D007371:Interferon-gamma / Q000096:biosynthesis; D016753:Interleukin-10 / Q000235:genetics; D008264:Macrophages / Q000378:metabolism; D016133:Polymerase Chain Reaction; D012333:RNA, Messenger / Q000032:analysis; D014396:Tuberculosis, Pleural / Q000276:immunology*
chemical_list: D016207:Cytokines; D012333:RNA, Messenger; D016753:Interleukin-10; D007371:Interferon-gamma; D004247:DNA