HLA-DPB1 allelic frequency of the Pumi ethnic group in south-west China and evolutionary relationship of Pumi with other populations.

A sequencing-based typing of the HLA-DPB1 gene was carried out in 51 unrelated healthy individuals from the Yunnan Pumi ethnic minority. A total of 18 DPB1 alleles, in which DPB1*0501 (52.0%) and DPB1*0402 (15.7%) greatly predominated, were found, of which alleles DPB1*20011, 2201, 3601, 3701, 3801, 4901, 5001 and 8001 were recorded for the first time in the Chinese population. This may be because the typing methods used in previous genotyping of Chinese populations were of lower sensitivity than that used in our study. A dendrogram constructed by the maximum likelihood method showed that the Pumi ethnic minority belongs to the Asian/Australasian cluster and has the closest relationship to Trobriander, implying an unusual relationship between Australasian and South China populations. However, the Yi ethnic minority, which also comes from the ancient Qiang, did not show a very close relationship with the Pumi. This is probably because the Pumi were historically assimilated by local south-west China populations.     

Polymorphism of the DPB1 locus in Hani ethnic group of south-western China

Polymorphism of HLA-DPB1 was revealed with a sequencing-based typing (SBT) method in 47 unrelated healthy individuals from Yunnan Hani ethnic minority. The alleles DPB1*5901 and DPB1*7001 were detected for the first time in Chinese populations. A dendrogram showed that the Hani ethnic group belongs to the southern group of Chinese.     

HLA-DPB1 polymorphism in Blang and Puyi ethnic groups of southwest China inferred from sequence-based typing

In the present study, DNA typing for HLA-DPB1 was performed using polymerase chain reaction (PCR)-sequence-based typing method in two isolated Chinese populations: the Blangs (n = 94) in Shuangjiang County and the Puyis (n = 76) in Luoping County from Yunnan province of Southwest China. These two populations exhibited certain similarity in their allelic distributions of the HLA-DPB1 gene. A total of 11 and 12 alleles at the DPB1 locus were found in the Blang and Puyi groups, respectively. In the Blang group, the most frequent alleles were DPB1*0501 (51.0%) and DPB1*1301 (17.0%). DPB1*030101 was also common with a frequency of 6.4%. In the Puyi group, the most frequent allele was also DPB1*0501 with a frequency of 47.5%, followed by DPB1*1301 (21.1%). Two alleles DPB1*2101 and DPB1*0202 followed, with frequencies ranging between 5% and 8%. The alleles DPB1*4101, DPB1*3301, DPB1*6801 and DPB1*8401 were found for the first time in Chinese populations. A dendrogram constructed by neighbor-joining method showed that the Blang and Puyi ethnic minorities, which had the closest relationship belonged to the southern Chinese.     

HLA-DPB1 allelic frequency of the Lisu ethnic group in the Southwest China and evolutionary relationship of Lisu with other populations

A sequencing-based typing of human leukocyte antigen-DPB1 (HLA-DPB1) gene was carried out in 37 unrelated healthy individuals from the Yunnan Lisu ethnic minority. A total of 12 DPB1 alleles, in which DPB1*1301 (33.3%), DPB1*0402 (16.6%), DPB1*040101 (13.8%), and DPB1*0501 (11.1%) were highly predominant, were found, and allele DPB1*200102 was found for the first time in a Chinese population. A dendrogram constructed by neighbor-joining method showed that the Lisu ethnic group belongs to East Asian cluster.     

Sequencing-based analysis of the HLA-DPB1 polymorphism in Nu ethnic group of south-west China

In the present study, DNA typing for HLA-DPB1 was performed using polymerase chain reaction-sequencing-based typing method in 72 randomly selected Nu ethnic individuals inhabiting the Yunnan province of south-west China. Among the 12 detected DPB1 alleles, the most frequent was DPB1*1301, with the percentage of 20.83%, followed by DPB1*0501 (19.44%), DPB1*040101 (16.67%) and DPB1*2801 (9.72%). The allele DPB1*1501 was found for the first time in the Chinese population. Neighbour-joining showed that the Nu ethnic minority belonged to East Asian cluster and was most closely related to Lisu, being consistent with the historical records. In addition, the results obtained in this study will also provide useful information on organ transplantation, forensic investigations and disease association studies.     

