The study of the HLA system was primarily initiated to understand the basis for the histocompatibility between recipients and tissue donors. HLA typing methods are being continuously improved and biochemical and molecular typing, in particular, are expected to provide precise typing of the HLA system. Conventional HLA typing methods can define antigen specificities, while biochemical and molecular methods will provide direct allele typing that is based on the actual sequence polymorphism. The precise tissue typing will definitely improve the outcome of transplantation. Structural studies have revealed the highly polymorphic nature of the HLA system and given insight to understanding the molecular basis of the HLA polymorphism. One big immunological puzzle remaining to be answered is how T-cell receptor molecules recognize peptide antigen in conjunction with the HLA molecule. The crystallization of the T-cell receptor molecule, an experiment currently underway, will eventually reveal the structural basis of the trimolecular interaction.
Animals ; Genes, MHC Class I ; Genes, MHC Class II ; Histocompatibility Antigens Class I/analysis/chemistry/*physiology ; Histocompatibility Antigens Class II/analysis/chemistry/*physiology ; Human ; Polymorphism (Genetics) ; Protein Conformation

OBJECTIVE: To detect loss of heterozygosity (LOH) at human leukocyte antigen (HLA) loci in a Chinese patient with leukemia after haploidentical hematopoietic stem cell transplantation. METHODS: HLA genotyping was carried out on peripheral blood, hair follicle and buccal swab samples derived from the patient after the transplantation as well as peripheral blood samples from his parents by using PCR-sequence specific oligonucleotide probe method and PCR-sequence based typing method. Short tandem repeat (STR) loci were detected by using a 23 site STR assay kit and a self-developed 6 STR loci assay for the HLA regions. RESULTS: After the transplantation, the HLA genotype of the peripheral blood sample of the patient was identical to his father. The patient was HLA-A*02:01,24:02, C*03:03,03:04, B*13:01,15:01, DRB1*08:03,12:02, DQB1*03:01,06:01 for his hair follicle specimen. However, homozygosity of the HLA loci was found in his buccal swab sample. Only the HLA-A*24:02-C*03:03-B*15:01-DRB1*08:03-DQB1*06:01 haplotype from his father's was present, while the HLA-A*02:01-C*03:04-B*13:01-DRB1*12:02-DQB1*03:01 haplotype from his mother was lost. After the transplantation, the alleles of the 23 STR sites in the patient's peripheral blood sample were consistent to his father, with no allelic loss detected in his buccal swab sample. However, at least 4 STR loci in the HLA region were lost in his buccal swab sample. CONCLUSION LOH at the HLA loci has been detected in the buccal swab sample of a patient with leukemia who received haploidentical hematopoietic stem cell transplantation.
HLA Antigens/genetics* ; HLA-A Antigens/genetics* ; Histocompatibility Antigens Class I/genetics* ; Humans ; Leukemia/genetics* ; Loss of Heterozygosity

Fresh rat corneas as well as corneas preserved in several different corneal preservation media were stained with Avidin-Biotin-peroxidase Complex method in order to evaluate major histocompatibility complex (MHC) antigen expression. In fresh corneas, class I antigen was identified in corneal epithelium, stroma and endothelium. Class II antigen was identified only in stroma. In corneas preserved in the media which contained chondroitin and dextran for 7 days, class I antigen was somewhat decreased but class II antigen was increased. In corneas preserved in the medium which contained insulin or epidermal growth factor for 7 days, class II antigens seemed to be increased compaired to the fresh cornea. Expression of MHC antigens of corneas in the medium with fetal bovine serum were similar to those of fresh corneas.
Animals ; Cornea/*metabolism ; Culture Media ; Histocompatibility Antigens Class I/*biosynthesis ; Histocompatibility Antigens Class II/*biosynthesis ; Immunoenzyme Techniques ; Major Histocompatibility Complex ; Organ Preservation/methods ; Rats ; Rats, Inbred Lew

