bluuebunny/crossref_metadata_2025 huggingface dataset:
A monoclonal antibody exhibiting reactivity with both X‐hapten‐ and lactose‐bearing glycolipids
Authors: Hogg Russell J. , Williams Keryn A. , Zola Heddy
Published: October 2008

Abstract: The tissue reactivity of a monoclonal antibody raised against human granulocytes, FMC 34, was shown to overlap with antibodies including FMC 10, which is directed against the X‐hapten, and with T5A7, which is directed against lactose‐bearing antigens. Reactivity of FMC 34 on sections of human cornea was more widespread than that of either FMC 10 or T5A7. High sensitivity fluorescence testing revealed that both FMC 34 and T5A7 reacted weakly with the T‐lymphocyte cell line JM, unlike the CD 15 antibody FMC 10. Competitive binding experiments showed that binding of FMC 34 to granulocytes was inhibited by preincubation with T5A7, while FMC 34 itself was able to inhibit the binding of both FMC 10 and T5A7. The binding of FMC 10 was not inhibited by preincubation of cells with T5A7. Saturation‐binding analyses revealed that each human granulocyte could bind 17.5 × 105 molecyles of FMC 34, which was more than three times the number of molecules of FMC 10 which could be bound to each cell. The additional reactivity of FMC 34 appears to be due to its capacity to bind fucosylated and non‐fucosylated lactose‐bearing structures.
Binding of IgG to B cell via HLA molecules
Authors: Chia David , Terasaki Paul I. , Chan Henry , Ozawa Miyuki
Published: October 2008

Abstract: Binding of immunoglobulins to major histocompatibility complex (MHC) molecules was demonstrated by two different assays: the binding of IgG to B cells by flow cytometry, and purified MHC antigens with an Elisa assay. Fc fragment from immune‐complex binds to the Fc receptor on B lymphocytes. Here, Fab was also shown to bind to B cells. This binding was inhibited by specific human alio anti‐HLA Class I and II sera directed at the polymorphic sites. Thus, in addition to the Fc receptor, MHC can also serve as a binding site for IgG. In an Elisa assay using purified antigens, IgG was shown to bind to HLA Class I and II molecules. Other proteins such as transferrin, human serum albumin, gelatin, etc., did not bind to the MHC proteins. Immunoglobulins bound to MHC molecules by sites on the Fab fragment independent of the hypervariable region. This was demonstrated by the retention of antibody activity even after binding of antibody (anti‐lactoferrin) to MHC. The relative avidity between Fab and HLA Class I and II was 4‐8xl05M−1.
Biochemical analysis of class I and class II MHC antigens in cynomolgus macaques by one‐dimensional isoelectric focusing
Authors: Manning Carolina H. , Heise Eugene R.
Published: October 2008

Abstract: To obtain a better estimate of major histocompatibility complex (MHC) polymorphism in the Old World macaque, Macaca fascicularis, class I and class II MHC proteins from 42 animals were analyzed by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS‐PAGE) and one‐dimensional isoelectric focusing (1D‐IEF). The panel represented both related and unrelated animals with a total of at least 30 serologically distinct haplotypes. Cells were sequentially immunoprecipitated with monoclonal antibody (mAb) W6/32 for class I and with mAb L243 for class II molecules, followed by neuraminidase treatment. Both sets of immunoprecipitates yielded 5‐7 major bands on IEF. All bands present in offspring were present in at least 1 parent. Siblings which were serologically identical for class I and which were non‐stimulatory in mixed lymphocyte culture (MLC) yielded identical IEF patterns for both class I and class II; in other sibling pairs which were serologically identical for class I antigens (Ag), IEF produced convincing evidence that the siblings were indeed nonidentical, or helped to verify that recombination had occurred within the MHC. Specific bands were found which correlated with class I specificities A8, A24, and B25 previously defined by serology. Comparison of serological and biochemical data will broaden our understanding of the MHC in Macaca fascicularis and will increase the potential use of this species in transplantation research, as a model of disease, and for comparative studies.
HLA‐A, ‐B, ‐DR, ‐DQ antigens in black North Americans
Authors: Lee T. D. , Lee A. , Shi W. X.
Published: October 2008

Abstract: HLA antigens of black North Americans are not as well‐characterized as those of other ethnic groups. We present results from HLA typing of black Americans using the 10th International Histocompatibility Workshop (IHW) sera. 1502 black Americans are typed for HLA‐A, ‐B antigens. 128 black Americans are typed for HLA‐DR, ‐DQ antigens. New specificities and splits established by the 10th IHW are identified in our data. Antigen frequency, gene frequency and haplotype frequency are computed for each phenotype observed. Haplotype A30 ‐Bw42 ‐ DRwl8 ‐ DQw4 and haplotype Aw36 ‐ Bw53 in strong linkage disequilibrium is characteristic of this black population. DR and DQ association patterns are similar to those of white North Americans. A major difference is the association of DRw8 with DQw7 in blacks versus an association of DRw8 with DQw4 in whites.
Distribution of HLA‐DRB1, ‐DQA1 and ‐DQB1 alleles and DQA1‐DQB1 genotypes among Norwegian patients with insulin‐dependent diabetes mellitus
Authors: Rønningen Kjersti S. , Spurkland Anne , Iwe Thomas , Vartdal Frode , Thorsby Erik
Published: October 2008

Abstract: We have studied 87 unrelated Caucasian insulin‐dependent diabetes mellitus (IDDM) patients and 181 healthy controls by oligotyping for 20 DRB1, eight DQA1 and 13 DQB1 alleles, and established their DR‐DQ haplotypes and DQ genotypes. An increase of DRB1 alleles encoding DR4 was found among IDDM patients, but the distribution of DR4 subtypes did not differ among DR4‐positive IDDM patients and controls. The frequency of certain DRB1‐DQA1‐DQB1 haplotypes and DQA1‐DQB1 genotypes was significantly increased among IDDM patients. Taken together, the data suggest that IDDM is primarily associated with several (at least five) different DQ αβ heterodimers.
Production and characterization of human monoclonal lymphocytotoxic autoantibodies from a renal dialysis patient
Authors: Taylor C. J. , Ting A. , Morris P. J.
Published: October 2008

