bluuebunny/crossref_metadata_2025 huggingface dataset:
THE H‐2 COMPLEX REGULATES BOTH THE SUSCEPTIBILITY TO MOUSE VIRAL LYMPHOMAGENESIS AND THE PHENOTYPE OF THE VIRUS‐INDUCED LYMPHOMAS
Authors: Zijlstra M. , Vasmel W. L. E. , Radaskiewicz T. , Matthews E. , Melief C. J. M.
Published: April 2007

Neonatal infection of C57BL/10 mice with cloned ecotropic and/or dual‐tropic mink cell focus‐inducing (MCF) mouse leukaemia viruses (MuLV), induces a wide spectrum of different lymphomas of T, B, and non‐T/non‐B cell types. The H‐2 complex has a marked influence on both the development of lymphoma incidence and lymphoma type. A study using the oncogenic MCF 1233 virus and a series of B10 congenic mice enabled the mapping of the following:

(a) Resistance to the early development of T cell lymphomas is controlled by the H‐21‐A locus.

(b) Susceptibility to early T cell lymphomagenesis is associated with an I‐A‐regulated low anti‐MCF 1233 envelope antibody response and persistent infection of the thymus.

(c) B10 (H‐2b) mice, which are resistant to early T cell lymphomagenesis induced by MCF 1233 or other MuLV isolates, have high anti‐MuLV envelope antibody responses which are I‐A‐regulated. These mice develop more B cell lymphomas late in life in contrast to the early development of T cell lymphoma in B10.A (H‐2a mice. The possible response mechanisms which underlie these observations, including: (1) I‐A ‐regulated immunoselection against MuLV antigens expressed by (pre) leukaemic T cells, (2) aberrant expression of class II MHC antigens on some B cell lymphomas and (3) I‐A‐regulated chronic immunostimulation of MuLV‐expressing (pre) leukaemic B cells, are discussed.
IMMUNOGENICITY OF H‐2 POSITIVE AND H‐2 NEGATIVE CLONES OF A MOUSE TUMOUR, GR9
Authors: Garrido M. L. , Pérez M. , Delgado C. , Rojano J. , Algarra I. , Garrido A. , Garrido F.
Published: April 2007

The GR9 tumour was induced with methylcholanthrene in a BALB/c mouse, adapted to tissue culture and cloned without any passage in vivo. GR9 clones were typed for H‐2 with three monoclonal antibodies that define H‐2Kd+ Dd, Kd and Dd antigens. A great heterogeneity of H‐2d expression was found from clones which were Kd and Dd positive to clones Kd and Dd negative. These results were confirmed for A7 and B9 clones using immunoprecipitation with anti‐H‐2D.4 and anti‐H‐2K.31 alloantisera and SDS‐PAGE analysis. In addition, the number of chromosomes per cell was heterogeneous amongst the clones, ranging from 38 ± 2 to pseudotetraploid clones which have 75 ± 2 chromosomes.

GR9 clones were injected into syngeneic BALB/c mice to measure local tumour growth. We found that the growth correlated with the amount of H‐2 antigen expressed, i.e. clones with low H‐2d expression were highly malignant while clones with normal H‐2d expression were highly immunogenic. Finally we found that BALB/c mice immunized against A7 (Kd, Dd‐positive) protected against A7, as expected, but surprisingly also against B9 (Kd, Dd‐negative).
EXPRESSION OF MHC CLASS II PRODUCTS ON HUMAN COLORECTAL CANCER AN IMMUNOHISTOLOGICAL AND FLOW CYTOMETRIC STUDY
Authors: Moore M. , Ghosh A. K. , Johnston D. , Street A. J.
Published: April 2007

The MHC status of epithelial cells from 32 primary colorectal neoplasms, villous adenomata (VA; 2) and inflammatory bowel disease (IBD; 3) were evaluated using a panel of monoclonal antibodies (mAbs). Class I antigens and β2 microglobulin (β2m) were expressed on all normal, benign, inflammatory and malignant epithelia with the exception of two carcinomas. A more complex pattern of reactivity was encountered with anti‐class II mAbs. Some expression was detected on normal glandular and luminal epithelium, particularly adjacent to the tumour. Inflammatory tissues, VA and 23/32 carcinomas were also antigen‐positive, the proportion of stained epithelial cells ranging from 5% to 90%. Expression was usually non‐coordinate, DR being the predominant specificity followed by DP and DQ, which is suggestive of independent D region gene regulation. The hypothesis that class II expression is induced in vivo by locally generated IFN γ was not confirmed by in vitro treatment with this agent of epithelial colorectal carcinoma‐derived cell lines. These provisional data suggest that although IFN γ may be a necessary stimulus for class II expression it is insufficient and that other factors also influence the responsiveness of tumour cells in this respect.
EXPRESSION OF IMMUNOGLOBULIN LAMBDA CHAINS IN THE LABORATORY RAT
Authors: Zhou D. , Leslie G. A. , Guo K. , Gutman G. A.
Published: April 2007

We immunized a BALB/c mouse with the lambda‐bearing rat IgG, myeloma IR31, fused its spleen cells with the hybridoma parent line P3.X63.Ag8.653, and isolated a monoclonal antibody (G33/11) directed against rat immunoglobulin lambda chains. We used this antibody to classify two existing rat hybridomas as lambda‐bearing proteins (D4.37HL.252 and PC61.5, and isolated one new lambda‐bearing rat IgM hybridoma, G36/1. All the normal inbred rat sera that were tested contained lambda‐bearing Ig as detected by G33/11, at levels ranging from 1.5% to 13% of the total serum Ig, the mean value being 7.9%. This antibody will be valuable for broadening our understanding of the immunogenetics of the rat, and for the characterization of monoclonal antibodies made in this species.
HETEROGENEITY OF SYSTEMIC LUPUS ERYTHEMATOSUS ELUCIDATED BY CLUSTER ANALYSIS
Authors: Stenszky V. , Kozma L. , Szegedi GY. , Sonkoly I. , Bear J. C. , Farid N. R.
Published: April 2007

