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PubMed1996

Analysis of HLA-DRB1 alleles using PCR-RFLP and PCR-MPH.

In this study we compare the results of HLA-DRB1 genotyping by PCR-RFLP and PCR-MPH. HLA-DR specificities were also performed by LCT. Samples were obtained from 20 Thai patients who were on the waiting list for kidney transplant. DNA was extracted by phenol-chloroform extraction. It was found that the results gave complete agreement with two methods of DNA typing, however, there were 3 discrepancies in assigning serologic DR specificities and DNA subtypes (p = 0.0001) which were due to the cross reactive antibodies and the lack of potent antisera to define proper HLA-DR subtypes by LCT. These PCR techniques can be applied to identify other alleles such as HLA-DPB1 and HLA-DQB1 which will improve the standard histocompatibility testing in the future.

Alleles↗
PubMed1995

[Allogeneic bone marrow transplantation in malignant blood diseases].

Allogeneic bone marrow transplantation has an established role in the treatment of malignant blood diseases. For some disorders it is at the moment the only curative treatment. As the complication risks of this treatment increase with age, the upper age limit for allogeneic transplantation is usually 50 to 60 years. The main indications are acute leukaemias, chronic myeloid leukaemia and myelodysplastic syndromes. Selected patients with chronic lymphatic leukaemia, multiple myeloma and lymphoma are also treated with allogeneic transplantation. The most common intensive conditioning regimen preceding the transplantation consists of total body irradiation and cyclophosphamide. The source of haematopoietic stem cells has routinely been bone marrow, but the number of transplantations with stem cells harvested from blood is rapidly increasing. The proportion of transplantations from unrelated donors is growing. Histocompatibility testing is becoming more precise, which is likely to improve the results of unrelated donor transplantations to the level achieved with sibling donor transplantations.

Bone Marrow Transplantation↗
PubMed1999

New high resolution typing strategy for HLA-A locus alleles based on dye terminator sequencing of haplotypic group-specific PCR-amplicons of exon 2 and exon 3.

In this study, a new sequencing-based typing strategy for the HLA-A locus is presented which involves group-specific separate amplification of exon 2 and 3 of HLA-A alleles in a first step. Conserved HLA-A locus-specific primers of intron 1 or 3 were combined in 10 primer-mixes with group-specific primers hybridizing to the 5'- or 3'-end of exon 3 or 2 for pre-typing of the HLA-A alleles in 14 allelic groups. Maximally four overlapping short amplicons are produced under identical polymerase chain reaction (PCR) conditions with individual separate amplification of exon 2 and exon 3 of the haplotypic alleles in most heterozygous combinations. Time- and money-saving one-directional Big Dye Terminator cycle sequencing is shown to provide reliable high resolution typing of the HLA-A alleles, even in a few cases of two amplicons in one primer reaction mixture. In comparison, to other sequencing-based typing (SBT) techniques the applied typing strategy minimizes the risk of unequal amplification or of drop-outs of one of the haplotypic alleles and allows unequivocal definition of the cis/ trans linkage of polymorphic positions of the complete exon 2 and exon 3 in most heterozygous cells. This also includes detection of new alleles differing in the polymorphic template generating primer annealing sites as well as in unusual combinations of known exon 2 and 3 sequences. With 10 primer sets working under identical conditions for pre-grouping and separate amplification of the haplotypic alleles our SBT procedure also could be implemented in clinical settings of large-scale stem cell donor histocompatibility testing for fast molecular HLA-A matching.

Alleles↗
PubMed1994

[Bone marrow transplantation from donors other than HLA matched siblings for hematological malignancies. Nagoya Bone Marrow Transplantation Group and Tokai Marrow Donor Bank].

