bluuebunny/crossref_metadata_2025 huggingface dataset:
AN EXTERNAL STIMULUS THAT MIMICS Mls LOCUS RESPONSES
Authors: Janeway C. A. , Chalupny J. , Conrad P. J. , Buxser S.
Published: April 2007

The response to a novel set of T cell mitogens has been analysed and compared to the response to Mls locus differences. These polyclonal T cell activators, staphylococcal enterotoxins A and B, stimulate T cells in a way that requires an antigen‐presenting cell bearing class II MHC products and involves the CD4:T cell receptor complex. However, the specificity of MHC recognition by the T cell receptor is lost in this response. Thus, these mitogens produce a response with characteristics similar to that induced by Mls locus differences. These mitogens can be used to analyse the immunobiology of this response, and may help in understanding and identifying the Mls locus product as well.
Mls AND THE HELPER T‐CELL REPERTOIRE: II. does theMlsgene product influence the formation of the T‐cell repertoire? studies withMlstolerance
Authors: Hoffmann M. K. , Chun M. , Dennig D. , Hämmerling U.
Published: April 2007

In the formation of a repertoire, T cells are selected through a window of autoreactivity which is defined by a low boundary representing the minimal autoreactivity required to enter the T‐cell compartment and a high boundary which determines the highest admissible autoreactivity. We propose that the Mls gene product down regulates autoreactivity and thus modifies the formation of the T‐cell repertoire. Given the polymorphism of Mls and the assumption that Mlsb is more inhibitory than Mlsd, it is conceivable that Mlsb (responder) mice admit into their T‐cell compartment T cells with higher autoreactivity than Mlsd mice. We suggest that it is this highly autoreactive fraction of Mlsb T cells which responds in the overt Mls response. We show that Mls tolerance eliminates T cells responding in the overt Mls response but not T cells responding in the latent Mls response. This is consistent with the finding that T cells responding in the latent Mls response occur in Mlsb as well as in Mlsd mice.
COMPLETE CODING REGION SEQUENCE OF Eβk cDNA CLONES: LACK OF POLYMORPHISM IN THE NH2‐TERMINUS BETWEEN Eβk AND Eβb MOLECULES
Authors: King L. B. , Sharma S. , Corley R. B.
Published: April 2007

Sequence analysis of two independently isolated Eβk cDNA clones revealed that the Eβk molecule is identical to the Eβb molecule at the NH2‐terminus. These data resolve a discrepancy between previously published amino acid and nucleotide sequences and indicate that the Eβ NH2‐terminus is not as polymorphic as was once believed.
ALPHA‐1‐ANTITRYPSIN (Pi) TYPES IN MULTIPLE SCLEROSIS AND LACK OF INTERACTION WITH IMMUNOGLOBULIN (GM) MARKERS
Authors: Francis D. A. , Klouda P. T. , Brazier D. M. , Batchelor J. R. , McDonald W. I. , Hern J. E. C.
Published: April 2007

Susceptibility to multiple sclerosis is thought to involve several genetic loci apart from the HLA region on chromosome 6. No Pi allotype nor phenotype was associated with the disease in 125 unrelated patients from the Grampian region of northeast Scotland. Despite a previously reported Gm‐HLA association in this population there was no apparent interaction between Pi alleles and Gm type which influenced disease susceptibility.
A MONOCLONAL ANTIBODY, R1, AND A POLYCLONAL SERUM, S10, RECOGNIZE THE SAME MOLECULES: A NOVEL USE OF DNA TRANSFECTANTS
Authors: Banting G. , Mondello C. , Hope R. , Goodfellow P.
Published: April 2007

The monoclonal antibody R1 defines the product of the MIC5 locus which has been localized to the long arm of the human X chromosome in the same region as the fragile site FRAXA. DNA transfectants, selected by R1 on the fluorescence‐activated cell sorter (FACS), have been used to demonstrate that a previously described polyclonal antiserum, S10 (Buck & Bodmer, 1976), recognizes the same cell surface molecule as R1. Immunoprecipitation from metabolically and surface‐labelled cells has shown that this molecule is a Mr 200 K phosphoprotein which is synthesized as a Mr 180 K precursor and subsequently modified to a Mr 195 K extracellular form. A Mr 150 K molecule is coprecipitated with the Mr 200 K phosphoprotein, but is only detected in lysates of surface‐labelled cells, raising the possibility that the 200 K molecule is a cell surface receptor and the 150 K molecule a ligand.
ROLE OF MHC, Mls AND TCR IN IMMUNE TOLERANCE
Authors: Anderson G. , David C.
Published: April 2007

MHC class II molecules and self antigens, such as Mls, influence T‐cell selection by clonal deletion of potentially self‐reactive T cells. In order to examine the role of various class II molecules in the T‐cell receptor‐self antigen interaction, class II transgenic and recombinant mice were analysed for TCR expression. Our studies indicate that the Aα and Eα chains can present Mls gene products for the clonal deletion of Vβ6‐bearing T cells, and that the Aαq chain is defective in this process. We have also shown that Eα Aβ heterodimer in transgenic and recombinant mice is expressed and functions to delete I‐E reactive Vβ11 T cells, demonstrating again the role of the Eα molecule.
Qa‐12—‐A NOVEL DETERMINANT OF ACTIVATED T AND B LYMPHOCYTES
Authors: Oudshoorn‐Snoek M. , Demant P.
Published: April 2007

Investigation of the antigenic phenotype of activated lymphocytes using the broadly cross‐reactive mAb 6.3.4 revealed two phenotypic alterations as compared with the resting lymphocytes. The Qa specificity Qa‐m208 disappears after lectin activation of Qa‐m208‐positive T lymphocytes. Analysis of Q7 and Q9 transfectants expressing the Qa‐2 polypeptides shows that Qa‐m208 is an epitope of the Qa‐2 antigen. Because the Qa‐2 antigens are still expressed on T lymphoblasts which have lost Qa‐m208, changes of the Qa‐2 molecules occur and result in the loss of certain epitopes.

