RED-BLOOD-CELL GLYCOPHORINS AS B-CELL AND T-CELL ANTIGENS IN CANINE AUTOIMMUNE HEMOLYTIC-ANEMIA
RED-BLOOD-CELL GLYCOPHORINS AS B-CELL AND T-CELL ANTIGENS IN CANINE AUTOIMMUNE HEMOLYTIC-ANEMIA
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DOI：
10.1016/0165-2427(94)05407-j
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发表时间：
1995-08-01
影响因子：
1.8
通讯作者：
ELSON, CJ
中科院分区：
文献类型：
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作者：
BARKER, RN;ELSON, CJ
来自两只患有自身免疫性溶血性贫血（AIHA）的狗的致病性自身抗体被证明与来自犬红细胞（RBC）膜的血型糖蛋白反应。在这两种情况下，自身抗体结合纯化的血型糖蛋白的酶联免疫吸附试验（ELISA），和主要的自身抗原免疫沉淀的抗体对应的表观分子量与血型糖蛋白。此外，神经氨酸酶处理的沉淀抗原，或犬血型糖蛋白，导致在十二烷基硫酸钠聚丙烯酰胺凝胶电泳（SDS-PAGE）中的表观分子量相同的变化。证明从血型糖蛋白中去除唾液酸会导致SDS-PAGE迁移的变化，这在RBC膜蛋白中是独特的。在另外两个AIHA病例中，自身抗体不免疫沉淀血型糖蛋白模式，ELISA显示RBC反应性IgG存在于血清和RBC洗脱液中，但这些抗体未能结合犬血型糖蛋白。因此，我们认为，血型糖蛋白特异性自身抗体存在于一些，但不是所有的，狗与AIHA. T细胞增殖的情况下AIHA在体外反应自体红细胞，或多个红细胞膜成分的SDS-PAGE分级。三个馏分，对应于主要的血型糖蛋白，红细胞阴离子通道带3，和血影蛋白从膜骨架，是刺激性的。相反，健康狗的T细胞对RBC或任何印迹组分均无反应，但在一只动物中，有血影蛋白的组分除外。这表明，具有多种特异性的自身反应性T细胞的激活可能是必要的，以提供足够的帮助致病性自身抗体的产生。
Pathogenic autoantibodies from two dogs with autoimmune haemolytic anaemia (AIHA) were shown to react with glycophorin from the canine red blood cell (RBC) membrane. Autoantibodies in both cases bound to purified glycophorin in enzyme-linked immunosorbent assays (ELISAs), and the major autoantigen immunoprecipitated by the antibodies corresponded in apparent molecular mass with glycophorin. Furthermore, neuraminidase treatment of the precipitated antigen, or of canine glycophorin, resulted in identical changes in apparent molecular mass in sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE). Such removal of sialic acid from glycophorins was demonstrated to cause shifts in SDS-PAGE migration that are unique among RBC membrane proteins. In two further cases of AIHA, where autoantibodies did not immunoprecipitate the glycophorin pattern, ELISAs revealed that RBC-reactive IgG was present in serum and RBC elutes, but that these antibodies failed to bind to canine,glycophorin. Thus, we consider that autoantibodies specific for glycophorin are present in some, but not all, dogs with AIHA.T-cells from a case of AIHA proliferated in vitro in response to autologous RBC, or to multiple RBC membrane components fractionated by SDS-PAGE. Three fractions, corresponding to major glycophorins, to the RBC anion channel band 3, and to spectrin from the membrane skeleton, were stimulatory. In contrast, T-cells from healthy dogs failed to respond to RBC, or to any blot fractions with the exception, in one animal, of the fraction bearing spectrin. It is suggested that activation of autoreactive T-cells with multiple specificities may be necessary to provide sufficient help for pathogenic autoantibody production.