Study on relationship between platelet transfusion efficacy and KIR-HLA receptor-ligand compatibility / 中国输血杂志

【Objective】 To study the correlation between platelet transfusion efficacy and KIR receptor-HLA ligand. 【Methods】 Thirty-three leukemia patients with positive HLA antibody were tested for cross-matching with donor platelets. Platelets from suitable donors were selected for transfusion, and the 24-hour platelet corrected count increment (CCI) was used to determine the transfusion effect. KIR and ligand genotyping were performed on blood samples from patients and donors by PCR-SSP method, and the relationship between platelet transfusion effects and KIR receptor-HLA ligand was analyzed. 【Results】 In 74 occasions of platelet transfusion, 42 were ineffective and 32 were effective. When the donor had C2 gene and HLA-B Bw4-80T gene, the frequency of ineffective platelet transfusion in the recipient was 69.0% (29/42) and 52.4% (22/35), respectively, which was significantly higher than that in the effective group [25.0% (8/32) and 25.0% (8/32)]. When the donor had C1 gene, and the frequency of effective platelet transfusion in the recipient was 100.0%(32/32), which was higher than that in the ineffective group [83.3%(35/42)]. When the recipient-donor matching mode was KIR2DL1-C2 and KIR3DL1-(HLA-B Bw4-80T), the frequency of ineffective platelet transfusion was 69.0%(29/42) and 40.5%(22/42),higher than that of the effective group [25% (8/32) and 18.8% (6/32)]. When the recipient-donor matching model was KIR2DL3-C1, the rate of effective platelet transfusion in 32 patients (100.0%), which was higher than that (35 patients 83. 3%) in the ineffective group. When the mismatch mode of recipient iKIR+donor HLA ligand receptor was KIR2DL1-C2, the frequency of effective platelet transfusion in the recipient was 78.1% (25/32), which was much higher than that in the ineffective group [31.0% (13/42)]. When the mismatch mode was KIR3DL1-(HLA-B Bw4-80T), the rate of effective platelet transfusion in the recipient was 68.8% (22/32), which was higher than that in the ineffective group (42.9%, 18/42). The difference between the above groups was statistically significant(P<0.05). 【Conclusion】 HLA-C1 and HLA-C2 genes are the key factors affecting the efficacy of platelet transfusion.For platelet refractorines, HLA-C1 is the protective gene, while HLA-C2 and HLA-B Bw4-80T are the susceptible genes. The recipient iKIR+donor HLA ligand receptor model may play an important role in platelet refractoriness.

Conceptos básicos en inmunología de la reproducción: revisión narrativa de la bibliografía / Basic concepts in reproductive immunology: a narrative review of the literature

Resumen: ANTECEDENTES: La inmunología de la reproducción no es un área nueva: siempre ha estado relacionada con el aborto recurrente y con la falla repetida en la implantación, sobre todo en el contexto de una fertilización in vitro. Recientemente emergieron nuevos conceptos importantes que los ginecoobstetras deben considerar. OBJETIVO: Interrelacionar los conceptos básicos de inmunología, embriología y reproducción asistida para comprender mejor lo que la primera puede resolver y lo que no. METODOLOGÍA: Estudio retrospectivo efectuado con base en la búsqueda electrónica, llevada a cabo en febrero de 2020 en las bases de datos: PubMed y Google Scholar con los siguientes términos (MeSH): abortion, spontaneous/immunology; embryo implantation/immunology; HLA-c antigens/immunology; immune tolerance/immunology; immunity, maternally-acquired/immunology; uterus/immunology; killer cells, natural/immunology; placentation/immunology; receptors, kir/immunology; antigen presentation/genetics; antigen presentation/immunology; maternal-fetal exchange/genetics; maternal-fetal exchange/immunology. RESULTADOS: Se reunieron 289 artículos y se eliminaron 248 por no cumplir con los criterios de inclusión; solo se analizaron 41. Los artículos identificados sirvieron de base para actualizar la situación de la inmunología en el contexto de la medicina de la reproducción. Durante el proceso se revisaron otros artículos que sirvieran de soporte bibliográfico a los conceptos descritos en esta revisión. CONCLUSIONES: Debido al destacado interés en el estudio de la genética de los embriones, la medicina de la reproducción se enfocó más en ella y dejó de lado a la inmunología. Sin embargo, como la genética sigue sin poder explicar de manera adecuada las fallas en la implantación, la inmunología de la reproducción vuelve a cobrar impulso.