The second phenotypic change that we observed is the appearance of a novel specificity, Qa‐12. Its expression is induced by lymphocyte activation and it is expressed on lymphoblasts of both T and B cell origin. The presence of this novel non‐ubiquitous antigenic specificity is determined by the Tla region.
ORGANIZATION OF THE AKR Qa REGION: STRUCTURE OF A DIVERGENT CLASS I SEQUENCE, Q5k
Authors: Weiss E. H. , Bevec D. , Messer G. , Schwemmle S. , Großhaus C. , Steinmetz M. , Schmidt W.
Published: April 2007

We established the organization of the AKR Qa region and determined the sequence of the 44 and Q5 genes. Restriction mapping and genomic Southern blot analysis revealed that the AKR strain codes for only three H‐2K homologous genes in this region. The AKR Q5 gene is not homologous to the Q5 gene of the C57BL strain, but is presumably allelic to the Q5 gene isolated from Balb/c. The organization and structure of the AKR Qa family is virtually identical to the Qa genes of the C3H mouse. The AKR Q5 gene, in contrast to other H‐2K homologous Qa region genes, codes for a typical transmembrane region, and upon transfection into BHK cells, a 1.6 kb Q5 transcript is detected.
H‐2Kb TRANSFECTION OF B16 MELANOMA CELLS RESULTS IN REDUCED TUMOURIGENICITY AND METASTATIC COMPETENCE
Authors: Porgador A. , Feldman M. , Eisenbach L.
Published: April 2007

The metastatic B16 mouse melanoma shows a low cell surface expression of H‐2Kb and H‐2Db class I antigens on cells of both the high‐metastatic line B16‐F10 and the low‐metastatic line B16‐F1. Similarly, newly generated clones of these lines, having different metastatic properties, all express low levels of major histo‐compatibility antigens. One of these clones, the high‐metastatic F10.9, was transfected with H‐2Kb genes to generate H‐2Kb‐expressing transfectants. The resulting clones showed reduced tumourigenicity and a low metastatic phenotype. Unlike the parental cells, H‐2Kb‐positive transfectants are potent inducers and sensitive targets of H‐2Kb‐restricted syngeneic cytotoxic T cells. Immunization of mice with H‐2Kb‐positive transfectants conferred protection against a subsequent challenge with Kb‐positive transfectants but had only a small effect on growth and metastatic spread of parental cells.
NK SENSITIVITY, H‐2 EXPRESSION AND METASTATIC POTENTIAL: ANALYSIS OF H‐2Dk GENE TRANSFECTED FIBROSARCOMA CELLS
Authors: Gopas J. , Rager‐Zisman B. , Har‐Vardi I. , Hammerling G. J. , Bar‐Eli M. , Segal S.
Published: April 2007

We have used the 3‐Methylcholanthrene induced T‐10 fibrosarcoma tumour cell system (H‐2b xH‐2k)F1 to elucidate the possible correlation between metastatic potential, expression of individual H‐2 antigens and susceptibility to NK cells.

Transfection of the non‐metastatic and NK sensitive IC9 cells (Db+, Dk‐, Kb‐, Kk‐) with the H‐2Dk gene, altered the metastatic phenotype of the parental cells, yet had no effect on the susceptibility of these tumour cells to lysis by NK and did not elicit a specific CTL response in syngeneic hosts. Variants of the metastatic and NK resistant IE7 clone (Db+, Dk+, Kb‐, Kk‐), lacking H‐2Dk, were selected by treatment with monoclonal anti H‐2Dk antibodies and complement. These variants were sensitive to NK and poorly or non metastatic. Retransfection of ‘Dk′loss’ variants with the H‐2Dk gene, resulted in the isolation of several clones expressing a wide range of metastatic phenotypes but maintained sensitivity to NK. These results indicate that the H‐2D region of the MHC and or closely linked genes may be involved in the complex interrelationship between target susceptibility to NK and metastasis.
GENERATION OF SYNGENEIC ANTI‐TUMOUR DOUBLE NEGATIVE CELLS (CD4− CD8−), WITH CYTOTOXIC ACTIVITY AGAINST CLONES OF DIFFERENT MHC CLASS I EXPRESSION.
Authors: Caballero A. , Garrido A. , Algarra I. , Perez M. , Garrido F.
Published: April 2007

In vivo immunization and secondary culture techniques were used to generate cytotoxic T lymphocytes against a tumour cell clone obtained from a methylcholantrene‐induced fibrosarcoma. Our CTLs differed from classical CTLs (Ly2 + and MHC restricted) in that our lymphocytes exhibited a non‐MHC restricted cytotoxic activity directed to the original tumour and to other MCA‐induced tumours, but have failed to demonstrate any killing activity against a wide range of tumour cell lines of diverse origin (including NK sensitive targets). Depletion and phenotypic studies demonstrated that these cells bear the Thy1.2 antigen but are negative for both Ly2 and L3T4 antigens. These CTLs may belong to a double negative subset (CD4−, CD8−) involved in the anti‐tumour response.
RADIATION LEUKEMIA VIRUS AND ITS EFFECT ON H‐2 GENE EXPRESSION
Authors: Brown G. D. , Meruelo D.
Published: April 2007

In this report we demonstrate that lowered expression of the H‐2 antigens on RadLV‐induced tumour cells is a result of depressed levels of stable mRNA in these cells. Whether this observation is a result of lowered transcription or of mRNA instability is under investigation. In an effort to determine which viral sequences are essential for mediating both the H‐2 regulatory function and the transforming function of RadLV, we have begun to assemble newly integrated proviral genomes from tumours. The restriction enzyme cleavage sites of four isolates are presented; these isolates differ substantially from RadLV genomes previously presented. One of these molecular clones is shown to encode a non‐defective B‐tropic, ecotropic virus which when reinjected into resistant mouse strains can mediate the up‐regulation of H‐2Dd antigen expression. Finally, possible mechanisms of H‐2 regulation are discussed.
FUNCTIONAL STUDIES OF H‐2k‐LIKE EPITOPES ON DTIC TREATED AND UNTREATED L1210 (H‐2d) CLONES
Authors: Marelli O.E.A. , Franco P. , Labeta M.O. , Festenstein H.
Published: April 2007