One hundred and fourteen patients with hematological malignancies received bone marrow transplantation from donors other than HLA-identical siblings. Sixty-three patients received transplantations from related donors; 20 were phenotypically identical for HLA-A, B, D/DR (RM0). 32 differed at one locus (RM1) and 11 differed at more than one loci (RM2). Fifty-one transplantations were from unrelated donors; 37 were phenotypically identical and mixed lymphocyte culture (MLC) negative (UR0) and 14 were MLC positive (UR1). One hundred and four patients had durable engraftment. Four (RM1(1), RM2(2), UR0(1)) failed to achieve engraftment. In terms of the probability of > or = Grade II acute graft-versus-host disease (GVHD), there was no significant difference among the groups according to HLA disparity (RM0:25%, UR0:33%, UR1:39%, RM1:47%, and RM2:50%). The probability of chronic GVHD was significantly higher in UR0 and UR1 than RM0 (71%, 75% vs 28%, p < 0.05). The disease-free survival at 3 years was 45% (RM0), 50% (RM1) and 42% (UR0). More than 50% of patients other than RM0 died of fatal complications including GVHD within 60 days after grafting. In conclusion, unrelated donor and related donor mismatched at one locus could be selected for marrow graft in the case of the absence of an HLA-matched related donor. However, more advances in post-transplant management and in histocompatibility testing should be required.

Adolescent↗
PubMed1995

Cadaver kidney transplantation in the north Italy transplant program in the nineties.

The most relevant changes which have taken place in the NITp in the nineties include the introduction of HLA-DRB1 matching, the extension of both recipient and donor selection criteria, and an increase in donor procurement and consequently, in transplantation activity. Some of the changes in policy resulted from extensive analysis of our previous experience. For the future years, the NITp has set the following priorities: Consolidate the increase of donor procurement activity registered in the past 15 months by educational campaigns for health workers and the public, and organizational measures aimed at strengthening ICUs, nominating transplant coordinators and introducing a system of reimbursement for organs procured. Improve the quality of results in terms of patient rehabilitation and cost-benefit through: continued evaluation of protocols for patient admission on the waiting lists and careful selection of donors; and prospective use of genomic HLA Class II matching and also consider genomic typing for HLA Class I which is almost a reality. Establish a single pool of patients on the waiting list evaluated according to common protocols with the possibility of performing the transplant in each of the authorized centers in turn, respecting the best HLA match and the local use of organs. Finally, standards for donor treatment, organ procurement, histocompatibility testing and transplantation must be established. For this purpose, accreditation programs which have begun to be applied in Europe seem to be the adequate tool.

Adolescent↗
PubMed1980

Nomenclature for factors of the HLA system, 1980.

This article outlines the decisions made by the WHO nomenclature committee on leukocyte antigens at a meeting held after the 8th International Workshop on Histocompatibility Testing. Particular attention is given to new designations for provisional HLA-B and HLA-DR specificities and the upgrading of certain HLA-D and HLA-DR specificities to full HLA status. The existence of supertypic cross-reacting specificities was confirmed and extended.

HLA Antigens↗
PubMed1984

The improving prognosis after kidney transplantation. New strategies to overcome immunologic rejection.

For the 70,000 patients with end-stage renal disease in the United States, renal transplantation offers the only hope of full recovery from chronic renal failure. However, transplantation has had only limited use, principally because of the risks of graft rejection and immunosuppression. The last ten years have witnessed striking improvements in the survival of patients and grafts resulting from advances in immunologic management, including restricted use of immunosuppression, better histocompatibility testing, HLA matching, blood transfusions, and new drugs for prevention and reversal of transplant rejection. Kidney transplantation now is safe and effective and should be considered for most young and middle-aged adults.

Adrenal Cortex Hormones↗
PubMed2000

Fourier-transform infrared spectroscopy as a tool for detecting early lymphocyte activation: a new approach to histocompatibility matching.