For 75 patients with systemic lupus erythematosus (SLE), 39 laboratory and clinical characteristics, including HLA‐A, B, C and DR typing, were analysed using a cluster analysis technique. Three groups were identified. Group I (46 patients) was characterized by infrequently severe disease, good response to therapy and infrequent multisystem involvement. Group I1 (24 patients) was characterized by a severe course of disease (although the tendency to remit after therapy was not unusual), and, frequently, renal involvement and pericarditis. Group 111 (5 patients) was characterized by more severe renal disease. Of the 75 patients studied, 38.7% possessed HLA‐DR3, compared to 17.4% of controls. Group I patients did not differ from controls but 80% of Group II patients and 4/5 Group III patients had DR3. Cluster analysis identifies subsets of SLE patients who show marked differences in disease course and severity, correlated with possession of the HLA B8, DR3 phenotype.
BENT‐LIMB DISEASE IN LAMBS
Authors: Millot P.
Published: April 2007

In crossbred Charmois lambs, Bent‐limb disease appears to be associated with the phenotype ‘z,i’ which corresponds to a doubly recessive genotype at the two loci OL‐Z (a minor lymphocyte antigen locus independent of OLA, the major histocompatibility complex of the sheep) and I (involved in genetic control of the expression of R and O antigens found in various body fluids and on erythrocytes). The disease seems to be controlled by a single autosomal recessive gene, provisionally named bl (its normal or healthy allele Bl being dominant) which is distinct from, though linked to, the genes at the OL‐Z and I loci (observed haplotype: bl,z,i). In the Lacaune breed, besides the already observed bl,r,i haplotype, there is an additional one: bI,z,I. In Vendéen sheep, and in animals which belong to two other breeds, the bl,ZII,I haplotype (bl linked to the two dominant alleles) appears to be relatively frequent. Further studies are needed in order to confirm the genetic hypothesis suggested by the present data.
MONOCLONAL ANTI‐LWab AND ANTI‐D REAGENTS RECOGNIZE A NUMBER OF DIFFERENT EPITOPES
Authors: Shaw M.‐A.
Published: April 2007

Monoclonal antibodies (mAbs) which detect antigens on human red cells are also suitable for testing cells of other species. Such studies may reveal previously unrecognized heterogeneity in antibodies which apparently detect the same antigen on the human red cell surface. Information is also provided on specificities shared amongst several species. Here three anti‐LWab and a variety of Rh‐related antibodies have been tested against the red cells of various primates. One monoclonal anti‐LWab antibody, BS46, reacted with the red cells of gorillas and rhesus monkeys but not those of orang‐utans, baboons or marmosets. In contrast, BS56 and NIM‐M8 reacted with the cells of all these species. Chimpanzee cells, however, reacted only with NIM‐M8. Use of primate cells has shown that all three monoclonal anti‐LWab antibodies recognize different epitopes. These observations may explain early conflicting data concerning primate cells. The difference between the monoclonal anti‐D, D4, and three other anti‐D antibodies, 8G2, 8D6 and 7D10, has been confirmed. The D antigen is apparently confined to the red cells of apes and humans. D4 recognizes a polymorphism in chimpanzees and 8G2, 8D6 and 7D10 recognize a polymorphism in gorillas. Two Rh‐related mAbs, R6A and K70, were also investigated. R6A fails to react with Rhnull cells and reacts more weakly with homozygous ‐D‐ cells than with cells of common Rh phenotypes. K70 reacts weakly with Rhnull and ‐D‐/‐D‐ cells. The antigen detected by R6A is confined to the red cells of humans, gorillas and chimpanzees, while the antigen detected by K70 shows a wider species distribution. In some primates LW antigens are expressed in the absence of the determinant recognized by R6A. This phenotype has never been known to occur in humans.
A GENE LOCUS CONTROLLING A SERUM PROTEIN MIGRATING ELECTROPHORETICALLY IN THE β REGION OF MICE AND DETECTED BY USING A STRAIN DERIVED FROM THE JAPANESE WILD MOUSE (MUS MUSCULUS MOLOSSINUS)
Authors: Harada Yoshi‐Nobu , Hayakawa Jun‐Ichiro , Noda Eiko , Tomita Takeshi
Published: April 2007

Antigenic specificities of serum proteins from the MOL‐ANJ strain of mice (a strain derived from Japanese wild mice, Mus musculus molossinus) were studied by gel precipitation with alloantisera produced by reciprocal alloimmunization between MOL‐ANJ and BALB/c mice. An alloantigen which migrates immuno‐electrophoretically in the β region of serum proteins has been identified. Evidence indicates that this antigenic specificity is controlled by a co‐dominant autosomal gene locus designated by the symbol Sas‐2. The evidence also suggests that Sas‐2 is genetically different from the previously described Sus‐1 which controls a serum protein in the mouse. Sas‐2 was located by linkage analysis between Zdh‐1 locus and Akp‐1 locus on chromosome 1.
IgA2 ALLOTYPES DETERMINED BY RESTRICTION FRAGMENT LENGTH POLYMORPHISM IN IgA DEFICIENCY RE‐EXPRESSION OF THE SILENT A2m(2) ALLOTYPE IN THE CHILDREN OF IgA‐DEFICIENT PATIENTS
Authors: Hammarström L. , Lange G. G. se , Smith C.I.E.
Published: April 2007

IgA2 allotyping was performed on DNA from 60 IgA‐deficient Caucasian individuals. The frequency of A2m(2) was not statistically different from that of normal controls. Two informative families were selected for further studies. In both families, the A2m(2) allotype (derived from the IgA‐deficient parent) was inherited by some of the children, as determined by Southern blotting experiments. In all cases the ‘silent’ IgA2m2 gene was re‐expressed, as judged by conventional serological allotyping of serum proteins. These data strongly argue against structural gene deletions or mutations as a cause of IgA deficiency.
ASSOCIATION OF HLA‐DRw9 WITH MYASTHENIA GRAVIS IN CHINESE
Authors: Chiu H.‐C. , Hsieh R.‐P. , Hsieh K.‐H. , Hung T.‐P.
Published: April 2007

