Synovial fluid T cell clones from oligoarticular juvenile arthritis patients display a prevalent Th1/Th0-type pattern of cytokine secretion irrespective of immunophenotype

Abstract

SUMMARYThe aim of the present study was to investigate the patterns of cytokine production by T cell clones raised from in vivo activated synovial fluid (SF) mononuclear cells (MNC) of five patients with oligoarticular juvenile arthritis (JA). Freshly isolated SF T cells were cultured in vitro with low dose recombinant IL-2 and subsequently cloned by limiting dilution. Sixty-four clones were obtained from the five patients studied. Fifty-nine clones were TCR a/b þ , either CD4 þ (n ¼ 43) or CD8 þ (n ¼ 15). The remaining five clones were TCR g/d þ , CD4 ¹ , CD8 ¹ . Clone immunophenotypes differed in the individual patients. Forty-four T cell clones were stimulated with phytohaemagglutinin (PHA) and phorbol myristate acetate (PMA) and supernatants tested for the presence of IL-2, IL-4, IL-5 and interferon-gamma (IFN-g) by ELISA or bioassays. Cytokine mRNA accumulation was tested by reverse transcriptase-polymerase chain reaction (RT-PCR). Most of 44 clones tested released large amounts of IFN-g irrespective of the immunophenotype. Of these, 27 were classified as Th1-type and 17 as Th0-type based upon the IFN-g/IL-4 ratio in culture supernatants. Finally, when 10 representative T cell clones were tested for pro-and anti-inflammatory cytokines, gene expression by RT-PCR, all of them were found to express the granulocyte-macrophage colony-stimulating factor (GM-CSF), tumour necrosis factor-alpha (TNF-a), IL-10 and transforming growth factor-beta 1 (TGF-b1) genes, and half of them IL-6 and IL-8 mRNA. In conclusion, T cell clones, that represent the progeny of in vivo activated SF T cells from oligoarticular JA patients, display heterogeneous immunophenotypes, but all share the ability to produce large amounts of IFN-g, with a predominant Th1/Th0 pattern. The expression of pro-and anti-inflammatory cytokine genes in these clones suggests that in vivo activated SF T cells modulate joint inflammation in a complex fashion.

Polyunsaturated fatty acids and T‐cell function: Implications for the neonate

“…During activation, sIL-2R is cleaved off and its concentration is used to estimate lymphocyte activation (91). In vitro, IL-2 is produced by activated Th1, and more recently, Th0-type CD4 cells [both CD45RA + and CD45RO + (40,92,93)]. The precise role of this cytokine in vivo is not known; however, a decreased production of IL-2 in response to an in vitro mitogen challenge has been interpreted to indicate a reduced capacity to recruit T cells (8,94).…”

Decreased in vitro production of interferon-gamma and interleukin-2 in whole blood of patients with schizophrenia during treatment

“…53 Also in rheumatoid arthritis, the in vitro production of IFN-␥ is lowered, 4,54 although increased IFN-␥ levels play a significant role at the inflammatory sites of the disease. 55,56 In multiple sclerosis, IFN-␥ seems to promote the disease process. 57 An elevated production was found which preceded the manifestation of clinical symptoms, 58 although this was not seen in acute episodes or remissions.…”

Regulation of Cytokine Production by γδ T Cells

“…This seems to hold true also for human γδ T cells. Synovial fluid γδ T cell clones derived from oligoarticular juvenile arthritis patients produce either IFN-γ only (Th1 pattern) or IFN-γ and IL-4 (Th0 pattern), whereas γδ T cells from the bronchoalveolar lavage fluid of untreated patients with allergic asthma displayed a Th2 profile when analysed ex vivo [83,84]. Additional studies performed with established γδ T cell clones have shown that individual human γδ T cells can be driven to a Th1-or Th2-profile [85].…”