Abstract
AbstractInterleukin-10 (IL-10), also known as cytokine synthesis inhibitory factor, has pleiotropic effects in immunoregulation and inflammation. It is capable of inhibiting synthesis of pro-inflammatory cytokines like interferon γ (IFNγ), IL-2, IL-3, tumor necrosis factor α(TNFα) and granulocyte macrophage colony stimulating factor (GM-CSF) made by cells such as macrophages and T helper Type 1 cells. We observed that normal human serum, derived from a healthy individual but containing large amounts of IL-10 (arbitrarily designated as "IL-10 serum"), inhibited cytotoxic activity and interfered with granzyme B release from alloreactive cytotoxic T cell (CTL) clones in vitro, but did not affect perforin release. The addition of normal human serum containing high levels of anti-IL-10 IgG (arbitrarily designated as "anti-IL-10 IgG serum") neutralized the inhibitory effects of IL-10 serum. Moreover, we have identified that cytotoxic activity and granzyme B release from an Epstein-Barr virus (EBV)-specific CTL clone was similarly inhibited in the presence of IL-10 serum, while perforin release was unaffected. Anti-IL-10 IgG serum also appeared to neutralize the inhibitory effect of IL-10 serum on an EBV-specific CTL clone.
Highlights
IL-3, tumor necrosis factor α (TNF α ) and granulocyte macrophage colony stimulating factor (GM-CSF) made by cells such as macrophages and T helper Type 1 cells
Exposure to IL-10 serum has a continued effect on alloreactive CTL clones To determine the recovery of CTL clones from suppression by IL-10 serum, the clones were mixed with a 10% volume of control or IL-10 serum followed by incubation at 37°C for 4 hours
After washing three times with RPMI 1640 medium, clones were cultured in fresh medium [RPMI 1640 + 10% fetal calf serum (FCS) + 50 u/ml recombinant IL-2] for 24 hours at 37°C, and subjected to a cytotoxic assay
Summary
IL-3, tumor necrosis factor α (TNF α ) and granulocyte macrophage colony stimulating factor (GM-CSF) made by cells such as macrophages and T helper Type 1 cells. We observed that normal human serum, derived from a healthy individual but containing large amounts of IL-10 (arbitrarily designated as “IL-10 serum”), inhibited cytotoxic activity and interfered with granzyme B release from alloreactive cytotoxic T cell (CTL) clones in vitro, but did not affect perforin release. Based on the information from the literature cited above, we analyzed serum containing high levels of IL-10 (arbitrarily designated “IL-10 serum”), derived from normal healthy blood donors, for its ability to inhibit the cytotoxic activity of a number of alloreactive, and one Epstein-Barr virus (EBV)-specific, cytotoxic T cell (CTL) clones. We have tested whether anti-IL10 IgG containing serum (arbitrarily designated “anti-IL-10 IgG serum”) derived from normal healthy blood donors can neutralize the inhibitory effects of IL-10 serum. Perforin, a serine protease that is released from cytotoxic T cells concomitantly with granzyme B, was not inhibited by the addition of IL-10 serum
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