HLA phenotypes were studied in 59 Chinese myasthenia gravis (MG) patients. Unfractionated peripheral blood mononuclear cells were used for typing of HLA‐A, B and C, and B‐enriched lymphocytes for HLA‐DR. Seventy‐nine healthy subjects were included as controls. When compared with healthy controls, the patients showed significant increases in HLA‐Bw46 (47·5% versus 14%, χ2= 18·7, P < 0·001), HLA‐DRw9 (59·3% versus 11%, χ2= 35·7, P < 0.001), while HLA‐DR3 was decreased (3·4% versus 32·9%, χ2= 18·2, P < 0.001). Further analysis showed that the primary association was with DRw9 and the increase in Bw46 was secondary to it. Both HLA‐Bw46 and ‐DRw9 were increased in all subgroups except where the age of onset was greater than 40 years. Finally, the HLA‐A2‐Bw46‐DRw9 combination was also significantly increased in patients (25.4% versus 5.1%, χ2= 11.5, P < 0.001), especially in the subgroups of MG with age of onset of less than 10 and males with age of onset of less than 40 years.
DEFICIENCY OF C4A IS A GENETIC DETERMINANT OF SYSTEMIC LUPUS ERYTHEMATOSUS IN THREE ETHNIC GROUPS
Authors: Dunckley H. , Gatenby P. A. , Hawkins B. , Naito S. , Serjeantson S.W.
Published: April 2007

Systemic lupus erythematosus (SLE) has shown associations with the major histocompatibility complex (MHC) class II DR antigens and class III complement components C2 and C4 in previous studies. The primary susceptibility locus has been difficult to identify, however, on account of linkage disequilibrium within the MHC. We have studied C4A and C4B distributions in 63 Caucasoid, 75 Chinese and 51 Japanese SLE patients. All three populations showed a statistically significant increase in C4A*Q0 (null) alleles when compared with 323 ethnically matched controls. We conclude that complete or partial deficiency of C4A is a genetic determinant of SLE common to these three ethnically distinct populations.
NEW HLA CLASS I‐LIKE ALLOANTIGENS EXPRESSED ON BLAST CELLS
Authors: Crepaldi T. , Peruccio D. , Lecchi M. , Resegotti L. , Guerra M. Grazia , Funaro A. , Richiardi P.
Published: April 2007

New β2‐microglobulin (β2‐m)‐associated, HLA‐linked alloantigens, selectively expressed on phytohaemagglutinin (PHA)‐activated lymphocytes, were identified using human alloantisera. Reactivity of the antisera against activated cells correlated with HLA‐A2, A10 and A28 specificities. The new alloantigens were undetectable on peripheral blood mononuclear cells, B lymphocytes and platelets using either the lymphocytotoxicity or the absorption techniques, and appeared on lymphocytes upon PHA activation, with time‐dependent kinetics. They were found on some, but not all, acute leukaemias of B, T and myeloid origin, being absent from chronic B lymphocytic leukaemias. The presence of the new β2‐microglobulin‐associated allospecificities was not correlated with the quantity of classical class I antigens present, as analysed on different cell types by flow cytometry. Thus, these antigenic determinants appear to be different from the classical HLA class I antigens and could be the human counterpart of the murine Qa system or of the second H‐2K gene.
HETEROGENEITY AND LINKAGE OF EQUINE C4 AND STEROID 21‐HYDROXYLASE GENES
Authors: Kay P. H. , Dawkins R. L. , Bowling A. T. , Bernoco D.
Published: April 2007

The fourth component of complement (C4) is polymorphic in most species studied, and is encoded by a gene or genes within the MHC. In man and mouse there are two closely linked C4 and steroid 21‐hydroxylase (21‐OH) genes. Therefore we have used Southern blotting to determine whether equine C4 and 21‐OH genes are linked. C4 restriction fragment length polymorphism (RFLP) was found with the enzymes EcoRI and BamHI. Comparison of the sizes of EcoRI‐digested fragments of genomic DNA hybridizing with C4 and 21‐OH probes revealed that equine C4 and 21‐OH genes are separated by no more than 13 kb. Further, there is no evidence of C4 and 21‐OH gene duplication in the horse. Segregation of ELA and different polymorphic forms of equine C4 suggest that C4 and 21‐OH genes are within the MHC. It is likely that equine MHC supratypes will provide improved markers of disease susceptibility.
DNA SEQUENCE ANALYSIS OF TWO BOVINE IMMUNOGLOBULIN CH GAMMA PSEUDOGENES
Authors: Symons D. B. A. , Clarkson C. A. , Milstein C. P. , Brown N. R. , Beale D.
Published: April 2007

A bovine calf liver DNA library in λ2001 bacteriophage has been screened with a human Igγ4 heavy‐chain constant‐region gene probe. Four hybridizing clones have been identified, and the DNA sequences in two of these, which have high homology with CHγ genes, are reported here. Within the bovine sequences, four separate exons can be identified, corresponding to the three CH domains and the hinge of gamma heavy‐chain genes. Both of these genes contain atypical sequences around one or more of their exon/intron boundaries with consequent loss of splice sites, indicating that these are probably γ pseudogenes. One sequence codes for a C‐terminal peptide which matches the 18‐mer C‐terminal heavy‐chain peptide of bovine serum IgG2, the other encodes a C‐terminal peptide unknown in the bovine. These results suggest that evolutionary duplication of CHγ genes has occurred in the bovine.
SEROLOGICAL EVIDENCE FOR A DEFECT IN RT1.B (I‐A) EXPRESSION BY THE BDIX RAT STRAIN
Authors: Male D. K. , Pryce G. , Butcher G. W.
Published: April 2007

Astrocytes, astrocytic cell lines and endothelium from BDIX rats were stimulated with recombinant interferon‐gamma (IFN‐γ) and the expression of MHC molecules quantified using an enzyme immunoassay (EIA). Using the two mouse anti‐RT1.B monoclonal antibodies MRC OX4 and OX6, previously described as recognizing a monomorphic determinant on RT1.B, as well as polyvalent rabbit anti‐rat class II antisera, we were unable to demonstrate any induction of RT1.B molecules on these cells under conditions that induced RT1.B expression in all other strains tested. In contrast, RT1.D locus class II molecules, detectable by the antibody MRC OX17, are more strongly expressed in BDIX than in other strains. In experiments using BDIX lymphocytes, this serologically detected defect in RT.1B expression was confirmed using four additional mouse anti‐mouse I‐Ak monoclonal antibodies, which cross‐reacted on all rat strains tested except BDIX. It appears likely that BDIX rats lack either a structural or controlling gene required for RT1.B expression.
T CELLS DEFINE NEW ALLOANTIGENS CONTROLLED BY A GENE(S) IN THE Mls REGION ON MOUSE CHROMOSOME 1
Authors: Kimura S. , Furze J. , Festenstein H.
Published: April 2007