Abstract: BACKGROUND: Reproductive immunology is not a new area in reproductive medicine, it has always been related to recurrent miscarriage and repeated implantation failure, especially in the context of IVF. Recently, new concepts have emerged that are important for OBGYN specialists to keep in mind. OBJECTIVE: Interrelating the basic concepts of immunology, embryology and assisted reproduction to better understand what the former can and cannot solve. METHODOLOGY: Retrospective study based on the electronic search, carried out in February 2020, in the databases: PubMed and Google Scholar with the following terms (MeSH) The following MeSH terms were used: Abortion, Spontaneous/immunology; Embryo Implantation/immunology; HLA-C Antigens/immunology; Immune Tolerance/immunology; Immunity, Maternally-Acquired/immunology; Uterus/immunology; Killer Cells, Natural/immunology; Placentation/immunology; Receptors, KIR/immunology; Antigen Presentation/genetics; Antigen Presentation/immunology; Maternal-Fetal Exchange/genetics; Maternal-Fetal Exchange/immunology. RESULTS: 289 articles were collected, and 248 articles were deleted because they did not meet the inclusion criteria; only 41 were analyzed. The articles identified served as a basis for updating the status of immunology in the context of reproductive medicine. During the process, other articles were reviewed to serve as bibliographic support for the concepts described in this review. CONCLUSIONS: Due to the outstanding interest in the study of embryo genetics, reproductive medicine focused more on it and left immunology aside. However, since genetics still cannot adequately explain implantation failures, reproductive immunology is gaining momentum again.

Human Leukocyte Antigen-C Genotype and Killer Immunoglobulin-like Receptor-Ligand Matching in Korean Living Donor Liver Transplantation

BACKGROUND: The interaction between killer immunoglobulin-like receptors (KIRs) and HLA class I regulates natural killer (NK) cell cytotoxicity and function. The impact of NK cell alloreactivity through KIR in liver transplantation remains unelucidated. Since the frequency of HLA-C and KIR genotypes show ethnic differences, we assessed the impact of HLA-C, KIR genotype, or KIR-ligand mismatch on the allograft outcome of Korean liver allografts. METHODS: One hundred eighty-two living donor liver transplant patients were studied. Thirty-five patients (19.2%) had biopsy-confirmed acute rejection (AR), and eighteen (9.9%) had graft failure. The HLA-C compatibility, KIR genotypes, ligand-ligand, and KIR-ligand matching was retrospectively investigated for association with allograft outcomes. RESULTS: Homozygous C1 ligands were predominant in both patients and donors, and frequency of the HLA-C2 allele in Koreans was lower than that in other ethnic groups. Despite the significantly lower frequency of the HLA-C2 genotype in Koreans, donors with at least one HLA-C2 allele showed higher rates of AR than donors with no HLA-C2 alleles (29.2% vs 15.7%, P=0.0423). Although KIR genotypes also showed ethnic differences, KIR genotypes and the number of activating KIR/inhibitory KIR were not associated with the allograft outcome. KIR-ligand mismatch was expected in 31.6% of Korean liver transplants and had no impact on AR or graft survival. CONCLUSIONS: This study could not confirm the clinical impact of KIR genotypes and KIR-ligand mismatch. However, we demonstrated that the presence of HLA-C2 allele in the donor influenced AR of Korean liver allografts.

A novel real-time quantitative PCR method for analyzing HLA-C allele expression level / 中国免疫学杂志

Objective: To establish real-time qPCR method to analyze each HLA-C allele expression level of individual.Methods:Database including exon 2&3 sequences of HLA class Ⅰalleles was built ,HLA-C allelic specific primers were designed and the real-time quantitative PCR method for analyzing HLA-C allele expression level was built .The allelic specificity of these primers were confirmed in database and 835 normal peripheral blood samples of Han population .The mRNA level of each HLA-C allele from 20 pairs of liver tumor tissues and non-tumor tissues was analyzed by the qPCR method we built .Results:20 pairs of allelic specific primers were designed to distinguish the two HLA-C alleles of each individual with frequency over 0.96%in 835 cases of Han population.Among 55%of the liver cancer tissues ,the expression levels of the two HLA-C alleles from the same tumor tissue changes differently compared to that of the relevant non-tumor tissue.Conclusion:This study provides method for HLA-C allele expression level analysis of Han population and each HLA-C allele expression level is inconsistent of liver cancer tissue .

