bluuebunny/crossref_metadata_2025 huggingface dataset:
HLA haplotype segregation in families with allergic asthma
Authors: Caraballo L R. , Hernandez M.
Published: October 2008

Abstract: In order to evaluate genetic linkage between Allergic Asthma (AA) and the HLA system, we studied 20 families having AA affected sib pairs and 8 families with Intrinsic Asthma (IA) affected sib pairs. All AA patients had a strong IgE immune response to the mite D. farinae. Serological HLA typing (A, B, C, DR, and DQ antigens) was performed by the standard microcytotoxicity assay. Genetic analysis was made by means of the “Affected sib pairs” method. Out of the 20 affected sib pairs, 14 shared two HLA haplotypes, five shared one HLA haplotype and one was HLA‐non‐identical. These results differed significantly from the random ratio 1:2:1 for sharing, 2, 1 or 0 haplotypes (p< 0.0001), and was very close to that expected for a recessive mode of inheritance. In contrast, among the IA sib pairs there was not an important distortion in the pattern of haplotypes segregation. However, a significant association between any of the HLA alleles and the two types of Asthma studied was not found. The results suggest the existence of an HLA‐linked recessive gene controlling the IgE immune responsiveness to mite allergens and conferring susceptibility to AA.
Restriction fragment length polymorphism of a lymphocyte surface antigen, Blast‐1, in Japanese and Caucasians, and in patients with rheumatoid arthritis
Authors: Matsui Y. , Shibano K. , Kashiwagi H. , Yamakawa‐Kobayashi K. , Inoko H. , Staunton D. E.
Published: October 2008

Abstract: Analysis of polymorphic systems, demonstrating differences among ethnic groups, provides a valuable tool for biology and medicine. Blast‐1 is a member of the immunoglobulin superfamily and an activation‐associated glycoprotein expressed on the surface of mononuclear cells. Blast‐1 demonstrates DNA polymorphism in healthy controls and patients with rheumatoid arthritis (RA). The sizes of polymorphic restriction endonuclease fragments of genomic DNA encoding Blast‐1 were 2.4 and 1.9 kb. In normal controls, the frequency of the homozygote for the 2.4 kb fragment (L‐L) was 0.69 and 0.47, and that for the 1.9 kb fragment (S‐S) was 0.04 and 0.11 in Caucasians and Japanese, respectively. The frequency of the heterozygote for both fragments (L‐S) was 0.27 and 0.42 in Caucasians and Japanese, respectively. The frequencies of the L and S alleles were 0.83 and 0.17 for Caucasians, respectively, and were 0.68 and 0.32 for Japanese, respectively. The difference in the allele frequency between Caucasians and Japanese was significant. In Japanese patients with RA, the frequency of the L‐L, L‐S and S‐S types was 0.45, 0.45 and 0.10, respectively. Lung fibrosis in Japanese RA patients was associated with an increase in the L‐S and S‐S types and a decrease in the L‐L type. The present study indicates that the investigation for gene polymorphisms of Blast‐1 among distinct ethnic groups is important because Blast‐1 appears to be a genetic marker for the manifestation associated with RA.
Shared SstI RFLPs by HLA‐Aw19, A23/24 and A3/11 crossreacting groups
Authors: Segurado Oscar G. , Iglesias‐Casarrubios Paz , Vicario Jose Luis , Gorell Alfredo , Regueiro Jose R. , Arnaiz‐Villena Antonio
Published: October 2008

Abstract: Three novel restriction fragment length polymorphisms (RFLPs) have been identified using a pan‐HLA class I probe and the endonuclease SstI. This study, in conjunction with previously reported SstI RFLPs, now allows the identification of the HLA‐crossreacting antigens Aw19 (A29/30/31/32/w33), A23/24 and A3/11 by specific hybridization patterns with a single enzyme/probe combination. Three of the corresponding polymorphic SstI restriction sites map within the HLA‐A gene and generate two allelic RFLPs (5.06, 5.92 kb) and one single RFLP (5.92 kb) that show an absolute correlation with HLA‐A23/24 and A29/32 crossreacting antigens, respectively. However, other SstI RFLPs (7.97, 9.4, 9.6, 9.8 and 13.34 kb), also linked to HLA‐A crossreacting antigens, map outside the HLA‐A gene and probably correspond to non‐HLA‐A,B,C class I genes in strong linkage disequilibrium with the HLA‐A gene. These data show that HLA‐A crossreacting antigens share more SstI RFLPs than non‐crossreacting antigens both within the HLA‐A gene and also in neighboring non‐HLA‐A,B,C class I genes or pseudogenes; also, this has raised the possibility that some crossreacting HLA‐A alloantisera might additionally recognize shared antigenic determinants in non‐HLA‐A,B,C proteins since the HLA‐Aw19 crossreactivity cannot be fully explained by analyzing the HLA‐A amino acid sequence.
New HLA‐DR2‐related specificities detected in South African Blacks and individuals of mixed ancestry
Authors: Oudshoorn M. , Du Toit E. D. , Martell R. W.
Published: October 2008

Abstract: This study describes a new HLA‐DR2‐related specificity DR2LUM (CT) present in South African Blacks and individuals of mixed ancestry (Cape Coloureds). It can be distinguished from the “classic” DR2 specificities, DRw15 and DRw16, using serological and Southern blot techniques. Although no HLA‐Dw specificity could be assigned to the DR2LUM(CT) cells, borderline typing reactions with Dw2 HTCs were observed. Southern blot analysis using a DRB probe and the TaqI enzyme has shown that DR2LUM(CT) shared a 1.6 kb fragment with DRwl5 and a 4.7 kb fragment with DR1 and DRw10, indicating sequence homology between DR2LUM(CT) and these alleles. In addition, another unusual HLA‐DR2 haplotype was found. The DR antigen was typed serologically as DRw16 but showed a combination of restriction fragments which are associated with both the DRwl5 and DRwl6 specificities. This study demonstrates the value of investigating non‐Caucasoid populations in further characterizing the polymorphisms of the HLA class II genes.
A new duplication C4B 1,12 at the C4B locus associated with BF S07 in a Tunisian population
Authors: Gorgi Y. , Arnold D. , Uring‐Lambert B. , Bardi R. , Ayed K. , Hauptmann G.
Published: October 2008