Two different types of T cell clones which recognize Mlsb haplotype gene products were established by using mice congenic at the Mls locus. The antigens defined by these T cells showed differences not only in strain distribution but also in the restriction element for antigen recognition by T cells (i.e. Kd or Dd). The antigens appear to be expressed on various types of lymphocytes.
GENETIC COMPLEXITY OF Mls
Authors: Click R. E. , Adelmann A.
Published: April 2007

For almost 20 years, little new has been described for Mls or the products it encodes. In the present report, data is presented which indicate that either the number of loci or the number of alleles per locus that control Mls‐like products is much larger than the two‐locus model or five allele‐model presently envisaged.
Mls ALLO‐DETERMINANTS ARE RECOGNIZED IN AN MHC CLASS II ANTIGEN‐DEPENDENT BUT UNRESTRICTED FASHION BY A DISCRETE SET OF T CELLS
Authors: Macphail S. , Stutman O.
Published: April 2007

Blocking studies carried out with anti‐H‐2 class II antigen antibodies show that H‐2 class II molecules are intimately involved in the recognition of Mlsa determinants by unprimed, specifically responsive T cells. The blocking of the anti‐Mlsa response by these antibodies was not due to inhibition of IL‐1 production by H‐2 class II antigen positive macrophages. A strain analysis indicates that the response to Mlsa is regulated by H‐2‐linked genes and that this effect is exerted at the level of the stimulator cells; however, the response to Mls epitopes does not appear to be H‐2 restricted. Finally, the ability of spleen cells from Mls‐incompatible mice to induce a state of Mls‐specific, clonal deletion type tolerance by neonatal injection, shows that Mls determinants exist in qualitatively allelic forms and suggests that these determinants are recognized by specific clones of T cells.
SPECIFICITY OF ANTI‐H‐2 CLASS I ANTIBODIES INDUCED BY SYNGENEIC IMMUNIZATION WITH SENDAI VIRUS‐TREATED CELLS IS REGULATED BY THE MOUSE MHC AND VIRAL ANTIGENS. NO EVIDENCE FOR MHC‐RESTRICTED VIRUS‐ SPECIFIC ANTIBODIES
Authors: Kievits F. , Van Bleek G. M. , Boerenkamp W. J. , Pla M. , Ivanyi P.
Published: April 2007

In a previous study, we searched for Sendai virus (SV)‐specific antibodies that were restricted in their binding by self‐major histocompatability complex (MHC) antigens. In C57BL/6 (B6; H‐2b) mice, most of the sera obtained after i.p. injections with syngeneic SV‐coated (SV+) spleen cells contained auto‐ and alloreactive lymphocytotoxic antibodies directed against H‐2 class I molecules, but no viral‐specific, MHC‐restricted antibodies.

Here we report that syngeneic immunization with SV+ cells regularly induced H‐2‐specific antibodies in various mouse strains. From a total of 12 strains tested, only the B10.S (H‐2s) strain appeared to be a low responder. The immune responses are of two types: (i) mice of some strains produce autoreactive antibodies and a broad variety of alloreactive antibodies; and (ii) mice of some strains produce only narrow or widely alloreactive antibodies. Because most of the strains differ only in the H‐2 region, the patterns observed are regulated by the MHC. To locate the genes involved in the induction of H‐2‐specific antibodies more precisely, two B6 mutant strains, bml (Kb mutant) and bm13 (Db mutant), were immunized with syngeneic SV+ cells. The results suggest that the H‐2Db region plays an important role in the induction and specificity of the lymphocytotoxic H‐2 class I‐specific antibodies present in sera of H‐2b mice after syngeneic immunization with SV+ cells.
TUMORIGENIC AND METASTATIC ABILITY OF SV40‐TRANSFORMED BALB/c CELL LINES AND MHC ANTIGEN EXPRESSION
Authors: Giovanni C. De , Nicoletti G. , Re B. Del , Scotlandi K. , Prodi G. , Lollini P.‐L. , Nanni P.
Published: April 2007

Tumorigenic and metastatic potential were studied in relation to class I MHC expression in four different SV40‐transformed BALB/c cell lines. All the lines studied, tumorigenic or not, expressed both H‐2Kd and Dd, so MHC antigens did not seem to be involved in the control of SV40‐transformed cells' growth in vivo. Lung metastases were observed in all tumour‐bearing mice. Cells cultured after in vivo passage, obtained either from tumour tissue or from individual lung metastases, still expressed similar levels of H‐2d antigens, thus suggesting that tumour growth and metastasis do not occur through the selection of variants with altered MHC expression.
A HYPOTHESIS ON THE DUAL SIGNIFICANCE OF ABH, LEWIS AND RELATED ANTIGENS
Authors: Pendu J. Le
Published: April 2007

ABH and related antigens appeared a long time ago in the evolution of vertebrates on tissues in contact with the external environment, which suggests that the polymorphism given by these antigens might play a role in the relationships of the species with pathogens. However, they are also oncodevelopmental markers and some recent experimental data suggest that they might play a role in cell‐cell recognition at some stages of development. This type of function is difficult to reconcile with the polymorphic nature of these markers unless one considers that the glycosyltransferases necessary for the synthesis of the active structures are encoded by various members of multigene families. Some non‐polymorphic members of the families would have their expression limited in time and space during development, leading to the same antigenic patterns in every individual, and these could reappear in some tumours, while the expression of other polymorphic members (A/B/O, H/h, Se/se, Le/le), leading to a variety of antigenic phenotypes, would be expressed at later stages and remain so during the whole life of the individual. The corresponding antigens could disappear from some cancer cells. It is argued that the ABH and related antigens would have primarily been involved in cell‐cell recognition phenomena. The polymorphism would have evolved later from gene duplication under environmental pressure, the expression on erythrocytes which occurred very late in evolutionary time probably being of very little biological significance.
MAGNETIC CELL SORTING USING COLLOIDAL PROTEIN—MAGNETITE
Authors: Owen C. S.
Published: April 2007

