Open AccessSynovial fluid T cell clones from oligoarticular juvenile arthritis patients display a prevalent Th1/Th0‐type pattern of cytokine secretion irrespective of immunophenotype
Author(s) -
GATTORNO M.,
FACCHETTI P.,
GHIOTTO F.,
VIGNOLA S.,
BUONCOMPAGNI A.,
PRIGIONE I.,
PICCO P.,
PISTOIA V.
Publication year - 1997
Publication title -
clinical & experimental immunology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.329
H-Index - 135
eISSN - 1365-2249
pISSN - 0009-9104
DOI - 10.1046/j.1365-2249.1997.4331330.x
Subject(s) - biology , cd8 , cytokine , microbiology and biotechnology , t cell , phytohaemagglutinin , clone (java method) , immunology , peripheral blood mononuclear cell , immunophenotyping , t lymphocyte , tumor necrosis factor alpha , lymphocyte , flow cytometry , antigen , in vitro , immune system , gene , biochemistry
The 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 α/β + , either CD4 + ( n = 43) or CD8 + ( n = 15). The remaining five clones were TCR γ/δ + , CD4 − , CD8 − . 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‐γ) 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‐γ irrespective of the immunophenotype. Of these, 27 were classified as Th1‐type and 17 as Th0‐type based upon the IFN‐γ/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‐α), IL‐10 and transforming growth factor‐beta 1 (TGF‐β1) 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‐γ, 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.