Molecular analyses of HLA-DRB1, -DPB1, and -DQB1 in Jing ethnic minority of Southwest China

In the present study, DNA typing for HLA-DRB1, DQB1 and DPB1 was performed using polymerase chain reaction-sequencing based typing (PCR-SBT) method in 144 random selected Jing ethnic individuals inhabiting in South China. Allele frequencies and two-locus haplotypes (DRB1-DQB1) were statistically analyzed and 20 DPB1 alleles, 27 DRB1 and 20 DQB1 were detected. The most frequent DPB1 allele was DPB1*0501 with the percentage of 36.9% followed by DPB1*1301 (15.7%), DPB1*0401 (11.0%) and DPB1*020102 (9.8%). Among the 27 detected DRB1 alleles, DRB1*120201 (13.8%) was most commonly observed followed by DRB1*150201, *030101 and *090102 alleles with the frequencies of 9.4%, 9.1% and 8.3%, respectively. Among the 20 detected DQB1 alleles the most predominant one was DQB1*030101/0309 (19.9%). DQB1*050201 (19.1%), DQB1*0201/0202 (16.1%) and DQB1*050101 (12.3%) were also frequently observed in Jing population. Statistical analysis of two-locus haplotypes showed that DRB1*120201-DQB1*030101/DRB1*120201-DQB1*0309 (HF = 9.4%, D = 6.65x10(-2)) was most predominant followed by DRB1*030101-DQB1*0201/DRB1*030101-DQB1*0202 (HF = 8.1%, D = 6.66 x 10(-2)). The comparison of HLA class II allele and haplotype frequencies in Jing with those in other populations all over the world and a dendrogram based on the DRB1, DQB1 and DPB1 genes suggested that Jing ethnic population has an origin of Southeast Asia and is belonged to the southern group of Chinese populations.     

Polymorphism of HLA-DRB1, -DQB1 and -DPB1 genes in Bai ethnic group in southwestern China

In this work, polymorphism of human leukocyte antigen (HLA)-DRB1, -DQB1 and -DPB1 genes was detected using polymerase chain reaction-sequence-based typing method in 128 healthy unrelated volunteers from the Bai ethnic group of Yunnan province of southwest China. Among all the 28 alleles detected for the DRB1 gene, the most common allele was DRB1*120201 with a frequency of 16.41%, followed by DRB1*090102, DRB1*080302, DRB1*1404, DRB1*150101, DRB1*140101 and DRB1*160201, with frequencies of 10.16%, 9.77%, 9.38%, 8.98%, 8.59% and 8.21%, respectively. Among 19 DQB1 alleles detected, the most frequent allele was DQB1*030101/0309 (35.94%), followed by DQB1*050201 (11.33%), DQB1*060101/060103 (10.54%) and DQB1*0401 (10.16%). For the DPB1 locus, the most common alleles were DPB1*0501 (42.19%), DPB1*1301 (13.28%), DPB1*020102 (10.93%) and DPB1*040101 (9.77%). The comparison of HLA class II allele frequencies of Bais with those of other Chinese populations suggested that the Bai ethnic group belonged to the southern group of Chinese.     

HLA antigen and gene frequencies in Eskimos of East Greenland

A population of 78 Ammassallik Eskimos was tested for HLA‐A, B, Cw, DR and DQ specificities using serological techniques and HLA‐Cw, DRB, DQB1 and DPB1 using three different genomic techniques. The application of two new genomic techniques for HLA‐Cw (PCR‐SSP phototyping) and HLA‐DPB1 (PCR‐RLFP) typing confirmed serological results and gave a higher resolution, with more heterozygotes than previously found. High gene frequencies of HLA‐A24 (0.80), B51 (0.21), B61 (0.30), B62 (0.21), Cw*0303 (0.27), Cw*0304 (0.51), DRB1*0401 (0.45), DRB1*1402 (0.24), DQ7 (0.88), DPB1*0201 (0.18), DPB1*0401 (0.40) and DPB1*0402 (0.30) were found. A limited number of alleles were found at all HLA loci. The Ammassallik Eskimos of East Greenland are genetically homogenous with little, if any, admixture with European populations. This contrasts with other Greenlandic populations formerly tested. The pattern of alleles indicates a close genetic relationship with other Eskimo populations.     