PURPOSE: Stem cells are considered promising candidates for cell replacement therapy in many devastating diseases. However, it is assumed that stem cells may be rejected on transplantation. Therefore, we introduced human cytomegalovirus (hCMV) US genes, which are known to be able to reduce MHC class I expression on the cell surface after infection, into two known stem cell lines in order to test the feasibility of modifying these cells to reduced MHC class I antigens by the introduction of hCMV US genes. METHODS: The MHC class I expressions of mock-transfected or hCMV US gene-transfected human embryonic neural stem cell line (HB1.F3) and human breast epithelial stem cell line (M13SV1) were examined by FACS. RESULTS: MHC class I expressions in HB1.F3 and M13SV1 cells were dramatically induced by IFN-gamma treatment. In FACS analysis, cells transfected with the hCMV US2, 3, 6 or 11 genes exhibited a dramatic reduction (40~60%) of MHC class I expression compared with mock-transfected cells. CONCLUSION: Our results suggest that human stem cells express high levels of MHC class I antigens, and thus may be rejected on transplantation unless they are odified. In addition introduction of hCMV US genes can be exploited for stemcell transplantation.
Breast ; Cytomegalovirus ; Down-Regulation* ; Histocompatibility Antigens Class I ; Humans* ; Neural Stem Cells ; Stem Cells*

PURPOSE: Due to their unique capacity to self-renew and for multiple differentiation, stem cells are considered potent candidates for cell replacement therapy in many devastating diseases. However, studies on immune rejection, which is a major problem facing successful stem cell therapy, are rare. Thus, we examined MHC expression of human stem cells and effects of IFN-gamma on the MHC class I expression of the cells in order to determine whether human stem cells might be rejected after transplantation. METHODS: The MHC antigen expressions of human embryonic neural stem cell line (HB1.F3) and human breast epithelial stem cell line (M13SV1) were examined by RT-PCR and FACS. The effects of varying concentrations of IFN-gamma and of varying incubation times with IFN-gamma on the expression of MHC class I antigens in these stem cell lines were also examined by FACS. RESULTS: The results show low expression levels of MHC class I antigens on surfaces of these cells. A dramatic induction of MHC class I expression was observed when the cells were treated with IFN-gamma. Maximal induction of MHC class I antigen expression in HB1.F3 and M13SV1 cells was observed at above the concentrations of 20 ng/mL and 5 ng/mL of IFN-gamma 48 h after treatment, respectively. Elevated MHC class I levels in HB1.F3 and M13SV1 cells were sustained for 48 h and 72 h after withdrawing IFN-gamma, respectively. CONCLUSION: These results suggest that human stem cells express high levels of MHC class I antigens, and thus may be rejected on transplantation unless they are modified. Therefore, in addition to studies on stem cell differentiation, studies on overcoming the immunological barriers to stem cell transplantation are prerequisite for successful clinical application of stem cell therapy.
Adult* ; Breast* ; Histocompatibility Antigens Class I ; Humans* ; Neural Stem Cells* ; Stem Cell Transplantation ; Stem Cells*

OBJECTIVETo review the role of polymorphism of major histocompatibility complex class I-related chain A (MICA) gene and antibodies against MICA antigens in transplant immunology.

DATA SOURCESThe data used in this review were mainly from our own results and from the relevant English language literatures published from 1999 to 2010. Some data presented in this review are in press.

STUDY SELECTIONArticles regarding MICA gene discovery and pioneering finding of antibodies against MICA antigen and allograft rejection were selected. This review chronicles the development of our understanding of the role that MICA antigens and antibodies may play in organ transplantation.

RESULTSPolymorphic glycoprotein MICA antigens were detected on freshly isolated human umbilical cord endothelial cells, but not on peripheral lymphocytes. Antibodies were found and typing of recipients and donors by sequencing the MICA alleles has established that de novo antibodies produced in kidney transplant recipients are directed at mismatched MICA epitopes and are associated with acute rejection and chronic transplant failure. The specificity of antibodies against the epitopes of MICA antigens were well characterized by donor MICA typing, single antigen array testing with antibody absorption and elution. Acute graft-versus-host disease was observed in stem-cell recipients who were mismatched for MICA.

CONCLUSIONSImmunization against mismatched MICA epitopes encountered in donor organs after transplantation may result in antibodies against MICA alleles. Testing for MICA donor-specific antibodies (DSA) which are associated with early failure of kidney transplants may be helpful for identifying some of the targets of antibodies against antigens other than the human leukocyte antigen (HLA) and for improving transplantation outcome.