Following ‘in vivo’ treatment with 5‐(3–3′‐dimethyl‐l‐triazeno)imidazole‐4‐carbox‐amide (DTIC), murine leukemic cells acquire new antigenic specificities not detectable on parental cells and responsible for the rejection of the tumour by syngeneic hosts.‘In vivo’ and ‘in vitro’ experiments pointed out an immunological cross reactivity between DTIC treated and untreated lines. Furthermore, specific CTLs raised against DTIC treated L1210 tumour cells (H‐2d) were cytotoxic for H‐2k target cells. The aim of this study is to investigate whether the H‐2k cross reactivity displayed by L1210/DTIC is related to the drug treatment rather than due to an antigen already present in the parental line and maintained after treatment. Cloned cells from L1210, obtained by limiting dilution ‘in vitro’, were recloned ‘in vivo’ and then treated with DTIC. Syngeneic and allogeneic CTLs raised ‘in vitro’ against parental and treated clones showed lytic activity against H‐2k target cells. Treated and untreated clones were then checked for the presence of H‐2k‐like determinants using monoclonal antibodies. One of these, HB‐53 (IgG2bKkDk was highly positive with all the clones tested in binding assay using iodinated Fab anti‐mouse Ig, fluorescence and FACS analysis. Others displayed a low reactivity against both treated and untreated clones without significant differences. After neuraminidase treatment of two clones (D and D/DTIC), the H‐100.5 (anti H‐2Kk)‐reactive epitope was dramatically exposed mi the DTIC tumour cells but not on the parental clones. These data suggest that the H‐2k cross reactivity is related to the presence of a TAA that is maintained after treatment. Nevertheless DTIC could enhance the expression of some H‐2k‐like determinants. So far it is not possible to rule out that this phenomenon can contribute to increase the antigenicity of DTIC clones.
MHC CLASS I AND II ANTIGENS ON GASTRIC CARCINOMAS AND AUTOLOGOUS MUCOSA
Authors: Ferron A. , Perez‐Ayala M. , Concha A. , Cabrera T. , Redondo M. , Oliva M. R. , Ruiz‐Cabello F. , Garrido F.
Published: April 2007

The expression of HLA class I and II antigens was analysed in 30 primary gastric carcinomas, 27 autologous lymph node metastases and 25 autologous gastric mucosae. We used an immune alkaline phosphatase technique on cryostatic sections and mAbs directed against HLA class I monomorphic determinants, HLA‐B locus‐specific products and HLA‐DR, ‐DP and ‐DQ molecules. In addition HLA class I genes were analysed in tumour tissue and compared by Southern blots with the RFLP from autologous mucosa using locus‐specific HLA probes. Finally the infiltrating mononuclear cells were studied on gastric tumours and adjacent mucosa with mAbs defining CD4, CD8 and CD11b differentiation antigens.

The results obtained showed that three out of 27 primary gastric carcinomas completely lack HLA‐ABC antigens (10%). In addition, two primary tumours presented a variable expression. The remaining 22 tumours presented a homogeneous positive HLA class I expression. Interestingly, when the autologous mucosa was analysed, only 12 out 25 specimens were homogeneously stained with mAbs against HLA class I antigens, suggesting that this tissue may lack the expression of HLA antigens before becoming malignant. Indeed, the majority of the gastric carcinomas studied presented a higher HLA‐ABC antigenic expression than autologous mucosa. Finally, the HLA expression observed in the primary tumour was similar to that observed in autologolous metastases.

As a second part of the study we have found a direct relationship between the expression of HLA‐DR antigens in mucosa and the intensity of inflamatory infiltration. This relationship was not maintained in the tumour tissue. In the mucosa the CD4‐positive T cell was the predominant lymphocyte, while it was CD8 in the HLA‐DR‐positive tumours.
TNF‐alpha GENE POLYMORPHISMS: ASSOCIATION WITH TYPE I (INSULIN‐DEPENDENT) DIABETES MELLITUS
Authors: Badenhoop K. , Schwarz G. , Bingley P. , Trowsdale J. , Usadel K. H. , Gale E. A. M. , Bottazzo G. F.
Published: April 2007

The localization of TNF genes on the short arm of chromosome 6 between HLA B and the complement genes focused attention to that genetic region which harbours many immunologically relevant genes and is also thought to hold susceptibility genes for a variety of autoimmune diseases that are linked to specific alleles of particular loci in the HLA D region. Since the recently established HLA‐DR‐DQ variation accounts only for part of the genetic susceptibility to insulin‐dependent diabetes mellitus (IDDM) we searched for genomic variation of the tumour necrosis factor (TNF) alpha. We have identified a TNF‐alpha restriction fragment length polymorphism (RFLP) with N coI and analysed diabetic patients including their families, controls and homozygous typing cell lines (HTC) defined by the 10th International Histocompatibility Workshop. Segregation analysis in families and HTC results show a strong linkage of the TNF‐alpha 5.5 kb allele with DR types in particular with AIB8DR3. This tight linkage of TNF‐alpha alleles with extended haplotypes and the significant increase of heterozygotes in patients could lead to some explanation of the DR3 association with a variety of autoimmune diseases particularly IDDM.
HLA‐DP ANTIGENS AND HTLV‐1 ANTIBODY STATUS AMONG JAPANESE WITH MULTIPLE SCLEROSIS: EVIDENCE FOR AN INCREASED FREQUENCY OF HLA‐DPw4.
Authors: ØDum N. , Saida T. , Ohta M. , Svejgaard A.
Published: April 2007

Previously, an association between multiple sclerosis (MS) and HLA‐DPw4 has been reported in Scandinavians. In the present study, the distribution of HLA‐DP antigens was studied in 34 Japanese MS patients, a11 of whom fulfilled the criteria for definite MS. HLA‐DP typings for DPwl through w6 and the local specificity, CDP‐HEI, were performed using the primed lymphocyte typing (PLT) technique. In addition, the patients were typed for a DR2 +, Dw2 +/Dw12–related, PLT defined specificity. The distribution of DPw1‐w5 in 121 healthy, unrelated Japanese controls were from Nishimura et al., 1984; Nishimura, personal communication). Sera from all 34 patients and 38 controls (both from the HTLV‐1 non‐endemic, Kyoto region) were examined for the presence of HTLV‐1 reacting antibodies by a highly sensitive radioimmuno assay (RIA) using two sources of HTLV‐1 antigens, namely total crude protein preparations from disrupted HTLV‐1 virions and affinity purified p24 HTLV‐1 core proteins.