Magnetic filtration of labelled cells as a way of separating leucocyte subpopulations was tested with a very simple and easy filtration device, using colloidal magnetite as the labelling reagent. In order to quantitate cell enrichment, a double label (both fluorescent and magnetic) was used, under conditions which labelled less than 10% of the cells in the initial sample. Up to 20 million cells were simply passed through a small magnetic filter with a hand‐held syringe. Depletion of labelled cells in the suspension that passed through was threefold, and enrichment of labelled cells in the wash of the filter after its removal from the magnet was approximately fivefold. Factors which limited the quality of separation are discussed. Other, more preliminary, experiments found enrichments of 15–to 30‐fold with the same colloidal magnetite and hand‐held apparatus when the cell labelling system was more selective.
IMMUNOMAGNETIC DEPLETION OF MALIGNANT CELLS FROM AUTOLOGOUS BONE MARROW GRAFT: FROM EXPERIMENTAL MODELS TO CLINICAL TRIALS
Authors: Combaret V. , Favrot M. C. , Chauvin F. , Bouffet E. , Philip I. , Philip T.
Published: April 2007

Using experimental models with normal allogeneic bone marrow (BM) contaminated with Burkitt or neuroblastoma cell lines, and a liquid culture assay, we demonstrated that, when used in optimal conditions, the immunomagnetic depletion technique permitted a reproducible elimination of 3–4 log malignant cells. Results were very similar to those obtained with the complement lysis procedure in Burkitt lymphoma. This immunomagnetic procedure was used in 123 cases of autologous bone marrow transplantation (ABMT) in children with neuroblastoma. The analysis of the cases demonstrated, first, that the procedure induced a significant loss of mononuclear cells but was not toxic for BM precursors. Delays to engraftment observed in a few patients were probably due to the combination of pejorative factors, especially the damage caused to the micro‐environment by previous heavy and prolonged chemotherapy or the double ABMT programme. Second, patients presented with profound T‐cell defect with undetectable IL2 secretion up to 1 year post‐graft but they all had normal NK functions from the first month post‐graft, these functions exceeding normal values on the second and third months post‐graft. Finally, in 20 cases, dual‐immunofluorescence staining permitted the demonstration that the autograft contained malignant cells before purging that were eliminated by the immunomagnetic depletion.
APPLICATIONS OF AUTOMATED SIMULTANEOUS DOUBLE FLUORESCENCE (SDF). II. HLA CLASS I PHENOTYPING USING IMMUNOMAGNETICALLY SEPARATED T LYMPHOCYTES
Authors: Laundy G. J. , Peberdy M. , Klouda P. T. , Bradley B. A.
Published: April 2007

HLA class I phenotyping was performed using T‐lymphocyte populations isolated by immunomagnetic beads (IMBs) coated with monoclonal antibodies with specificity for CD2, CD4 or CD8. The results were compared to those obtained using density gradient‐separated lymphocytes (PBL). The typing trays were read by the automated simultaneous double‐fluorescence (SDF) technique previously established in our laboratory using an Astroscan 2100 system. The aims of the present study were to establish whether the advantages of IMB lymphocyte separation and automated plate reading by SDF were complementary and whether the results obtained by IMB‐SDF and PBL‐SDF were concordant.

Similarity coefficients for paired results obtained by IMB‐SDF and PBL‐SDF varied between 0.825 using anti‐CD8‐coated IMBs and 0.914 using anti‐CD4‐coated IMBs with a consistent excess of stronger results observed with the PBL‐SDF technique. The variations observed did not result in incorrect phenotype assignment but would significantly influence a cross‐matching test.

These results illustrate the feasibility of using IMB‐separated lymphocytes for HLA phenotyping by SDF.
POST‐TRANSCRIPTIONAL DOWNREGULATION OF MHC CLASS I EXPRESSION IN ONCOGENE‐TRANSFORMED CELLS IS REVERTED BY IFN‐GAMMA AND TNF‐ALPHA
Authors: Seliger B. , Pfizenmaier Klaus
Published: April 2007

Transformation of murine NIH3T3 fibroblasts with retroviral vectors carrying the mos, myc and the Ha‐ras oncogene, respectively, was associated with a strong reduction of H2 antigen expression in the cell membrane. Analysis of H‐2K and β2‐microglobulin promoter‐driven CAT activity in these oncogenic transformants and normal NIH3T3 fibroblasts revealed unchanged promoter activity, suggesting post‐transcriptional control of MHC class I expression by these oncogenes. Treatment with IFN‐gamma and TNF‐alpha caused 2‐ to 3‐fold enhancement of H‐2K and β2‐microglobulin promoter activity, as well as a normalization (TNF‐alpha treatment) or enhancement (IFN‐gamma treatment) of H2 membrane expression. These data suggest that IFN‐gamma as well as TNF‐alpha can counteract downregulation of H‐2 genes by interference with an oncogene‐induced, post‐transcriptional block as well as by a direct enhancement of H‐2 gene transcription.
MHC‐RESTRICTED AUTOANTIGEN‐REACTIVE T CELL CLONES IN MULTIPLE SCLEROSIS
Authors: Rohowsky‐Kochan C. , Troiano R. , Cook S. D.
Published: April 2007

Multiple sclerosis is a demyelinating disease of the central nervous system with genetic, viral and autoimmune characteristics. Myelin basic protein (MBP) is a suspected target autoantigen since it induces experimental autoimmune encepha‐lomyelitis, an animal model closely resembling multiple sclerosis. The disease is mediated by Class II restricted, MBP‐reactive T cells possessing the T helper/inducer phenotype. In the present study, we have isolated MBP‐reactive T cell clones from the peripheral blood of a chronic progressive multiple sclerosis patient. The clones displayed blastogenic memory responses when rechallenged with the autoantigen and irradiated autologous lymphocytes. MBP recognition by the autoantigen‐reactive T lymphocytes was restricted by major histocompatibility complex Class II antigens. Both CD4+8−‐ and CD4–8+ MBP‐reactive T cell clones were obtained.
GLYCOSURIA AND INSULITIS IN NOD MICE EXPRESSING THE HLA‐DQw6 MOLECULE
Authors: Fukui Y. , Nishimura Y. , Iwanga T. , Kimura A. , Inamitsu T. , Hanaoka Y. , Kitigawa T. , Sasazuki T.
Published: April 2007

