Formby B, Hosszufalusi N, Chan E, Miller N, Teruya M, Takei S, Charles M A
Sansum Medical Research Foundation, Laboratory of Immunology, Santa Barbara, California 93105.
Autoimmunity. 1992;12(2):95-102. doi: 10.3109/08916939209150315.
Cytofluorometric analysis using specific monoclonal antibodies directed against the T cell antigens Thy-1.2, CD4, CD8, CD4V beta(8.1 + 8.2 + 8.3), and the antigen Mac-1 expressed by mature macrophages and NK cells were used to characterize and quantify the phenotypes of (1) unfractionated and Percoll gradient fractionated in situ islet immune cells isolated from prediabetic and diabetic female NOD mouse spleens. We found in prediabetic female mice that the majority (approximately 70%) of the in situ islet immune cells were Thy-1.2 positive T cells. CD4 positive T cells (approximately 40%) were the most abundant phenotype together with double negative T cells (approximately 20%). The percentage of CD8 positive T cells were approximately 10%, and only approximately 4% of the immune cells were Mac-1 positive. The percentages of CD4V beta (8.1 + 8.2 + 8.3) positive and double negative T cells in diabetic spleens were significantly higher in comparison to prediabetic spleens. In C57B1/6J control nondiabetic mice the percentage of double negative T cells in the spleens was significantly 4-fold lower when compared to diabetic NOD spleens. The specific cytolytic activity mediated by in situ islet immune cells against 51Cr-labeled dispersed syngeneic single-cell islet cells at an effector to target ratio of 20 was twenty- to thirty-fold higher than that mediated by prediabetic splenic lymphoid cells. It is concluded that prediabetic NOD mouse in situ islet immune cells are mostly CD4 positive and double negative T cells, and that CD4 and CD8 positive T cells in the intra-islet infiltrate warrants further evaluation as potential effector T cells in target beta-cell destruction.
使用针对T细胞抗原Thy-1.2、CD4、CD8、CD4Vβ(8.1 + 8.2 + 8.3)的特异性单克隆抗体以及成熟巨噬细胞和NK细胞表达的抗原Mac-1进行细胞荧光分析，以表征和量化从糖尿病前期和糖尿病雌性NOD小鼠脾脏中分离出的（1）未分级和经Percoll梯度分级的原位胰岛免疫细胞的表型。我们发现在糖尿病前期雌性小鼠中，大多数（约70%）原位胰岛免疫细胞是Thy-1.2阳性T细胞。CD4阳性T细胞（约40%）与双阴性T细胞（约20%）是最丰富的表型。CD8阳性T细胞的百分比约为10%，只有约4%的免疫细胞是Mac-1阳性。与糖尿病前期脾脏相比，糖尿病脾脏中CD4Vβ(8.1 + 8.2 + 8.3)阳性和双阴性T细胞的百分比显著更高。在C57B1/6J对照非糖尿病小鼠中，脾脏中双阴性T细胞的百分比与糖尿病NOD脾脏相比显著低4倍。原位胰岛免疫细胞以20的效应细胞与靶细胞比例对51Cr标记的分散同基因单细胞胰岛细胞介导的特异性细胞溶解活性比糖尿病前期脾淋巴细胞介导的活性高20至30倍。结论是，糖尿病前期NOD小鼠原位胰岛免疫细胞大多是CD4阳性和双阴性T细胞，胰岛内浸润的CD4和CD8阳性T细胞作为靶β细胞破坏中的潜在效应T细胞值得进一步评估。
