HLA‐DPB1*612:01 differs from HLA‐DPB1*13:01:01:01 by a single nucleotide substitution at codon 37 (CGC‐ > TGC).

Thalassemia is an autosomal recessive disorder associated with gene mutations blocking the synthesis of a-or β-chain of globin.Hematopoietic stem cell transplantation (HSCT) is the only curative therapy for patients with Thalassemia.However,HSCT has been limited by human leukocyte antigen (HLA) matching degree.Moreover,the relatively probability of graft rejection and regimen-related adverse reactions in many patients receiving HSCT for Thalassemia are associated with the preconditioning regimens.It is important to choose suitable donors and pretreatments for treatment of thalassemia.In this review,the authors aimed to discuss the alternate HSCT approaches for Thalassemia,including unrelated donor HSCT,related donor HSCT,or umbilical cord blood transplantation(UCBT) and the preconditioning regimens.

OBJECTIVE:To study the genetic polymorphisms of human leukocyte antigen (HLA)- A, B, C, DRB1, DQA1, DQB1, DPA1and DPB1among ethnic Hans from southern China.METHODS:481 randomly selected individuals were genotyped using a polymerase chain reaction (PCR) sequence-based typing (SBT) method for the above genes. Their allele frequencies were determined by direct counting.RESULTS:In total, 28 HLA-A, 57 HLA-B, 28 HLA-C, 40 HLA-DRB1, 18 HLA-DQA1, 17 HLA-DQB1, 6 HLA-DPA1and 21 HLA-DPB1alleles were identified. Among these, common alleles (with allelic frequencies > 0.05) included A*1101, A*2402, A*0207, A*3303, A*0201, B*40:01, B*46:01, B*58:01, B*13:01, B*15:02, C*01:02, C*07:02, C*03:04, C*03:02, C*08:01, C*03:03, C*04:01, DRB1*09:01, DRB1*15:01, DRB1*12:02, DRB1*08:03, DRB1*03:01, DRB1*04:05, DRB1*11:01, DQA1*01:02, DQA1*03:02, DQA1*03:03, DQA1*06:01, DQA1*01:03, DQA1*05:05, DQA1*01:04, DQA1*03:01, DQA1*05:01, DQB1*03:01, DQB1*03:03, DQB1*06:01, DQB1*05:02, DQB1*03:02, DQB1*02:01, DQB1*03:02, DQB1*06:02, DPA1*02:02, DPA1*01:03, DPA1*02:01, DPB1*05:01, DPB1*02:01, DPB1*13:01, DPB1*04:01and DPB1*02:02.For each of the locus, the overall frequencies of common alleles were 75.57%, 52.81%, 78.28%, 62.16%, 86.70%, 77.23%, 95.32% and 81.59%, respectively.CONCLUSION:The allelic frequencies of the 8 selected HLA loci among ethnic Hans from southern China may served as a reference for anthropology, legal medicine, transplantation and disease association studies.

KIR2DL1*00602 differs from KIR2DL1*00302 by a non‐synonymous mutation in exon 7.

OBJECTIVE:To explore the association of KIR-HLA gene polymorphism with chronic myeloid leukemia (CML) among ethnic Hans from southern China.METHODS:A total of 172 adult CML patients and 480 unrelated healthy controls were screened for the presence of KIR with sequence-specific primers-PCR (PCR-SSP) and sequence-based typing (SBT) of HLA-A, -B and -C loci. Polymorphisms of the KIR-HLA system were analyzed at 4 levels, and the frequencies of KIR framework genes and KIR profiles, classⅠHLA ligands, matched KIR+HLA pairs and KIR-HLA compound profile were compared between the two groups. P values were calculated using SPSS 13.0 software.RESULTS:For the CML group, the frequencies of HLA-C2 ligand, 2DL1+HLA-C2 pair and HLA-B Bw4-80I were significantly lower than those of the control group, suggesting a protective effect against CML (HLA-C2: OR=0.386, 95%CI:0.240-0.620, P<0.01; 2DL1+HLA-C2: OR=0.316, 95%CI:0.191-0.525, P<0.01; HLA-B Bw4-80I: OR=0.576, 95%CI:0.384-0.862, P<0.01). The frequencies of KIR2DL1 ligand (HLA-C2) and KIR3DL1 ligand (HLA-B Bw4-80I) in the CML group were significantly lower than that of the control group, suggesting that the HLA-C2 and HLA-B Bw4-80I expression is probably decreased in the CML patient group, which led to reduced inhibitory signal and enhanced activating signal of KIR2DL1+ and/or KIR3DL1+ NK cells. Notably, the frequency of KIR-HLA compound profiles ID2 (KIR AA1-HLA-C1/C1-Bw6/Bw6-A3/11) in CML patients significantly increased in the CML patient group compared with the control group, suggesting that the KIR-HLA compound profiles ID2 may be a risk factor for CML (OR=2.163, 95%CI 1.198-3.906, P<0.01).CONCLUSION:Above analysis has identified certain protective and risk factors for CML from the KIR-HLA system, which may provide a clue for the pathogenesis of leukemia and development of individualized immune therapy.