Abstract: We have produced human monoclonal lymphocytotoxic autoantibodies from a renal dialysis patient by the generation of a mouse/human heterohybridoma. The antibodies are of the IgM class and react with the patient's autologous cells, the B‐Iymphoblastoid cell line producing the antibody, normal T and B lymphocytes, B cells from chronic lymphatic leukemia patients (CLL cells), and the autoantibody‐sensitive cell line K562. Screening of the monoclonal antibodies (mAb) against panels of normal T and B cells and CLL cells demonstrated that different reactivity profiles could be generated at different dilutions of the mAb. These profiles were identical to those seen with autoantibodies from different renal patients and this suggests that these profiles do not imply different antibody specificities but differing target cell sensitivity. Reactivity profiles seen in the fluorescence binding assays suggest that the target cell sensitivity is dictated not by antigen density alone but also by antibody/ antigen affinity. The results from studies of enzyme treatment of target cells and lectin inhibition of the molecular specificity suggest that the autoantibodies are polyreactive, capable of binding sialic acid‐dependent epitopes and other negatively‐charged cell surface molecules.
Immunogenetic studies of soluble Fc receptor (CD 16) antigens in human seminal plasma
Authors: Thaler Christian J. , McIntyre John A. , Faulk W. Page
Published: October 2008

Abstract: Two forms of CD 16 Fc receptors (FcRMIII) have been defined on human leukocytes. Type 1, present on polymorphonuclear neutrophils (PMN), expresses the NA alloantigen system and type 2, present on natural‐killer (NK) cells and macrophages, has no known polymorphic variations. We have described the presence of soluble FcRMIII antigens in human seminal plasma (SP) (see Ref. 9). These SP antigens retain an affinity for IgG‐Fc and are biochemically distinct from leukocytic FcRMIII. Their origin within the male reproductive tract and how they relate to FcRMIII types 1 and 2 were not known. By using monoclonal antibodies CLB/Gran 11 and Leu lie that differentially react with FcRMIII type 1 and type 2 from different individuals, we studied how SP FcRMIII antigens relate to leukocytic FcRMIII. CLB/Gran 11 and Leu 11c reactivities with PMN and NK cells from a selected panel of male donors were compared with enzyme‐linked immunosorbent assay reactivities of CLB/Gran 11 and Leu lie with SP samples from these donors. The reactivity pattern with SP samples was shown to be different from findings with donors’PMN, but it corresponded with their NK cells. Our results indicate that SP FcRMIII antigens do not manifest the polymorphic variations that are detected by CLB/Gran 11 and Leu 11c on PMN. These findings suggest that SP FcRMIII antigens do not originate from PMN.
Increased frequency of HLA‐DRwl4b(w6)‐associated RFLP in Greenlanders of Eskimo origin
Authors: Grunnet N. , Steffensen R. , Jersild C.
Published: October 2008

Abstract: HLA‐DR typing by the restriction fragment length polymorphism (RFLP) technique of 42 Greenlanders living in northern Jutland, Denmark, revealed a phenotype frequency of 33.3% for HLA‐DRwl4b(w6), which is significantly different from the frequency of 0.0% observed among 98 Danes of Caucasian origin. When comparing the two populations, the frequency of other HLA‐DRB allogenotypes show insignificant variations. Since HLA‐DRwl4b(w6) is carried by approximately one‐third of the Greenlanders tested, this allogenotype may serve as a useful marker in further anthropological and immunogenetic studies.
The HLA‐DQβ non‐Asp‐57 allele: a predictor of future insulin‐dependent diabetes mellitus in patients with autoimmune Addison's disease
Authors: Boehm Bernhard O. , Manfras Burkhard , Seidi Siegfried , Holzberger Gerhard , Kuhnl Peter , Rosak Christoph , Schoffllng Karl , Trucco Massimo
Published: October 2008

Abstract: HLA‐DR specifities in 72 Addison's (AD) patients and 808 local controls were compared. We confirmed earlier reports that the HLA‐DR3 specificity is significantly increased in AD patients. In our study a relative risk of 3.4 X2= 22.5; pc= 0.01) for the disease was calculated. Analysis of HLA‐DQB1 alleles in DR4+ Addison's patients with diabetes mellitus (N = 6) and without IDDM (14 of 18 individuals tested) revealed that the HLA‐DQw8 allele (DQB1*0302) was significantly increased in AD patients with IDDM (x2= 13.5; p=0.001); conversely, a clustering of the HLA‐DQw7 allele was detected in DR4+ Addison's patients without IDDM. We thus conclude that particular polymorphic alleles corresponding to non‐charged amino acids at position 57 of the HLA‐DQ β‐chain [non‐Asp‐57 alleles] are associated with IDDM also in Addison's patients.
Polymorphism of C4 and CYP21 genes in various primate species
Authors: Bontrop R. E. , Broos L. A. M. , Otting N. , Jonker M. J. , Bontrop Ronald E.
Published: October 2008

