Jarolim P, Murray J L, Rubin H L, Coghlan G, Zelinski T
Department of Biomedical Research, St. Elizabeth's Medical Center, Tufts University School of Medicine, Boston, Massachusetts, USA.
Transfusion. 1997 Apr;37(4):398-405. doi: 10.1046/j.1537-2995.1997.37497265340.x.
Recent family studies established that the low-incidence red cell antigen WARR is not part of the MNS, Lutheran, Lewis, Duffy, Kidd, Xg, Chido/ Rodgers, Kx, or Gerbich blood group systems. Continued serologic and genetic studies of WARR suggest that it is carried on erythroid band 3.
To test the hypothesis that expression of WARR is controlled by the anion exchanger 1 gene (AE1), AE1 intronic primers that flank the exons encoding the membrane domain of band 3 were prepared. Polymerase chain reaction-amplified products corresponding to exons 11-20 of AE1 were analyzed for single-strand conformational polymorphism (SSCP) in DNA from WARR-positive and WARR-negative individuals.
An SSCP was detected in exon 14. Subsequent sequencing revealed a C-->T mutation in codon 552 that leads to a Thr-->Ile substitution. Because the C-->T mutation eliminates a Bbs I restriction site, it was possible to confirm the phenotypes of all family members. To study the possible effect of the Thr552-->Ile substitution on the expression and function of band 3, polymerase chain reaction-amplified reverse-transcribed reticulocyte mRNA was digested with Bbs I. Both alleles of band 3 mRNA were detected in similar quantities, which suggests that the substitution underlying WARR did not interfere with mRNA stability. Comparison of sodium dodecyl sulfate-polyacrylamide gel electrophoretic mobility and size patterns revealed no difference between proteins isolated from WARR-positive and WARR-negative red cells. Further, the presence of WARR did not alter the di-isothiocyano-dihydrostilbene disulfonate (DIDS)-inhibitable influx of radiolabeled sulfate.
Although it appears inconsequential to the function of band 3, the red cell polymorphism known as WARR is controlled by AE1. WARR should be therefore included in the Diego blood group system.
近期的家族研究证实，低发生率红细胞抗原WARR并非MNS、路德维希、刘易斯、达菲、基德、Xg、奇多/罗杰斯、Kx或杰尔比希血型系统的一部分。对WARR持续进行的血清学和遗传学研究表明，它由红系带3携带。
为检验WARR的表达受阴离子交换蛋白1基因（AE1）控制这一假说，制备了位于编码带3膜结构域的外显子两侧的AE1内含子引物。对WARR阳性和WARR阴性个体的DNA中与AE1外显子11 - 20相对应的聚合酶链反应扩增产物进行单链构象多态性（SSCP）分析。
在外显子14中检测到一个SSCP。随后的测序显示密码子552处有一个C→T突变，导致苏氨酸→异亮氨酸替代。由于C→T突变消除了一个Bbs I限制性酶切位点，因此可以确认所有家族成员的表型。为研究苏氨酸552→异亮氨酸替代对带3表达和功能的可能影响，用Bbs I消化聚合酶链反应扩增的逆转录网织红细胞mRNA。检测到带3 mRNA的两个等位基因数量相似，这表明WARR相关的替代并未干扰mRNA稳定性。十二烷基硫酸钠 - 聚丙烯酰胺凝胶电泳迁移率和大小模式的比较显示，从WARR阳性和WARR阴性红细胞中分离的蛋白质之间没有差异。此外，WARR的存在并未改变二异硫氰酸二氢芪二磺酸（DIDS）抑制的放射性标记硫酸盐内流。
尽管WARR对带3的功能似乎无关紧要，但这种红细胞多态性由AE1控制。因此，WARR应纳入迭戈血型系统。
