The method of deriving human T‐cell clones alters the proportion of IL‐10‐producing cells

SBA Cohen, LMC Webb, M Feldmann - Immunology, 1996 - Wiley Online Library
SBA Cohen, LMC Webb, M Feldmann
Immunology, 1996Wiley Online Library
It is now well documented that the mode of primary stimulation in the mouse determines the
ratio of Th1‐, Th0‐or Th2‐type T cells obtained. In this paper we determine whether driving
and cloning human peripheral blood mononuclear cells (PBMC) non‐specifically with
interleukin‐2 (IL‐2) and/or IL‐4 and mitogen, will differentially select T cells for IL‐10
production. The presence of IL‐2 during culture (with or without IL‐4) yielded predominantly
Th1‐type clones (81% of clones with IL‐2 alone and 77% with IL‐2+ IL‐4) and a low …
It is now well documented that the mode of primary stimulation in the mouse determines the ratio of Th1‐, Th0‐ or Th2‐type T cells obtained. In this paper we determine whether driving and cloning human peripheral blood mononuclear cells (PBMC) non‐specifically with interleukin‐2 (IL‐2) and/or IL‐4 and mitogen, will differentially select T cells for IL‐10 production. The presence of IL‐2 during culture (with or without IL‐4) yielded predominantly Th1‐type clones (81% of clones with IL‐2 alone and 77% with IL‐2 + IL‐4) and a low frequency of Th0‐type clones (19% of clones with IL‐2 and 10% of clones with IL‐2 + IL‐4), whereas IL‐4 alone increased the yield of the IL‐4 producing Th0 (35%) clones. The proportions of IL‐2‐producing clones did not alter with treatment; however, the combination of IL‐2 + IL‐4 altered the proportions of IL‐10‐producing clones. 47% of IL‐2‐cultured cells and 51% of IL‐4‐cultured cells produced IL‐10 respectively, whereas only 22% of clones driven with IL‐2 + IL‐4 produced detectable levels of IL‐10. Thus in the preliminary experiments described here we have demonstrated that the cytokines used to initially drive out human T cells and maintain the T‐cell growth can skew the Th1/Th2/Th0 cytokine profiles of the clones obtained from mitogen‐stimulated cultures. Moreover and unexpectedly the proportion of IL‐10‐producing cells is altered. These results are important in interpreting analysis of the cytokine profiles of human T cells and raise questions concerning how we should derive T‐cell clones for a cytokine analysis that truly reflects the in vivo situation.
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