Abstract: To study the genetic heterogeneity of the C4 and CYP21 genes in selected primate species we used the technique of restriction fragment length polymorphism (RFLP). Genomic DNA was digested using several restriction endonucleases and filters were hybridized with a 500 bp BamHI/ Kpnl fragment derived from the 5’section of a human C4‐cDNA and with a 1700 bp BamHI obtained from a human CYP21 gene. Abundant RFLP heterogeneity was observed for the C4 genes within a rhesus monkey population but not for the chimpanzee colony analyzed. Duplicated C4‐CYP21 clusters can be traced back in humans, but also in chimpanzees, orang‐utans and rhesus monkeys. Thus, duplication of the basic C4‐CYP21 cluster in primates may have happened more than 30 million years ago. Non‐duplicated C4‐CYP21 regions were found for the gorilla and orangutan. Apart from this, shortened C4A and C4B genes were observed in chimpanzees, orangutans and rhesus monkeys, whereas the so‐called long variety of the C4A gene appears to be present in humans and orang‐utans. This ancestral modification, resulting from an insertion of a 6.5 kb intron in the C4A gene, therefore predates at least speciation of human and orang‐utan which is estimated to have taken place more than 12 million years ago.
No association between susceptibility to multiple sclerosis and HLA‐DPB1 alleles in the French Canadian population
Authors: Howell W. M. , Sage D. A. , Evans P. R. , Smith J. L. , Francis G. S. , Haagert D. G.
Published: October 2008

Abstract: HLA‐DPBl typing was performed using polymerase chain reaction DNA amplification and sequence‐specific oligonucleotide probing (PCR‐SSOP) which permitted identification of 17 distinct DPB alleles using 15 oligonucleotide probes. The accuracy of this approach was confirmed in an initial study of 26 human B‐lymphoblastoid cell lines which demonstrated close agreement between PCR‐SSOP and PLT assigned types. A cohort of 47 adult French Canadians was then studied to provide an estimate of DPB 1 allelic frequencies in an ethnically homogeneous population. DPB1*0401 was the most frequent phenotype (61.5%) and only DPB1*0101, 0301 and 0402 were also present at frequencies greater than 10%. HLA‐DPw4 has been reported to be associated with multiple sclerosis (MS) but our PCR‐SSOP analysis of 52 French Canadian MS patients showed no association with either the DPB 1*0401 or DPB 1*0402 splits of DPw4 or with any other DPB1 allele.
Novel HLA‐DR2 and ‐DR3 haplotypes among Norwegian Caucasians
Authors: Ranningen Kjersti S. , Spurkland Anne , Iwe Thomas , Sollid Ludvig M. , Vartdal Frode , Thorsby Erik , Renningen Kjersti S.
Published: October 2008

Abstract: In the Northern European population, all DR2 haplotypes encoded by DRB1*1501 have previously been found to carry the DQA1*0102 and DQB1*0602 alleles, and DR3 haplotypes have been found to carry the DQA 1*0501 and DQB 1*0201 alleles. Here we report a novel recombinant DR2 haplotype carrying the DRB1*1501, DQA1*0102 and DQB1*0603 alleles as well as a novel recombinant DR3 haplotype carrying the DRB1*0301, DRB3*0101, DQA1*0102 and DQB 1*0602 alleles.
HLA Class I nucleotide sequences, 1991
Authors: Zemmour Jacqueline , Parham Peter , Zemmour Jacqueline
Published: October 2008

The HLA Class I sequences included in this compilation are taken from publications listed in the papers: Nomenclature for factors of the HLA system, 1990 (1) and Nomenclature for factors of the HLA system, 1989 (2). Where discrepancies have arisen between reported sequences, the original authors have been contacted where possible, and necessary amendments to published sequences have been incorporated into this alignment. Future sequencing may identify errors in this list and we would welcome any evidence that helps to maintain the accuracy of this compilation. In the sequence alignments, identity between residues is indicated by a hyphen (‐). An unavailable sequence is indicated by a period (‐). Gaps in the sequence are inserted to maintain the alignment between different alleles showing variation in amino acid number.
HLA Class II nucleotide sequences, 1991
Authors: Marsh Steven G. E. , Bodmer Julia G.
Published: October 2008

Abstract: The HLA Class II sequences included in this compilation are taken from publications listed in the papers: Nomenclature for factors of the HLA system, 1990 (1) and Nomenclature for factors of the HLA system, 1989 (2). Where discrepancies have arisen between reported sequences, the original authors have been contacted where possible, and necessary amendments to published sequences have been incorporated into this alignment. Future sequencing may identify errors in this list and we would welcome any evidence that helps to maintain the accuracy of this compilation. In the sequence alignments, identity between residues is indicated by a hyphen (‐). An unavailable sequence is indicated by an asterisk (*). Gaps in the sequence are inserted to maintain the alignment between different alleles showing variation in amino acid number.
Association of four HLA class III region genomic markers with HLA haplotypes
Authors: Pei Ji , Choo S. Yoon , Spies Thomas , Strominger Jack L. , Hansen John A.
Published: October 2008

Abstract: We have studied restriction fragment length polymorphism (RFLP) in the region 300 kb centromeric to the HLA‐B locus. Four probes were used: one was genomic DNA derived from the tumor‐necrosis factor (TNF)‐β gene, one was a cDNA for the BAT3 gene, and two single‐copy genomic probes, R5A and M20A. The order of these markers from HLA‐B towards the centromere is M20A, R5A, TNF and BAT3. The BAT3 and TNF‐β probes each detected two allelic bands with Taq I and Nco I digestion, respectively; the R5A and M20A probes each detected three polymorphic allelic bands with BstEII digestion. To determine if these restriction polymorphisms are preferentially associated with certain HLA‐B and ‐DR haplotypes, a total of 153 HLA haplotypes was analyzed. The haplotypes Al, B8, DR3 and A3, B7, DR2 were each associated with a distinct combination of polymorphisms identified at these four sites, thereby demonstrating that the strong linkage disequilibrium characteristic of these haplotypes extends also to this segment of the class III region. In contrast, haplotypes that are not in positive linkage disequilibrium, such as A1,B8,DR4 and A2,B7,DR3, showed ho preferential association with any of these polymorphisms. The antigens HLA‐B27 and B35 were also found to be in positive linkage disequilibrium with RFLP patterns at three of these sites, and HLA‐B14,B35,B44,Bw57 and Bw62 were found preferentially associated with polymorphisms at one or two of these sites, independent of the DR antigen present. These data further demonstrate that genetic linkage disequilibrium in the HLA class III region is complex and variable among different HLA halpotypes.
HLA‐DRB101 subtyping by allele‐specific PCR amplification: A sensitive, specific and rapid technique
Authors: Olerup Olle , Zetterquist Henrik
Published: October 2008