The frequency of DPw4 was significantly increased to 35.3% in Japanese MS patients compared to 16.5% in controls (Relative Risk, RR = 2.8, p = 1.9 × 10−2). 41.6% of the MS patients gave clear typing responses with a PLT reagent which recognized a Dw2+ related specificity, which is higher than the frequency of Dw2 (6.8%) in Japanese.

Fourteen of the 34 patient sera contrasting to none of the sera from 38 controls contained antibodies of IgG and/or IgM subclasses reacting with the HTLV‐1 derived antigens. This difference is highly significant (P < 1 × 10−5). However, there was no association between DP or Dw2 phenotypes and antibody status.

These data provide evidence that genes within (or closely linked to) the HLA‐DP region confer susceptibility to MS in two genetically different ethnic groups: Japanese and Scandinavians.
VARIANTS OF Mta, THE MOUSE MATERNALLY TRANSMITTED ANTIGEN, DETERMINED BY THREE ALLELES OF THE EXTRACHROMOSOMAL GENE Mtf
Authors: Lindahl K. Fischer , Hausmann B. , Guénet J.‐L.
Published: April 2007

Two new forms of Mta, the maternally transmitted antigen of the mouse, were defined by cytotoxic T lymphocytes. One is found in laboratory mice carrying the β allele of the maternally transmitted factor, Mtf, which were previously thought to be Mta‐negative. The other is associated with a new allele, Mtfγ, and is found in wild mice from Toulouse. Like the common form of Mta, determined by the extrachromosomal Mrfα, the new forms are also dependent on the α allele of Hmt, a chromosomal gene linked to H‐2. and are associated with β2‐microglobulin. Killers could be raised in all six combinations of a, β, and γ mice, showing that each form of Mtf determines a distinct (set of) epitope(s). We propose that the product of the Hmt gene, a class I MHC antigen, presents a peptide derived from a mitochondrial gene product on the cell surface and thereby creates the target antigen, Mta.
POST‐TRANSLATIONAL POLYMORPHISM OF HUMAN IgA IDENTIFIED BY IMMUNOISOELECTROFOCUSING
Authors: Benedictis G. De , Rose G. , Brancfti C.
Published: April 2007

Immunoisoelectrofocusing (IIEF) reveals a microheterogeneity of human serum IgA controlled by an autosomal polymorphic gene, termed S. The microheterogeneity disappears when sialic acid is removed from serum glycoproteins by neuraminidase treatment. It can be postulated, therefore, that S encodes a sialyltransferase which attaches sialic acid at the outer prosthetic chains of IgA.
IDENTIFICATION OF A RECOMBINATIONAL SIGNAL SEQUENCE‐SPECIFIC DNA‐BINDING PROTEIN(S) OF Mr 115,000 IN THE NUCLEAR EXTRACTS FROM IMMATURE LYMPHOID CELL LINES
Authors: Miyake S. , Sugiyama H. , Tani Y. , Fukuda T. , Kishimoto S.
Published: April 2007

Rearrangements of immunoglobulin genes are mediated by highly conserved heptamer and nonamer recombinational signal sequence. Using a protein‐blotting procedure, a heptamer and nonamer recombinational signal sequence‐specific DNA‐binding protein(s) was examined in the nuclear extracts from lymphoid and nonlymphoid cell lines. Nuclear extracts were subjected to SDS‐polyacrylamide gel electrophoresis, and transferred by electroblotting to nitrocellulose filters. Then the filters were hybridized to *P‐labelled synthetic double‐stranded heptamer‐23bp‐nonamer or nonamer‐ l2bp‐heptamer recombinational signal sequence probes. A relatively large amount of a DNA‐binding protein(s) of M, 115,000 for both probes was detected in the nuclear extracts from immature B and immature T cell lines. No DNA‐binding proteins were detected in a myeloma cell line. Interestingly, this DNA‐binding protein(s) might be able to recognize both heptamer and nonamer. Recombinational signal sequence‐specific DNA‐binding activity of the protein(s) and the presence of the protein(s) in a stage‐specific manner strongly suggest that the protein(s) of M, 115,000 detected here may play an important role in the recombination of Ig and TCR genes.
BIOCHEMICAL ANALYSIS OF THE RAT MHC CLASS I ANTIGENS. RT1.Aa, RT1.Fa and Pa
Authors: Misra D. N. , Kunz H. W. , Saito M. , Gill T. J.
Published: April 2007

In DA strain rats, there are two other MHC class I loci (Pa and RT1.Fa) in the vicinity of the classical class I locus RT1.Aa. The Pa antigen is the pregnancy‐associated antigen, and it was detected by antibodies elicited in WF females pregnant by DA males without any other immunization. The Fa antigen was detected by a monoclonal antibody raised by alloimmunization. In the present work, the Aa, Fa and the Pa antigens have been compared by HPLC peptide mapping and by isoelectric focusing after their isolation by appropriate monoclonal antibodies. All the three antigens are identical in primary structure with respect to lysine, methionine, asparagine and the aromatic amino acid residues, but they differ from one another with respect to glutamic acid and/or aspartic acid residues. The PI values of the antigens differ slightly. All three antigens have two identical N‐linked glycans, but the Fa antigen has an additional N‐linked glycan. Based on the available amino acid sequence of the Pa antigen, it can be concluded that both Aa and Pa antigens are devoid of glycosylation in the second domain. This lack of glycosylation of the classical antigen Aa is unique for the rat, since classical class I antigens of the mouse show glycosylation in the first and second, and sometimes in the third domain, and those in the human, in the first domain only. The high degree of similarity among the Aa, Fa, and Pa molecules that this study indicates is also unique for the rat, since antigens encoded by different class I genes of the same haplotype are quite disparate in the mouse and human.
EXPRESSION OF I‐Aαk AND ALLELIC RESTRICTION ON Aα/Aα PAIRING IN TRANSGENIC MICE
Authors: Martin J. , Wei B‐Y. , Savarirayan S. , David C. S.
Published: April 2007