Allelic and haplotype diversity of HLA‐A,HLA‐B and HLA‐DRB1 gene at high resolution in the Nanning Han population

The distribution of human leucocyte antigen (HLA) allele and haplotype varied among different ethnic populations. In this study, we investigated the allele and haplotype frequencies of HLA‐A, HLA‐B and HLA‐DRB1 loci in the Nanning Han population who live in Guangxi province of China. We identified 26 HLA‐A, 56 HLA‐B and 31 HLA‐DRB1 alleles in 562 Nanning individuals of Han ethnic group by sequence‐based typing method. Of these, the three most common alleles in HLA‐A, HLA‐B and HLA‐DRB1 loci, respectively, were A*11:01 (32.12%), A*02:07 (12.54%), A*24:02 (12.01%); B*46:01 (14.41%), B*15:02 (13.61%), B*40:01 (11.48%); DRB1*15:01 (14.15%), DRB1*16:02 (11.57%) and DRB1*12:02 (10.14%). With the exception of HLA‐DRB1, the p values of the HLA‐A and HLA‐B loci showed that the HLA allelic distribution in this population was in accordance with Hardy–Weinberg expectation (p > 0.05). A total of 173 HLA~A‐B~DRB1 haplotype with a frequency of >0.1% were presented and the three most common haplotype were HLA‐A*33:03~B*58:01~DRB1*03:01 (6.12%), HLA‐A*11:01~B*15:02~DRB1*12:02 (3.39%) and HLA‐A*11:01~B*15:02~DRB1*15:01 (3.22%). The phylogenetic tree and the principal component analysis suggested that Nanning Han population had a relative close genetic relationship with Chinese Zhuang population and a relative distant genetic relationship with Northern Han Chinese. The information will be useful for anthropological studies, for HLA matching in transplantation and disease association studies in the Chinese population.     

Polymorphism of HLA class II genes in Miao and Yao nationalities of Southwest China

In the present study, the polymorphism of human leucocyte antigen class II genes was investigated by the sequence-based typing method in two Chinese populations: the Miaos (n = 85) from Guizhou province and the Yaos (n = 66) from Yunnan province. These two populations exhibited certain similarity in their allelic distributions. Among 24 DRB1 alleles detected, DRB1*150101, DRB1*140101, DRB1*160201 and DRB1*090102 in Miao and DRB1*120201, DRB1*140101, DRB1*150101 and DRB1*090102 in Yao were highly predominant. Sixteen DQB1 alleles in total were found in these two populations among which DQB1*050201, DQB1*060101/060103 and DQB1*030101/0309 in both Miao and Yao and DQB1*050301 in Yao were commonly observed. In the 13 DPB1 alleles detected, the most frequent allele was DPB1*0501 in Miao and Yao followed by DPB1*02 and DPB1*1301. Frequent comparisons with other Chinese populations suggested the southern Chinese feature for both the Miao and Yao nationalities.     

Evaluation of Ion Torrent sequencing technology for rapid clinical human leucocyte antigen typing

