bluuebunny/crossref_metadata_2025 huggingface dataset:
WHAT IS THE SIGNIFICANCE OF THE PRESENCE OF MHC MOLECULES ON THE SURFACE OF PARASITES IN HUMAN NEUROCYSTICERCOSIS?
Authors: Trejo V. , Talamas O. , Granados G. , Castro L. , Rabiela M. T. , Sotelo J. , Gorodezky C.
Published: April 2007

Sixteen cysticerci excised from 15 surgery patients were examined for the presence of HLA molecules on their surface, to confirm the role of these molecules in parasite damage and to investigate if HLA products are host specific or perhaps host‐like antigens synthesized by the parasite. MoAbs against monomorphic and polymorphic HLA were chosen according to the patients HLA phenotypes. MoAbs against host and non‐host antigens were selected and tested on cyst slides by indirect immunofluorescence assays. Host molecules were present in 43.7% of the cysts, but non‐host antigens were also apparent in 62.5%. These results suggest mimicry as a possible mechanism to explain the presence of MHC products on the surface of the parasite; inflammation may also induce the expression of HLA that could become associated with the parasite. In vitro cellular immune response to specific antigens was also performed and positive responses correlated with the presence of HLA molecules on the cyst's surface. Moreover, damaged parasites had host molecules as well. Parasites from responder patients had all kind of HLA molecules or at least, antigenic determinants while the cysts from non‐responders did not have molecules on their surface. These data support the role of HLA in cyst destruction.
GENETIC ANALYSIS OF THE ANTIGENS DEFINED AT THE THIRD INTERNATIONALBoLAWORKSHOP
Authors: Stear M. J. , Pokorny T. S. , Fryda‐Bradley S. , Lie Ø. , Bull R. W.
Published: April 2007

A comparison of lymphocyte antigens showed that 32 of the 33BoLAantigens defined at the third internationalBoLAworkshop (Bullet al., 1989) corresponded to previously defined local antigens (Stearet al., 1988). The third workshop antigen w18 had no locally defined equivalent. All 32 antigens were shown in family studies to be expressed by autosomal co‐dominant genes, and all 32 workshop antigens were shown to be products of theBoLAsystem. After excluding the supertypic antigens, nearly all animals tested possessed only one or two antigens and there were no observed recombinants in family studies. These results do not exclude the possibility that the 32 workshop antigens are the products of one locus (BoLA‐A).
METHYLATION PATTERN OF THE HLA‐DRα GENE IN HUMAN TISSUES
Authors: Barbieri R. , Nastruzzi C. , Volinia S. , Villa M. , Piva R. , Giacomini P. , Natali P. G. , Gambari R.
Published: April 2007

The CCGG and GCGC sites of the human HLA‐DRα gene are hypermethylated in human tissues (including B‐lymphocytes, T‐lymphocytes, muscle, brain, sperm, skin, kidney, suprarenal and mammary glands) and three B‐lymphoid cell lines. Therefore, the HLA‐DRα gene can be transcribed even though extensively methylated. The only exception to the hypermethylated state of the HLA‐DRα gene is represented by one or both of the two HhaI sites (H1 and H2) localized in the 5’portion of the gene. Analysis of the computer‐generated secondary structure of the HLA‐DRα mRNA suggests that the H1 and H2 sites belong to a region (5′‐GAGCGCCCA‐3′/5′‐UGAGCGCUC‐3′) exhibiting extensive base pairing. Therefore, unmethylation of these CG sites can contribute in preventing mCG→TG/CA changes in this region, which would lead to extensive alterations of the secondary structure of the 5’portion of the HLA‐DRα MRNA.

On the other hand, the selective pressure to maintain unaltered the methylated CG dinucleotides in the coding regions of the HLA‐DRα gene could be due to codon restrictions, since the majority of the methylation‐related CG→TG or CG→CA variations would generate aminoacid changes.