HLA-B*15:374 has one nucleotide change from HLA-B*15:02:01 at nucleotide 553 bp where G → A in exon 3.

目的 研究T、B淋巴细胞亚群在双份机采血小板献血者献血前后及一般对照人群中的表达变化,探讨双份机采血小板对献血者淋巴细胞亚群的影响.方法 应用流式细胞术检测50例双份机采血小板献血前、后及50例未献血对照者外周血淋巴细胞亚群的表达水平,包括CD3+、CD3+CD4+、CD3+CD8+、CD19+淋巴细胞百分比及CD3+ CD4+/CD3+CD8+比值.结果 双份机采血小板献血者献血前、后CD3+T淋巴细胞百分比、CD3+CD4+T淋巴细胞百分比、CD3+CD8+T淋巴细胞百分比、CD3+CD4+/CD3+CD8+比值及CD19+B淋巴细胞百分比与正常对照组四项指标相比,差异均无统计学意义(P＞0.05).结论 双份机采血小板对献血者T、B淋巴细胞亚群均无明显影响.

The novel KIR2DL2*013 allele differs from the closest allele KIR2DL2*00302 by two non-synonymous mutations.

The inhibitory KIR3DL1 and the activating KIR3DS1 segregate as alleles of the same locus. KIR3DL1 is highly diversified at the allele level and KIR3DL1 alleles exhibit varied levels of expression and ligand binding affinity resulting in varied degrees of NK cell inhibition. Previous studies have shown that the KIR3DL1/3DS1 polymorphism associated with viral infection, cancer and transplantation. However, little is known about the population distribution of KIR3DL1/3DS1 alleles in Chinese. The present study examined allelic diversity of KIR3DL1/3DS1 in a southern Chinese population (N=306) using PCR-SSP and sequencing based typing. The presence of KIR3DL1 and KIR3DS1 were detected in 97.1% and 34.0% of the tested individuals respectively. A total of 10 KIR3DL1 alleles (including 2 novel ones) and 6 KIR3DS1 alleles (including 5 novel ones) were identified. Common KIR3DL1 alleles (>10%) were KIR3DL1*01502 (74.8%), KIR3DL1*00501 (23.9%) and KIR3DL1*00701 (15.7%). KIR3DS1*01301 was the predominant KIR3DS1 allele with other KIR3DS1 alleles only sporadically observed. The knowledge of the allelic polymorphism of KIR3DL1/3DS1 may help to better understand the role played by KIR3DL1/3DS1 in associated diseases and clinical transplantation in southern Chinese.

Objective To investigate the number and ratio of ambiguous allele combinations from human leukocyte antigen (HLA) confirmatory test by sequencing-based typing for unrelated donor marrow transplantation,and to establish an efficient strategy for identifying such ambiguities.Methods A total of 650 donor-receipt samples were genotyped for 5 loci of the HLA gene using an Atria SBT commercial kit.Exons 2,3 and 4 of HLA-A,-B and-C,exon 2 of HLA-DRB1 and exons 2 and 3 of HLA-DQB1 were tested by routine HLA genotyping.The ratio of usual ambiguous allele combination was calculated.The ambiguities were subjected to further confirmatory test by PCR-SSP or PCR-SBT retest at outside of the routine sequencing region.Results Among the 650 tested samples,the ratio of ambiguity at HLA-A,B,C,DRB1 and DQB1 were 76.31％ (496/650),91.08％ (592/650),97.69％ (635/650),88.62％ (576/650) and 43.38％ (141/650),respectively.A total of 36 ambiguous allele combinations inside the routine sequencing region and 22 ambiguous allele combinations outside of the routine sequencing region were discovered.After removing rare alleles based on the Chinese common and well-documented (CWD) Allele Table (Version 1.01),9 ambiguous CWD allele combinations inside the routine sequencing region,including 3 located in HLA-B,HLA-C and 1 located in other three HLA loci were found.Ten ambiguous CWD allele combinations outside of the routine sequencing region,including 4 located in HLA-C,-DRB1 and 1 in HLA-A,-B respectively were determined.All samples with ambiguous CWD allele combinations could be distinguished by high-resolution PCR-SSP commercial kits or PCR-SBT retest at outside of the routine sequencing region.Conclusion The common and well-documented allele combinations in sequencing-based typing at five HLA loci have been analyzed.Our strategy may provide valuable information for more efficient,low-cost and accurate method for high-resolution genotyping of HLA genes.

A novel HLA-DQB1 allele, DQB1*05:10, was detected in a Chinese individual by cloning and sequencing.

A novel HLA‐DQB1 allele, DQB1*05:10, was detected in a Chinese individual by cloning and sequencing.

A novel HLA-DQB1 allele, DQB1*05:10, was detected in a Chinese individual by cloning and sequencing.

HLA-A*24:02:09 shows one nucleotide difference from HLA-A*24:02:01:01 at position 408 in exon 3 (codon 112 GGG>GGC).