Abstract: The two DR1‐associated cellular specificities Dwl and Dw20, as well as DR'Br’(Dw'BON'), cannot be unequivocally assigned by serological typing or restriction fragment length polymorphism (RFLP) analysis. We have developed and compared two polymerase chain reaction‐based (PCR) typing methods for distinguishing these DRB1 alleles; allele‐specific amplification of DRB1*01 alleles followed by an agarose gel electrophoresis detection step and group‐specific DRBI*01 amplification followed by hybridization with sequence‐specific oligonucleotide probes. The two typing strategies gave completely concordant results in the 33 DRB1*01‐positive and the 46 DRB1*01‐negative individuals and cell lines studied. No false‐negative or false‐positive typing results were obtained. All possible heterozygous combinations of the DRB1*0101‐0103 alleles could be distinguished by both typing methods. DRB1*01 subtyping by allele‐specific PCR amplification was performed in less than 3 hours, including PCR amplification, detection and interpretation steps. The technique will be a valuable complement to DR typing by serology and RFLP analysis. Allele‐specific DRB1 amplifications or group‐specific amplifications followed by directed allele‐specific amplifications of DRB1 alleles, typing based on the absence or presence of amplified products, may well prove to be the technical innovation that will firmly establish PCR‐based DR typing in routine clinical tissue typing.
HLA‐DRB3 typing by restriction digestion of locus‐specific amplified DNA
Authors: Smrzka Oskar W. , Faé Ingrid , Pickl Winfried F. , Fischer Gottfried F. , Smrzka Oskar W.
Published: October 2008

Abstract: Locus HLA‐DRB3 codes for the serologically defined supertypic specificity DRw52 in HLA‐DR3, ‐5 and ‐w6 haplotypes. Three specificities of DRw52 (DRw52a, ‐b and ‐c) can further be distinguished by cellular techniques or by DNA typing with allele‐specific oligonucleotide probes. These specificities were recently reported to have significant importance in antigen presentation. To avoid a time‐consuming hybridization procedure, we have developed a simple typing system using PCR and subsequent digestion by allele‐specific restriction endonucleases. A system was established with locus‐specific amplification of HLA‐DRB3 and digestion by the enzymes KpnI, Seal and HinfI which recognize unique restriction sites within the amplified region. This allowed HLA‐DRB3 typing on agarose gel by determining whether the amplification product has been digested or not. This typing system was compared to conventional oligotyping by analyzing 145 RFLP‐typed individuals for their DRw52 specificity using both methods. Agarose typing correlated well with oligotyping and was shown to be more simple and practical even in heterozvaous individuals.
Probabilistic assessment of the HLA sharing of recurrent spontaneous abortion couples in the Japanese population
Authors: Koyama Masayasu , Saji Fumitaka , Takahashi Shunichi , Takemura Masahiko , Samejima Yoshihiro , Kameda Takashi , Kimura Tadashi , Tanizawa Osamu , Koyama Masayasu
Published: October 2008

Abstract: In spite of a number of investigations, the concept of human leukocyte antigen (HLA) sharing in recurrent spontaneous abortion (RSA) couples remains controversial. We introduced the basal antigen sharing rate (BSR) by the original mathematical approach using the gene frequency of all HLA specificities derived from our regional control population and applied this parameter for the comparison with RSA couples. RSA couples were classified into three subgroups; primary (3 or more consecutive abortions), secondary (3 or more consecutive abortions after 1 live birth), and potential (2 consecutive abortions) aborters. No significant differences between the HLA class I sharing rates (one or more antigens shared on a single locus) of the all RSA subgroups and the calculated BSRs were observed. In the HLA‐DR and DQ loci, on the other hand, the antigen sharing rates of primary aborters were significantly higher than BSRs (p<0.01/DR, p<0.05/DQ). While potential aborters showed a result similar to that of the primary aborters, no significant antigen sharing of HLA class II was observed in secondary aborters. Our data suggest that BSR is a useful parameter for detection of significant HLA sharing in regional populations and the consecutive abortions that occurred primarily are certainly relevant to HLA class II sharing.
Aspartic acid at position 57 of the HLA‐DQβ chain in insulin‐dependent diabetes mellitus: An association with one DRw9–DQw9 subtype in the Chinese population
Authors: Ju L. Y. , Sun Y. P. , Semana G. , Gu X. F. , Krishnamoorty R. , Fauchet R. , Charron D.
Published: October 2008

Abstract: Eighteen unrelated Chinese patients with insulin‐dependent diabetes mellitus (IDDM) were analyzed for HLA Class II genes using a variety of molecular biological techniques including restriction fragment length polymorphism (RFLP), polymerase chain reaction with allele‐specific oligonucleotides (PCR‐ASO) and direct DNA sequencing. The high frequency of DR3/DR4 heterozygotes found in the Chinese with IDDM strengthens the importance of this combination of haplotypes in IDDM susceptibility since it is present in two genetically distant populations ‐Chinese and Caucasians. The frequency of DRw9, a rare allele in the Caucasian population, is much higher in the Chinese. Moreover, the DQβ chain linkage of DRw9 was different in IDDM patients compared with control subjects. In contrast with previous results, codon 57 of the DQβ chain was aspartic acid in DRw9 Chinese IDDM patients. Furthermore, one particular DRw9–DQw9 haplotype may be associated with IDDM susceptibility in the Chinese population.
Modified PCR‐RFLP method for HLA‐DPB1 and ‐DQA1 genotyping
Authors: Ota Masao , Seki Takeshi , Nomura Nobuhiro , Suglmura Kazuhlto , Mizuki Nobuhisa , Fukushima Hirofuml , Tsuji Kimiyoshi , Inoko Hidetoshi
Published: October 2008