Accordingly, the analysis of different HLA‐DRα genomic sequences indicates that variations of the CpG dinucleotides occur only in the non‐coding portions of the HLA‐DRα gene.
INACTIVATION OF THE H‐2K1k GENE COULD INVOLVE THE SUBSTITUTIONS OF METHYLATED CpGs
Authors: Sim B. C. , Grosveld F. G. , Hui K. M.
Published: April 2007

By the isolation of overlapping cosmid clones and ‘chromosome walking’ studies from the H‐2Kk gene, we have obtained cosmid clones encoding the H‐2K1k gene from two separate cosmid libraries. The nucleotide sequence of one of the clones was determined. The cloned H‐2K1k gene could be transcribed in vitro to give a normal H‐2 class I mRNA of 1.7 kb. However, the deletion of four nucleotides in exon 3 of the H‐2K1k gene results in a translation termination codon at the beginning of exon 4. In agreement with this, when expressed in human cells, the H‐2K1k gene gave a truncated, cytoplasmic polypeptide of Mr 36,000. Therefore, although the H‐2K1k gene is homologous to other class I MHC genes in its molecular organization and nucleotide sequence, it is a pseudogene. When compared to the nucleotide sequence of the H‐2Kk gene, the H‐2K1k gene has undergone many substitutions of methylated CpG residues (meCpG). This represents further evidence to suggest that this gene is inactive.
CHARACTERIZATION OF SOLUBLE HLA MOLECULES IN SWEAT AND QUANTITATIVE HLA DIFFERENCES IN SERUM OF HEALTHY INDIVIDUALS
Authors: Zavazava N. , Westphal E. , Muller‐Ruchholtz W.
Published: April 2007

Soluble class I molecules were immunoprecipitated from human sweat and serum using the BB7.7 monoclonal antibody (mAb) coupled to immunomagnetic beads. Molecules were analysed biochemically on SDS‐PAGE gels and finally by 1D‐isoelectric‐focusing (IEF). Serum‐ and sweat‐HLA IEF‐band patterns of the same individual were fully identical, showing that HLA excreted in sweat possess polymorphic structures like those in serum. Quantitatively, we used a highly sensitive competitive enzyme‐linked immunosorbent (ELISA) assay to determine soluble class I concentrations. The first group was that of non‐HLA‐A9 and ‐Bw62 sera, which were found to contain HLA levels with a mean concentration of 0.82 ± 0.63 μg/ml (n= 44). However, sera that were HLA‐A23 or −24 (splits of HLA‐A9) contained higher levels, with a mean of 3.2 ± 0.94 μg/ml (n= 20). Similarly, HLA‐Bw62 individuals had a higher mean of 2.05 ± 0.65 μg/ml (n= 10). The difference of the HLA‐A9 group to the first group was statistically highly significant, P < 0.0001, and that of the HLA‐Bw62 to the first was also significant, P < 0.004. Individuals who were both HLA‐A9 and ‐Bw62 (n = 5) did not express significantly higher levels than those who only had one of these specificities. Sweat HLA levels had a mean of 0.42 ± 0.4 μg/ml (n= 10). These results show for the first time that soluble class I peptides are excreted in relatively high concentrations in sweat and possess polymorphic structures identical to those of serum HLA and that serum HLA levels are allotype dependent.
PCR‐SSO TYPING FOR HLA‐DRB ALLELES
Authors: Vaughan R.W.
Published: April 2007

Polymerase chain reaction (PCR) amplification of the second exon of DRB genes from genomic DNA is described. Sequence‐specific oligonucleotide (SSO) probes have been designed to allow for the typing of DRB alleles. The methods are described in detail and the results of a trial on both homozygous typing cells and restriction fragment length polymorphism (RFLP)‐typed local caucasoid controls are given. The ease of use of this form of DRB typing is emphasized and potential complications are discussed.
PCR‐SSO TYPING IN HLA‐DISEASE ASSOCIATION STUDIES
Authors: WORDSWORTH P.
Published: April 2007