The role of KIR2DL3/HLA-C*0802 in Brazilian patients with rheumatoid vasculitis

OBJECTIVES: Rheumatoid arthritis is a polygenically controlled systemic autoimmune disease. Rheumatoid vasculitis is an important extra-articular phenotype of rheumatoid arthritis that can result in deep cutaneous ulcers. The objective of this study was to establish a correlation between the frequency of major histocompatibility complex class I/II alleles and killer immunoglobulin-like receptor genotypes in patients with cutaneous rheumatoid vasculitis. METHODS: Using the Scott & Bacon 1984 criteria to diagnose rheumatoid vasculitis and after excluding any other causes such as diabetes, atherosclerosis, adverse drug reactions, infection, and smoking, patients who met the criteria were selected. All of the selected rheumatoid vasculitis patients presented deep cutaneous ulcers. Identification of the major histocompatibility complex class I/II and killer immunoglobulin-like receptor genotypes was performed by polymerase chain reaction assays of samples collected from the 23 rheumatoid vasculitis patients as well as from 80 controls (40 non-rheumatoid vasculitis RA control patients and 40 healthy volunteers). RESULTS: An association between the presence of the HLA-DRB1*1402 and HLA-DRB1*0101 alleles and cutaneous lesions in rheumatoid vasculitis patients and a correlation between the inhibitor KIR2DL3 and the HLA-C*0802 ligand in rheumatoid vasculitis patients were found. CONCLUSION: An association was found between the presence of the HLA-DRB1*1402 and HLA-DRB1*0101 alleles and the development of cutaneous lesions in rheumatoid vasculitis patients. Additionally, the HLA-C*0802 ligand protects these individuals from developing cutaneous lesions. .

Analysis of related gene of the HLA system class I in women with cervical intraepithelial neoplasia grades II and III / Análise de gene relacionado ao sistema HLA de classe I em mulheres com neoplasia intraepitelial cervical de grau II e III

Objective: To evaluate the HLA-C gene in the progression and regression of cervical lesions caused by HPV infection, the present study aimed to analyzethe genetic diversity and allelic variants of HLA-C gene in women CIN 2 and CIN 3 diagnosed, comparing with those without abnormalities on cervicalcytology with and without the presence of infection HPV. Materials and Methods: The study group consisted of 84 women with CIN 2 and 90 women withCIN 3; the control group had no abnormality in cervical cytology consisted of 102 women negative for the presence of the HPV and 76 women with positiveHPV infection. Samples were obtained from the metropolitan region of Curitiba-PR, Brazil, aged between 15 and 45 years old. The study group wastreated in the Department of Pathology of the Cervical Erasto Gaertner Hospital, Curitiba and the control group was recruited in cervical cancer preventioncampaigns promoted by public entities in association with the Department of Obstetrics and Gynecology, Hospital de Clínicas, Federal University of Paranaand Histocompatibility and Immunogenetics Laboratory, Department of Genetics, UFPR. The HPV detection was performed by the Hybrid Capture 2(CH2®) methodology. The DNA was extracted from peripheral blood samples and HLA-C genotyping was performed PCR-SSOP method...