Abstract: Twenty‐five Tunisian families were analyzed for their complement alleles in order to detect duplications at the C4 loci. In this population, the most characteristic duplications are C4A2, B1,12 or C4A1, B1,12 always associated with BFS07 and C2C. This previously undescribed C4B1,12 duplication was found in seven families, five times in association with HLA‐A2, B50.
Characterization of a novel, human cytotoxic lymphocyte‐specific serine protease cDNA clone (CSP‐C)**
Authors: Klein Jared L. , Selvakumar A. , Trapani Joseph A. , Dupont Bo
Published: October 2008

Abstract: A human cDNA clone encoding a novel serine protease, cytotoxic serine protease‐C(CSP‐C), has been isolated from a cDNA library prepared from recombinant interleukin‐2 (IL‐2)‐activated lymphocytes of a patient with a large granular lymphoproliferative disorder. The clone has a 741‐base pair open reading frame encoding a putative 246‐amino acid protein. The protein sequence contains the catalytic charge relay system characteristic of a serine protease and the conserved N‐terminal amino acid sequence of the mature cytotoxic lymphocyte serine proteases found in both mouse and human. The amino acid sequence of CSP‐C has 71% identity with the previously reported cytotoxic serine protease‐B(CSP‐B)/human lymphocyte protease (HLP)/SECT and 57% identity with the granulocyte‐specific serine protease cathepsin G. The homology with another lymphocyte‐specific serine protease, human Hanukah factor (HF)/Granzyme A was 41%. The transcript is expressed in lymphocytes stimulated with IL‐2 or IL‐2 plus phytohemagglutinin (PHA). CSP‐C is not expresed in B‐lymphoblastoid cell lines or in the T‐leukemia cell line MOLT4. The cDNA sequence suggests that the protein is expressed as a prepropeptide, as has been found in the other murine and human serine proteases of lymphocyte origin. It has recently been reported that human chromosome 14q 11, in addition to containing the genes encoding cytotoxic serine protease B (CSP‐B), cathepsin G, and the T‐cell receptor alpha and delta genes, also includes an additional genomic DNA clone which cross‐hybridized with CSP‐B and cathepsin G, cathepsin‐like gene‐2 (CGL‐2). It is likely that the CSP‐C cDNA clone reported in this study corresponds to CGL‐2.
Two‐disease‐locus model: segregation analysis using information on two markers in nuclear families. Application to IDDM *
Authors: Dizier M. H. , Clerget‐Darpoux F. , Hochez J.
Published: October 2008

Abstract: In the present paper, an extension of segregation analysis is proposed using information on the joint segregation of two unlinked markers conditional on the disease status in nuclear families, in order to consider two‐locus models with one locus linked to the first marker and the other linked to the second marker. We propose tests for examining evidence for the effect of genes located at these two loci and whether this effect is multiplicative or not. This method is then applied to a sample of IDDM families typed for the HLA and Gm markers to test, in addition to a factor of the HLA region, the potential involvement of the Gm system in the susceptibility to IDDM. The analysis does not provide evidence for such an involvement.
Immunohistological patterns of blood group ABO and type 1 chain (Lewis a Lewis b) and type 2 chain (H‐2, Y) antigens in normal uterine cervix
Authors: Torrado Julie , Hoyos Ariel Gutierrez , Blasco Eduardo , Larraz Juan , Rivas Jose Luis Fernandez
Published: October 2008

Abstract: Ten monoclonal antibodies and one lectin were used to study the localization and distribution of Blood Group ABH, type 1 chain (Lewis a, Lewis b) and type 2 chain (H‐2, Y) antigens in 22 cases of normal uterine cervix, with known ABO and Lewis phenotype and secretor status. The results showed that ABH isoantigen expression is clearly related to the secretor status. It is positive in the endo‐ and exocervical epithelium of secretor individuals and negative in non‐secretors. Lewis antigen expression in both endocervical and exocervical epithelium is related to the patient's Lewis phenotype and is clearly controlled by the secretor gene. Indeed, the expression of the Lewis a antigen is limited to the tissue of people of Lewis (a+b‐) phenotype (non‐secretors), and the expression of Lewis b antigen is limited to the tissue of people of Lewis a‐b+) phenotype (secretors). The Y antigen showed a focal expression in endocervical mucosa in all the cases, independently of secretor status or Lewis phenotype. The same pattern was observed for H‐type 2 antigen, though only in secretor individuals.
Mitogenic activity of anti‐CD28 MoAb CLB‐CD28/1 on peripheral blood mononuclear cells and its cooperation with other anti‐T cells MoAb in the activation of purified T lymphocytes
Authors: De Felice M. , Giarrusso P. C. , Lamberti A. , Turco M. C. , Valerio G. , van Lier R. A. W. , Yang S. Y. , Ferrone S. , Venuta S.
Published: October 2008

Abstract: Human T lymphocyte proliferative response induced via CD28 molecule is analyzed. An anti CD28 MoAb, CLB‐CD28/1, induces the proliferation of human peripheral blood mononuclear cells in the absence of other stimuli, indicating that CD28 molecule can directly mediate a mitogenic signal in this system. The mitogenic activity of MoAb CLB‐CD28/1 on PBMC does not require MoAb interaction with monocyte Fc receptors, since F(ab′)2 fragments from the MoAb are mitogenic to the same extent as whole IgG. Nevertheless, the activity depends on the presence of accessory cells, since purified T lymphocytes require addition of irradiated monocytes and interleukin 2 to proliferate when incubated with MoAb CLB‐CD28/1. On the other hand, MoAb CLB‐CD28/1 induces response to IL‐2 in thymocytes in the absence of accessory cells. Cooperation of MoAb CLB‐CD28/1 with three other MoAbs, recognizing CD3, CD5 and HLA Class I antigens, respectively, induces Tac antigen expression and IL‐2 responsiveness in purified T lymphocytes. This effect is obtained without cross‐linking of the MoAb. It does not rely on a physical association between CD28 and CD3, CD5 or HLA Class I molecules, as demonstrated by co‐modulation experiments. These data indicate that expression of IL‐2 receptor on T lymphocytes can result from interaction of multiple activation pathways and that some of them, such as those mediated by CD5 and HLA Class I antigens, previously reported to serve as modulatory circuits, can instead act as essential elements in the onset of T lymphocyte proliferation.
Large haplotype‐specific differences in inter‐genic distances in human MHC shown by pulsed field electrophoresis mapping of healthy and Type 1 diabetic subjects
Authors: Niven M. J. , Hitman G. A. , Pearce H. , Marshall B. , Sachs J. A.
Published: October 2008

