King M J, Kosanke J, Reid M E, Poole J, Banks J, Hemming N J, Smythe J, Martin P G
International Blood Group Reference Laboratory, Bristol, UK.
Transfusion. 1997 Oct;37(10):1027-34. doi: 10.1046/j.1537-2995.1997.371098016440.x.
Antigens of the human blood group system Gerbich (Ge) are located on sialoglycoproteins glycophorin C (GPC) and glycophorin D (GPD).
The Ge+ proposita (RW) produced an alloanti-Ge after receiving 2 units of red cells (RBCs) during surgery. Further studies were carried out to characterize the antibody specificity, RBC GPC and/or GPD (GPC/GPD), and the glycophorin C gene (GYPC) from RW and her compatible siblings. RW's serum contained an alloanti-Ge that did not react with RBCs from RW or four of her siblings or with RBCs with Ge-negative phenotypes. An eluate of RW's antibody reacted weakly with GPC in Western blotting. RW's RBCs were positive with 20 alloanti-Ge2, 1 autoanti-Ge2, 4 alloanti-Ge3, and 1 alloanti-Ge4. Titrations revealed weak expression of these antigens on her RBCs and those of her compatible siblings as compared with controls. In contrast, titrations of mouse and rat monoclonal antibodies specific for GPC/GPD showed no differences. Western blotting of RBC membranes using GPC/GPD specific monoclonal antibodies showed a broad diffuse band corresponding to GPC.Ge in addition to GPC and GPD. Blotting of membranes from trypsin-treated RBCs from these individuals revealed an increase of 1500 in M(r) of membrane-bound tryptic fragment over that in the membranes from typsin-treated RBCs from persons with normal GPC/GPD. In RT-PCR, two products were obtained for RW and her compatible siblings: one had a complete deletion of exon 3 and the other had a base change (A-->T) in nucleotide 173 in exon 3 (confirmed by genomic DNA sequencing of exon 3). This point mutation has resulted in the loss of restriction enzyme Tth111 I-sensitive site in the mutant GYPC.
The specificity of antibody in RW's serum was serologically anti-Ge2. Two genetic events occurred in exon 3 in GYPC of RW and her compatible siblings. The exon 3 deletion confirmed a Ge:-2,-3,4 haplotype. The abnormal tryptic fragment obtained was due to the (A173-->T) base change in exon 3 that resulted in Asp58-->Val in the deduced amino acid sequence at the membrane boundary.
人类血型系统杰尔比希（Ge）抗原位于唾液酸糖蛋白血型糖蛋白C（GPC）和血型糖蛋白D（GPD）上。
Ge阳性的先证者（RW）在手术期间输注2单位红细胞（RBC）后产生了一种同种抗-Ge。开展了进一步研究以鉴定抗体特异性、RBC的GPC和/或GPD（GPC/GPD）以及来自RW及其相合同胞的血型糖蛋白C基因（GYPC）。RW的血清含有一种同种抗-Ge，该抗体不与RW及其4名同胞的RBC或具有Ge阴性表型的RBC发生反应。RW抗体的洗脱液在蛋白质印迹中与GPC发生微弱反应。RW的RBC对20种同种抗-Ge2、1种自身抗-Ge2、4种同种抗-Ge3和1种同种抗-Ge4呈阳性。滴定显示与对照相比，这些抗原在她的RBC及其相合同胞的RBC上表达较弱。相比之下，针对GPC/GPD的小鼠和大鼠单克隆抗体的滴定未显示差异。使用GPC/GPD特异性单克隆抗体对RBC膜进行蛋白质印迹分析，除了GPC和GPD外，还显示出一条对应于GPC.Ge的宽弥散带。对这些个体经胰蛋白酶处理的RBC的膜进行印迹分析，结果显示膜结合胰蛋白酶片段在相对分子质量（M(r)）上比正常GPC/GPD个体经胰蛋白酶处理的RBC的膜增加了1500。在逆转录聚合酶链反应（RT-PCR）中，RW及其相合同胞获得了两种产物：一种产物外显子3完全缺失，另一种产物外显子3的核苷酸173处有碱基变化（A→T）（通过外显子3的基因组DNA测序证实）。该点突变导致突变型GYPC中限制性内切酶Tth111 I敏感位点的缺失。
RW血清中抗体的特异性在血清学上为抗-Ge2。RW及其相合同胞的GYPC外显子3发生了两个遗传事件。外显子3缺失证实了一种Ge:-2,-3,4单倍型。获得的异常胰蛋白酶片段是由于外显子3中的（A173→T）碱基变化，导致膜边界处推导氨基酸序列中的天冬氨酸58→缬氨酸。