OObjetivo: Para avaliar os genes HLA-C na progressão/regressão das lesões cervicais causadas pela infecção do HPV, o presente estudo teve como objetivo analisar a diversidade genética e variantes alélicos do gene HLA-C de mulheres com NIC 2 e NIC 3, comparando com aquelas sem anormalidades na citologia cervical com e sem a presença da infecção pelo HPV. Materiais e Métodos: O grupo caso foi composto por 84 mulheres com NIC 2 e 90 mulheres com NIC 3; e o grupo controle, por 102 casos controle negativo para a presença do vírus HPV e 76 casos controle positivo para a presença do HPV, sem a normalidade na citologia cervical. As amostras foram obtidas da região metropolitana de Curitiba com faixa etária entre 15 e 45 anos. O grupo de estudofoi tratado no Departamento de Patologia Cervical do Hospital Erasto Gaertner, Curitiba-PR e o grupo controle foi recrutado a partir de campanhas deprevenção do câncer cervical promovido por entidades públicas em associação ao Departamento de Tocoginecologia do Hospital de Clínicas da Universidade Federal do Paraná e Laboratório de Imunogenética e Histocompatibilidade do Departamento de Genética da UFPR. A detecção do HPV foi realizado pela metodologia da Captura Híbrida 2 (CH2®). O DNA foi extraído de amostras de sangue periférico e a genotipagem HLA-C foi feita pelo método PCR-SSOP...

Prevalence of HR-HPV and risk factors associated with the development of lesion cervical precousor of cervical cancer, a sample of the Curitiba metropolitan region, case-control study / Prevalência do HPV-AR e fatores de risco associados ao desenvolvimento de lesões cervicais precursoras do câncer cervical, numa amostra da região metropolitana de Curitiba, estudo caso-controle

Objective: Characterize the risk factors associated with the development of cervical intraepithelial neoplasia (CIN) and determine the prevalence of Highrisk Human Papillomavirus (HR-HPV) infection in women without cytological abnormalities and women diagnose with CIN 2 and 3, living in metropolitan region Curitiba, Parana state, Brazil. Material and Methods: Case-control study consisting of 382 women without cytological abnormalities, 233 women with high-grade intraepithelial lesions, which were subdivided in 131 women with CIN 2 and 102 women with CIN 3 diagnose by histological analysis of the cervical segment conical excision material. The age range of the women in the study was 15-45 years old, collected in the period of 2009 to 2012. The detection of HR-HPV DNA was performed by Hybrid Capture 2 test (CH2)...

Objetivo: Caracterizar os fatores de risco associados ao desenvolvimento de neoplasias intraepiteliais cervicais (NIC) e verificar a prevalência da infecçãopelo HPV de alto risco (HPV-AR) em mulheres sem anormalidade citológicas e com NIC 2 e 3, residentes na Região Metropolitana de Curitiba, Paraná,Brasil. Materiais e Métodos: Estudo caso-controle, composto por 382 mulheres sem anormalidades citológicas, 233 mulheres com lesão intraepitelial de alto grau, sendo estas subdivididas em 131 mulheres com NIC 2 e 102 com NIC 3 após a análise histológica do material da excisão cônica do segmento do colo do útero. A faixa etária das mulheres do estudo foi de 15 a 45 anos, coletadas no período de 2009 a 2012. A detecção do DNA do HPV-AR foi realizada pelo teste Captura Híbrida (CH2)...

The genetic study on a rare inter-allelic recombination of HLA-C locus in a Chinese Han family / 中华检验医学杂志

Objective To study the inter-allelic recombination event occurring in the HLA-C locus in a family of Chinese Han nationality, and to evaluate the molecular genetic background of the new HLA allele.Methods Peripheral blood samples were collected from a Chinese leukemia woman patient, as well as her healthy parents and two brothers.HLA-A, C, B, DRB1 and DQB1 alleles were typed by high-resolution PCR-sequence-based typing (SBT) method using Atria Genetic AlleleSEQR HLA SBT kits.The Protrans S4 HLA-C single allele-specific sequencing strategy was used to separate the two HLA-C alleles and to determine novelty of the allele.The full length sequences of HLA-C alleles of the patient and her parents were further analyzed using cloning and haplotype sequencing method. The HLA five loci linked haplotypes and the recombination site were analyzed by family study, meanwhile the full length sequences of the five HLA-C alleles were compared with the IMGT/HLA database by the program BLAST.Results The two haplotypes of the father and mother were a:A*0207-C*010201-B*550201-DRB1*090102-DRQ1*030302 and b:A*240201-C*120202-B*5201-DRB1*1502-DRQ1*0601, c:A*300101-C*060201-B*130201-DRB1*0405-DRQ1*0401 and d:A*110101-C*070201-B*4001-DRB1*080302-DRQ1*0601,respectively.The two brothers inherited their parent′s haplotypes a, d and b, c respectively.The two haplotypes of the patient were the maternal c and paternal recombinant a/b haplotype.The recombinant a/b haplotype A*240201-C*new-B*550201-DRB1*090102-DRQ1*030302, A*240201 came from the paternal haplotype b,while B*550201-DRB1*090102-DRQ1*030302 came from the other paternal haplotype a.When comparing the full length sequences of the HLA-C new allele with the father′s allele C*010201 and C*120202, it could deduce that the recombinant a/b haplotype derived from a recombination event occurring between the paternal chromosome 6 during meiosis.The crossover site was between genomic nt273 and nt330 of HLA-C alleles, which created a HLA-C new allele and the fifth haplotype of the family, and inherited it to the patient.The full length sequences of the new allele had been submitted to Genbank, and officially named C*0121 by WHO nomenclature committee.Conclusion This study demonstrates a rare inter-allelic recombination event occurring in the HLA-C locus within a Chinese Han family and illustrates the process of novel allele and haplotype, and provides direct theory for further studying the mechanisms of gene recombination and HLA polymorphism.

