No abstract available.
Histocompatibility Testing* ; Spondylitis, Ankylosing*

BACKGROUND: The flowcytometric analysis of HLA-B27 gives more objective results and is performed more rapidly than traditional serologic methods. We have used a flowcytometric method using only anti-HLA-B27 monoclonal antibody, but it gave frequently borderline mean fluorescence intensity (MFI) results. The authors compared the method using anti-HLA-B27 antibody (HLA- ABC-m3, Serotec) with the Becton Dickinson (BD) method which uses different HLA-B27 antibody (GS145.2) with CD3 antibody. METHODS: The 59 patients that requested HLA-B27 testing were measured by two methods. In the former method, the mononuclear cells were stained with HLA-B27-FITC and the MFIs were determined in lymphocytes. In the BD method, the whole blood was directly stained with CD3-PE and HLA-B27-FITC. The MFIs were determined in the CD3+ cells, and compared with the MFI of the standard. For the cases showing discrepancy in the two methods or borderline values, the HLA-ABC typing was done. RESULTS: Of 21 showing discrepancy, 10 samples had undergone HLA typing. Among nine samples that were positive by the Serotec method but negative by the BD method, four samples were B7, one B40, one B54 and three B7 CREG negative. One that was negative by the Serotec method but positive by the BD method was confirmed as HLA-B27. CONCLUSIONS: The Serotec method showed significant overlap between the MFIs of HLA-B27 and non B27 samples that resulted in a relatively low efficiency compared with the BD method. The discrepant results of the two methods seem to be due to maily the specificity of the antibody used.
Fluorescence ; Histocompatibility Testing ; HLA-B27 Antigen* ; Humans ; Lymphocytes ; Sensitivity and Specificity

BACKGROUND: The most common problem in HLA typing is unsatisfactory quality of the antisera, or a lack of understanding of their reactivities. Therefore, commercial antisera must be verified under the conditions applied in a particular tissue typing laboratory. METHODS: We evaluated the antisera reactivities of a commercial HLA-yping tray, Lymphotype HLA-BC 72 oriental, the lot 7220999, 7230100 (Biotest, Germany), in about 300 samples from organ transplant recipients and healthy potential donors. RESULTS: The relatively weak antisera were those that defined A26, A33, Cw5, Cw14, B46, B58, B64 and B71 etc. Some of these antisera were not indicated as 'weak reaction' in the test result catalogue. The reactivities of each antisera indicated as 'extra reaction' or 'sometimes missing' were various. CONCLUSIONS: As for antisera reactivities, the data obtained by a laboratory itself are necessary in addition to those in the test result catalogue. These data will be helpful for the correct interpretation for laboratories using same commercial kits.
Histocompatibility Testing* ; Humans ; Immune Sera* ; Tissue Donors ; Transplants

BACKGROUND: The most common problem in HLA typing is unsatisfactory quality of the antisera, or a lack of understanding of their reactivities. Therefore, commercial antisera must be verified under the conditions applied in a particular tissue typing laboratory. METHODS: We evaluated the antisera reactivities of a commercial HLA-yping tray, Lymphotype HLA-BC 72 oriental, the lot 7220999, 7230100 (Biotest, Germany), in about 300 samples from organ transplant recipients and healthy potential donors. RESULTS: The relatively weak antisera were those that defined A26, A33, Cw5, Cw14, B46, B58, B64 and B71 etc. Some of these antisera were not indicated as 'weak reaction' in the test result catalogue. The reactivities of each antisera indicated as 'extra reaction' or 'sometimes missing' were various. CONCLUSIONS: As for antisera reactivities, the data obtained by a laboratory itself are necessary in addition to those in the test result catalogue. These data will be helpful for the correct interpretation for laboratories using same commercial kits.
Histocompatibility Testing* ; Humans ; Immune Sera* ; Tissue Donors ; Transplants

Human lymphocyte antigen (HLA) is the most complicated human dominant polymorphic genetic system. Accurate HLA genotyping is clinically important for hematopoietic stem cell (HSC) transplantation, also important for research on many human diseases. Polymerase chain reaction-sequence based typing (PCR-SBT) provides the highest resolution level and defines new alleles, so it is widely used for HLA typing. One great disadvantage of PCR-SBT method is the fact that it cannot resolve sequences of heterozygous samples in diploid genomes, leading to ambiguous typing results which make much trouble to the accurate definition of HLA genotype. This article reviewed the occurring reasons and solution method of ambiguous allele combinations in the HLA high resolution genotyping as well as the research prospect in this field.
Genotype ; HLA Antigens ; genetics ; immunology ; Histocompatibility Testing ; Humans

OBJECTIVETo apply the single cell nested multiplex polymerase chain reaction (PCR) to HLA typing, and analyze the influence factors on the amplification results.