Hematopoietic Stem Cell Transplantation ; Histocompatibility Antigens Class I ; genetics ; Humans ; Polymorphism, Genetic ; Transplantation, Homologous

OBJECTIVETo investigate the genetic polymorphism of microsatellite in the exon 5 of MICA gene and the intron 1 of MICB gene in Guangdong Han population.

METHODSOne hundred and six samples of Guangdong Han population were genotyped by polymerase chain reaction and fluorescent technique (6-FAM). Gene frequency, power of discrimination, expected heterozygosity, polymorphism information content and probability of paternity exclusion were calculated.

RESULTSThe genotype distributions of MICA and MICB microsatellite met Hardy-Weinberg equilibrium. MICA A5 was the most common allele (0.2877), whereas A4 was the least popular one (0.1321). The genotype distribution frequencies of A5-5.1 (14.15%) and A5-5 (10.38%) are high. MICB CA14 was the most common allele (0.3255), and CA19,28 was the least popular one (0.0047). CA27 was not observed. The genotype distribution frequency of CA14-CA14(14.15%) is high.

CONCLUSIONThe microsatellite of the exon 5 of MICA gene and the intron 1 of MICB gene could be used as the genetic markers of Chinese population in the studies of anthropology, linkage analysis of genetic disease genes, individual identification and paternity test in forensic medicine.

OBJECTIVE: The explore the molecular basis of iron-overload in Tibet nationality population of Tibet. METHODS: The inpatients with iron-overload in our department from Dec. 1st 2014 to Jul.31st 2016 were enrolled in this study. Abdominal MRI and the mutation sites C282Y and H63D in HFE exon were examined. For HFE mutation-negative patients, the non-HFE mutation was detected, including 5 HJV mutations of G320V, p.Q312X, p.D249H, p.I281T, p.C321X and 2 TFR2 mutations: (Y250X, I238M), and 2 SLC40A1 mutations: (V162del, N144H). RESULTS: Among 113 iron overload patients, only one showed homozygous p.H63D mutation, and one showed heterozygosis p.H63D mutation. In 73 patients accepted non-HFE gene detection, only one was heterozygosis p.D249N mutation in HJV, and one was heterozygosis p.I238M mutation in TFR2. CONCLUSION Currently, the pathogenic gene for Tibetan iron-overload has not yet been found.
Genotype ; Hemochromatosis Protein ; Histocompatibility Antigens Class I ; Humans ; Iron Overload ; Mutation ; Tibet

OBJECTIVE: To investigate the influence of oridonin on the killing activity of NK-92 MI cells targeting THP1 and the related mechanism. METHODS: The killing activity of NK-92 MI to THP1 before and after oridonin treatment was detected by LDH release assay; the expression of natural killer cell ligands activating receptor D (NKG2D, including MICA, MICB, ULBP1, ULBP2 and ULBP3) was detected by real-time quantitative polymerase chain reaction (qRT-PCR) and Western blot respectively; the expression of cytokine TNF-α, TNF-β and IFN-γ in the co-culture supernatant of NK-92 MI cells and THP1 cells were measured by ELISA. RESULTS: The killing efficiency after oridonin treatment at different effector-target ratio (1:1, 5:1, 10:1) was all significantly up-regulated in comparison with that before oridonin treatment (P＜0.05). QRT-PCR and Western blot showed that the expressions of mRNA and protein levels of MICB, ULBP1, ULBP2 increased to varying degree (P＜0.05), but the expression levels of MICA and ULBP3 were not statistically significant between experimental group and control group (P＞0.05). ELISA results indicated that IFN-γ and TNF-β release were significantly increased after oridonin treatment (P＜0.05), however, the TNF-α release was not statistically different in comparison with control group (P＞0.05). CONCLUSION Oridonin can significantly improve killing efficiency of NK-92 MI on THP1, that might be related with up-regulation of MICB, ULBP1 and ULBP2 expression and promotion of IFN-γ and TNF-β release.
Cell Line, Tumor ; Diterpenes, Kaurane ; pharmacology ; GPI-Linked Proteins ; Histocompatibility Antigens Class I ; Humans