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目的 观察在条件性环磷酰胺免疫抑制反应中小鼠耳廓重量和白细胞移动抑制因子 (L MIF)活性的变化 ,并分析条件性反应小鼠血清中是否产生了某种介导物质。方法 以樟脑气味为条件刺激 ,以腹腔注射环磷酰胺为非条件刺激 ,以 2 ,4 -二硝基氯苯 (DNCB)作为致敏原诱发耳廓皮肤肿胀和同时诱发淋巴细胞释放L MIF。在条件性反应训练结束后 ,取耳称重并同时取肠系膜淋巴结细胞制备 L MIF。以左 /右耳重量之比和游出细胞光密度作为迟发过敏 (DTH)反应的检测指标。结果 (1) DTH皮肤反应和以 L MIF活性为指标的DTH反应在条件反应组均明显减弱 ,与不经过条件性训练的对照组相比差异显著 ,而与非条件反应组相比则没有差异。 (2 )条件性反应组小鼠的血清在透析前能够抑制正常小鼠皮肤 DTH反应 ,但经过透析处理后 (阻滞相对分子质量 10 0 0 0 )则不表现抑制 DTH的作用。结论 本工作建立了稳定的环磷酰胺条件性抑制 DTH皮肤反应和 L MIF活性的模型 ;并发现条件性反应小鼠血液中存在一种 (类 )小分子物质 ,介导了 DTH的抑制效应。条件性抑制 LMIF活性的模型 ,为进一步分析血清中这种介导物质的生化特性提供了一个十分方便的检测手段。
Objective To observe the changes of the auricle weight and leukocyte migration inhibitory factor (L MIF) in conditioned cyclophosphamide immunosuppression, and to analyze whether certain mediators of serum are produced in the conditioned reaction mice. Methods Campylobacter was used as stimulus. Cyclophosphamide was intraperitoneally injected as a nonconditioned stimulus and 2,4,6 - dinitrochlorobenzene (DNCB) as sensitizer was used to induce auricular skin swelling and lymphocyte release of L MIF. After conditioned training, L MIFs were prepared by weighing ears and taking mesenteric lymph node cells simultaneously. The ratio of left / right ear weight and the cell optical density were used as indexes of DTH reaction. Results (1) The DTH response and DTH response to L MIF activity in the conditioned reaction group were significantly decreased compared with the control group without the conditioned training, but no significant difference compared with the non-conditioned group . (2) Serum of conditioned reaction mice could inhibit the skin DTH response of normal mice before dialysis, but did not show DTH inhibition after dialysis treatment (blocking relative molecular mass of 100 000). CONCLUSIONS: A stable model of cyclophosphamide that inhibits DTH skin response and L MIF activity is established in this work. It is found that there is a kind of small molecule substance in the conditioned reaction mice, which mediates the inhibitory effect of DTH. The model of conditional inhibition of LMIF activity provides a very convenient means of detection for further analysis of the biochemical properties of this mediator in serum.