酶联免疫斑点法研究小鼠急性移植物抗宿主病模型中产生干扰素γ细胞的水平

目的：探讨异基因小鼠混合淋巴细胞反应（MLR）和急性移植物抗宿主病（aGVHD）模型中，供者针对受者细胞反应后产生干扰素-γ细胞（IFN-γ PC）的水平。方法：在近交系MHC不合的异基因小鼠脾细胞MLR和aGVHD模型中，用酶联免疫斑点法（ELISPOT）对供者针对受者细胞反应后IFN-γ PC的水平及其影响因素进行了研究。结果：在异基因小鼠脾细胞MLR后，用ELISPOT检测到IFN-γ PC显著升高，具有针对受者细胞的特异性。在异基因小鼠aGVHD模型中，重度aGVHD小鼠脾细胞与未移植的异基因受鼠脾细胞在MLR后，IFN-γ PC较中度aGVHD明显升高，无aGVHD药物预防组较预防组IFN-γ PC明显升高，并具有针对受者细胞的特异性，与aGVHD的临床评分相一致。结论：用ELISPOT可以快速而敏感地检测异基因小鼠脾细胞MLR中的同种异体反应，并和aGVHD的程度以及病程具有相关性。

Evaluation of interferon-γ producing-cells using enzyme linked immunospot assay in mice model of acute graft versus host disease

OBJECTIVE: To investigate IFN-gamma producing-cells (IFN-gamma PCs) in allogeneic mixed lymphocyte reaction (MLR) and acute graft versus host disease (aGVHD) model of mice. METHODS: Enzyme linked immunospot assay (ELISPOT) was applied to study IFN-gamma PCs in MHC mismatched mice spleen cell MLR and aGVHD model of mice. RESULT: IFN-gamma PCs increased significantly in MLR after allogeneic mice spleen cell stimulation. In the experimental mice aGVHD model, IFN-gamma PCs were significantly higher in the severe aGVHD group than those in the moderate aGVHD. In the moderate aGVHD group, mice with GVHD prophylaxis regimen demonstrated significantly lower level of IFN-gamma PCs, compared with those without prophylaxis. IFN-gamma PCs were significantly correlated with the GVHD clinical scores in the group with moderate aGVHD and prophylaxis regimen. CONCLUSION: ELISPOT is a fast, sensitive and specific approach to evaluate alloresponse in allogeneic mice MLR and IFN-gamma PCs are correlated closely with the severity of aGVHD and prophylaxis regimen in the MHC-mismatched mice model.