The development of techniques to define the human leucocyte antigen (HLA) region has proven to be challenging due to its high level of polymorphism. Within a clinical laboratory, a technique for high‐resolution HLA typing, which is rapid and cost effective is essential. NGS has provided a rapid, high‐resolution HLA typing solution, which has reduced the number of HLA ambiguities seen with other typing methods. In this study, the One Lambda NXType NGS kit was tested on the Ion Torrent PGM platform. A total of 362 registry donors from four ethnic populations (Europeans, South Asians, Africans and Chinese) were NGS HLA typed across 9‐loci (HLA‐A, ‐B, ‐C, ‐DRB1,‐DRB345 ‐DQB1 and ‐DPB1). Concordance rates of 91%–98% were obtained (for HLA‐A, ‐B, ‐C, ‐DRB1, ‐DQB1 and ‐DPB1) when compared to historical PCR‐SSO HLA types, and the identification of uncommon alleles such as A*24:07:01 and C*04:82 were observed. A turnaround time of four days was achieved for typing 44 samples. However, some limitations were observed; primer locations did not allow all ambiguities to be resolved for HLA Class II where Exon I and IV amplification are needed (HLA‐DRB1*04:07:01/04:92, HLA‐DRB1*09:01:02/*09:21 and HLA‐DRB1*12:01:01/*12:10). This study has demonstrated high‐resolution typing by NGS can be achieved in an acceptable turnaround time for a clinical laboratory; however, the Ion Torrent workflow has some technical limitations that should be addressed.     

HLA-DRB1, DQB1 and DPB1 polymorphism in the Naxi ethnic group of South-western China

Polymorphism of HLA-DRB1, DQB1 and DPB1 was revealed with a sequencing-based typing (SBT) method in unrelated healthy volunteers from the Naxi ethnic group. Among the 43 DRB1 alleles detected, the most common allele was DRB1*12021 with a frequency of 17%, followed by DRB1*08032, DRB1*09012 and DRB1*1404 with frequencies of 8.5%, 7.4% and 7.4%, respectively. Among 23 DQB1 alleles detected, the most frequent DQB1 allele was DQB1*03011/0309 (21.9%), followed by DQB1*0502 (16.4%) and DQB1*05031 (9.6%). For the DPB1 locus, the most common alleles were DPB1*0501 (25.5%), DPB1*0402 (14.6%) and DPB1*02012 (12.0%). The most common DRB1-DQB1-DPB1 haplotype was DRB1*1404-DQB1*05031-DPB1*0402 with a frequency of 5.26%, followed by the DRB1*08032-DQB1*06011-DPB1*1301 (3.51%). The distribution characteristics of the HLA class II alleles revealed that the Naxi ethnic group belonged to the Southern group of Chinese.     

HLA-DPB1, -DRB1, and -DQB1 polymorphism defined in Ewenki ethnic minority of China Inner Mongolia Autonomous Region

In the present study, DNA typing for human leucocyte antigen (HLA)-DPB1, -DRB1, and -DQB1 was performed using polymerase chain reaction-sequence-based-typing (PCR-SBT) method on 94 randomly selected, healthy, unrelated individuals from the Ewenki ethnic population in Inner Mongolia Autonomous Region of China. A total of 64 alleles: 25 in DRB1, 19 in DQB1 and 20 in DPB1, were found. Among the 25 detected DRB1 alleles, DRB1*090102, DRB1*030101, DRB1*040101, DRB1*070101, and DRB1*120101/1206 were commonly observed, with frequencies of 16.0%, 13.3%, 10.1%, 7.4%, and 7.4%, respectively. The most predominant DQB1 allele was DQB1*030101/0309 with the frequency of 27.7%, followed by DQB1*0201/0202 (19.7%), DQB1*030302 (12.8%), DQB1*060101/060103 (6.4%), and DQB1*050201 (5.9%). Of the 20 detected DPB1 alleles, DPB1*020102 was the most frequent allele with the frequency of 25.5%. DPB1*0402 (21.3%), DPB1*0401 (20.2%), DPB1*0501 (10.6%) and DPB1*4101 (3.7%) were also very frequent alleles. The most frequent two-locus haplotypes observed in the Ewenki were: DRB1*030101-DQB1*0201/0202(10.7%), DRB1*090102-DQB1*03032(9.8%), DRB1*070101-DQB1*0201/0202 (5.5%), DRB1*070101-DQB1*030302 (5.2%) and DRB1*120101/1206-DQB1*030101/0309 (4.6%). The distribution of the HLA class II alleles and haplotypes frequencies as well as the dendrogram showed that the Ewenki population belongs to the northern group of Chinese.     

HLA antigen and gene frequencies in Eskimos of East Greenland.