Fourier transform infrared (FT-IR) spectroscopy due to its speed and sensitivity is becoming an increasingly powerful tool in the study of cell composition. We outline the potential of FT-IR microspectroscopy in monitoring mitogenic and cell mediated lymphocyte activation. We demonstrate the potential of FT-IR spectroscopy in histocompatibility testing by showing that significant spectral differences (p < 0.001, for the mean integrated intensity of phosphodiester band located in the 1142-996 cm(-1) region) exist between lymphocyte cocultures from pairs of HLA matched and mismatched volunteers after only 90 min of incubation. The preliminary results indicate that early spectral changes measured are due to HLA differences between individuals, although the relative contribution of class I and class II differences has yet to be determined. FT-IR spectroscopy represents a novel approach to histocompatibility matching and the rapidity and sensitivity of the technique indicates a potential role in matching protocols for clinical use, particularly in allogeneic bone marrow transplantation.

Analysis of Variance↗
PubMed1977

HLA determination in renal transplantation with living donor.

During the period Feb. 1967--Aug. 1976 94 first renal transplantations were performed using living related donors and histocompatibility tests. Eight transplantations were performed on children under 15 years old and four on patients over 50 years old. The rest of the patients were between 15 and 50 years old. No exclusions were performed. 22 patients expired, seven of these with a well functioning graft. The patient survival (P.S.) was 90% at 1 year, 87% at 2 years and 77% at 5 years, the graft survival (G.S.) was 81% at 1 year, 80% at 2 years and 68% at 5 years. The clinical results showed a fairly good correlation with the histocompatibility degree. In the A-B-match groups the 1, 2 resp. 5 years P.S. was 95, 96 resp. 89% and G.S. 90, 90 resp. 89%. In 43% of the patients rejection of variable degree developed. The rejection led to graft loss in 12%.

Adolescent↗
PubMed1990

HLA histocompatibility affects cardiac transplant rejection and may provide one basis for organ allocation.

Prospective human lymphocyte antigen (HLA) typing is not performed for heart transplantation, and the relation between HLA matching and cardiac graft rejection is unclear. Recipient and donor HLA matching were analyzed retrospectively in 51 patients undergoing orthotopic cardiac transplantation. Immunosuppression was based on cyclosporine and prednisone. During the mean follow-up of 34 months (range, 16 to 63 months), the 46 operative survivors had an average of 3.95 rejection episodes (range, zero to 11 episodes). Twenty-one patients had steroid-resistant rejection requiring treatment with polyclonal or monoclonal antithymocyte globulin. Human lymphocyte antigen typing was available for 44 patients, and antigens were grouped in broad specificities. Patients with two or more HLA-A or HLA-B matches had a reduced number of rejection episodes (3/10 versus 19/34) and a lower incidence of steroid-resistant rejection (1/10 versus 18/34; p = 0.01). Inclusion of HLA-DR matches did not alter the findings. There was a strong correlation between the increased frequency of rejection and the incidence of steroid-resistant rejection (p less than 0.0001). Four of six late deaths occurred in patients with steroid-resistant rejection; four were due to acute rejection and two to graft atherosclerosis. Although not currently done, prospective HLA matching is feasible with present typing methods. Our results suggest a rationale for prospective histocompatibility testing in cardiac transplantation with allocation of donor hearts to patients with two or more HLA matches.

Adult↗
PubMed1992

[Indications, problems and future perspectives of bone marrow transplantation in pediatrics].

Bone marrow transplantation (BMT) is an established therapy in pediatric oncology and is increasingly used as curative approach in the treatment of congenital, non-oncologic diseases of the lymphohematopoietic system. There is increasing evidence, however, that BMT can be followed by severe long term effects including neuroendocrine, ophthalmologic, dental and central nervous system abnormalities, particularly in children. Therefore the indication to BMT depends on the results obtained by conventional therapy. Due to the high cure rates of leukemia with conventional therapy BMT is only warranted following relapse except for certain forms with poor prognosis factors. For patients with chronic myelogeneous leukemia, however, BMT is the only chance of cure. In solid tumors the role of BMT is more difficult, because there is no clear evidence that BMT is superior to conventional therapy with regard to long term survival. In severe aplastic anemia, however, the long term results of BMT are clearly better than those obtained by conventional therapy. Other undisputed indications for BMT are severe combined immuno deficiencies and other congenital diseases for which BMT is currently the only curative therapy. Progress with matched unrelated donor transplantations by better histocompatibility testing and more specific immunosuppressive therapy to reduce graft-versus-host disease, still a major problem of allogeneic BMT, as well as the perspectives of gen therapy in the future will offer a chance of cure to many patients without a matched sibling donor.