Associations between a large number of diseases and markers within the major histocompatibility complex (MHC) have been described. In particular, susceptibility to several autoimmune disorders, including type I diabetes mellitus and rheumatoid arthritis, is linked to genes within the MHC and strong population associations are demonstrable between certain HLA class II alleles and these conditions. Genetic mapping of HLA susceptibility loci has traditionally relied on the use of phenotypic markers defined by alloantisera, cellular typing reagents and biochemical analysis of histocompatibility antigens. Polymerase chain reaction sequence‐specific oligonucleotide (PCR‐SSO) typing combines the ability to define the finest of HLA specificities, by analysis of the corresponding DN A sequences, with the possibility of studying large populations of normal and affected individuals. The applications of this technology to characterizing precisely the MHC loci associated with susceptibility to autoimmune diseases such as rheumatoid arthritis, type I diabetes mellitus, coeliac disease and pemphigus vulgaris are reviewed here.
ANALYSIS OF Ly5 CHROMOSOME 1 POSITION USING ALLELIC DIFFERENCES AND RECOMBINANT INBRED MICE
Authors: Zebedee S.L. , Barritt D. , Epstein R. , Raschke W.C.
Published: April 2007

Differences between the mouse Ly5a and Ly5b alleles can be distinguished on the basis of polymerase chain reaction (PCR)‐restriction enzyme analysis and differential monoclonal antibody reactivities. To more precisely map the Ly5 gene on the mouse chromosome 1, analytical DNA and protein tests were performed on recombinant inbred strains of mice prepared from SJL/J (Ly5a) and B ALB/cke (Ly5b) progenitor strains. Each recombinant inbred strain was characterized to determine whether it carried the Ly5a or Ly5b allele. Both assays, DNA‐PCR and protein‐immunofluorescence, yielded identical results for each strain examined. Placement of the Ly5 gene with respect to other characterized markers of mouse chromosome 1 for these recombinant inbred mouse strains shows a gene order of Idh‐1.Ity:Pep3:/Ly5, CfhJ.
PHAGOCYTOSIS OF FLUORESCENT LATEX MICROBEADS BY PERITONEAL MACROPHAGES IN DIFFERENT STRAINS OF MICE: A FLOW CYTOMETRIC STUDY
Authors: ÁBel G. , Szollösi J. , Fachet J.
Published: April 2007

The phagocytosis of uniform fluorescent latex particles by resident and thiogiycollate‐elicited macrophages was analysed by flow cytometry. The percentage of phagocytos‐ing macrophages and the number of internalized microspheres per cell was determined from cell size and fluorescence histograms. Results were corrected for the adherence of microbeads to the cells in the presence of sodium azide in the medium. Human C3b‐or murine monoclonal IgG‐coated microspheres were applied to assess receptor‐mediated phagocytosis in different inbred strains of mice. Phagocytic activity of thioglycollate‐elicited macrophages was consequently higher than that of resident macrophages. A decreasing gradient of C3b and Fc receptor‐mediated phagocytosis was established in the following order: B10.BR, BIO, C3H/Di and C3H.SW strains. Our results indicate that the phagocytic function of murine macrophages is under control of both the somatic (non‐H‐2) and H‐2 genes.
CHARACTERIZATION OF BOVINE MHC CLASS II POLYMORPHISM USING THREE TYPING METHODS: SEROLOGY, RFLP AND IEF
Authors: Davies C. J. , Andersson L. , Joosten I. , Mariani P. , Gasbarre L. C. , Hensen E. J.
Published: April 2007

Various methods, with different strengths and weaknesses, are currently used to define polymorphism of the bovine major histocompatibility complex (MHC) class II genes. A more complete characterization of bovine lymphocyte antigen (BoLA) haplotypes can be achieved by combining several of these methods. In this study BoLA class II polymorphism was characterized using three typing methods: serology, restriction fragment length polymorphism (RFLP), and isoelectric focusing (IEF). Twenty six Holstein‐Friesian and 15 Angus cattle that carried an array of serologically defined BoLA haplotypes were selected for the study. The panel included 12 BoLA complex homozygotes. The three class II typing methods recognized polymorphism associated with the same or very tightly linked genes in the DQ‐DR class II subregion. In total 25 BoLA‐A locus (class I) —DQ‐DR subregion (class II) haplotypes were defined. Three of the serological class II specificities, Dx1, Dx3, and Dx4, were associated with more than one RFLP defined DQ‐DR haplotype. The other 4 class II specificities behaved as private specificities. One BoLA haplotype was found in both Holstein and Angus cattle. Two other BoLA haplotypes defined here have previously been described in other breeds. This suggests that these haplotypes exist in strong linkage disequilibrium.
NUCLEOTIDE SEQUENCING OF HLA‐DQ GENE SECOND EXONS IN CHINESE HOMOZYGOUS CELLS
Authors: Wang* Y. , Lu P. , Zhou K. , Clay T. , Wood N. , Bradley B. , Bidwell J.
Published: April 2007

