几种结核分支杆菌重组蛋白诱发豚鼠迟发型超敏反应的研究

目的　了解结核分支杆菌重组抗原ESAT6 (E)、MPT64 (M)、Ag85A(A)及两两混合物诱发豚鼠产生迟发型超敏 (DTH)反应的能力。方法 常规皮肤实验法 :灭活H₃₇R_v全菌体致敏豚鼠，皮肤试验以生理盐水为阴性对照，5UPPD标准品为阳性对照，0.8μgE、0.8μgM、0.8μgA、0.4μgA+0.4μgM、0.4μgA+0.4μgE、0.4μgE+0.4μgM分别于背部皮内注射，注射后24、48、72h分别测量硬结反应横、纵直径 (mm)，取二者的平均值。皮肤试验轮圈法 :生理盐水、5UPPD及0.8μgE、0.8μgM、0.8μgA、0.4μgA+0.4μgM、0.4μgA+0.4μgE、0.4μgE+0.4μgM，以轮圈法注射于同一只豚鼠背部两侧，每批6只，共3批。注射后24、48、72h分别测量硬结反应横、纵直径 (mm)，取二者的平均值。结果 常规皮肤试验0.8μgM、A、M+E及E+A诱发豚鼠DTH的平均直径与PPD相比无显著性差异 (P＞0.05)，而0.8μgE抗原及M+A诱发豚鼠DTH的平均直径与PPD相比有显著性差异(P＜0.05)。时间效应 :重组抗原及其混合物诱发豚鼠DTH的平均直径24h平均直径显著高于48h平均直径 (P＜0.05)，24h测量效果较好。轮圈法皮肤试验排除个体差异后，结果与常规皮肤试验结果一致。结论 从试验结果看，重组ESAT6抗原及MTP64与Ag85A混合物诱发豚鼠DTH反应强于PPD。

Study on DTH reaction of guinea-pig induced by several recombinant antigen of M.tuberculosis

Objective To observe the DTH reaction induced by recombinant antigen of M.tuberculosis MPT64(M)、Ag85A(A)、ESAT6(E) and the mixture of two of them.Methods Normal skin test:The guinea-pig were sensitived by all body of dead H₃₇R_v.We used water as negative control,5U PPD as positive control,0.8μgE、0.8μgM、0.8μgA、0.4μgA+0.4μgM、0.4μgA+0.4μgE、0.4μgE+0.4μgM were injected on guinea-pig’s back respectively.The Results were observed 24、48、72h after injection.Alternative skin test:water、5U PPD、0.8μgE、0.8μgM、0.8μgA、0.4μgA+ 0.4μgM、0.4μgA+ 0.4μgE、0.4μgE+ 0.4μgM were injectecd on the back of the same guinea-pig.The Results were observed 24、48、72h respectively after injection.Results Normal skin test showed:the DTH reaction of guinea induced by 0.8μgM、A、M+E、E+A mixture respectively was no real difference with the DTH reaction induced by PPD (P＞0.05).But the DTH reaction induced by 0.8μgE and M+A mixture respectively was stronger than the Results induced by PPD (P＜0.05).24h after injection was the best observing time of the test.The Results of alternative skin test were coincidence with normal skin test.Conclusion The DTH reaction induced by ESAT6 and A+M compounds was stronger than the DTH Results induced by PPD.