Anemia, Aplastic↗
PubMed1975

An analysis of the 4c complex of HL-A based on Indian populations.

Sera detecting part or all of the 4c group of HL-A antigenic specificities were compared in two American Indian populations, the Ixils and the Pimas, and in one Spanish-Indian hybrid population, the Mestizos of Peru. Several new variants appeared. These were not identical to HL-A5, W5 or W18. Two of the variants appeared to represent specificities Z57 and Z51 as described during the 1972 Histocompatibility Testing Workshop. In addition, three new specificities could be detected. These were Z42, Ao88 and Ao93. Z42, Ao88 appeared to belong to the 4c complex and Ao93, a new specificity of the second locus, may belong to this same cross-reactive group.

Antigen-Antibody Reactions↗
PubMed1978

Leukapheresis and granulocyte transfusion.

Granulocytes for transfusion can now be obtained from normal donors by one of four techniques that involve either centrifugation or reversible adhesion of granulocytes to nylon fibers. The leukapheresis process appears to be safe for donors and standards for the selection and care of donors are being formulated. It appears desirable to transfuse granulocytes that are compatible in a leukoagglutination crossmatching, however, better methods for histocompatibility testing must be developed. Granulocyte transfusions clearly are of benefit to patients with Gram-negative sepsis and granulocyte counts of less than 500/cu mm for at least ten days. They may be valuable for granulocytopenic patients with other severe infections; however, there is no indication that granulocyte transfusions are indicated prophylactically or for febrile granulocytopenic patients without evidence of infection.

Agranulocytosis↗
PubMed1985

Use of SEOPF regional crossmatch trays to share kidneys for sensitized patients. Local experience of three centers.

Regional organ procurement (ROP) crossmatch trays are used by members of the South-Eastern Organ Procurement Foundation (SEOPF) to facilitate sharing of cadaver kidneys for highly sensitized patients. ROP trays carry a single representative serum sample from over 900 patients with panel reactive antibody (PRA) levels of greater than or equal to 60%. Trays are centrally prepared periodically and distributed to member laboratories where they are used for preliminary crossmatching against locally obtained donors. Crossmatching results are used in conjunction with the SEOPF computer match program for sharing of kidneys. Proficiency testing studies by the 40-member laboratories show a greater than 90% concordance rate on results with these highly and broadly reactive sera. Data were available from 3 centers on 74 kidneys shared between SEOPF-member institutions on the basis of a remote, preliminary, negative ROP tray crossmatch. Of these, 44 (59%) crossmatched negative locally with the same and other sera, and were thus considered acceptable to be transplanted to the intended highly sensitized patients. Sixteen (22%) donors had a positive crossmatch locally, but with sera other than that present on the ROP tray used for screening. In 14 cases (19%) the same serum as on the ROP tray gave a positive crossmatch. The majority of ROP tray inconsistencies appeared to be due to use of more sensitive crossmatching techniques at the recipient center. Of the 27 patients transplanted at these 3 centers with kidneys received on the basis of ROP tray results, none experienced hyperacute or early irreversible rejection and actual graft survival at 6-48 months is 74%. These studies indicate that regional sharing of patient sera by the existing network of histocompatibility testing laboratories is an effective and reliable mechanism to identify crossmatch-negative donors for highly sensitized patients.

ABO Blood-Group System↗
PubMed2002

Immunogenetic selection of donors for haematopoietic stem cell transplantation: an approach.