To investigate HLA‐linked genes controlling the susceptibility and resistance to insulin dependent diabetes mellitus (IDDM), HLA‐DQ alleles of 45 Japanese patients with IDDM were analysed, using sequence specific oligonucleotide (SSO). DQA1*0301 and DQBI*04 were positively associated (R.R = 6.6, Pc<0.05 and R.R. = 4.7 Pc<0.01) and DQAI*0103 and DQBI*0104 were negatively associated (R.R. =0–2, Pc<0.01) with IDDM. DQAI*0103 and DQB1*0104 were in strong linkage disequilibrium to encode for DQw6 molecule. Therefore, in a Japanese population, the DQw6 molecule seems to control the resistance to IDDM. To determine whether or not the DQw6 molecule itself can protect against glycosuria and insulitis in NOD mice, these animals were mated with HLA‐DQw6 transgenic‐C57BL/6 mice (DQw6‐B6) and the F1 progeny expressing the DQw6 molecule were backcrossed with NOD mice. Eighty‐five female backcross progenies were classified into four groups, according to the MHC classII phenotype; I‐Anod/I‐Anod DQw6(‐), I‐Anod/I‐Anod DQw6(+), I‐Anod/I‐Ab DQw6(‐) and I‐Anod/I‐Ab DQw6(+). At the age of 16 weeks, 9.1% of the DQw6(‐) I‐Ab(‐) mice had a glycosuria whereas none of the DQw6(+) I‐Ab(‐) mice had a glycosuria. At the age of 30 weeks 13.6% of the DQw6(‐) I‐Ab(‐) mice had a glycosuria and 7.7% of the DQw6(+) I‐Ab(‐) mice had a glycosuria. Histological examinations of the pancreas were performed in the 30 week old mice or after the development of glycosuria. About 50% of I‐Ab(‐) mice developed insulitis, regardless of the expression of the DQw6 molecule. Thus, the DQw6 molecule seemed to delay the onset of glycosuria but did not protect against glycosuria and insulitis in the NOD mice despite a functional expression.
PROTECTION OF NEWBORN MICE FROM GRAFT VERSUS HOST DISEASE BY MATERNAL PRE‐IMMUNIZATION
Authors: DeGiorgi L. , Matossian‐Rogers A. , Festenstein H.
Published: April 2007

Alloimmunization of BALB/c (H‐2d) female mice with allogeneic spleen cells from C57BL/6 (H‐2b) or CBA/H (H‐2k) mice protects BALB/c offspring from graft‐versus‐host disease (GVH‐D) following neonatal intraperitoneal inoculation of high doses of spleen cells respectively of C57BL/6 or CBA/H strains of mice.

The mice survived GVH‐D over one year after the allogeneic inoculum 24–48 h after birth and they did not show any signs of GVH reaction nor splenomegaly. We show that this phenomenon is antibody mediated and affects the developing immune system of the foetus.

Repeated immunization of virgin female BALB/c with allti‐H‐2b or anti‐H‐2k antisera (Abl) can equally abrogate GVH‐D in their newborn offspring challenged at 24–48 h after birth with allogeneic spleen cells of H‐2b or H‐2k phenotype.

Our results demonstrate that protection from GVH‐D is not specific to the immunizing strain and occurs when the neonatal mice are challenged with C57BL/6 or CBA/H spleen cells. There is thus crossreactivity of tolerance against H‐2 specificities. In this study we also report on the in vitro cellular immune responses of the surviving GVH‐resistant mice and demonstrate that these responses against both the challenge and third party lymphocytes are impaired.
IMMUNOGENETIC ASPECTS OF HUMAN THYROGLOBULIN‐REACTIVE T CELL LINES AND HYBRIDOMAS
Authors: Krco C. J. , Gores A. , David C. S. , Kong Y. M.
Published: April 2007

The in vitro proliferative response of T cells primed with human thyroglobulin (Tg) was compared in 11 independent haplotypes on B10 background. B10.K and B10.S mice were the most responsive, whereas, with the exception of B10.PL (H‐2u, all other B10 congenics were intermediate responders. The two best responders to in vitro challenge with human Tg, of the k and s haplotype, are the same as those showing H‐2‐linked susceptibility to induction of experimental autoimmune thyroiditis (EAT) with mouse Tg. Since shared epitopes on human and mouse Tgs have been shown to be thyroiditogenic by adoptive transfer studies in CBA (H‐2k) mice, the findings indicate that shared epitopes may be studied in appropriate (i.e. EAT‐susceptible) strains of mice. Therefore, we proceeded to develop methods to produce T‐cell lines and hybridomas to human Tg in B10.K and B10.S mice, test their cross‐reactivity to heterologous Tgs and their Ia restriction patterns. By using antigen‐presenting cells from recombinant strains, we identified restriction elements encoded by the I‐A subregion alone and a combinatorial molecule from the I‐AI/I‐E subregions.
VHDJH TO JH JOINING IS NOT BLOCKED IN μ‐CHAIN‐PRODUCING PRE‐B CELLS, IMPLYING THE BREAKDOWN OF ALLELIC EXCLUSION
Authors: Sugiyama H. , Komori T. , Minami Y. , Sakato N.
Published: April 2007

Intracytoplasmic μ‐positive pre‐B cells (μ+VH315− cells) transformed with Abelson virus continuously produced cells (μ+VH315+ cells) that were double‐stained by anti‐μ and anti‐VH315 antibodies which specifically recognized the immunoglobulin heavy chain variable region identical or closely related to that of MOPC315 myeloma protein. Southern blot analysis indicated that the μ+VH315+ cells were generated from the μ+VH315‐ cells by the VH315·D·JH2 to a germline JH3 joining on the non‐productive VH315·D·JH2 allele, which resulted in the production of μ‐chains with variable region detectable by anti‐VH315 antibodies. Therefore, it is strongly indicated that VHDJH to JH joining is not blocked in μ‐chain‐producing pre‐B cells, and thus is free from allelic exclusion machinery.
IEF ANALYSIS OF HLA MOLECULES IMMUNOPRECIPITATED BY PUTATIVE ANTI‐CLASS I‐LIKE ALLOANTISERA
Authors: Crepaldi T. , D'Alfonso S. , Richiardi P.
Published: April 2007

Two human alloantisera, previously described as possibly detecting new beta 2‐microglobulin associated proteins, selectively expressed on HLA‐A2 and HLA‐A1 phytohaemagglutinin (PHA)‐activated lymphocytes, immunoprecipitated only molecules with the same isoelectric point as HLA‐A1 and A2 products. This result suggests that the selected alloantisera do not react with the products of an HLA class I locus different from ABC but probably recognize a new epitope arising on HLA‐A molecules upon conformational changes consequent to cell activation.
PCR‐SSO TYPING FOR DR4‐Dw SUBTYPES: APPLICATION TO UNRELATED BONE MARROW TRANSPLANT DONOR SELECTION
Authors: Clay T. M. , Howard M. R. , Bidwell J. L. , Bidwell E. A. , Raymond P. A. , Evans J. E. , Bradley B. A.
Published: April 2007