Six HLA class I and class II‐homozygous Chinese cell lines with unique HLA‐Dw types were studied. Since the majority of HLA class II nucleotide sequence polymorphism is localized within the second exons of the genes, we used the polymerase chain reaction (PCR) to amplify these regions in HLA‐DQA and DQB genes and subsequently determined the nucleotide sequences. No unique DQA1 or DQB1 alleles were found. However, a new haplotype of DQA1*601‐DQB1*0301‐DRB1*1202 was found in two cells; and DQA1*03011 was found in association with DR9 in another two cells. This indicates that new DR‐DQ associations may explain the observed new HLA‐Dw types. The DQB2 sequences were identical in all six cells and were identical to a sequence previously reported in a DR6 haplotype. The DQA2 sequences from two clones obtained from two cells differed from each other and from previously reported sequences. The results show that the DQA1 and DQB1 alleles in the Chinese individuals studied are as previously reported in Caucasian populations and as such may be typed by restriction fragment‐length polymorphism (RFLP) or PCR‐sequence‐specific oligonucleotide typing (PCR‐SSO) or PCR‐RFLP using conventional probe or restriction enzyme sets.
POLYGENIC INFLUENCES ON THE LENGTH OF OESTROUS CYCLES IN INBRED MICE INVOLVE MHC ALLELES*
Authors: Lerner S. P. , Anderson C. P. , Harrison D. E. , Walford R. L. , Finch C. E.
Published: April 2007

Genetic influences on female reproductive cycles were analysed in histocompatibilitycongenic strains of mice. Oestrous cycles of young, virgin mice of inbred‐congenic strains, hybrid crosses (F1, and parental‐hybrid backcrosses (F2) were monitored for 3 months. Oestrous cycles were categorized by length (inter‐oestrous interval): 4,5,6, or 7–14 days. Mice with the following H‐2 haplotypes had a greater proportion of 5‐day oestrous cycles: H‐2b, H‐2r, H‐2h2, H‐2h4, and H‐2i5. In contrast, the H‐2k and H‐2d haplotypes had mostly 4‐day oestrous cycles. Influences of H‐2 haplotype were seen on two genetic backgrounds, C57BL/10Sn and C3H. Non‐H‐2 alleles were also implied by different patterns of cycles between strains with the same H‐2b haplotype: C57BL/10Sn with predominantly 5‐day cycles vs. C57BL/6J with a mix of 4‐ and 5‐day cycles.

The genetic basis for strain differences was investigation in F1 hybrids and their backcrosses. F1 hybrids of an H‐2b (C57BL/10Sn; 5‐day cycles) and an H‐2k (B10.BR; 4‐day cycles) strain had mostly 5‐day cycles, indicating dominance of an H‐2b allele(s). However, F1 hybrids from the reciprocal B6 × B10 cross (both H‐2b) also display a preponderance of 5‐day cycles, indicating dominance of a non‐H‐2 autosomal allele from the C57BL/10Sn strain. Among F2 mice, a ‘4‐day’ phenotype segregated with homozygosity for the k haplotype (P < 0.05, χ2). These findings demonstrate the influence of genetic differences at the major histocompatibility complex on oestrous cycles.
C5 DEFICIENCY IN A/J MICE IS NOT ASSOCIATED WITH RESISTANCE TO THE DEVELOPMENT OF SECONDARY AMYLOIDOSIS
Authors: Coutinho M. , Zahedi K. , Whitehead A. S. , Davis A. E.
Published: April 2007