Abstract: We previously developed a new technique for HLA class II genotyping by digestion of polymerase chain reaction‐amplified genes with restriction endonucleases (PCR‐RFLP method). This PCR‐RFLP method is an efficient and convenient typing technique for class II alleles. However, small fragments or bands located close to each other on polyacryl‐amide gels sometimes prevent precise analysis of the RFLP bands. Furthermore, the restriction enzymes we have reported in the previous papers are not sufficient to identify the genotypes of all heterozygous individuals. Here, we report an improved PCR‐RFLP method using some informative restriction enzymes which have either a single cleavage site or, alternatively, no cleavage site in the amplified DNA region, depending on the HLA alleles, making reading of RFLP band patterns much easier. Each second exon of the HLA‐DQA1 or ‐DPB1 gene was selectively amplified from genomic DNAs of 70 HLA‐homozygous B‐cell lines and 100 healthy Japanese by PCR. Amplified DNAs were digested with restriction endonucleases and then subjected to electrophoresis assaying simply for cutting, or no cutting, of the DNA. ApaLI, HphI, BsaJI, Fokl, MboII and Mnll can discriminate eight alleles of the DQA1 gene. Similarly 19 alleles of the DPB1 gene can be discriminated with Bsp1286I, Fokl, Ddel, BsaJI, BssHII, Cfr13I, Rsal, EcoNI, and AvaII enzymes. This modified PCR‐RFLP method can be successfully applied to heterozygotes. Thus, the method is technically simpler and more practical for routine HLA typing work than our previous PCR‐RFLP method.
HLA‐DR molecules enhance signal transduction through the CD3/Ti complex in activated T cells
Authors: Ødum Niels , Martin Paul J. , Schieven Gary L. , Masewicz Susan , Hansen John A. , Ledbetter Jeffrey A.
Published: October 2008

Abstract: Crosslinking HLA‐DR molecules by monoclonal antibodies (mAb) induces protein tyrosine phosphorylation and results in a secondary elevation of free cytoplasmic Ca2+ concentration ([Ca2+]i) in activated human T cells. Here we have studied the effect of DR on CD3‐induced signal transduction in allospecific T‐cell clones and T‐leukemia (HUT78) cells. Co‐crosslinking of DR with CD3 produced an enhanced [Ca2+]i response compared to that seen with CD3 alone. In contrast, CD2 responses were not enhanced by co‐crosslinking with DR. Co‐crosslinking CD45 in a tri‐molecular complex of CD45, CD3, and DR completely abrogated the enhancing effects of DR on CD3‐induced [Ca2+]i responses. In contrast, the enhancing effect of co‐crosslinking CD4 on CD3 responses was not inhibited by co‐crosslinking CD45. Thus, the DR‐mediated accessory signals appear to be regulated differently from those provided by CD4 accessory molecules. The present data confirm, at the level of second messengers, recent findings suggesting that DR molecules have accessory functions in CD3/Ti‐mediated T‐cell responses.
HLA‐DR generic typing by AFLP
Authors: Tunis Ivan , Salazar Marcela , Yunis Edmond J.
Published: October 2008

Abstract: We have described a practical and inexpensive method whereby any individual can be typed and assigned to any of the 14 generic HLA‐DR types: DR1, DRwl5, DRwl6, DRwl7, DRwl8, DR4, DRwll, DRwl2, DRwl3, DRwl4, DR7, DRw8, DR9 and DRw10. Previous methods to type these specificities include ‐ among others ‐ serology, conventional RFLP, PCR‐oligonucleotide typing, and a PCR‐RFLP method useful for typing homozygous individuals. In the method reported here, DNA is amplified with a set of group‐specific primers and then restricted with a number of endonucleases. Six group‐specific pairs of primers have been chosen to avoid cross‐amplification with other DRB alleles, including DRB3, DRB4 and DRB5 alleles, and to anneal at uniform temperature: 61deg;C. Endonucleases were chosen to generate unique patterns of easily recognizable bands that led to unequivocal assignments of HLA‐DR generic types in heterozygous as well as homozygous individuals. This technique involves two steps: 1) Amplification of DNA with six different pairs of primers where DR1, DR4 and DRw10 types can be assigned at once, and 2) Endonuclease digests of amplified DNA to assign DR7, DRw8, DR9, DRw11, DRw12, DRw13, DRw14, DRw15, DRw16, DRw17 and DRw18 types. Individuals carrying any combination of all alleles published so far can be typed by this method. The ease, low cost and reliability of this method are discussed.
Dwl4(DRBl*0404) is a Dw4‐dependent risk factor for rheumatoid arthritis Rethinking the “shared epitope” hypothesis
Authors: Nelson J. Lee , Mickelson Eric , Masewicz Susan , Harrington Rob , Dugowson Carin , Koepsell Thomas , Hansen John A.
Published: October 2008

Abstract: HLA‐DR4 has been shown to be associated with risk for developing rheumatoid arthritis (RA) in multiple populations and racial groups. The allelic variants of DR4 share the DR4 serologic specificity but differ by 1 to 3 amino acids in the third hypervariable region (positions 67 to 74) and at positions 57 and 86 of the DRβ1 chain. We have examined DR4 variants in 61 DR4+ RA cases and 55 DR4+ healthy controls. Dwl4 was not associated with RA risk in DR4 heterozygous (DR4,X) cases. Only 15% of DR4, × cases had the Dwl4 allele compared with 28% of DR4,X controls. In homozygous (DR4, 4) individuals who also expressed Dw4, however, Dwl4 was associated with increased RA risk. Moreover, the relative risk for Dw4, Dwl4 (16.1, p = 0.001) actually exceeded that of Dw4, Dw4 (2.2, p = ns). Thus Dwl4 is not an independent risk factor for RA but is a synergistic risk factor for individuals who also have the Dw4 allele.
Gene conversion in the evolution of the human and chimpanzee MHC class I loci
Authors: Kuhner Mary K. , Lawlor David A. , Ennis Peter D. , Parham Peter
Published: October 2008