Cell surface expression of class II major histocompatibility complex encoded (la) molecules depends on association of the component α and β chain into a stable heterodimer. Studies on cell lines transfected with MHC class II genes have revealed important limitations on the assembly of haplotype‐mismatched Aα:Aβ complex. In order to study α: β chain pairing restrictions in vivo a number of lines of transgenic mice carrying the Aαk gene were generated. The transgene was studied in the context of H‐2b, H‐2s, H‐24, H‐2u, H‐2f and H‐2v haplotypes. Initial FACS analysis of spleen and peripheral blood cells showed no Aαk expression. Spleen cells stimulated with LPS and analysed by FACS showed Aαk expression in three different H‐2b strains as well as H‐2u, but not in others. These data indicate that Aαk can associate with Aβb and Aαk, but not with Aβs,Aβq, Aβf, Aβv.
MOLECULAR ANALYSIS OF TWO RECOMBINANT MOUSE STRAINS WITH CROSSOVERS IN THE Eα RECOMBINATION HOT SPOT
Authors: Lafuse W. P. , Lee S. T. , David C. S.
Published: April 2007

Recombination in the Eα gene of the mouse major histocompatibility complex has been found to occur only in mice derived from crosses between strains with the k and p haplotypes. In the present paper the crossover sites of six additional strains were examined by RFLP analysis. These recombinant strains were derived from crosses between strains (B10.RPD1, B10.RPD2) with crossovers in the Eα gene and B10.M (H‐2f). Four have crossover sites between the Eα gene and the S region. One recombinant (B10.RFD7) also crossed over within the Eα gene and a second (B10.RPF1) crossed over within a segment located to the left of the Eα gene. This study suggests that the DNA sequence responsible for recombination in the Eα gene is also present in the B10.RPD1 and B10.RPD2 recombinant strains.
HLA IN JUVENILE DERMATITIS HERPETIFORMIS: CLINICAL HETEROGENEITY CORRELATED WITH DNA AND SEROLOGICAL POLYMORPHISM
Authors: Amoroso A. , Mazzola G. , Canale L. , Borelli I. , Dallomo A. M. , Curtoni E. S. , Ansaldi N. , Fusco P. , Elia G. , Barbera C.
Published: April 2007

A group of 30 Italian children affected by Dermatitis Herpetiformis (DH) was analysed for HLA region polymorphisms with both serological and DNA methods. Serological typing was performed on HLA‐A, B, C, DR, DQ antigens and C4A, C4B, Bf polymorphisms. DNA RFLPs obtained with TaqI enzyme were investigated with cDNA probes specific for DR beta, DQ alpha and DQ beta genes. The results were correlated with intestinal involvement and age at onset of the disease. The following observations were made: (1) the intestinal biopsies revealed a direct correlation between degree of lesions and age at onset of DH; (2) a significantly increased relative risk (RR) was found for the following HLA antigens: A1 (RR = 2.2), B8 (RR = 6.2), Cw7 (RR = 3.9), C4AQO (RR = 7.4), DR3 (RR = 5.2), DR7 (RR = 4.4), DRw53 (RR = 4.7), DQw2 (RR = 6.0); (3) B8 and DR3 were significantly more frequent in patients with severe intestinal lesions; and (4) of the two DR3 subtypes revealed by RFLP typing, only 3.1 showed an increased frequency in DH patients (RR = 8.4). It is suggested that the susceptibility to Juvenile DH is determined by the same genes, within the HLA region, that are associated with Coeliac Disease.
THE EUROTRANSPLANT HLA‐DRB OLIGONUCLEOTIDE TYPING SET
Authors: Giphart M.J. , Verduijn W.
Published: April 2007

Several sequence‐specific oligonucleotides (SSO) were designed based upon available sequences of HLA‐DRB 1 genes which are associated with HLA‐DR serotypes. Targets for these SSO were polymerase chain reaction (PCR) products using primers specific for the second exon of DRB genes. In view of practicality, a hybridization system was developed which was independent of temperature or composition of the hybridization buffers, hence uniformly applicable to all SSO, yet allowing discrimination based upon one nucleotide mismatch only. The SSO typing results were compared with HLA serological typing and a high correlation was observed (99%).
HLA‐DRB AND DQB TYPING BY A COMBINATION OF SEROLOGY, RESTRICTION FRAGMENT LENGTH POLYMORPHISM ANALYSIS AND OLIGONUCLEOTIDE PROBING
Authors: Doherty D.G. , Donaldson P.T.
Published: April 2007

DNA sequencing has revealed extensive polymorphism within the HLA‐DRB and DQB genes of the major histocompatibility complex. At least 43 alleles of the DRB loci and 13 alleles of the DQB1 locus are currently recognized. Identification of these alleles can be performed phenotypically by cellular and serological techniques or genotypically by restriction fragment length polymorphism (RFLP) analysis and sequence‐specific oligonucleotide (SSO) probing of DNA amplified by the polymerase chain reaction (PCR). However, each method has its technical limitations so the tissue typing laboratory must therefore choose from the range of available techniques to maximize the efficiency and accuracy of HLA typing. In this paper we describe a scheme for the accurate determination of all the serologically defined DRB and DQB allotypes using a combination of serology, RFLP analysis and PCR‐SSO probing. The efficiencies of serology versus RFLP analysis for the DR typing of 800 individuals, and RFLP analysis versus PCR‐SSO probing for the DQB typing of 317 individuals, are compared and the merits of each technique discussed. This scheme, which types for both HLA‐DRB and DQB with an accuracy approaching 100%, is now routinely employed in all HLA studies of disease and transplantation in our laboratory.
COMBINED USE OF RFLP AND PCR‐ASO TYPING FOR HLA‐DR‐Dw AND DQw TYPING
Authors: BIGNON J.D. , BlDWELL J.L.
Published: April 2007