Molecular analysis of human leukocyte antigen class I and class II allele frequencies and haplotype distribution in Pakistani population.

AIM: Distribution of HLA class I and II alleles and haplotype was studied in Pakistani population and compared with the data reported for Caucasoid, Africans, Orientals and Arab populations. MATERIALS AND METHODS: HLA class I and II polymorphisms in 1000 unrelated Pakistani individuals was studied using sequence-specific primers and polymerase chain reaction and assay. RESULTS: The most frequent class I alleles observed were A*02, B*35 and CW*07, with frequencies of 19.2, 13.7 and 20%, respectively. Fifteen distinct HLA-DRB1 alleles and eight HLA-DQB1 alleles were recognized. The most frequently observed DRB1 alleles which represented more than 60% of the subjects were DRB1 *03, *07, *11 and *15. The rare DRB1 alleles detected in this study were HLADRB1 *08 and *09, having frequencies of 0.9 and 1.7%, respectively. In addition, at DRB1-DQB1 loci there were 179 different haplotypes and 285 unique genotypes and the most common haplotype was DRB1*15-DQB1*06 which represented 17% of the total DRB1-DQB1 haplotypes. In our population, haplotype A*33-B*58-Cw*03 comprised 2.8% of the total class I haplotypes observed. This haplotype was seen only in the oriental populations and has not been reported in the African or European Caucasoid. CONCLUSION: Our study showed a close similarity of HLA class I and II alleles with that of European Caucasoid and Orientals. In Pakistani population, two rare loci and three haplotypes were identified, whereas haplotypes characteristic of Caucasians, Africans and Orientals were also found, suggesting an admixture of different races due to migration to and from this region.

Sequencing analysis of exons 1,5, 6, 7 of HLA-Cw gene located outside of the routine testing region and its application in clinical matching / 中华检验医学杂志

Objective To study the molecular genetic polymorphism of exons 1,5, 6, 7 of HLA-C gene in Chinese population and evaluate the significance of additional sequencing based typing at exons 1,5, 6, 7 of HLA-Cw gene in clinical HLA matching, Methods A total of 324 individuals were typed at exons 2,3, 4 of HLA-C gene by sequence-based typing. If ambiguities appeared outside of exons 2 -4, we designed a total of 5 in-house sequencing primers and optimized the sequencing reaction, additional sequencing based typing at exons 1,5, 6, 7 was performed to solove the emerging ambiguities. Results In the three hundred and twenty-four samples typed by PCR-SBT at exons 2, 3 and 4 of HLA-Cw gene, 23.8 % (77/324) of the typed samples were assigned the conclusive genotype in four digital level 76. 2% (247/324) of the typed samples were given with the ambiguous allele combination results, in which 73 kinds of ambiguous allele combinations were detected. Increasing the additional sequencing analysis at exons 1, 5, 6, 7 of HIA-C gene, ten frequent ambiguities including Cw* 030201/030202, Cw* 070201/0750, Cw* 040101/0409N/0430, Cw* 0403/0409N/0430, Cw* 080101/0822 could be distinguished. ConclusionsIncreasing the sequencing anlysis at exons 1, 5, 6 and 7 of HLA-Cw gene will help to make clear the ambiguous SBT results and also improve the accuracy of HLA-Cw typing. It shows important significance in clinical histoeompatibility matching.