A population of 78 Ammassallik Eskimos was tested for HLA-A, B, Cw, DR and DQ specificities using serological techniques and HLA-Cw, DRB, DQB1 and DPB1 using three different genomic techniques. The application of two new genomic techniques for HLA-Cw (PCR-SSP phototyping) and HLA-DPB1 (PCR-RLFP) typing confirmed serological results and gave a higher resolution, with more heterozygotes than previously found. High gene frequencies of HLA-A24 (0.80), B51 (0. 21), B61 (0.30), B62 (0.21), Cw*0303 (0.27), Cw*0304 (0.51), DRB1*0401 (0.45), DRB1*1402 (0.24), DQ7 (0.88), DPB1*0201 (0.18), DPB1*0401 (0.40) and DPB1*0402 (0.30) were found. A limited number of alleles were found at all HLA loci. The Ammassallik Eskimos of East Greenland are genetically homogenous with little, if any, admixture with European populations. This contrasts with other Greenlandic populations formerly tested. The pattern of alleles indicates a close genetic relationship with other Eskimo populations.     

Y-STR polymorphisms among five Chinese minorities, Mosuo, Mongolian, Naxi, Pumi and Tibetan, in Yunnan Province, PR China

Two hundred and thirty samples from five minority populations resident in Yunnan Province, PR China (the Mosuo, Mongolian, Naxi, Pumi and Tibetan communities) were analysed at eight polymorphic Y chromosome short tandem repeats (Y-STRs) loci. A total of 59 alleles were identified, with the largest number (n = 44) in the matriarchal Mosuo community. Two or three Y alleles were specific to each population. One hundred and thirty-five haplotypes were constructed, with 45 haplotypes in Tibetans, 34 in the Mosuo, 29 in Mongolians, 26 in the Pumi and 17 in the Naxi. The Tibetan and Mosuo peoples were closest in the neighbour-joining tree, whereas the Mongolians differed from the other four populations, reflecting their origins and present-day geographical location in Yunnan.     

Short ReportY-STR polymorphisms among five Chinese minorities,Mosuo, Mongolian,Naxi, Pumi and Tibetan,in Yunnan Province,PR China

Two hundred and thirty samples from five minority populations resident in Yunnan Province, PR China (the Mosuo, Mongolian, Naxi, Pumi and Tibetan communities) were analysed at eight polymorphic Y chromosome short tandem repeats (Y-STRs) loci. A total of 59 alleles were identified, with the largest number (n = 44) in the matriarchal Mosuo community. Two or three Y alleles were specific to each population. One hundred and thirty-five haplotypes were constructed, with 45 haplotypes in Tibetans, 34 in the Mosuo, 29 in Mongolians, 26 in the Pumi and 17 in the Naxi. The Tibetan and Mosuo peoples were closest in the neighbour-joining tree, whereas the Mongolians differed from the other four populations, reflecting their origins and present-day geographical location in Yunnan.     

Frequency of HLA‐A alleles in the Syrian population genotyped by sequence‐based typing

HLA‐A molecules are highly polymorphic. Their accurate typing at a high‐resolution level is crucial for successful organ, bone marrow and cord blood transplantation. Furthermore, several HLA alleles have been involved in susceptibility to autoimmune diseases, allergies, cancers and inflammations. In order to determine common HLA‐A alleles in Syria and their frequencies, sequence‐based typing (SBT) was used to genotype HLA‐A alleles at high resolution (four digit level) among one hundred and thirty randomly selected Syrian individuals. Exons 2, 3 and 4 of the HLA‐A gene were amplified by PCR and sequenced. The sbt ‐engine software was used for allele assignment. Ambiguities were solved using group‐specific sequencing primers (GSSPs). We could identify 32 different HLA‐A alleles which were divided into 3 groups: high frequency (approximately 10%, A*01:01; A*24:02; A*03:01; A*02:01), moderate frequency (approximately 3%, such as A*02:05, A*31:01 and A*33:01), and low frequency (approximately 1%, such as A*02:11, A*29:01, A*02:02 and A*36:01). Homozygosity rate was higher than expected (11.5% vs. 7.15%). For high frequency alleles, our results show similarity to neighbouring countries. However, 15 alleles (such as A*02:04, A*02:06, A*02:11 and A*02:17) found in our cohort in low frequencies were never reported in some or all neighbouring countries. This is the first report on HLA‐A allele frequencies in Syria. In spite of the relatively low number of tested subjects, our results revealed a high degree of diversity, with 32 different alleles, reflecting the high ethnic heterogeneity of the Syrian population. The identification of alleles rarely or never reported in neighbouring countries indicates a higher genetic diversity in Syria.     