Abstract: Type 1 diabetes is associated with extended haplotypes defined by combinations of specific alleles of genes in the MHC. We have used pulsed field gel electrophoresis mapping to examine the gross structure of the Class II region of the MHC and its relationship to susceptibility to Type 1 diabetes. We have studied heterozygous members of a family in which susceptibility to Type 1 diabetes is associated with an A1/B8/DR3 haplotype and resistance with A2/B7/DR2, an unrelated diabetic DR3,4 patient and a healthy DR4,w10 subject and a DR2/Dw2 cell line. Digestion was performed with the enzymes Sst II, Mlu I, and Pvu I and hybridization with 21‐hydroxylase, DRA, DQB, DOB and DPA probes. Within the DQ/DR region the DR4‐ and DR7‐bearing haplotypes studied contain insertions of 140‐150kb relative to the DR3 haplotypes whilst the DR2 haplotype in the family was smaller than the DR3 haplotypes by 130kb, whilst that in the cell line was smaller by up to 220kb. This cell line, previously thought to be homozygous by consanguinity, was also shown to be heterozygous in the DP region. Although no differences between diabetic and healthy subjects were observed within the family, these differences in long‐range structure may be of importance to the etiology of Type 1 diabetes, as well as to the evolution of the MHC.
A novel human leucocyte surface membrane antigen defined by murine monoclonal antibody
Authors: Henniker A. J. , Bradstock K. F. , Grimsley P. , Atkinson. K.
Published: October 2008

Abstract: A murine monoclonal antibody has been produced which identifies a novel human leucocyte differentiation antigen. The antibody, designated WM‐66, of IgM subclass, was cytolytic with human complement. WM‐66 was shown to react with virtually all normal T and B lymphocytes from peripheral blood and lymphoid tissues, as well as blood monocytes and approximately 40% of bone marrow mononuclear cells. The antibody also bound to the majority of cases of chronic B‐cell malignancies, including chronic lymphatic leukaemia and non‐Hodgkin's lymphoma, but not to cases of acute leukaemia or to the majority of leukaemic and lymphoblastoid cell lines. WM‐66 also reacted with epithelium of bronchus and salivary gland ducts. A single band of relative molecular mass 65,000 Daltons was immunoprecipitated from membrane extracts of normal lymphocytes and the B‐cell line Daudi. Treatment of a number of WM‐66‐negative B‐cell lines with neuramidindase resulted in WM‐66 binding, indicating that the antigen exists in a covert form masked by sialic acid residues on a wider spectrum of cell types than was initially apparent. The reactivity pattern of WM‐66 indicates that it recognises a previously undescribed surface membrane molecule with broad non‐lineage‐specific distribution on leucocytes. This has recently been confirmed at the Fourth International Workshop on Human Leucocyte Differentiation Antigens. Although the biological function of the molecule recognised by WM‐66 is unknown, the lytic properties of the antibody suggest a possible in vivo therapeutic role as an immunosuppressant or for treatment of lymphoid malignancy.
Familial alopecia areata: No linkage with HLA
Authors: Zlotogorski Abraham , Weinrauch Louis , Brautbar Chaim
Published: October 2008

Abstract: HLA antigens of two Jewish families with alopecia areata were studied. There was no linkage between the disease and the HLA complex.
Restriction fragment length polymorplusm (RFLP) of two HLA‐B‐associated transcripts (BATS)‐ genes in healthy Danes
Authors: Fugger L. , Morling N. , Ryder L. P. , Jakobsen B. K. , Svejgaard A. , Spies T. , Strominger J. L.
Published: October 2008

Abstract: The restriction fragment length polymorphism (RFLP) of the two human HLA‐B‐associated transcripts (BATs) genes, BATl and BAT2, was investigated using 5 different restriction enzymes and two human BATl and BAT2 cDNA probes. Two of the enzymes, NcoI and RsaI, revealed polymorphic patterns which were investigated in healthy Danes. The cDNA/restriction enzyme combination BATl/NcoI identifies polymorphic bands at 12 kb, 8 kb, 2.5 kb, and 1.1 kb, while the BAT2/RsaI combination identifies polymorphic bands at 3.3 kb, 2.7 kb, 2.3 kb, and 0.9 kb. The frequencies of these markers were determined in 90 unrelated Danes. Co‐dominant segregation and allelic behavior was seen for the BATl/NcoI 12 kb and 8 kb bands and the BAT2/RsaI 2.7 kb and 2.3 kb bands, respectively. It is possible that the BAT2IRsaI 3.3 kb band represents a rare allele of the BAT2/RsaI system. The BAT2/RsaI 2.3 kb marker was strongly negatively associated with HLA‐B8 and HLA‐DR3 while there were no strong associations between the BATl and BAT2 markers, but the BAT2/RsaI 2.7 kb marker was strongly positively associated with the TNFα/NcoI 5.5 kb marker, which in turn is positively associated with HLA‐B8 and HLA‐DR3. The negative associations between the BAT2/Rsa1 2.3 kb marker and HLA‐B8 and HLA‐DR3 raise the question as to whether this BAT allele may play a role in protecting against certain autoimmune diseases.
Functional properties of HLA‐DR‐BON alleles associated with DQw5(w1) and with DQw7(w3): A family study
Authors: Bignon J. D. , Tkaczuk J. , Coppin H. , Essakte S. , Cesbron A. , Cheneau M‐L. , Herfray P. , Muller J. Y. , Huchenq A. , Cambon‐Thomsan A. , De Préval C. , Thomsen M.
Published: October 2008