Histocompatibility between the patient and his donor is a prerequisite for the success of transplantation of haematopoietic stem cells (HSCT). Histocompatibility testing is clinically performed by HLA typing. HLA typing can be highly informative if performed in families. In this case, comparatively little typing effort often allows to predict the identity of whole HLA haplotypes. HLA typing becomes more demanding, however, if unrelated individuals have to be considered as donors. Given the huge diversity of HLA molecules the answer to the question, whether patient and donor possess 'matched' HLA types requires considerable typing effort. Interaction between the HLA typing laboratory and the clinician is highly needed. It has to be agreed upon what loci are considered relevant and what degree of resolution is necessary. Furthermore, a strategy has to be developed, which ensures that a maximum of potential donors are tested in a reasonable time frame avoiding at the same time a work-overload of the laboratory and an excessive uneconomical typing effort. At present, experience with HSCT with unrelated donors is limited. So there is no consent on the 'correct' way of choosing unrelated donors. Each transplantation centre will have its own characteristic needs, resources, manpower and skills; therefore, the ways vary between centres. I want to present the approach in Vienna.

Alleles↗
PubMed1985

Allogeneic bone marrow transplantation with related donors other than HLA MLC-matched siblings, and the use of antithymocyte globulin, prednisone, and methotrexate for prophylaxis of graft-versus-host disease.

Fifteen patients ranging in age from 1-29 years (median age 9 years) had bone marrow transplantations (BMT) from related donors other than HLA mixed lymphocyte culture (MLC) identical siblings. Donors were selected on the basis of HLA similarity and low reactivity in the MLC. Posttransplant immunosuppression consisting of methotrexate (MTX), antithymocyte globulin (ATG), and prednisone was used in an effort to decrease graft-versus-host disease (GVHD). Seven children were treated for aplastic anemia, 7 for hematologic malignancy, and 1 for osteopetrosis. Primary engraftment failure contributed to death in 3 patients, all of whom had aplastic anemia. Nine of 12 engrafted patients developed moderate-to-severe acute graft-versus-host disease. Of the 15 patients, 7 developed interstitial pneumonitis. Three patients demonstrated mixed chimerism (at or beyond 3 months posttransplant). Two of the seven patients treated for aplastic anemia are currently alive six months and more than five years posttransplant; the latter patient has chronic GVHD. Four of the seven patients treated for hematologic malignancy are currently alive more than 500 days posttransplantation. Three have chronic GVHD. Analysis of patient outcome according to the degree of similarity in histocompatibility testing revealed that patients with low reactivity in the MLC (less than 5% relative response in both directions) had a better prognosis (5/6, 83% long-term survival) than patients with maximum donor vs. recipient mixed lymphocyte culture relative response greater than 5% (1/9, 12% long-term survival), P = .016.

Adolescent↗
PubMed1987

Progress report on the ASHI/CAP Histocompatibility Survey Program, 1981-1986.

Two histocompatibility testing surveys were conducted over a five-year period by the American Society for Histocompatibility and Immunogenetics and the College of American Pathologists. More than 200 laboratories participated in the HS survey, which consisted of four shipments of cells and serum samples to be tested for ABO type, HLA-A and -B types, antibody identification, and lymphocytotoxicity crossmatching. About 100 laboratories participated in the DR survey, which consisted of four annual shipments to be tested for HLA-DR and -DQ types, antibody identification, and crossmatching. The results of the analysis of the HLA-typing results showed high consistency (generally greater than 90%) among laboratories in the definition of most recognized HLA antigens. In contrast, the HLA antibody screening was generally less consistent between laboratories. On the basis of 90% or greater consensus among participants, it was possible to develop a performance grading system.

ABO Blood-Group System↗
PubMed1992

Diagnostic use of molecular probes before and after marrow transplantation.

The application of molecular techniques to disease diagnosis and histocompatibility testing before marrow transplantation and to the evaluation of engraftment and immune reconstitution, and to the identification of infectious and neoplastic complications following transplantation has revolutionized the approach to the care of marrow transplant patients. The techniques of restriction fragment length polymorphism analysis using informative restriction enzymes, the Southern blot technique, and of polymerase chain reaction-amplified DNA and oligonucleotide probe hybridization methods now enable the study of donor and host cells at the nucleotide level.

Amino Acid Sequence↗