Thirty‐seven DR4‐positive patient‐unrelated bone marrow donor pairs previously DR/DQ restriction fragment length polymorphism (RFLP) typed and tested in mixed lymphocyte culture (MLC), have been DR4‐Dw subtyped retrospectively using sequence specific oligonucleotide probes. We found that DR4‐Dw subtyping substantially increased the accuracy of pre‐MLC matching and could potentially accelerate donor searches by avoiding unnecessary MLC tests on Dw‐mismatched donors.
DISCRIMINATION BETWEEN HLA‐DR1 (DRB1*0101) AND DR'Br’(DRB1*0103) USING SEQUENCE‐SPECIFIC OLIGONUCLEOTIDE PROBES
Authors: Bidwell J.L. , Bidwell E.A.
Published: April 2007

Serological identification of the HLA‐DQw1(w5)‐associated or HLA‐DQw3(w7)‐associated DR ‘Br’(DRB1*0103) allele cannot be accomplished in the presence of a second DQw1(w5)‐positive or DQw3(w7)‐positive haplotype, respectively. DNA‐restriction fragment length polymorphism (RFLP) analysis assists in identification of DR ‘Br’, though not in the presence of DR1. We describe an alternative or complementary method for identification of DR ‘Br’ using two oligonucleotide probes which target HLA‐DRB1 gene HV3 regions.
INVESTIGATION OF MINK MHC (MhcMuvi) CLASS I MOLECULES BY ISOELECTRIC FOCUSING (IEF)
Authors: WIENBERG L. , Aasted B.
Published: April 2007

Mink (Mustela vision) class I leucocyte antigens, here abbreviated MhcMuvi according to the proposal given by Kleinet al.(1990), were characterized by isoelectric focusing (IEF), using Triton X‐114‐extracted and neuraminidase‐treated membrane proteins from spleen cells, followed by immunoblotting and development with a murine monoclonal antibody raised against denatured major histocompatibility complex (MHC) class I antigens.

Muvi class I antigens were investigated in a group of 97 Danish mink (including five families), of six types (Standard, Wild, Pastel, Pearl, Violet and Sapphire), and a group of 110 French ‘Wild’ mink. For each mink type maximally six protein bands in IEF were identified. Standard, Pastel and Sapphire mink only exhibited from two to four bands. This to us indicated restricted polymorphism of the Muvi class I antigens for these mink. The restriction patterns varied from farm to farm. In the family material the Muvi antigens were found to segregate as expected.

The French Wild mink were all naturally infected with Aleutian disease virus (ADV), a parvovirus, and their disease status classified as progressive or non‐progressive. There were approximately 50% in each group. The mink were grouped into one of five class I Muvi profiles. When the profiles were compared to progressive versus non‐progressive disease status, we found that mink with Muvi profile 4 and 5 almost exclusively (13 out of 15) were classified as having progressive Aleutian disease.
EXTENDED HLA‐DQw2 HAPLOTYPES: MOLECULAR ANALYSIS
Authors: Avoustin P. A. , Tkaczuk J. , Coppin H. L. , Cambon‐Thomsen A. , de Préval C.
Published: April 2007

The HLA‐DQw2 specificity, homogeneous in serology, is strongly associated to two HLA‐DR specificities: DR3 and DR7. These alleles are found mainly on DQw2 bearing extended haplotypes with strong linkage disequilibrium. We describe, with BamHI, HindIII and RsaI, two restriction fragments length polymorphisms (RFLP) for the A gene of DQw2. These two subtypes correlated with the DR3 and DR7 specificities. Interestingly, by non‐equilibrium pH gradient electrophoresis (NEPHGE), two DQα chains were also found, respectively correlated with the same DR specificities. In addition, Hindi polymorphism allowed us to distinguish several patterns of B genes for (DR7) DQw2 haplotypes but without any detectable association with another HLA marker. However, only one DQ(3 chain was found by NEPHGE in the (DR7) DQw2 haplotype. Furthermore, MncII discriminated the B genes of the two extended haplotypes: (B8, DR3) DQw2 and (B18, DR3) DQw2. The same result was found by NEPHGE: two DQβ chains were described, corresponding to the same extended haplotypes. The use of exon‐specific DQB probes showed that the genomic polymorphism in DOw2 haplotypes is located, at least, at the 3’end of the gene. These data add new characteristics to the different DQw2 extended haplotypes.
THE MOUSE FACTOR H ALLOTYPES WITH MULTIPLE AMINO ACID REPLACEMENT, H.l AND H.2 SHOW INDISTINGUISHABLE CO‐FACTOR ACTIVITY FOR FACTOR I
Authors: Natsuume‐Sakai S. , Okada M. , Seya T. , Nonaka M. , Nonaka M. , Harada Y. , Moriwaki K.
Published: April 2007

The authors report the functional analysis of the purified mouse factor H allotypes H. 1 and H.2, which were clearly distinguished from each other by an immunodiffusion test. Both allotypes acted as a co‐factor for factor I in cleaving mouse C3b and we found no significant difference between their activities. The results strongly suggest that the function of mouse factor H for the co‐factor activity has been well conserved between two allotypes.
SUSCEPTIBILITY TO MURINE HEPATITIS VIRUS (Type 3)‐INDUCED PARALYSIS IS INFLUENCED BY CLASS I GENES OF THE MHC
Authors: Oth D. , Lussier G. , Cainelli‐Gebara V. C. B. , Dupuy J.‐M.
Published: April 2007

Using H‐2 recombinant congenic strains of mice, genetic analysis of resistance to murine hepatitis virus type 3 (MHV3)‐induced paralysis was performed. It appeared that both H‐2K and H‐2D, two class I gene regions of the mouse major histocompatibility complex (MHC), can play independent significant roles in the establishment of such resistance.
NOVEL HLA‐DR2‐RELATED HAPLOTYPES IN HONG KONG CHINESE IMPLICATE THE DQB1*0602 ALLELE IN SUSCEPTIBILITY TO MULTIPLE SCLEROSIS
Authors: Serjeantson S.W. , Gao X. , Hawkins B.R. , Higgins D.A. , Yu Y.L.
Published: April 2007