Abstract: Among the DR specificities undefined by serology, DR‐BON is peculiar because RFLP cannot distinguish it from the well‐defined allele DR1 even if the two specificities are very different functionally. The occurrence of two DR‐BON‐like alleles in the same family, one associated to the DQw5 split of DQwl and the other associated to the DQw7 split of DQw3, enabled us to compare the properties of these alleles. The RFLP analysis showed a typical DR1‐like picture for both alleles when probed with DR beta, but for one of the alleles the DQ beta probe gave a DQw7 pattern. Primary mixed lymphocyte cultures showed weak to moderate stimulation between cells from individuals identical for one haplotype and differing for the DR‐blank haplotypes, but by test with cloned reagents we were not able to define differences between the two DR‐blank molecules. Two‐dimensional gel electrophoresis and spot‐test with a probe covering the third hypervariable region of the DRBl gene showed no difference between the two alleles. We thus think that the two DR alleles are identical and that the stimulation observed in primary cultures probably is caused by incompatibility for DQ and DP or class I.
HLA‐Bw54‐DR4‐DRw53‐DQw4 haplotype controls nonresponsiveness to hepatitis‐B surface antigen ‐via CD8‐positive suppressor T cells
Authors: Watanabs Hiroshi , Okumura Makoto , Hirayama Kenji , Sasazuki Takehike
Published: October 2008

Abstract: We have previously reported that in nonresponders to hepatitis‐B (HB) vaccine there was an HLA‐linked immune suppression gene for hepatitis‐B surface antigen (Is‐HBsAg) controlling the nonresponsiveness to HBsAg, through HBsAg‐specific suppressor T cells, and that the Is‐HBsAg was in strong linkage disequilibrium with the HLA‐Bw54‐DR4‐DRw53 haplotype (1). We have now done the HLA typing on an additional 6 nonresponders, and using the system of T‐cell proliferative response to HBsAg we found that the Is‐HBsAg controlled the nonresponsiveness to HBsAg through HBsAg‐specific suppressor T cells in nonresponders to HB vaccine who have HLA‐Bw54‐DR4‐DRw53‐DQw4 haplotype. T‐and B‐cell recognition of HB vaccines might play an important role at 3 to 5 weeks after the last immunization. Use of an anti‐HLA monoclonal antibody has shown that the HLA‐DR molecule plays an important role in helper T‐cell proliferation in nonresponders, although the role of HLA‐DQ molecule in nonresponders was unclear.
Microcytotoxicity assay using mouse L cells transfected with human MHC class II genes. A method and its application
Authors: Schreuder Geziena M. Th. , Marsh Steven G. E. , Hayes Judith H. , Moses Jonathan H. , Krausa Peter , Bodmer Julia G.
Published: October 2008

Abstract: Modifications of the standard microcytotoxicity assay make it possible to use this technique to screen both alloantisera and monoclonal antibodies with mouse L cells transfected with Class II genes. It is necessary to maintain a protein‐rich environment in order to prevent nonspecific complement lysis. Selection of the complement itself is also an important factor, the best results being acheived using a commercially available complement that had previously been absorbed with mouse cells and used at a dilution of 1/8. Using this modified method with transfectants of Dw2 origin we could show that alloantisera against DRw15 recognize the DRB1*1501 gene product, whereas broad DR2 sera react only with the DRB5*0101 product. This technique can be applied succesfully to study the fine specificity of polymorphic monoclonal antibodies, as shown by the reactivity of HU‐30 which binds to the LDR2b transfectant and not to the LDR2a, indicating that the antibody recognizes an epitope present on the DRBl chain and not the DRB5 chain of DR2 cell lines.
The HLA‐DR4‐associated DQw8 allele is cod1.ned to HLA‐DR3/DR4 heterozygous type I (insulin‐dependent) diabetics
Authors: Beebn Bernhard O. , Schifferdecker Ekkehard , Rosak Chrhtopk , Kuehnl Peter , Driese Albert J. , Schëffling Karl
Published: October 2008

Abstract: The HLA‐DR4 specificity revealed a relative risk of 8.5 (x2= 99.6; p< 0.0001) when 193 type I diabetics were compared to 305 controls. Prevalence of the HLA‐DR4‐associated DQ types, i.e. DQw7 and DQw8, were determined, using a restriction fragment length polymorphism (RFLP) typing that combines the probe/enzyme combinations DQB/Taq I and DQB/Bam HI. The HLA‐DQw8 specificity was confined to HLA‐DR3/DR4 heterozygous patients when compared to controls (x2= 4.9; p < 0.025) or to all other DR4‐heterozygous patients (x2= 6.7; p < 0.01). No association with HLA‐DQw8 was seen in HLA‐DRl/DR4 or HLA‐DR“X”/DR4 (X ≠ 1,3,4) heterozygous patients. Due to the excess of HLA‐DR3/DR4 patients the DQw8 allele is a risk factor in type I diabetics, but in HLA‐DRl/DR4 and DRX/DR4 heterozygotes one might suggest that DOB1 and DRB combinations confer HLA‐associated susceptibility.
HLA Class II typing by digestion of PCR‐amplified DNA with allele‐specific restriction endonucleases will fail to unequivocally identify the genotypes of many homozygous and heterozygous individuals
Authors: Olerup Olla
Published: October 2008

Abstract: Recently, a new technique for HLA class II genotyping has been introduced, the polymerase chain reaction‐restriction fragment length polymorphism (PCR‐RFLP) method, claimed to be a practical alternative to conventional serological and cellular HLA class II typing (1‐3). The PCR‐RFLP technique is ingenious, relatively rapid and does not require hybridization with sequence‐specific oligonucleotide probes. However, analysis of whether homozygous and heterozygous combinations of PCR‐RFLP patterns for the various investigated HLA class If loci are unique or not unfortunately shows that only 19% of DRB homozygous and heterozygous combinations are unique. The figures for the DQA1, DQB and DPB loci are 56%, 29% and 65%, respectively. As not all nucleotide sequences analyzed in the above‐mentioned studies (1‐3) gave rise to unique PCR‐RFLPs and as more sequences now are known, for the DRB1, DRB3, DRB5 and DPB1 loci (4), the frequencies of unique PCR‐RFLP patterns for the different HLA class II loci will be reduced even further. Thus, the present analysis demonstrates that the PCR‐RFLP technique, performed as described in references 1‐3, is not yet ready to be used for routine HLA class II genotyping. The resolution of the PCR‐RFLP method can be improved by various modifications. However, the role of a modified PCR‐RFLP technique in HLA class II typing still remains to be shown.
Kinetics of Class II MHC expression on cytotoxic T cells generated by skin allograft
Authors: Fernandes I. , Jiuan K. H. , Tanji M. M. , Sato M. N. , Duarte A. J. S.
Published: October 2008