Due to some limitations of restriction fragment length polymorphism (RFLP) analysis in HLA‐DR‐DQ typing, we present a combined use of RFLP and polymerase chain reaction (PCR)‐allele‐specific oligonucleotide (ASO) typing. This scheme consists in selectively amplifying the few RFLP ill‐defined genes (DR1/DR ‘Br’ and DR4‐Dw subsets) using PCR with allele specific primers to avoid cross‐hybridization.
HLA CLASS I NUCLEOTIDE SEQUENCES, 1991
Authors: Zemmour J. , Parham P.
Published: April 2007

The HLA class I sequences included in this compilation are taken from publications listed in the accompanying paper, Nomenclature for factors of the HLA system, 1990 (Bodmer et al., 1991), and also in Nomenclature for factors of the HLA system, 1989 (Bodmer et al., 1990). 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 full point (.). Gaps in the sequence are inserted to maintain the alignment between different alleles showing variation in amino acid number.
ANALYSIS OF HLA SPECIFICITY OF HUMAN MONOCLONAL ANTIBODIES BY CYTOFLUORIMETRY AND CELL ELISA
Authors: Pistillo M. P. , Tazzari P. L. , Mazzoleni O. , Urlacher A. , Falco M. , Vitale M. , Karr R. W. , Ferrara G. B.
Published: April 2007

In this study we report on the characterization of cytotoxic human monoclonal antibodies (HmAb) detecting polymorphic HLA class II specificities using cytofluorimetric analysis in combination with micro cell ELISA. In both techniques, five anti‐HLA HmAb were tested against HLA‐transfected murine L cells as target cells and the bound antibody was detected, either by cytofluorimetry or by cell ELISA reader, after addition of fluoresceinated or peroxidase‐conjugated anti‐human IgG+IgM antibodies, respectively. The results demonstrate that HmAb directed against HLA‐DR, ‐DQ and ‐DP molecules can be efficiently discriminated by cytofluorimetry and cell ELISA, which appear to be highly sensitive and perfectly comparable to the standard cytotoxicity assay.
A DR7 SPECIFIC MONOCLONAL ANTIBODY TAL13.1, RAISED AGAINST A TRANSFECTANT DETECTS IL‐4 UPREGULATED ANTIGEN ON PERIPHERAL B‐LYMPHOCYTES
Authors: Heyes J.M. , Marsh S.G.E. , Sadler A.M. , Madrigal J.A. , Bodmer J.G.
Published: April 2007

A monoclonal antibody TAL13.1 was raised against mouse L cells transfected with the human HLA‐DRB 1*0701 gene. This antibody was found to be polymorphic recognizing a determinant expressed by the DR7, DRβ1*0701 and DRB1*0702 gene products. Four polymorphic sites unique to this specificity have been identified within the DR pi domain. These are residues 11–14, 25, 30 and 71–74, one or a combination of which is postulated as being responsible for conferring the specificity of the antibody. In Western blot analysis TALI3.1 was found to react with the DRαβ dimer, but not with the free α or β chains. However, in flow cytometry it failed to bind a DRα/DQβ mixed pair transfectant confirming that it recognizes an epitope on the DRβ not the DRα chain. Although TAL13.1, a low affinity antibody is negative or only weakly positive on resting peripheral blood lymphocytes (PBLs), we have demonstrated that by interleukin‐4 (IL‐4) stimulation we can up‐regulate the levels of antigen already present and gain a level of binding comparable to that found on B lymphoid cell lines (B‐LCLs) where it has been found to be a valuable reagent in their characterization.
COMPARISON OF TCR‐αβ SEQUENCES OF CROSS‐REACTIVE ANTI‐DR ALLOREACTIVE T‐CELL CLONES: IDENTIFICATION OF POSSIBLE CONTACT RESIDUES BASED ON CHARGE COMPLEMENTARITY BETWEEN TCR CHAINS AND DR DETERMINANTS
Authors: Champagne E. , Essaket S. , Huchenq A. , Fabron J. , Coppin H. L. , Sevin J. , Thomsen M.
Published: April 2007

We analysed alloreactive T‐cell clones selected for their differential recognition of DR variants differing in the third hypervariable region (hvr) of the DRB1 gene (amino acid positions 67–70–71). This polymorphism leads to two main hvr3 types: a basic form (Leu67‐Gln70‐Arg/Lys71) and an acidic form (Ile67‐Asp70‐Glu71) where residue 70 is probably directly accessible to the TCR on DRβ chains. The TCRs have been sequenced. Three DRw13‐reactive clones use similar Va2 and Vβ13 gene family members but differ mainly by their cross‐reactivity towards acidic or basic DR4 variants and by the sequence of CDR3 on their TCRα and/or β chains. One anti‐DRw13 clone cross‐reacts with most specificities sharing the DRw13 type of hvr3 and reciprocally one anti‐DRBon (DRB1*0103) clone cross‐reacts with DRwl3. These two clones use similar V0 genes and share negative charges in CDR2α at position 56. They also share these negative charges in CDR2α with two other clones reacting specifically with DRBon, the acidic variant of DR1. We hypothesized that the selective recognition of positively or negatively charged residues on the DR(3 chain would necessitate reciprocal charges on the TCR complementarity determining regions (CDRs) responsible for this interaction. This facilitated identification of those residues of the TCR that possibly interact with the hvr3 determinant of HLA‐DR. From these observations the mechanisms allowing the recognition of alloantigens by these T‐cell clones are discussed.
DAMAGE IN B2m GENES AND DNA METHYLATION OF H‐2 GENES ARE INVOLVED IN LOSS OF EXPRESSION OF CLASS I MHC PRODUCTS ON THE MEMBRANE OF LR.4, A CELL LINE DERIVATIVE OF THE T‐CELL LYMPHOMA L5178Y
Authors: Nava G. , Ocadiz R. , Ortega V. , Alfaro G.
Published: April 2007