Multiple sclerosis (MS) is strongly associated with the HLA‐Dw2 haplotype DRw15.DQw6 in Caucasoids, although the relative contributions of DR and DQ loci to disease susceptibility are unknown. The situation is further complicated by the apparent lack of an association between DR2 and MS in Orientals. This study examined 42 DR2‐positive chromosomes in healthy Chinese and 12 DR2‐positive chromosomes in MS patients from Hong Kong, using oligonucleotide hybridizations of DQA1, DQB1, DRB1, and DRB5 polymerase chain reaction (PCR) products. There was marked heterogeneity in DR2‐related haplotypes in controls (ten types), where the most frequent haplotype, confirmed in one family, involved the novel arrangement DRB1*1501, DQB1*0601. Another common haplotype had the unusual combination of DRB1*1602, DRB5*0101 as confirmed by DNA sequencing of the DRB5 allele. In contrast, the most common DR2‐related haplotype in MS patients was the ‘classical’ Dw2 haplotype DRB1*1501, DQB*0602, with a frequency of 50% compared with 12% in controls (P= 0.01). Novel DR, DO linkage disequilibrium relationship in Hong Kong Chinese have permitted recognition of DQB1*0602 as a susceptibility allele in DR2‐positive MS patients, although a role for the DRB1*1501 allele in MS pathogenesis has not been excluded by this study.
COMPLEXITY IN THE MAJOR HISTOCOMPATIBILITY COMPLEX
Authors: Trowsdale J. , Campbell R. D.
Published: April 2007

The human major histocompatibility complex (MHC) is one of the most intensively studied regions of the human genome, containing over 70 known genes and spanning about 4 million base pairs (4Mbp) of DNA on chromosome 6p21.3 (Klein, 1986). It can be divided up into three regions: the class I region (telomeric), the class II region (centromeric), and the class HI region (between class I and II), which includes the complement component genes C2, C4, and Bf (Trowsdale & Campbell, 1988). The MHC has been mapped in detail using pulse field gel electrophoresis (PFGE) and by cloning in yeast artificial chromosome (YAC) and cosmid vectors, revealing long stretches of DNA between the regions as well as between individual class I and class II genes. Novel genes, that have no sequence relationships with class I, class II or complement components, have recently been found in these areas, and we will present an update on these after reviewing the more established loci.
TWELVE HLA‐DR6‐RELATED DRB1 ALLELES AND ASSOCIATED DR, DQ HAPLOTYPES IN TRADITIONAL AUSTRALIANS AND OTHER POPULATIONS OF ASIA‐OCEANIA
Authors: Gao X. , Serjeantson S. W.
Published: April 2007

The relative distributions of 12 HLA‐DR6‐related HLA‐DRB1 alleles in indigenous populations of Australia, Melanesia, Polynesia, Micronesia, and northern and southern China have been determined by analysis of nucleotide sequence polymorphisms in 364 examples of HLA‐DR6 positive chromosomes. Oligonucleotide hybridizations of polymerase chain reaction products of HLA‐DQA1, DQB1, DRB1 and DRB3 genes generated 24 HLA‐DR6‐related haplotypes. The study aimed to determine the regional distribution of DR DQ haplotypes associated with three novel HLA‐DR6 alleles, namely DRB1*1408, 1409, and 1410, known to occur in Australian Aborigines, to gain further insights into the molecular phylogeny of these alleles. DRB1*1408 was the most common HLA‐DR6 subtype in Oceania, although it was not detected in Chinese. In Australian Aborigines and Papua New Guinean highlanders, DRB1*1408 was associated with DRB3*0202, while in Polynesians and Micronesians it was associated with DRB3*0101. The different haplotype arrangements, together with the near absence of DRB1*1408 in coastal Melanesians, suggest the possibility that two independent mutations have generated DRB1*1408 in Australia and Oceania. DRB1*1409 and 1410 alleles were confined to Australian Aborigines, while DRB1*1407 was found exclusively in Melanesians; DRB1*1401 was the only HLA‐DR6 allele represented in all study populations. The population‐specific HLA‐DR6 alleles and haplotypes have important implications for unrelated bonemarrow donor registries in Australia and Oceania.
DIVERSITY IN DNA REARRANGEMENTS AND IN RNA EXPRESSIONS OF IMMUNOGLOBULIN GENE ON COMMON VARIABLE IMMUNODEFICIENCY
Authors: Mori S. , Kondo N. , Motoyoshi F. , Kaneko H. , Inoue Y. , Orii T.
Published: April 2007

Six heterogeneous common variable immunodeficiency (CVID) patients were analysed for germ‐line DNA, DNA rearrangements, and RNA expressions of immunoglobulin (Ig) gene by Southern or northern blotting using appropriate probes. We detected no polymorphism in neutrophil DNA hybridized to a C μ and a C γ probe. In three patients, both serum Ig and Ig‐bearing cells were scarcely detected, and by northern hybridization methods, neither μ mRNA, γ mRNA, α mRNA nor k mRNA was detected. However, one Epstein‐Barr virus‐transformed B lymphoblastoid cell line (LCL) of these three patients was different from the germ line in the region of JH, Cγ, and Ck, and expressed μ mRNA at a higher level. The B cell defects of these three patients lay on the B cell maturation stage similar to X‐linked agammaglobulinaemia (XLA). In two others among the six CVID patients, serum IgM and IgM‐bearing cells were detected to a certain degree, and by northern hybridization, μ mRNA was detected at a lower level, but neither μ mRNA, α mRNA, nor k mRNA was detected. One LCL of these two patients could express μ mRNA at the normal level. In the last patient, the serum IgM was normal, serum IgG and IgA were somewhat low, Ig‐bearing cells were normal, μ mRNA and k mRNA were detected at the normal level, and γ mRNA and k mRNA were detected at a lower level. The defect of this patient affected the class switch stage. These results showed that primary B cell defects in CVID occurred at several B cell differentiation stages which could be classified by expression of the Ig gene, and at the degree of clonal diversity in the B cell repertoire. Furthermore, this study provides support for the idea that the CVID defect is related to a more generalized cellular function, such as regulating the proliferation and/or clonal expansion of cells of the B lymphoid lineage.