Abstract: In the present work the kinetics of class II MHC expression on OX8 + lymphocytes generated by skin allograft and its relationship to the lytic activity were studied. Mononuclear cells from the spleen of LEW (RT11) rats bearing BN (RTln) skin graft for 3, 5 or 7 days were sorted out by sequential immune affinity using columns of Degalan‐V26 beads treated with anti‐rat or anti‐mouse Ig. After depletion of B cells, T cells were precoated with W3/25 MoAb (anti‐CD4 equivalent) and sorted out using an anti‐mouse Ig column. The W3/25‐/0X8 + cells (CD8 equivalent) were then coated with OX4 MoAb (anti‐RT1.B) or murine A.TH anti‐A.TL alloantiserum (anti I‐E, cross‐reacts with RTl.D) and were passed through a new anti‐mouse Ig column in order to obtain the four subpopulations, RTl. B +,. B ‐,. D + and. D ‐. Their specific lytic activity against BN Con A‐stimulated cells increased from the 3rd to the 7th d after the skin graft. The lyhc activity observed on the 3rd and 5th d was associated with all four subpopulations analyzed. In contrast, on the 7″ d, the lytic activity was concentrated in the RTI.B+ subpopulation. These results, associated with the increase in the number of OX8 +/RT1.B + cells along with days after graft, suggest that RT1.B expression is not essential but is associated with the effectiveness of the cytotoxic activity. It is also possible that RT1.B expression is a marker of cytotoxic T‐cell differentiation.
The production of a human monoclonal antibody defining a split of HLA‐DRw13 (DRw13b)
Authors: Bunce M. , Sutton P. M. , Ting A. , Morris. P. J. , Bunce M.
Published: October 2008

Abstract: By use of the heterohybridoma technique we have produced a human monoclonal antibody (NDS40) which detects a split of HLA‐DRwl 3 (DRwl3b) which is in linkage with HLA‐DQwl. In addition, the antibody reacts with cells positive for HLA‐DRw8 and DRwl 1, but does not react with the commonly found split of DRw1 3 (DRw1 3a) which is associated with DQwl. NDS40 is cytotoxic and is of the lambda IgM class.
Familial Mediterranean Fever ‐ linkage studies with genetic markers on chromosome 6
Authors: Shohat T. , Shohat M. , Tyan D. B. , Wang S. J. , Sparkes R. S. , Schwabe A. D. , Rotter J. I.
Published: October 2008

Abstract: Familial Mediterranean Fever (FMF) is an autosomal recessive disease manifested by recurrent short episodes of fever associated with polyzerositis. Although the biochemical defect is unknown, there are several immunologic abnormalities which have been described in this disease. To examine critically whether there is linkage between FMF and the immunogenetic region (major histocompatibility complex‐MHC) on chromosome 6, including the HLA, BF, and GLOl loci, blood samples from members of 13 nuclear Armenian families were tested for these genetic markers. These 13 families included 28 affected and 42 unaffected family members. Linkage could be excluded at 7.5% recombination with the HLA ABC and D loci. Linkage could be excluded at 2.5% recombination for GLOl. Linkage could not be excluded with BF individually, but can be rejected based on the haplotype data. No association was found with either BF, GLO1, and HLA DQ alleles. HLA DR4 was found more often in affected cases than in controls; however, after adjusting for the number of antigens tested this was not significant. Our results would appear to exclude the immunogenetic region on chromosome 6 from linkage with FMF in the Armenian population.
Functional analysis of cis‐linked regulatory sequences in the HLA DRA promoter by transcription in vitro
Authors: Hums Clifford R. , Lee Janet S.
Published: October 2008

Abstract: Two consensus sequences, called × and Y boxes, capable of binding nuclear proteins and regulating expression in B cells have been defined within the immediate upstream region of major histocompatibility complex (MHC) class II promoters. Unlike other class II promoters, the HLA‐DR a (DRA) promoter also contains one element identical to the “octamer” motif of immunoglobulin variable region promoters that is responsible for B cell‐specific transcription. This “octamer” in the context of DRA appears capable of binding both the ubiquitous (OTF‐I) and lymphoid‐specific (OTF‐2) “octamer” binding proteins, but at least one other distinct “octamer” complex was found. In order to characterize the function of cis‐acting elements, we have developed an in vitro system in which a DRA promoter construct is transcribed more efficiently in extracts from B cells than in extracts from class II‐negative HeLa cells. 5′ deletion constructs which lacked the Y box, but retained the “octamer” the Y box reduced transcription by 95%. Using supercoiled, but not linear templates, we observed differences in transcription efficiencies from templates lacking or disrupting the × consensus element that reflect effects of random replacement of × box sequences in transient expression assays. Demonstration of the complete dependence on the Y box in this system suggests that, despite its demonstrated importance in the DRA promoter, the DRA “octamer” does not utilize OTF‐2 in a manner analogous to immunoglobulin promoters in B cells.
HLA frequencies in Black Zimbabweans
Authors: Du Toit E. D. , Emmanuel J. G. , We G. , Taljaard D. G. , Oudshoorn M.
Published: October 2008

