ANTIGENIC CROSS-REACTIVITY BETWEEN MEDIA SUPPLEMENTS FOR CULTURED KERATINOCYTE GRAFTS
ANTIGENIC CROSS-REACTIVITY BETWEEN MEDIA SUPPLEMENTS FOR CULTURED KERATINOCYTE GRAFTS
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DOI：
10.1097/00004630-199107000-00005
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发表时间：
1991-01-01
期刊：
影响因子：
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通讯作者：
MEYER A A
中科院分区：
文献类型：
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作者：
JOHNSON L F;DESERRES S;MEYER A A
移植了培养的人类角质形成细胞的患者已被证明能够产生针对异种蛋白质的抗体，这些抗体补充了培养基。对这些抗原的免疫反应可能导致晚期移植物丢失或随后培养的角质形成细胞移植物的加速排斥。消除这一潜在问题的方法包括在无血清培养基中培养细胞，并用牛垂体提取物作为血清替代品。为了确定牛垂体提取物是否会产生类似于胎牛血清引起的免疫反应，我们研究了它们的抗原交叉反应性。通过十二烷基硫酸钠-聚丙烯酰胺凝胶电泳进行比较时，发现两种补充剂之间存在相当大的相似性。用牛垂体提取物和胎牛血清的抗体进行免疫染色证实了抗原交叉反应。抑制性酶联免疫吸附测定研究表明特异性交叉反应性范围为 60% 至 68%。当测试与牛垂体提取物的抗体交叉反应性时，来自 5 名接受人角质形成细胞移植物的患者的血清样本显示出 30% 至 141% 的交叉反应性，这些移植物在胎牛血清补充培养基中生长，并且对胎牛血清表现出阳性抗体滴度。牛垂体提取物含有与胎牛血清发生交叉反应的蛋白质。因此，在补充有牛垂体提取物的培养基中培养的人角质形成细胞将受到类似于在胎牛血清中培养的人角质形成细胞所引起的免疫反应。
Patients who were grafted with cultured human keratinocytes have been shown to make antibodies to the xenogeneic proteins, which supplement the culture media. An immume response to these antigens may lead to late graft loss or accelerated rejection of subsequent cultured keratinocyte grafts. Methods to eliminate this potential problem include growing cells in a serum-free medium with bovine pituitary extract as a serum substitute. To determine whether bovine pituitary extract would be subject to an immune response similar to that which is evoked by fetal bovine serum, we investigated their antigenic cross-reactivity. When compared by sodium dodecylsulfate-polyacrylamide gel electrophoresis, considerable similarity was identified between the two supplements. Immunostaining with antibodies to both bovine pituitary extract and fetal bovine serum confirmed antigenic cross-reactivity. Inhibitory enzyme-linked immunosorbent assay studies demonstrated specific cross-reactivity that ranged from 60% to 68%. When tested for antibody cross-reactivity to bovine pituitary extract, serum samples from five patients who were receiving human-keratinocyte grafts that were grown in fetal bovine serum-supplemented media and that exhibited a positive antibody titer to fetal bovine serum showed cross-reactivity that ranged from 30% to 141%. Bovine pituitary extract contains proteins that cross-react with fetal bovine serum. Human keratinocytes that are cultured in media that is supplemented with bovine pituitary extract would therefore be subject to immune response similar to that which is evoked by human keratinocytes that have been cultured in fetal bovine serum.