Kelly J L, Lyons A, Soberg C C, Mannick J A, Lederer J A
Department of Surgery, Harvard Medical School/Brigham and Women's Hospital, Boston, Mass 02115, USA.
Surgery. 1997 Aug;122(2):146-52. doi: 10.1016/s0039-6060(97)90003-9.
Studies have shown that susceptibility to sepsis after severe injury correlated with reduced production of T-helper 1 (Th1) cytokines (interleukin-2 [IL-2] and interferon-gamma [IFN-gamma]) and a persistence of T-helper 2 (Th2) cytokines (IL-4 and IL-10). The mechanisms responsible for this effect are not clear. We used a T-dependent antigen to study both the effect of burn injury on antigen-specific Th functions in vivo and the effect of anti-IL-10 antibody on these functions.
Male A/J mice were anesthetized and given a 25% scald burn or a sham burn. On day 0 all mice were immunized with 100 micrograms trinitrobenzene sulfonic acid (TNP) haptenated ovalbumin (TNP-OVA) in complete Freund's adjuvant. Mice (10 per group) were given 250 micrograms monoclonal rat antimurine IL-10 antibody (anti-IL-10 MAB) or control rat immunoglobin G (IgG) on day 0 and 100 micrograms anti-IL-10 MAB or IgG on day 2. On day 10 the mice were killed to obtain serum and spleen cells. TNP-specific serum antibody isotype titers were determined by enzyme-linked immunosorbent assay (ELISA). Splenocyte proliferation and cytokine-production in response to TNP-OVA or to anti-CD3 MAB were determined by tritiated thymidine incorporation and by ELISA, respectively.
Burn injury resulted in depressed levels of the TNP-specific IgG2a antibody isotype (Th1 dependent), whereas TNP-specific IgG1 and IgE (Th2 dependent) levels were not decreased in burn versus sham burn mice. Anti-IL-10 MAB but not IgG restored the IgG2a response. Burn injury also resulted in reduced TNP-OVA-specific proliferation of splenocytes, whereas anti-CD3 proliferation was equivalent in burn and sham mice. TNP-OVA-specific IL-2 and IFN-gamma production were significantly reduced by burn injury. Anti-IL-10 MAB restored TNP-OVA-specific proliferation and antigen-specific IL-2 and interferon-gamma production by splenocytes from burn mice.
Burn injury induces the loss of antigen-specific Th1 cell function, and IL-10 acts as a trigger to down-regulate Th1 activity after injury.
研究表明，严重创伤后脓毒症易感性与辅助性T细胞1（Th1）细胞因子（白细胞介素-2 [IL-2]和干扰素-γ [IFN-γ]）产生减少以及辅助性T细胞2（Th2）细胞因子（IL-4和IL-10）持续存在相关。造成这种效应的机制尚不清楚。我们使用一种T细胞依赖性抗原研究烧伤对体内抗原特异性Th功能的影响以及抗IL-10抗体对这些功能的影响。
将雄性A/J小鼠麻醉后给予25%体表烫伤或假烫伤。在第0天，所有小鼠均用100微克三硝基苯磺酸（TNP）偶联卵清蛋白（TNP-OVA）加完全弗氏佐剂进行免疫。小鼠（每组10只）在第0天给予250微克单克隆大鼠抗小鼠IL-10抗体（抗IL-10单克隆抗体）或对照大鼠免疫球蛋白G（IgG），并在第2天给予100微克抗IL-10单克隆抗体或IgG。在第10天处死小鼠以获取血清和脾细胞。通过酶联免疫吸附测定（ELISA）测定TNP特异性血清抗体亚型滴度。分别通过氚化胸腺嘧啶核苷掺入法和ELISA法测定脾细胞对TNP-OVA或抗CD3单克隆抗体的增殖和细胞因子产生情况。
烧伤导致TNP特异性IgG2a抗体亚型（依赖Th1）水平降低，而与假烧伤小鼠相比，烧伤小鼠中TNP特异性IgG1和IgE（依赖Th2）水平未降低。抗IL-10单克隆抗体而非IgG可恢复IgG2a反应。烧伤还导致脾细胞TNP-OVA特异性增殖减少，而抗CD3增殖在烧伤和假烧伤小鼠中相当。烧伤显著降低了TNP-OVA特异性IL-2和IFN-γ的产生。抗IL-10单克隆抗体可恢复烧伤小鼠脾细胞的TNP-OVA特异性增殖以及抗原特异性IL-2和干扰素-γ的产生。
烧伤诱导抗原特异性Th1细胞功能丧失，且IL-10在损伤后作为下调Th1活性的触发因素。