Abstract: One‐hundred‐and‐nineteen normal Black Zimbabweans were typed for HLA antigens. We found a close similarity between the HLA antigen frequencies in Zimbabweans and Black South Africans (Xhosa), another Bantu‐speaking group. The only significant differences noted were for HLA‐Aw36, ‐Bw53, ‐Cw6, ‐DR4 and ‐DQw3 (p<0.0l). The most common HLA haplotypes in significant positive linkage disequilibrium in the two groups were different. The differences that were noted may be due to gene flow from the San to the Black South Africans, since it is thought that the Zimbabweans had little if any contact with the San.
A nucleotide substitution in a Bgl II site is responsible for the RFLP discrimination between DPw4 and DPa
Authors: Dormoy Anne , Urlacher Arlette , Tongio Marie Marthe
Published: October 2008

Abstract: In a previous work we showed that the two functionally different specificities DPw4 and DPa could only be differentiated by RFLP analysis using two mutually exclusive fragments (respectively, Bgl II 5.29 kb for DPw4 and 7.24 kb for DPa). The DP Workshop synthetic analysis localized these fragments in the DPA2 pseudogene region. Our results demonstrate, however, that they are located between the A1 and B1 genes; the Bgl II restriction site responsible for the 5.29 kb fragment was localized between the first and second exon of the DPBl gene and inside the 7.24 kb fragment. A single mutation point inside this restriction site is responsible for the absence of the 5.29 kb fragment, changing the specificity attributed by RFLP typing.
An MHC (HLA‐A, ‐B, C2, BF, HLA‐DR, GLO1) haplotype study of 497 Danish normal families with 1970 children including 97 twin pairs
Authors: Nielson Lillian Staub , Elberg Hans , Fengrr Kirsten , Mohr Jan
Published: October 2008

Abstract: Extended MHC haplotypes comprising HLA‐A, ‐B, ‐DR, C2, BF and GLOl loci observed in the parents of 497 Danish normal families are presented, with particular regard to the haplotypes that include BF variants or the C2*2 allele. The known association of HLA‐B35, ‐DR1 with both ‐A3 and ‐A11 appeared to depend upon the BF type: HLA‐B35, BF*S, ‐DR1 is strongly associated with ‐All, whereas ‐B35, BF*F,‐DRl is strongly associated with ‐A3. Further, in the present material DZ twins of the same sex shared HLA‐haplotypes more often than did twin pairs of different sex.
The application of oligonucleotide probes to HLA Class II typing of the DRB sub‐region
Authors: Vaughan Robert W. , Lanchbury Jerry S. S. , Marsh Steven G. E. , Hall Margaret A. , Bodmer Julia G. , Welsh Ken I.
Published: October 2008

Abstract: A plan for DRB typing at the sequence level is detailed. Only one polymerase chain amplification reaction is needed and the application of a limited number of short oligonucleotide probes allows an almost complete definition of DRB alleles. The scheme was tested on 40 homozygous cell‐lines selected to cover a wide range of specificities, and 40 RFLP‐typed controls. The results are presented and discussed. The simplicity and accuracy of this scheme are emphasized.
Polymorphism of HLA‐DR2,DQwl haplotypes in Asian Indians
Authors: Singal D. P. , D'Souza M. , Sood S. K.
Published: October 2008

Abstract: We examined the polymorphism of DR2,DQw1 haplotypes in Epstein‐Barr virus‐transformed B‐lymphoblastoid cell lines (HTCs) and unrelated (32 Canadian Caucasians and 24 Asian Indians) individuals by restriction fragment length polymorphism (RFLP) and oligonucleotide typing. The data demonstrate that three subtypes of DR2,DQwl haplotypes, DRwl5(B1.1501).DQw6a(A1.0102,B1.0602), DRwl5(B1.1502). DQw6b(Al.0l03,Bl.0601), DRwl6(B1.1601).DQw5(A1.0102,B1.0502) are present in HTCs and Canadian Caucasians. Of these, DRwl5(Bl.l501).DQw6a (A 1.0102,B 1.0602) haplotype was present in majority (81.3%) of Caucasians. Among Asian Indians, this haplotype was present only in one DR2,DQw1‐positive individual. In addition, three new haplotypes representing different combinations of DRB1, DQAl and DQBl genes were demonstrable in Asian Indians. These new haplotypes are DRwl5(BI.1501).DQw6b(AI.0103,B1.0601), DRwl5(B1.1501). DQw5(A1.0102,B1.0502), and DRwl5(B1.1501).DQw6c(A1.0102, B1.0601). The most frequent haplotypes among Asian Indians were DRwl5(Bl.1502).DQw6b(A1.0103,B1.0601) and DRwl5(B1.1501). DQw6b(A1.0103,B1.0601). The distribution of subtypes of DR2,DQwl haplotypes in Asian Indians was significantly different from that in Canadian Caucasians. The results in the present study have important implications for HLA and for HLA‐disease associations.
Mapping of an HLA‐DRw52‐associated determinant on DRβ1 molecules
Authors: Ballas M. , Elermann T. H. , Wölpl A. , Goldmann S. F.
Published: October 2008

Abstract: The molecular reaction patterns of the DRw52‐specific mouse monoclonal antibodies UL‐52 and 7.3.19.1 were investigated. Upon immunoprecipitation and two‐dimensional IEF‐SDS polyacrylamide gel electrophoresis analysis (2D‐PAGE) mAb UL‐52 selectively isolated DRβ1 molecules from DRw52‐positive cell lines, whereas mAb 7.3.19.1 predominantly precipitated DRβ3 molecules. Reduced mAb UL‐52 binding affinity was observed to DRw8‐ and DRwl2‐positive cells, potentially resulting from structural modifications within the antibody binding site. Comparison of mAb UL‐52 reactivity with published DRβ chain amino acid sequences demonstrates that the amino acid residues ‐S‐ in positions 11 and 13 on DRβ1 molecules essentially contribute to the formation of the antibody binding site. mAb 7.3.19.1 reactivity, on the other hand, correlates with the expression of DRβ3 chain amino acid residues K, G and N, in positions 71, 73 and 77, respectively. In contrast to other DRw52 monoclonal antibodies described so far, mAb UL‐52 demonstrates a similar reactivity to DRw52 allosera, suggesting that mAb UL‐52 and DRw52 allosera possibly recognize the same or a similar determinant on DRβ1 molecules.
HLA‐DP incompatibilities induce significant proliferation in primary mixed lymphocyte cultures in HLA‐A, ‐B, ‐DR and ‐DQ compatible individuals: Implications for allogeneic bone marrow transplantation
Authors: Olerup Olle , Möller Erna , Persson Ulla
Published: October 2008