METHODSSingle cell DNA templates were prepared with different methods. The exon 2, 3 and intron 2 of HLA-A, B, and exon 2 of DRBI were amplified using multiplex PCR. The second round of SSP-PCR HLA typing was carried out according to the large scale routine HLA typing results.

RESULTSEnzyme lysis method was the most efficient procedure for preparing the single cell DNA template, with a success rate (SR) of 93.3%, while the SRs of alkali lysis and freezing-thaw lysis methods were 83.3% and 73.3%, respectively. The second round amplification using enzyme lysis and SSP-PCR in 20 samples obtained a 95% success rate and a 15% allele drop out rate. The time for performing the whole procedure was less than 6 hours.

CONCLUSIONThe modified nested multiplex PCR technique is efficient for single cell HLA typing and might be applied to clinical preimplantation genetic diagnosis.

Histocompatibility Testing ; methods ; Humans ; Polymerase Chain Reaction ; methods

Graft Rejection ; HLA Antigens/analysis* ; Histocompatibility Testing ; Humans ; Quality Control

OBJECTIVETo study the histocompatibility of three bio-derived bones.

METHODSAfter treatment with different physical and chemical method, three bio-derived bones, the composite fully deproteinized bone (CFDB), partially deproteinized bone (PDPB) and partially decalcified bone (PDCB) were implanted into rabbits. The toxicity, immune response and subperiosteum osteogenesis of CFDB, PDPB and PDCB were studied through gross observation, serum antibody measurement, evaluation of local cellular immune response and HE staining.

RESULTSThe study showed that CFDB, PDPB and PDCB had no toxicity. They could conduct peripheral tissue to grow into them and had no harmful effect on subperiosteum osteogenesis. They could also promote cartilage and osteoid tissue derived from periosteum to calcify to new bone, and combine with the peripheral bone. The degree of immune response caused by them was in the sequence of PDCB > PDPB > CFDB.

CONCLUSIONSThe three bio-derived bones, CFDB, PDPB and PDCB have good histocompatibility.

Animals ; Antibodies ; blood ; Bone Transplantation ; Bone and Bones ; immunology ; Female ; Histocompatibility ; Histocompatibility Testing ; Male ; Rabbits ; Tissue Engineering

Detection of significant alloimmune response, which affects graft function and survival by effective immune monitoring, is critical for treatment decision making. However, there is no consensus regarding immune monitoring (IM) for kidney transplantation (flow KT) in Korea. The IM protocol may be affected by the level of immunological risk, the methods of desensitization and the availabilities of resources such as laboratory support and cost of tests. Questionnaire surveys designed to identify the current practices regarding immune monitoring of KT among transplant clinicians and clinical pathologists in Korea and eventually provide a basis for the establishment of harmonized immune monitoring guidelines in KT were administered as part of a Korean Society for Transplantation Sponsored Research Project. The survey results revealed significant variations in IM protocols and interpretation of tests affecting treatment decisions between institutes. Moreover, the results revealed a need to expand the histocompatibility tests into high resolution HLA typing in multiple loci and non-HLA antibody tests that facilitate the epitope analysis and eventually virtual crossmatching. The results of the questionnaire survey from clinical pathologists are addressing the urgent need for the standardization of interpretation and harmonization of results reporting in single antigen bead based HLA antibody identification. Finally, communication between clinicians and clinical pathologists to meet the clinical expectations regarding various immune monitoring tests is needed.
Academies and Institutes ; Consensus ; Decision Making ; Histocompatibility ; Histocompatibility Testing ; Kidney Transplantation ; Korea* ; Monitoring, Immunologic* ; Transplants

Here, we report the results of the first histocompatibility proficiency testing (PT) performed by the Korean Association of External Quality Assessment Service in 2018. The directly prepared PT specimens of whole blood, sera, and mononuclear cell suspensions were distributed to participants biannually. The number of participants was comparable to that in the previous external PT program, and the response rate was 88%–100%. The accuracy rates for human leukocyte antigen (HLA) A, B, C, DR, and DQ low and high resolution typing were 100%/100%, 100%/98%, 100%/99%, and 99%/98%, respectively; HLA-B27 typing, 99.1%; T cell and B cell crossmatching, 3.1% and 6.0%, respectively; and HLA antibody screening and identification, 100% and 100%, respectively. The results of HLA crossmatching were not reported from four participants due to poor cell viability. Further improvements of the specimen delivery process, grading criteria for crossmatching, and format of participant summary are warranted.
Cell Survival ; Histocompatibility Testing ; Histocompatibility ; HLA-B27 Antigen ; Humans ; Leukocytes ; Mass Screening ; Suspensions