Abstract: Sixty‐five DNA sequences from human and chimpanzee major histocompatibility complex class I loci were searched for statistical evidence of past gene conversion. Twenty‐four potential conversions were detected: they were distributed across both variable and conserved portions of the gene, and involved both classical and non‐classical loci. The majority spanned less than 100 bp, comparable in length to the conversions observed in spontaneous mutatations in mice. Both within‐locus and between‐locus conversions were observed. Certain areas of the antigen recognition site appear to have been the target for multiple conversion events. The implications of these findings for the evolution of the class I multigene family are discussed.
Hetero and homozygosity of MHC class II gene products in systemic lupus erythematosus*
Authors: Hartung K. , Coldewey R. , Krapf F. , Specker B. Lang. Ch. , Schendel D. , Schneider P. , Seuchter S. , Stangel W. , Albert E. , Deicher H.
Published: October 2008

Abstract: An analysis of HLA class II antigens in 356 white patients with systemic lupus erythematosus (SLE) showed that all HLA‐DR and ‐DQ homozygous and heterozygous combinations appear with frequencies expected from the observed gene frequencies. HLA‐DR2 and HLA‐DR3 gene frequencies were both increased in SLE, as were the odds ratios of all DR2 and DR3 hetero‐ and homozygous combinations. HLA‐DR2/ C4AQ0 heterozygotes were also not increased over expected values. Therefore, gene complementation at MHC loci does not contribute to susceptibility to SLE, but rather one or two MHC allele(s) in linkage with HLA‐DR2 and HLA‐DR3.
Polymorphism of HLA‐DRw52‐associated DRB1 genes as defined by sequence‐specific oligonucleotide probe hybridization and sequencing
Authors: Petersdorf Effie W. , Smith Anajane G. , Haase Anne M. , Martin Paul J. , Hansen John A.
Published: October 2008

Abstract: We have used group‐specific DNA amplification and sequence‐specific oligonucleotide probe (SSOP) hybridization to study DRB1 sequence polymorphisms associated with DR3, DRw11(5), DRwl2(5), DRwl3(w6), DRwl4(w6) and DRw8 alleles. Group‐specific amplification of DRw52‐associated DRB1 alleles was achieved using a 5′ amplification primer designed to hybridize with a first hypervariable region (HVR) sequence common to all known alleles in this group, together with a 3′ intron primer. Prospective SSOP typing of DR3, DRw11, DRwl2, DRwl3, DRwl4 and DRw8 alleles was performed in 318 individuals, including 124 patients, 46 family members and 148 unrelated marrow donors. Among the 395 DRw52‐associated DRB1 alleles tested in our study, a subtype corresponding to the previously defined alleles DRB1*0301–2 (DR3), DRB1*1101–4 (DR5), DRB1*1201–2 (DR5), DRB1*1301–5(DRw6),DRB1*1401–2 and 1404 (DRw6), and DRB1*0801–4 (DRw8) could be assigned in all but 6 individuals (1.9%) tested. In addition to the 22 known alleles, we identified two new DRw6‐associated alleles, DRB1*13.MW(1) and DRB1*14.GB(1). DRB1*13.MW typed serologically as DRwl3 and was identical to DRB1*1301 except at codon 71 where AGG encodes arginine instead of GAG encoding glutamic acid. DRB1*14.GB represents a DRB1*1402 variant whose sequence at codon 86 encodes valine (GTG) instead of glycine (GGT). These results demonstrate that SSOP methods represent an efficient and precise approach for typing DRB1 alleles and for identifying potential novel variants previously unrecognized by conventional typing methods.
HLA‐DPB1 oligonucleotide typing of a Southwest German Caucasian population
Authors: Elermann Thomas H. , Fakler Josef , Müller Carlheinz R. , Ballas Manfred , Goldmann Shraoa F.
Published: October 2008

Abstract: HLA‐DP genotyping with sequence‐specific oligonucleotides can detect known sequence variations in the polymorphic segments of the DPB1 second exon. Since the allelic polymorphism of the 22 published alleles is based on recombination of sequence motifs from six variable regions, DPB1 typing depends on the reactivity pattern of many different probes rather than from typing with single allele‐specific probes. By computer simulation, we have previously shown that the minimal set of probes to define the 22 different alleles and most of the heterozygous combinations is 18. Here we describe HLA‐DPB1 typing results and allele frequencies in a panel of 200 unrelated Caucasians from Southwest Germany. The result confirmed the power of the new HLA‐DPB1 typing method, but we failed to detect three of the previously described alleles in our panel. To accommodate with the observed 19 different alleles, the sequence and hybridization conditions of 17 oligonucleotide probes are given, which are able to differentiate all except two, resolved by group‐specific amplification, of the 190 possible heterozygous phenotypes.
Expression of CD31 epitopes on human lymphocytes: CD31 monoclonal antibodies differentiate between naive (CD45RA+) and memory (CD45RA‐) CD4‐positive T cells
Authors: Ashman Leonie K. , Aylett Gabriella W.
Published: October 2008