Abstract: The major part of the proliferative response in primary mixed lymphocyte cultures (MLC) is caused by HLA‐DRB1 incompatibilities. In DRB1‐matched pairs the proliferation induced by HLA‐DRB3, ‐DQ and ‐DP mismatches may be unmasked. In most previous studies the influence of HLA‐DP incompatibilities in primary MLC has been investigated in homozygous typing cells representing only a few Dw specifities. We were interested in determining the stimulatory capacity of isolated HLA‐DP mismatches, ascertained by RFLP analysis, in primary MLC in HLA‐A, ‐B, ‐DR and ‐DQ compatible, unrelated heterozygous individuals of many different Dw specificities. Thirty‐eight MLCs performed with cells from related pairs and 67 with cells from unrelated pairs were evaluated. All but nine of the MLCs were analyzed in both directions, giving a total of 201 investigated reactions. The relative responses (RR) in the three MLCs performed between DP incompatible, related pairs were all positive (RR ≥ 8%). Eighty of 82 MLCs performed with cells from DP incompatible, unrelated individuals were positive, whereas 37 of 46 MLCs between DP compatible, unrelated pairs were negative (RR < 8%) (p < 10−10). The magnitude of the RR was influenced by the number of DP mismatches. Thus, the mean RR was approximately twice as high in MLCs in which responder and stimulator cells differed by two DP antigens (mean RR 60.5%) compared with reactions with only one DP mismatch (mean RR 35.4%) (p < 10−3). RFLP‐defined HLA‐DP incompatibilities predict a positive primary MLC in HLA‐A, ‐B, ‐DR and ‐DQ matched individuals with a high degree of accuracy (98%). We suggest that in the search for matched unrelated bone marrow donors HLA‐DP typing might preferably precede and in many instances even replace MLC. This may well facilitate the search for compatible unrelated donors by reducing the number of MLCs.
Involvement of the CDw50 molecule in allorecognition
Authors: Vilella Ramon , Mila Jordi , Lozano Francisco , Alberola‐Ila Jose , Places Lourdes , Vives Jordi
Published: October 2008

Abstract: The CDw50 differentiation antigen is defined by 101–1D2 and 140–11 monoclonal antibodies (mAb), both produced and characterized in our laboratory. This molecule is broadly expressed on hematopoetic cells but not on other cells. In this report we show that these 2 mAb recognize different epitopes of the same molecule, which are resistant to neuraminidase and proteases. We also demonstrate that the CDw50 antigen is expressed on thymocytes and T lymphocytes as an N‐glycosylated glycoprotein monomer with a relative molecular weight (Mr) of 130000 daltons with intrachain disulfide bonds, and that this molecule is resistant to treatment with phosphatidylinositol (PI) phospholipase C and therefore probably not PI‐anchored to the membrane. CDw50 is a poorly or non‐constitutively phosphorylated molecule that becomes phosphorylated by treatment with phorbol 12‐myristate 13‐acetate (PMA) of peripheral blood mononuclear cells (PBMC). The addition of affinity‐purified CDw50 mAb inhibits primary mixed lymphocyte culture (MLC) but not secondary MLC, cytotoxicity or proliferation induced by mitogens. The inhibition of alloreactivity is mediated at the level of both responding and stimulator cells.
Canine leukocyte integrins: Characterization of a CD 18 homologue
Authors: Moore P. F. , Rossitto P. V. , Danilenko D. M.
Published: October 2008

Abstract: The major characteristics of the canine CD18 leukocyte antigen family, defined by a murine monoclonal antibody CA1.4E9, are described. The overall molecular organization of this family of leukocyte integrins was similar to the human counterpart, and consisted of a common beta subunit of 95 kD (CD18) and non‐covalently linked alpha subunits of 150 kD (CD11c), 165 kD (CD11b) and 180 kD (Cd11a). Conservation of CD18 epitopes between dog, human, feline, equine and bovine was observed. CD18 expression in lymphoid tissue had a characteristic pattern which was based on differences in the intensity of expression observed in different cell types. The expression of LeuCAMs in canine epidermotropic lymphosarcoma (mycosis fungoides) was also explored since LeuCAMs might be expected to play a major role in the tissue tropism of this tumor. Intense CD18 expression was observed in the malignant T cells which infiltrated the epidermis; these cells differed phenotypically from dermal T cells in that they often lacked Thy‐1 expression. The potential pathogenetic significance of LeuCAM expression in canine mycosis fungoides is discussed.
Distribution of MHC class II antigens in feline tissues and peripheral blood
Authors: Rideout Bruce A. , Moore Peter F. , Pedersen Niels C.
Published: October 2008

Abstract: A new monoclonal antibody raised against gradient‐purified feline immunodeficiency virus was found to recognize a bimolecular complex, comprising 27–29 kD and 32–35 kD subunits, on feline peripheral blood lymphocytes. Immunoperoxidase staining of feline tissues with this antibody, designated 43.2H2, demonstrated a reactivity pattern similar to that described for MHC II antigens of the dog, horse, and pig, but differed from human and mouse in having staining of T‐cell zones in spleen and lymph nodes. Flow cytometric analysis revealed that 42.3H2 reacted with 88.97%± 16.00% of feline peripheral blood lymphocytes (n = 20). This high level of reactivity was found to be consistent by repeated sampling over a 4‐month period. Two‐color flow cytometric analysis was used to determined the reactivity pattern on lymphocyte subsets: 88.92%± 7.30% of CD4+ lymphocytes were 42.3H2‐positive, while 85.99%± 11.46% of CD8+ cells were positive (n = 11 for both). B lymphocytes had the highest reactivity (99.47%± 0.45; n=9) and also had the highest fluorescence intensity. By gating based on light scatter properties, 95.06%± 7.35% of monocytes were 42.3H2‐reactive (n= 18), while granulocytes were negative.
Rheumatoid arthritis may be primarily associated with HLA‐DR4 molecules sharing a particular sequence at residues 67–74
Authors: Rønningen Kjersti S. , Spurkland Anne , Egeland Torstein , Iwe Thomas , Munthe Eimar , Vartdal Frode , Thorsby Erik
Published: October 2008