Analysis of hlgh-resolution human leukocyte antigen matching status for the unrelated donor-receipt pairs matched at low-resolution for HLA-A, HLA-B and HLA-DRB1 / 中华检验医学杂志

le full matching for unrelated donor-receipt pairs. However, HLA-Cw mismatching at antigen level could no longer be ignored. The results indieated that HLA-Cw genotyping should be incorporated into future CMDP unrelated marrow donor routine HLA test.

Distribution of HLA-A, B, C Allele and Haplotype Frequencies in Koreans / 대한진단검사의학회지

BACKGROUND: The HLA system is known to be the most polymorphic gene cluster in the human genome. HLA allele and haplotype distribution varies widely among different ethnic groups. In this study, we examined the frequency of HLA class I alleles and haplotypes in 309 healthy Koreans. METHODS: We typed HLA-A, -B, and -C genes at the allelic level in 109 unrelated Korean individuals using a sequence-based typing. With the additional data of 200 healthy Koreans from dbMHC (http: //www.ncbi.nlm.nih.gov/mhc/), allele and haplotype frequencies were estimated by the maximum likelihood method. Serological typing results of 49 individuals were compared with the results highly resolved. RESULTS: A total of 22 HLA-A, 41 HLA-B, and 21 HLA-C alleles were found in this study. Alleles showing frequencies of more than 10% in each HLA locus were A*2402 (22.5%), A*0201 (15.7%), A*3303 (14.4%), A*1101 (11.0%), B*5101 (12.1%), Cw*0102 (18.8%), and Cw*1402 (10.2%). The most common A-B-C haplotypes at a frequency of more than 3% were A*3303-B*5801-Cw*0302 (5.2%), A*2402-B*5101-Cw*1402 (4.5%), A*1101-B*1501-Cw*0401 (4.3%), A*3303-B*4403-Cw*1403 (4.0%), A*3001-B*1302-Cw*0602 (3.7%), and A*0207-B*4601-Cw*0102 (3.2%). Misassignment of HLA-C antigen by serotyping was detected in 11 (22.4%) of 49 individuals. CONCLUSIONS: Our results will be useful as a basic data for studies on anthropology, disease association, and bone marrow transplantation. Misidentification of HLA-C by serotyping is so high that it would be desirable to perform a DNA typing especially in unrelated bone marrow transplantation.

Molecular genetics (HLA) of Behcet's disease

Behcet's disease (BD) has been known to be strongly associated with the human leukocyte antigen (HLA) B51. This B51 association has been confirmed in many different ethnic groups between the Middle East and Japan, and it has been proposed that BD is prevalent in those ethnic groups along the old Silk Route. The hypothesis could be made that B51 molecules are primarily involved in BD development through specific antigen presentation. However, polymorphic analyses of the TNFB gene and Tau-a microsatellite between the HLA-B and TNF genes indicate that the pathogenic gene of BD is not the HLA-B51 gene itself but another gene located around the HLA-B gene. HLA-C genotyping by the PCR-SSP method also suggests that the BD pathogenic gene is not the HLA-C gene itself but other gene located near the HLA-B gene. Recently we sequenced a single contig of 236,822 bp from the MICA gene (58.2 kb centromeric of HLA-B) to 90.8 kb telomeric of HLA-C and identified 8 novel genes designated NOB1-8 (NOB: new organization associated with HLA-B). During the course of the genomic sequence analysis we clarified the genetic structure of the MICA (MHC class I chain-related gene A) gene and found a triplet repeat microsatellite polymorphism of (GCT/AGC)n in the transmebrane (TM) region. Furthermore, the microsatellite allele consisting of 6 repetitions of GCT/AGC (MICA A6 allele) was present at a significantly higher frequency in the BD patient group than in the control group and a significant fraction of B51-negative patients were positive for this MICA A6 allele. These results suggest the possibility of a primary association of BD with MICA rather than HLA-B.