Abstract: The CD31 antigen, a member of the immunoglobulin superfamily with a possible cell adhesion function, is expressed on approximately 50% of periperal blood lymphoid cells at relatively low intensity (10–20% of the level on monocytes). In the accompanying paper we showed that a mAb, 5A2.G5, which identifies a glycosylation‐dependent epitope of the CD31 antigen, bound to fewer lymphocytes than two other CD31 mAb, B2B1 and 2BD4, although the 3 antibodies bound equally well to monocytes. We have now analyzed the pattern of expression of epitopes of the CD31 antigen on lymphoid cell subpopulations using two‐color immunofluorescence and flow cytometry. Large granular lymphocytes (CD16+), CD8‐positive T cells and B cells (SMIg+) were mostly CD31‐positive as indicated by the binding of mAb B2B1 and 2BD4. Single populations displaying some overlap with the negative control were obtained in each case. In contrast, CD4‐positive T cells fell into two discrete populations with respect to CD31 antigen expression. mAb 5A2.G5 displayed weaker binding to all lymphoid cell types, indicating that the pattern of glycosylation of the CD31 antigen differs between lymphocytes (of all types) and cells of the myeloid lineages. The heterogeneity of CD31 antigen expression by CD4‐positive cells was further examined by dual‐labelling of purified CD4 cells with mAb B2B1 and CD45RA or CD29 mAb which identify naive and memory T cells respectively. The CD31 antigen was found to be preferentially expressed by the CD45RA‐positive, naive cell population.
Inhibition of cell proliferation with an HLA‐A‐specific monoclonal antibody
Authors: Scheinberg David A. , Bull Marcia K. , Yang Soo Young , Glass Kenneth A. , Minniti J. Gerald
Published: October 2008

Abstract: A new mouse IgG2A monoclonal antibody ‐ JD 12 ‐ is described, which was selected for its ability to inhibit peripheral blood mononuclear cell (PBMC) proliferation. Serologic, immunoprecipitation and immunoelectrophoretic studies showed JD12 to be monomorphically reactive with HLA‐A molecules with an affinity of 109 M‐1. JD12 was capable of inhibiting growth in resting or stimulated PBMC with an ID5o of 300 ng/ml in a time‐dependent and dose‐dependent manner without cytotoxicity. IgG, F(ab)2 and F(ab) were active. Growth stimulation by phorbol ester was not inhibited, suggesting that JD12 action occurred prior to protein kinase C activation. DNA and RNA synthesis were both slowed with a resulting G0/G1 block. Analysis of separated PBMC components showed that adherent cells displayed increased activity (clumping and RNA synthesis) upon JD12 binding. In addition, the inhibitory activity of JD12 could be transferred to new cultures by cell‐free, IgG‐free supernatant from JD 12‐treated PBMC, suggesting that the antiproliferative effects could be mediated, at least in part, in an indirect manner through an inhibitory factor. Therefore, potent, noncytotoxic inhibition of cell proliferation, DNA and RNA synthesis via MHC molecules can be mediated specifically through effects initiated by binding to HLA‐A molecules alone.
Oligotyping of celiac multiplex families with the 11th International Histocompatibility Workshop reagents*
Authors: Petronzelli F. , Ferrante P. , Triglione P. , Bonamico M. , Mazzilli M. C.
Published: October 2008

Abstract: Using PCR and SSO probes from the 11th International Histocompatibility Workshop, we oligotyped for HLA‐DRB1 gene and DQA1*0501, DQB1*0201 alleles 10 celiac families each with 2 affected children. All families belong to the Italian population except for one, whose mother is originaly from Cape Verde island. 8/10 sibling pairs share the DQA1*0501/B1*0201 heterodimer, inherited in cis or in trans arrangement. All the dimer‐negative patients were DR4‐positive.
Sequences of four previously undescribed human T‐cell receptor β chain variable genes
Authors: Hansen Torbjarn , Qvigstad Erik , Lundin Knut E. A. , Thorsby Erik
Published: October 2008

ABSTRACT: T‐cell receptor β chain mRNA from 4 human T‐lymphocyte clones was reverse transcribed, amplified by PCR, cloned in M13 and sequenced. Four hitherto undescribed variable genes were found. One of them (Vβ‐IW22) showed the highest homology (74.5%) to Vβ1. Two others (Vβ‐IW6 and Vβ‐IW10) belonged to the recently described vβ21 family. The last gene (Vβ‐VW114) belonged to the Vβ9 family. However, the latter gene a is pseudogene because of an in‐frame stop codon.
HLA association of idiopathic Peyronie's disease: An indication of autoimmune phenomena in etiopathogenesis?
Authors: Rompel Rainer , Weidner Wolfgang , Mueller‐Eckhardt Gertrud
Published: October 2008

Abstract: HLA typing for class I and class II antigens was done in 52 unrelated patients suffering from idiopathic Peyronie's disease. The controversially discussed association with the HLA‐B7 cross‐reacting group could not be confirmed. Marked deviations of antigen frequencies were observed for HLA‐A1, B8, Cw7, DR3 and DQw2 compared to healthy local controls. After correction of p‐values, Al (pc < 0.05) and DQw2 (pc < 0.01) remained significant. A possible association of Peyronie's disease with markers of the HLA‐A1, B8, Cw7, DR3, DQw2 haplotype, as first described here, would suggest autoimmunological factors in this disorder of otherwise unknown etiopathogenesis.
HLA‐DP and susceptibility to insulin‐dependent diabetes mellitus in Japanese
Authors: Yamagata Kazuya , Hanafusa Toshiaki , Nakajima Hirumn , Sada Masaharu , Amemiya Hiroshi , Tomita Koji , Miyagawa Jun‐Ichiro , Noguchi Tamlo , Tanaka Takebiko , Kono Norio , Tarui Seiichiro
Published: October 2008

ABSTRACT: Human leukocyte antigen (HLA) genes are candidates for susceptibility genes in insulin‐dependent diabetes mellitus (IDDM). Recently, the association of DR and DQ with IDDM has been reported, but the role of HLA‐DP genes remains uncertain. To address the question, we analyzed the DPB1 gene of 20 Japanese IDDM patients and 30 control subjects using a combination of polymerase chain reaction (PCR) and restriction fragment length polymorphism (RFLP) analysis (PCR‐RFLP method). DPB1*0501 was the most frequent allele both in Japanese patients and control subjects. There was no appreciable association between IDDM and the DPB1 allele in Japanese. The absence of association between IDDM and DP, in spite of the known association between this disease and both DR and DQ, suggests that the HLA locus (loci) telomeric to DP encodes susceptibility to IDDM.
Clinical applicability of the immunomagnetic beads technique for serological crossmatching in renal transplantation
Authors: Povlsen Johan V. , Madsen Melvin , Rasmussen Allan , Strata Michael , Graugaard Bodil H. , Birkeland Sven A. , Hansen Hans E. , Fjeldborg Ole , Lamm Lars U.
Published: October 2008

