Chen Qing, Srivastava Kshitij, Ardinski Stefanie C, Lam Kevin, Huvard Michael J, Schmid Pirmin, Flegel Willy A
Department of Transfusion Medicine, Clinical Center, National Institutes of Health, Bethesda, Maryland.
Transfusion. 2016 Mar;56(3):729-36. doi: 10.1111/trf.13300. Epub 2015 Oct 5.
Human neutrophil antigen-3a (HNA-3a) alloantibodies can cause severe transfusion-related acute lung injury. The frequencies of the single-nucleotide polymorphisms (SNPs) indicative of the two clinically relevant HNA-3a/b antigens are known in many populations. In this study, we determined the full-length nucleotide sequence of common SLC44A2 alleles encoding the choline transporter-like protein-2 that harbors HNA-3a/b antigens.
A method was devised to determine the full-length coding sequence (CDS) and adjacent intron sequences from genomic DNA by eight polymerase chain reaction amplifications covering all 22 SLC44A2 exons. Samples from 200 African American, 96 Caucasian, two Hispanic, and four Asian blood donors were analyzed. We developed a decision tree to determine alleles (confirmed haplotypes) from the genotype data.
A total of 10 SNPs were detected in the SLC44A2 CDS. The noncoding sequences harbored an additional 28 SNPs (one in the 5'-untranslated region [UTR]; 23 in the introns; and four in the 3'-UTR). No SNP indicative of a nonfunctional allele was detected. The nucleotide sequences for 30 SLC44A2 alleles (haplotypes) were confirmed. There may be 66 haplotypes among the 604 chromosomes screened.
We found 38 SNPs, including one novel SNP, in 8192 nucleotides covering the CDS of the SLC44A2 gene among 302 blood donors. Population frequencies of these SNPs were established for African Americans and Caucasians. Because alleles encoding HNA-3b are more common than non-functional SLC44A2 alleles, we confirmed our previous postulate that African American donors are less likely to form HNA-3a antibodies compared to Caucasians.
人类嗜中性粒细胞抗原-3a（HNA-3a）同种抗体可导致严重的输血相关急性肺损伤。在许多人群中，已知指示两种临床相关HNA-3a/b抗原的单核苷酸多态性（SNP）的频率。在本研究中，我们确定了编码携带HNA-3a/b抗原的胆碱转运样蛋白-2的常见SLC44A2等位基因的全长核苷酸序列。
设计了一种方法，通过覆盖所有22个SLC44A2外显子的8次聚合酶链反应扩增，从基因组DNA中确定全长编码序列（CDS）和相邻内含子序列。分析了200名非裔美国人、96名白种人、2名西班牙裔和4名亚洲献血者的样本。我们开发了一个决策树，以从基因型数据中确定等位基因（确认的单倍型）。
在SLC44A2 CDS中共检测到10个SNP。非编码序列中还存在另外28个SNP（5'非翻译区[UTR]中有1个；内含子中有23个；3'UTR中有4个）。未检测到指示无功能等位基因的SNP。确认了30个SLC44A2等位基因（单倍型）的核苷酸序列。在筛选的604条染色体中可能存在66种单倍型。
我们在302名献血者中覆盖SLC44A2基因CDS的8192个核苷酸中发现了38个SNP，包括1个新的SNP。确定了这些SNP在非裔美国人和白种人中的群体频率。由于编码HNA-3b的等位基因比无功能的SLC44A2等位基因更常见，我们证实了我们之前的假设，即与白种人相比，非裔美国献血者形成HNA-3a抗体的可能性较小。