Abstract: Genomic typing of in vitro amplified DNA with sequence‐specific oligonucleotide (SSO) probes was performed for DRB1, DQA1, DQB1, DPA1 and DPB1 alleles in 54 random Norwegian rheumatoid arthritis (RA) patients and 181 healthy controls. DRB1 alleles encoding the serological specificity DR4 were found in 80% of the patients, compared to 34% of the controls (relative risk = 7.9, p < 0.0001). All DR4‐positive RA patients carried either DRB1*0401 (Dw4), 0404 (Dw14), or 0405 (Dw15), while no patients were found to carry DRB1*0402 (Dw10) or 0403 (Dw13). The frequency of the DRB1*0101 allele encoding DR1 was not increased, even among DR4‐negative RA patients, and we were unable to detect any sharing of other class II alleles among DR4‐negative patients. No contribution of any DQA1, DQB1, DPA1 or DPB1 alleles to RA susceptibility could be detected. The results suggest that in the Norwegian population RA is primarily associated with a shared sequence at residues 67‐74 of the DRβ1 chain, but only when this sequence is expressed on DR4 molecules.
Absence of DRwl5/3 and of DRwl5/7 heterozygotes in Caucasian patients with systemic lupus erythematosus
Authors: Reinharz Daniel , Tiercy Jean‐Marie , Mach Bernard , Jeannet Michel
Published: October 2008

Abstract: Polymorphic MHC class II molecules determine immune responsiveness towards pathogens and also contribute to susceptibility or resistance to a number of different autoimmune diseases, including systemic lupus erythematosus (SLE). The HLA‐DR and ‐DQ alleles of 52 patients with SLE were analyzed by serology and, for 42 patients, HLA‐DRB1, ‐B3 and DQB1 allelic polymorphism was determined by oligotyping on PCR‐amplified DNA. While we confirm the increase of DR3 (44.2% versus 16% in controls; p < 0.001) reported by others, we observed a complete absence of DRwl5(2)/DR3 and DRwl5(2)/DR7 heterozygotes among Caucasian patients. Moreover HLA‐DQB1 oligotyping revealed the absence of DQB 1*0602/0201 heterozygotes in our panel of Caucasoid SLE patients. Since both DR3 and DR7 haplotypes share the same DQB 1*0201‐encoded DQβ chain, and since DRwl5 is known to be in linkage disequilibrium with DQA1*0102, it can be predicted that DQA1*0102/DQB1*0201 combinations are absent in Caucasian patients. We therefore propose that a DQA 1*0201/DQB 1*0201‐encoded HLA‐DQ transdimer formed in these heterozygotes might function as a supressor‐inducer molecule that confers resistance against SLE.
Partial DNA sequences derived from two previously unknown HLA class I genes
Authors: Hansen Torbjsrn , Markussen Gunnar , Paulsen Gunnar , Thorsby Erik
Published: October 2008

Abstract: Genomic DNA of HLA class I gene segments was amplified by the polymerase chain reaction (PCR). The amplified 0.8 kb gene segment encompassed sequences corresponding to an exon 2 (α1 domain), an intron 2 and an exon 3 (α 2 domain). The PCR product was cloned in M13 and sequenced. Two previously unknown sequences were found. One of them (DAN2) had an open reading frame and intact intron splice sites, but we did not find evidence for transcription. Best homology (87.5%) was found with HLA‐BeWo C,1, a recently described HLA‐C gene. The other sequence (DAN4) is derived from a pseudogene, because the putative 3′ splice site of intron 2 was changed from AG to ATG and the sequence corresponding to exon 3 had a shift in reading frame resulting in a stop codon.
Recognition of monocyte‐associated antigens in the dog
Authors: Krumbacher K. , Happel M. , Grosse‐Wilde H.
Published: October 2008

Abstract: Canine pregnant sera were investigated for monocyte‐specific cytotoxic antibodies against a cell panel from 50 unrelated dogs. Contaminating DLA and DLB reactivity was removed by absorption on a random pool of purified lymphocytes. Two monocyte‐associated antigens were recognized by two independent clusters of antisera; the existence of further antigens is suggested. In preliminary segregation studies, the inheritance of these determinants in linkage with DLA could not be observed.
Activation of peripheral CD8+ T lymphocytes via CD28 plus CD2: Evidence for IL‐2 gene transcription mediated by CD28 activation
Authors: Carabasi Matthew H. , DiSanto James P. , Yang Soo Young , Dupont Bo
Published: October 2008

Abstract: It is well established that peripheral CD8+ and CD4+ T cells display different requirements for in vitro activation by mitogenic mAb. Most CD4+ T cells can be activated by anti‐CD3 or mitogenetic combinations of anti‐CD2. In contrast, CD8+ T cells display minimal responses to CD3 activation, and no proliferation is observed via CD2 activation. Purified peripheral blood CD8+ T cells, stringently depleted of APC, have been studied for their capacity to respond to mAb directed against CD3, CD2 and CD28, used alone or in combination. It is demonstrated that proliferation can be induced by co‐stimulation of CD2 and CD28. This does not require autologous APC. CD8+ T cells can also be activated by the combination of anti‐CD3 plus anti‐CD28 in the presence of APC, but only minimal cell proliferation is obtained in the absence of APC. The response via CD2 plus CD28 is IL‐2‐dependent, as demonstrated by the ability of mAb against the IL‐2 receptor to block proliferation, and is almost completely inhibited by cyclosporine A (CsA). These results suggest that the signal generated by stimulation of CD28 in combination with CD2 differs from that seen with CD28 activation combined with either PMA or CD3. Induction of IL‐2 gene activation in CD8+, CD28+ peripheral T cells may therefore require additional “second signals”, which are not necessary for activation of CD4+ cells. One such signal might be the interaction between CD28 and its natural ligand.