HLA-B*15 subtypes distribution in Han population in Beijing,China, as compared with those of other populations

To identify HLA-B*15 subtypes distribution in Han population in Beijing, People’s Republic of China, 826 unrelated healthy individuals were typed using the polymerase chain reaction-sequence-based typing method. Within the 246 HLA-B*15 positive individuals, 29 HLA-B*15 alleles were identified, the most predominant of which is B*1501 (40.07%), followed by B*1502 (12.87%), B*1511 (12.87%), B*1518 (9.19%) and B*1532 (3.31%). The distribution of HLA-B*15 subtype frequencies was compared between the Beijing Han, eight other Chinese ethnic minorities and six Chinese populations covering the mainland of China, Taiwan, Hong Kong and Singapore. A neighbor-joining phylogenetic tree was constructed and revealed that the Beijing Han population clustered into the northern populations group and had a closer relationship with northern Han and Hui than with southern Han or other ethnic minorities. These results thus provide useful information that can be used in anthropology, selection for bone marrow transplantation as well as in disease-association study, such as in carbamazepine (CBZ)-induced Stevens–Johnson syndrome and toxic epidermal necrolysis.     

HLA-DPA1 and DPB1 polymorphism in four Pacific Islands populations determined by sequencing based typing

Class II HLA-DP antigens are heterodimers comprised of alpha and beta chains coded by HLA-DPA1 and HLA-DPB1 genes. Both genes are polymorphic with substantial variation between different populations world wide. This work describes DPA1 and DPB1 polymorphism in four Pacific Island populations of Cook Islands, Samoa, Tokelau and Tonga, living in New Zealand. Using sequencing based typing four DPA1 alleles and twelve DPB1 alleles were observed in total among the four populations. There are two predominant DPA1 alleles DPA1*01031 and DPA1*02022 and three predominant DPB1 alleles DPB1*02012, DPB1*0401 and DPB1*0501. Fourteen DPA1-DPB1 haplotypes in total are present in these four populations with three predominant haplotypes: DPA1*02022-DPB1*0501, DPA1*01031-DPB1*02012, and DPA1*01031-DPB1*0401. Strong positive and negative disequilibrium was observed for individual DPA1-DPB1 haplotypes. Significant differences in DPA1 and DPB1 allele and haplotype frequencies were observed between Tokelauan and other three populations. Phylogenetic analysis of genetic distances between the four Pacific Island populations and other Asian Oceanian populations have shown that Cook Islanders, Samoans and Tongans are more closely related to Asian populations whereas Tokelauans cluster towards non-Austronesian populations of Papua New Guinea Highlanders and Australian Aborigines.     

HLA-DP polymorphism in Sudanese controls and patients with insulin-dependent diabetes mellitus

Human leukocyte antigen (HLA) genes are candidates for susceptibility to insulin-dependent diabetes mellitus (IDDM). The association of IDDM with particular DR and DQ alleles has been reported in all populations studied, but its association with HLA-DP alleles has been controversial. To address this question we analyzed 19 DPB1 and 2 DPA1 alleles and their associations in well-characterized Sudanese (an admixture of Arab and Black) IDDM patients (n = 71) and ethnically matched controls (n = 86) using polymerase chain reaction-sequence specific oligonucleotide (PCR-SSO) typing. There were no significant differences between the patient and control groups in the DPB1 frequencies. DPB1*0201, *0401 and DPA1*01 were the most frequent alleles in both IDDM patients and control subjects. Significant positive and negative associations between DPB1 and DPA1 alleles were detected in both groups. A novel DPB1 allele included in DPB1*1701 was identified.