We have isolated an H‐2 deficient cell line (LR.4) from the T‐cell lymphoma L5178Y which grew without restrictions in the peritoneal cavity of different inbred strains of mice. The use of polyclonal anti‐H‐2 antiserum and complement indicated that LR.4 cells did not express class I determinants on the cell membrane. Southern blots of genomic DNA of LR.4 cells showed that B2m genes were severely damaged and that class I H‐2 genes were extensively methylated. Consequently, LR.4 cells failed to transcribe mRNAs for both B2m and class I H‐2 genes. On the other hand, specific immunity to LR.4 was demonstrated in C57BL/6J mice since, in subsequent challenges with either LR.4 or EL4.4, LR.4 did not grow, whereas EL4.4 grew and killed the mice. In C57BL/6J mice, rejection of LR.4 was accompanied by the production of cytotoxic antibodies. The immune response induced in C57BL/6J mice was determined by non‐H‐2 antigenic determinants in LR.4 cells.
FINE ANALYSIS OF RECOGNITION SITES OF MONOCLONAL ANTIBODIES, 1B7 AND 1E4, USING L CELL TRANSFECTANTS OF RECOMBINANT MOUSE MHC CLASS II GENES
Authors: Itoh Y. , Ogasawara K. , Onoé K.
Published: April 2007

Recognition sites of two monoclonal antibodies (mAbs), 1B7 and 1E4, specific for murine MHC class II were determined, using eleven L cell transfectants which express recombinant class II (I‐A) molecules. 1B7 positive cells possessed a haplotype k in the α‐helix of the β‐chain, whereas 1E4 positive cells had haplotype b at the α‐helix of the β‐chain. The α‐helix of the β‐chains is regarded as an important site for antigen binding. Thus, the 1B7 and 1E4 mAbs may be useful for analysis of functional sites on class II molecules involved in antigen recognition.
HLA CLASS II NUCLEOTIDE SEQUENCES, 1992
Authors: Marsh S. G. E. , Bodmer J. G.
Published: April 2007

The HLA Class II sequences included in this compilation are taken from publications listed in the papers: Nomenclature for factors of the HLA system, 1991 (Bodmer et al., 1992), Nomenclature for factors of the HLA system, 1990 (Bodmer et al., 1991) and Nomenclature for factors of the HLA system, 1989 (Bodmer et al., 1990). 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.
QUANTITATION OF HLA CLASS II ANTIGENS ON B‐CELL LINES
Authors: BUX J. , SPENGEL U. , MUELLER‐ECKHARDT G.
Published: April 2007

By quantitation of elutable immunoglobulin G in a recently described elisa, the number of HLA class I and II molecules on B‐cell lines was determined using monomorphic monoclonal antibodies. An average number of 183 000 binding sites per cell for HLA class I‐, 99000 for HLA‐DR‐, 63000 for DQ‐ and 42000 for DP‐specific monoclonal antibodies were determined. The small amount of HLA class II molecules can in part explain the difficulties observed in HLA class II typing or in detection of class II antibodies using the lymphocytotoxicity text.
HLA CLASS II POLYMORPHISM AND IDDM SUSCEPTIBILITY IN THE GREEK POPULATION
Authors: Khalil I. , Spyropoulou M. , Mallet C. , Loste M. N. , Douay C. , Laperriere J. , Bartzokas C. , Lepage V. , Charron D. , Stavropoulos C.
Published: April 2007

The frequencies of HLA‐DQA1, DQB1 and DRB1 alleles were compared between 50 Insulin‐Dependent Diabetes Melitus (IDDM) patients and 49 healthy controls in the Greek population. Statistically significant difference in the frequencies of HLA‐DQA1*0501‐DQB1*0201 (P = 10‐4), DQA1*0301‐DQB1*0201 (P= 0.01) and DQA P0301‐DQB 1*0302 (P= 0.001) were observed. The DRB1*0405‐DQA1*0301‐DQB 1*0201 was the only DR, DQ combination significantly associated with the disease. The unexpected increase of DRB1*0405 observed in the Greek IDDM may suggest as reported in Chinese and Japanese IDDM a contribution of DRβ and DQα in susceptibility. Moreover, in contrast to the Asians, in the Greek, the DRβ, DQα are found with the usual DQβ 57‐ve.
T‐CELL RECEPTOR α, δ, AND γ CHAIN GENES IN INSULIN‐DEPENDENT DIABETES MELLITUS
Authors: Martianez‐Naves E. , Penta M. , Loapez‐Larrea C.
Published: April 2007

We have studied the genotypic, haplotypic, and allelic distribution of germline Restriction Fragment Length Polymorphism (RFLP) of T‐cell receptor (Tcr) α, γ, and δ loci in 75 insulin‐dependent diabetes mellitus (IDDM) patients and 84 healthy blood donors as control population. The restriction endonuclease PvuII produces three allelic fragments of Tcr C‐γ (TcrCG) gene segment of 16,13, and 11.3 Kb respectively. Our observations revealed that PvuII/TcrCG RFLP allelic distributions were not significantly different in the IDDM and the control group. However, 85% of IDDM patients carried HLA DR3 and/or DR4 haplotypes, and when comparing these patients with a second group of HLA DR3+ and/or DR4+ healthy individuals, the 11Kb/PvuII fragment of TcrCG gene was found to be associated with IDDM patients (χ= 11.4, P= 0.003). 54.9% of IDDM patients carried at least one 11.3 Kb allele vs. 21% in controls (χ= 10.77, P = 0.004). No significant association was found between RFLP in Tcr, Cα, Cδ, Vγ9 loci and IDDM.
INVASIVE SQUAMOUS CELL CARCINOMA OF THE CERVIX: IS HLA‐DQ A DISEASE MARKER IN JEWISH PATIENTS?
Authors: Amar A. , Battat S. , Anteby S.O. , Brautbar C. , Reubinoff B.E.
Published: April 2007

