title: Human peripheral blood CD4+ and CD8+ T cells express Th1-like cytokine mRNA and proteins following in vitro stimulation with heat-inactivated Brucella abortus.
abstract: Defining the pattern of lymphokine production associated with Brucella abortus is critical for advancing the development of B. abortus as a vaccine carrier. In the present study we investigated the ability of heat-inactivated B. abortus or lipopolysaccharide from B. abortus to induce lymphokine production from purified human T cells in vitro. Gamma interferon (IFN-gamma), interleukin-2 (IL-2), IL-4, and IL-5 induction was assayed by mRNA-specific PCR and by enzyme-linked immunosorbent assay and bioassay for protein production. Following depletion of monocytes and B cells, B. abortus increased IFN-gamma and IL-2 mRNA expression in purified T cells compared with expression in unstimulated cells. In contrast, no IL-5 mRNA expression and only transient low-level IL-4 mRNA expression and no IL-4 protein secretion were detected. Phytohemagglutinin or phorbol myristate acetate plus ionomycin induced mRNA and protein for all these cytokines. Similar results were obtained with LPS purified from B. abortus. Removal of NK cells did not reduce lymphokine production, and enriched NK cells did not express IFN-gamma mRNA or secrete IFN-gamma protein in response to B. abortus, indicating that NK cells were not the responding population. Both CD4+ and CD8+ populations produced IFN-gamma and IL-2 in response to B. abortus. Preincubation of resting T cells with B. abortus or LPS from B. abortus for 7 days induced their differentiation into Th1-like cells as judged by their subsequent lymphokine response to phorbol myristate acetate plus ionomycin. These results suggest that B. abortus can induce differentiation of Th0 into Th1-type cells.
authors.affiliation: Laboratory of Retrovirus Research, U.S. Food and Drug Administration, Bethesda, Maryland 20892, USA.
authors.forename: M B
authors.initials: MB
authors.lastname: Zaitseva
authors.forename: H
authors.initials: H
authors.lastname: Golding
authors.forename: M
authors.initials: M
authors.lastname: Betts
authors.forename: A
authors.initials: A
authors.lastname: Yamauchi
authors.forename: E T
authors.initials: ET
authors.lastname: Bloom
authors.forename: L E
authors.initials: LE
authors.lastname: Butler
authors.forename: L
authors.initials: L
authors.lastname: Stevan
authors.forename: B
authors.initials: B
authors.lastname: Golding
mesh_terms: D001483:Base Sequence; D002003:Brucella abortus / Q000276:immunology*; D015496:CD4-Positive T-Lymphocytes / Q000276:immunology*; D018414:CD8-Positive T-Lymphocytes / Q000276:immunology*; D016207:Cytokines / Q000378:metabolism*; D017931:DNA Primers / Q000737:chemistry; D015870:Gene Expression / Q000187:drug effects; D006358:Hot Temperature; D006801:Humans; D007371:Interferon-gamma / Q000378:metabolism; D007376:Interleukin-2 / Q000235:genetics; D015847:Interleukin-4 / Q000096:biosynthesis; D015759:Ionomycin / Q000494:pharmacology; D007694:Killer Cells, Natural / Q000276:immunology*; D008070:Lipopolysaccharides / Q000276:immunology; D008969:Molecular Sequence Data; D012333:RNA, Messenger / Q000235:genetics; D016176:T-Lymphocyte Subsets / Q000276:immunology*; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology; D018417:Th1 Cells / Q000276:immunology*
chemical_list: D016207:Cytokines; D017931:DNA Primers; D007376:Interleukin-2; D008070:Lipopolysaccharides; D012333:RNA, Messenger; D015847:Interleukin-4; D015759:Ionomycin; D007371:Interferon-gamma; D013755:Tetradecanoylphorbol Acetate