ABSTRACT:The aim of the present prospective study was to investigate the clinical applicability of the immunomagnetic (IM) beads technique for serological crossmatching (XM) in renal transplantation. The IM XM were read after various periods of incubation, and the results were compared with those obtained by the conventional Kissmeyer‐Nielsen (KN) technique. A total of 132 sera from 96 potential recipients were tested against cells from 62 donors. Eighty‐nine KN T‐cell XM‐negative renal allograft transplantations were performed, and the IM XM results were related to clinical 3‐month follow‐up data (incidence of primary nonfunction, never functioning grafts, graft losses and rejection episodes). The IM technique was clearly more sensitive than KN, and sensitivity increased markedly with increasing duration of incubation. KN ‐, IM + reactions were predominantly found among sera from patients with panel‐reactive antibodies (PRA, 2p<0.01), and thus probably caused by HLA antibodies. However, positive IM XM, appearing after more than 35 min of incubation, did not influence the overall clinical outcome in the observation period. With reading after exactly 35 min of incubation, XM results obtained by IM and KN techniques correlated well. Thus, we believe, that the IM XM technique will be as safe and effective in avoidance of hyperacute rejections as the conventional assay. In the present material, the incidence of primary nonfunction was significantly (2p = 0.0023) higher among PRA+ recipients compared to PRA ‐ patients. To conclude, we recommended the IM technique with reading after exactly 35 min of incubation for easy, fast (70 min) and reliable XM, that is always possible to perform using peripheral blood.
Induction of CD 14 antigen on the surface of U937 cells by an interleukin‐6 autocrine mechanism after culture with formalin‐killed Gram‐negative bacteria
Authors: Ikewaki Nobunao , Inoko Hidetoshi
Published: October 2008

ABSTRACT: In order to better understand the regulation of CD 14 antigen on the surface of the monocyte‐like cell line U937 in response to bacteria, the expression and regulation of CD 14 antigen on these cells when cultured with formalin‐killed bacteria were determined using the monoclonal antibody MY‐4 and analyzed by means of the indirect immunofluorescence method. CD14 expression was induced on the U937 cells after about 48 hours of culture with all of the formalin‐killed Gram‐negative bacteria used in this study but with none of the Gram‐positive bacteria. Maximum expression was obtained after culture with formalin‐killed Salmonella enteritidis strain 116–54. Various cytokines such as interleukin‐1β, interleukin‐2, interleukin‐6, interferon‐γ and tumor‐necrosis factor‐α were assayed in the culture supernatant of U937 cells cultured with or without formalin‐killed Salmonella enteritidis 116–54 using an enzyme‐immunoassay or radioimmunoassay system. The U937 cells were found to produce a large amount of interleukin‐6 in response to formalin‐killed Salmonella enteritidis 116–54. On the other hand, culture supernatant (referred to as conditioned medium) obtained from the U937 cells after 72 h of culture with formalin‐killed Salmonella enteritidis 116–54 also induced strong expression of CD 14 antigen 48 to 72 h later, and this was blocked by the addition of anti‐human interleukin‐6 antibody. These findings suggest that the expression of CD 14 antigen on the surface of U937 cells cultured with formalin‐killed Gram‐negative bacteria is induced by interleukin‐6 and can be explained on the basis of the autocrine mechanism of interleukin‐6.
HLA class II‐associated genetic susceptibility in multiple sclerosis: A critical evaluation
Authors: Olerup Olle , Hillert Jan
Published: October 2008

Abstract: Multiple sclerosis (MS) has, since the 1970s, been known to be associated with the HLA‐Dw2 and ‐DR2 specificities in Caucasian Europeans and North Americans. By the use of genomic typing techniques, the association has been specified to be with the DRwlS,DQw6,Dw2, i.e. the DRBI* 1501‐DQAI*0102‐DQBl*0602 haplotype. A significant DPw4 association in Scandinavian MS patients has been described in one report. However, this association has not been confirmed in several subsequent studies with patients from the same and other ethnic groups. During the last few years several reports, based on serological, RFLP and PCR‐SSO data, have suggested that the HLA class II‐associated MS susceptibility gene(s) may be more closely associated with the DQ than with the DR subregion. The observations that the HLA‐DQBI genes of MS patients share long stretches of sequence motifs and also carry DQA1 alleles encoding glutamine at position 34 of the DQ α chain have received considerable attention. It has been suggested that the susceptibility to develop MS might be determined by the corresponding DQ α‐β heterodimers either encoded in cis or in trans. We have investigated these issues in a large group of Swedish MS patients (n= 179). We found that the associations with the suggested DQBI sequences and position 34 of the DQ α chain were due to linkage disequilibrium and secondary to the association with the DRw15,DQw6,Dw2 haplotype (p < 10‐9 and p < 10‐8, respectively). No overrepresentation of the implicated DQ α‐β heterodimers was observed in DRw1S,DQw6,Dw2‐negative patients. We conclude that available data does not allow the localization of the HLA class II‐associated MS susceptibility genes within the DRw15,DQw6,I>w2 haplotype. We have previously described immunogenetic differences between different clinical forms of MS. In the present study of a new group of MS patients compared with a new control panel, these differences were partly confirmed. The DRw17,DQw2 association in relapsing/remitting MS was affirmed. However, in patients with primarily chronic progressive MS a negative association with the DQw7 allele was not substantiated and a positive association with the DR4.DQw8 haplotype could neither be confirmed nor rejected.