HLA class I and class II were investigated in 30 Israeli patients with invasive squamous cell carcinoma of the cervix and compared to healthy controls. None of the studied serological specificities were found to be associated with the disease. Genomic DNA from the patients was amplified by PCR, dot‐blotted and hybridized with sequence specific oligonucleotide probes defining the known DQA1 and DQB1 allelic variants. Fifteen out of the 30 patients tested (50%) were found to carry the DQA1*0501 allelic variant, which is common in the local healthy population (67%). DQB 1*0302 was found in eight out of 30 patients (27%) while this allele was present in 17% of the healthy population, a difference which is not statistically significant. Our data indicate that there is no apparent association between invasive squamous cell carcinoma of the cervix and the HLA antigens and alleles studied including the alleles of the DQ A and DQB loci in the Israeli population. Our findings indicate that MHC genes could not be useful in the diagnosis of squamous cell carcinoma of the cervix.
ABSENCE OF STRONG HLA‐DR/DQ‐DP LINKAGE DISEQUILIBRIUM IN THE BRITISH AND FRENCH CANADIAN CAUCASOID POPULATIONS
Authors: Howell W.M. , Evans P.R. , Devereux S.A. , Sage D.A. , Smith J.L. , Haegert D.G.
Published: April 2007

HLA‐DR/DQ‐DP linkage disequilibrium was investigated in healthy, unrelated British (n= 150) and French Canadian (n= 67) caucasoid subjects. HLA‐DR and DQ typing was performed by Taq I DNA‐RFLP analysis, while DPB1 typing was performed by PCR‐SSOP. χ2 and Fisher's exact tests were performed for all 2‐locus biallelic comparisons and coefficients of linkage disequilibrium determined.

In the British population, only one example of linkage disequilibrium, significant at P= 0.05 (after correction for the number of comparisons made) was seen (DPB1*0101‐DRB1*0301[171]). Additional associations, significant at P = 0.05 before correction for the number of comparisons were also seen, including DPB1*0401‐DRB1*15, DPB1*1101‐DRB1*0701(71), DPB1*1701‐DRB1*0701/ 2(72), DPB1*0101‐DQA1*0501, DPB1*0401‐DQA1*0102, DPB1*0501‐DQA1*0102, DPB1*0101‐DQB1*0201, DPB1*0401‐DQB1*0602/0603 and DPB1*1101‐DQB1*0201. With one exception (DPB1*1101‐DQB1*0201), none of these associations was seen in the French Canadian group.

These results indicate that although more frequent than thought hitherto, HLA class II linkage disequilibrium involving DPB1 alleles is generally weak, and can differ even between different caucasoid populations. This may have implications for HLA and disease studies.
GENOMIC MHC HAPLOTYPING OF MUTANT CELL LINES USING A NOVEL MARKER
Authors: Degli‐Esposti M.A. , Williamson J.F. , Tay G.K. , Salmon B. , Joseph G.A. , Morley A.A. , Dawkins R.L.
Published: April 2007

The aim of this study was to characterize MHC mutant cell lines by studying haplospecific markers within the MHC and specifically in the 250 kilobase (kb) region between the HLA B and TNF loci. This region has been difficult to define because of the lack of appropriate markers. Spontaneous MHC mutants were isolated after immunoselection with an anti‐HLA A2 monoclonal antibody and complement. Ten mutants were characterized using serological or allelic and genomic DNA markers within the HLA A to HLA DQ region of the MHC. Most mutants lost at least the 3 megabases of DNA from HLA A to HLA DQ viz the whole haplotype carrying HLA A2. Variants which have lost either HLA A alone or HLA A and HLA B were also found. The results show that it is possible to map the extent of the deletion between HLA B and TNF. Haplospecific scanning patterns for the CL region appear particularly useful.
ROLE OF A GENETIC REGION ON CHROMOSOME 4 IN THE REGULATION OF NATURAL KILLER CELL ACTIVITY IN MICE
Authors: Dubey D.P. , Mirza N.M. , Zaharian B.I. , Yunis E.J.
Published: April 2007

Natural killer cell (NK) activity is regulated by both the H‐2 and non‐H‐2 genes. Using bilineal congenic HW26C and HW13 mice which differ from the background strain C57BL/6By (B6) in a region of chromosome 4, we investigated the role played by a gene/genes in a segment of chromosome 4 of BALB/cBy on NK cell activity. Percoll separated low density spleen cells from young HW26C and HW13 mice showed a 3.5 fold higher NK activity than the B6. We also observed that the increase in NK activity of HW26C was not due to an increase in the number of NK cells. Using five other bilineal congenics containing different regions of chromosome 4 of BALB/cBy, we observed that the putative gene(s) regulating NK activity may be located between b and IFN‐α/β genes of chromosome 4. The level of NK activity of (B6xHW26C)F1 ranked between the HW26C and B6 suggesting that the gene product described is inherited in an incompletely dominant fashion.
PERIPHERAL TOLERANCE TO ALLOANTIGEN: STRATEGIES FOR THE FUTURE
Authors: Wood K. J.
Published: April 2007

The mature, adult immune system is specifically designed to eliminate any foreign material that may enter the body, but not to respond to the body's own tissues and molecules. Indeed, during development, the potential of the immune system to respond to self antigens is removed, by eliminating or effectively silencing any autoreactive cells. These features are well adapted under normal circumstances, as they result in the efficient elimination of potentially harmful agents thereby protecting the body from infection and malignancy. However, in the context of transplantation, this ‘normal’ response is diametrically opposed to the desired clinical outcome, which is clearly the long term function and survival of the transplanted tissue. To prevent graft rejection the immune system of the transplant recipient has to be manipulated to ensure that it is incapacitated. Immunosuppressive drugs can be used for this purpose and are undoubtedly effective; indeed they have had a dramatic impact on success rates in clinical organ transplantation. However, as the mechanism of action of these chemical immunosuppressants is immunologically non‐specific, any immune response the recipient may need to make after transplantation, as well as the rejection response, is suppressed. In addition, to maintain graft survival the drugs have to be taken indefinitely after transplantation and therefore their use is not only associated with immunological complications such as increased risks of infection and malignancy, but also numerous non‐immunological side‐effects. One way to overcome these problems would be to develop strategies for specific immunosuppression, such that only leukocytes capable of responding to the foreign histocompatibility or alloantigens expressed by the transplanted tissue would be affected. The ability to manipulate or reprogramme the adult immune system in such a way as to induce specific immunological unresponsiveness or tolerance to the alloantigens of the organ donor would offer many advantages over conventional immunosuppressive therapy. Only leukocytes reactive with donor alloantigens would be affected, thus allowing transplant recipients to respond effectively to other immunological stimuli, such as virus infections.