Danilovs J A, Brown J, Terasaki P I, Clark W R
Tissue Antigens. 1983 Apr;21(4):296-308. doi: 10.1111/j.1399-0039.1983.tb00175.x.
In anticipation of clinical trials of fetal pancreas transplantation we have investigated the feasibility of performing HLA-DR and HLA-A, B, C typing on fetal lymphoid cells other than PBL. Using the standard NIH microcytotoxicity test modified for HLA-DR typing it was possible to demonstrate HLA-DR antigens on subpopulations of bone marrow cells and splenocytes but not on thymocytes or hepatocytes. In contrast, HLA-A, B, C antigens could be detected on all four tissues. Excellent HLA-DR typing, confirmed by maternal typing, was obtained for 19 fetuses (14 to 23 weeks old) using bone marrow cells isolated by two-fold purification on discontinuous Percoll buoyant density gradients. Similar purification of splenocytes resulted in weak reactions with anti-DR sera; however, adherent splenocytes recovered from nylon wool columns proved to be primarily DR-bearing and also provided excellent DR typing. As a corollary to these results, non-adhering splenocytes depleted of DR-bearing cells were ideal for HLA-A, B, C typing since spurious reactions due to DR antigens were greatly diminished, whereas strong specific reactions were obtained with anti-HLA-A, B, C sera. Despite weaker reactions with HLA-A, B, C antisera obtained for thymocytes, reliable HLA-A, B, C typing could be obtained when results from thymocytes were evaluated together with typing from bone marrow cells or splenocytes. The possible benefits of fetal HLA typing for fetal pancreas transplantation are discussed.
在预期进行胎儿胰腺移植临床试验之际,我们研究了对除外周血淋巴细胞(PBL)之外的胎儿淋巴细胞进行HLA - DR以及HLA - A、B、C分型的可行性。使用针对HLA - DR分型改良的标准国立卫生研究院(NIH)微量细胞毒性试验,有可能在骨髓细胞和脾细胞亚群上显示HLA - DR抗原,但在胸腺细胞或肝细胞上则无法显示。相比之下,在所有这四种组织上都能检测到HLA - A、B、C抗原。对于19例胎儿(14至23周龄),使用在不连续Percoll浮力密度梯度上通过两步纯化分离的骨髓细胞,经母本分型确认,获得了出色的HLA - DR分型。对脾细胞进行类似的纯化导致与抗DR血清的反应较弱;然而,从尼龙毛柱回收的贴壁脾细胞被证明主要带有DR抗原,并且也提供了出色的DR分型。作为这些结果的一个推论,去除了带有DR细胞的非贴壁脾细胞对于HLA - A、B、C分型是理想的,因为由DR抗原引起的假反应大大减少,而与抗HLA - A、B、C血清获得了强烈的特异性反应。尽管胸腺细胞与HLA - A、B、C抗血清的反应较弱,但当将胸腺细胞的分型结果与骨髓细胞或脾细胞的分型结果一起评估时,仍可获得可靠的HLA - A、B、C分型。本文讨论了胎儿HLA分型对于胎儿胰腺移植可能带来的益处。