OBJECTIVE:To explore the reason for HLA-DQB1 allele dropout during routine sequence-based typing(SBT) in order to improve the accuracy of typing.METHODS:Two thousand samples derived from HLA high-resolution typing laboratory were typed for HLA-DQB1 locus using an AlleleSEQR HLA-DQB1 SBT kit. Non-conclusive results and "abnormal" sequencing samples were retyped using a LABType rSSO HD HLA-DQB1 kit and further analyzed with both sequence-specific primers and group-specific primers and sequenced for haplotype analysis.RESULTS:Among the 2000 samples, 2 samples with no conclusive result were identified. The heterozygosity was confirmed with both the LAB Type SSO HD HLA-DQB1 kit and PCR-SBT in house method. Subsequent HLA-DQB1 cloning and haplotype sequencing have elucidated that HLA-DQB1*02:02 dropped out at exon 2 for the first sample and HLA-DQB1*02:01:01 dropped out at exon 2 for the second sample during PCR amplification. No novel nucleotide mutation was found.CONCLUSION:Our results indicated that preferential amplification at exon 2 of DQB1 may result in allele dropout in exon 2 sequences during HLA-DQB1 SBT test. This may provide useful information for HLA genotyping.

Objective To explore the polymorphisms of human leukocyte antigen (HLA)-DPA1 and-DPB1,and determine the common and well-documented (CWD) alleles in southern Chinese Han population.Methods From February 2010 to September 2011,a total of 1 730 cases of peripheral blood samples from unrelated healthy voluntary blood donors of Chinese Han population in Shenzhen blood center,were collected in this study by compater random number table.All these samples were subjected to HLA-DPA1 and-DPB1 sequencing-based typing at exon 2 in both directions.In-house group specific primers were designed and utilized to identify the ambiguous allele combinations.Products of sequencing reactions subjected to sequence cleaning by EtOH/ethylene diamine tetraacetic acid (EDTA) method.The purified products were run electrophoresis on the ABI 3730 DNA sequencer,then obtained sequences were imported into the Assign 3.5 software,and the HLA-DPA1 and-DPB1 allelic genotype were assigned.Results In 1 730 unrelated healthy southern Chinese Han individuals,a total of 7 HLA-DPA1 alleles were identified,including 5 common alleles of which the frequencies were more than 0.1％.Frequencies of each common allele were 54.65％ of DPA1 * 02 ∶ 02,31.24％ of * 01 ∶ 03,10.26％ of * 02 ∶ 01,3.64％ of * 04 ∶ 01,and 0.12％ of * 01 ∶ 04.Two rare novel alleles were observed in less than 3 times.Thirty three HLA-DPB1 alleles including 20 common alleles with a frequency more than 0.1％,and 2 well-documented alleles with a frequency less than 0.1％ but were detected more than 3 times and 11 rare alleles of which two were novel alleles were determined.Conclusions Technique for simultaneous HLA-DPA1 and-DPB1 sequencebased typing at exon 2 and assay for determining ambiguous allele combinations using the in-house group specific primers have been established in this study.The information of allele frequencies and CWD alleles for HLA-DPA1 and-DPB1 in southern Chinese Han population were obtained.

KIR and their HLA ligands are encoded by two of the most diverse gene families in the human genome. The function of KIR on the NK cell is highly dependent on the normal expression of class I HLA on the target cell. Previous population studies in southern Chinese have been focused on the KIR framework genes and genotypes but little is known about the compound profiles of KIR/HLA. The present study examined 503 unrelated individuals from southern Chinese Han population for the polymorphism of KIR and class I HLA genes. All 16 KIR genes were detected in the study population and the four framework genes KIR3DL2, 3DL3, 3DP1, and 2DL4 were present in all individuals. Thirty unique KIR gene profiles were found reflecting a rather limited number of KIR haplotypes in this population. KIRAA1 was the most common profile observed in 54.7% of the samples. Among the AA1 individuals, 15.6% were homozygous for the deleted KIR2DS4. Haplotype A (74.8%) was more common than haplotype B (25.2%). HLA-C1 was a much more common ligand for 2D KIRs than C2. Bw4-80I, Bw4-80T, and the Bw4-bearing HLA-A alleles were detected at similar frequencies. The matched KIR+HLA pairs 2DL2/3+C1 (98.1%), 3DL1+Bw4 (73.3%), 3DL2+A3/11 (60.0%) were the most common ones whereas 3DS1+Bw4-80I was the least common (9.4%). A total of 193 unique compound profiles of KIR–HLA were identified in 480 informative individuals, 130 of the profiles being detected only once. The study provided a comprehensive analysis of the KIR/HLA profiles in southern Chinese in regards of the presence/absence of KIR genes, HLA ligands, matched KIR+HLA pairs, and KIR/HLA compound profiles. The results could help to better understand the role played by KIR/HLA interaction in associated diseases and clinical transplantation in southern Chinese.

The novel KIR2DL3*025 allele differs from the closest allele KIR2DL3*0010101 by a non-synonymous mutation at CDS nt280 C>A in exon 4.

KIR2DL3*026 differs from the closest allele KIR2DL3*00101 by a nonsynonymous mutation in exon 4.