McManus K, Pongoski J, Coghlan G, Zelinski T
Rh Laboratory, Departments of Pediatrics and Child Health and of Human Genetics, Faculty of Medicine, University of Manitoba, Winnipeg, Manitoba, Canada.
Transfusion. 2000 Mar;40(3):325-9. doi: 10.1046/j.1537-2995.2000.40030325.x.
The low-incidence red cell antigens NFLD (700.37) and BOW (700.46) were first described in 1984 and 1988, respectively. Recent investigations showed that antigens of the Diego blood group system (including a number of low-incidence antigens) are coded by SLC4A1 (solute carrier family 4, anion exchanger member 1 gene). Among these newly characterized Diego system antigens is Wu (designated DI9). Because a serologic relationship among Wu, NFLD, and BOW has been established, a series of genetic and molecular investigations of SLC4A1 in relation to NFLD and BOW were undertaken.
By the use of exon-specific primers, single-strand conformational polymorphism (SSCP) analysis of SLC4A1 was performed on DNA isolated from an NFLD+ person from Japan, from the members of a Canadian kindred segregating for NFLD, and from two unrelated BOW+ persons. Exons displaying SSCPs were subjected to genetic linkage analysis (for NFLD only) and DNA sequencing.
SSCPs in DNA amplified from exons 12 and 14 of SLC4A1 were observed for all NFLD+ subjects. Linkage between each of these polymorphisms and NFLD was established with peak lods = 4.82 at theta = 0.00 for combined paternal and maternal meiosis. DNA sequencing of exons 12 and 14 of SLC4A1 from NFLD+ persons identified A-->T and C-->G mutations that underlie Glu429Asp and Pro561Ala substitutions in human erythroid band 3 protein (band 3). DNA from the two unrelated BOW+ persons only exhibited an SSCP in exon 14 of SLC4A1. Subsequent DNA sequencing revealed a C-->T mutation that accounts for a Pro561Ser substitution in band 3.
SLC4A1 codes for the low-incidence red cell antigens NFLD and BOW. In light of these findings, both antigens have been assigned to the Diego blood group system.
低发生率红细胞抗原NFLD（700.37）和BOW（700.46）分别于1984年和1988年首次被描述。最近的研究表明，迭戈血型系统的抗原（包括一些低发生率抗原）由SLC4A1（溶质载体家族4，阴离子交换成员1基因）编码。这些新鉴定的迭戈系统抗原中包括Wu（命名为DI9）。由于已确定Wu、NFLD和BOW之间存在血清学关系，因此对SLC4A1与NFLD和BOW相关的一系列遗传和分子研究展开。
使用外显子特异性引物，对从一名日本NFLD阳性个体、一个加拿大NFLD分离家系的成员以及两名无关的BOW阳性个体中分离的DNA进行SLC4A1的单链构象多态性（SSCP）分析。显示SSCP的外显子进行遗传连锁分析（仅针对NFLD）和DNA测序。
在所有NFLD阳性受试者，在从SLC4A1外显子12和14扩增的DNA中观察到SSCP。这些多态性与NFLD之间的连锁关系得以确立，父母本减数分裂合并时，在θ = 0.00处峰值lod值为4.82。对NFLD阳性个体的SLC4A1外显子12和14进行DNA测序，鉴定出A→T和C→G突变，这些突变导致人红细胞带3蛋白（带3）中Glu429Asp和Pro561Ala替换。两名无关的BOW阳性个体的DNA仅在SLC4A1外显子14中表现出SSCP。随后的DNA测序揭示了一个C→T突变，该突变导致带3中Pro561Ser替换。
SLC4A1编码低发生率红细胞抗原NFLD和BOW。鉴于这些发现，这两种抗原已被归入迭戈血型系统。