The aim of the study was to determine whether C5 deficiency in the mouse is associated with resistance to the development of secondary amyloidosis. Chronic inflammation was induced in the F2 progeny, derived from matings between amyloid‐susceptible and amyloid‐resistance mice, by daily injections of azocasein for thirty days. Using a restriction fragment length polymorphism generated by digestion of genomic DNA with the restriction enzyme HindIII, C5 sufficient and deficient DNA can be clearly differentiated. Eight mice were found to be C5 sufficient, 32 were heterozygotes and 14 were found to be C5 deficient. Grading of the splenic amyloid load from negative to 4+ was performed after staining tissue squashes with Congo red and viewing them under a polarizing microscope. Seventeen mice were noted to have negative to trace, 18 had moderate (1+‐2+) and 19 had heavy (3 H+‐4+) amyloid deposition. There was no correlation between splenic amyloid load and C5 deficiency. Based on these results it is clear that C5 deficiency and resistance to secondary amyloidosis are not associated.
HLA‐DRB1* GENE SEQUENCES IN HLA‐DR4 POSITIVE AND NEGATIVE PATIENTS WITH RHEUMATOID ARTHRITIS
Authors: Becking A. , Pluschke G. , Krawinkel U. , Melchers I. , Peter H.H. , Lang B.
Published: April 2007

The second exon of the DRB1 gene encoding for the first domain of the HLA‐DR β‐chain was sequenced in 16 patients (10 DR4/DR1 positive, 6 DR4/DR1 negative) with seropositive rheumatoid arthritis (RA). We could confirm the strong association of susceptibility to RA with functionally equivalent conformations on otherwise distinct MHC molecules. At least one HLA‐DR allele in all of the analysed DR4 or DR1 positive patients showed such an epitope with a minimal variability limited to residue 71.

However, in HLA‐DR4 and ‐DR1 negative patients such a similar epitope could not be detected.
Lp(a) LIPOPROTEIN AND HLA‐DR GENOTYPE IN EARLY CORONARY ARTERY DISEASE
Authors: Dahlén G.H. , Slunga L. , Holmlund G. , Langö A. , Lindblom B.
Published: April 2007

The interest in Lp(a) lipoprotein [Lp(a)] has increased dramatically during the last few years due to the documented strong association between high Lp(a) levels and early atherosclerosis and its sequelae and the probable additional thrombogenic effect of inherited high Lp(a) levels.

However, some paradoxes still remain to be solved in Lp(a) research. This pilot study was performed to test whether some criteria could be found for an association between Lp(a) levels, HLA genotype, and early coronary heart disease. If so, it could help to explain why some individuals with high Lp(a) levels get atherosclerosis while others with comparable lipid levels do not. It would also indicate that immunological processes could be involved in Lp(a) associated atherosclerosis.

In this case‐control study the HLA‐DR genotypes 13 or 17 were significantly more frequently encountered in 30 male patients with early coronary heart disease than in 30 sex and age matched healthy controls (P = 0.012). These HLA‐DR specificities were especially frequent in male patients with high Lp(a) levels. The same HLA‐DR genotype pattern was not seen in 30 female patients, although there was a trend towards more frequent 13a genotype.
SHARED EPITOPES OF THE HLA‐DR10 MOLECULE RECOGNIZED BY MURINE AND HUMAN mAbs
Authors: PISTILLO M.P. , SUN P.F. , MANTERO S. , FERRARA G.B.
Published: April 2007

In order to produce mAbs directed specifically against HLA‐DR10 molecule, transfected mouse L cells, expressing the DRB 1*1001 allele, were used to immunize C3H mice over a period of 4 weeks. Two mAbs, 2C12 and 4B6, derived from this fusion were found to recognize, with different affinity, polymorphic epitopes of DR10 that are shared with DR1, 3, 7, and 9. These mAbs were screened on a large panel of homozygous B lymphoblastoid cell lines using microlymphocytotoxicity and the results were confirmed by flow cytometry. The reactive pattern of 2C12 and 4B6 was compared to that of MP10 human mAb also recognizing the DR10 specificity in addition to DR1, 2 and 9.

Based on serologic specificity and cellular absorption experiments, we conclude that the epitopes the murine and human mAbs respectively recognize on the DR10 molecule, are probably different.
MULTIPLEX ARMS‐RFLP: A SIMPLE AND RAPID METHOD FOR HLA‐DR4 SUBTYPING
Authors: Jawaheer D. , Ollier W. E. R. , Thomson W.
Published: April 2007

We have developed a simple and rapid non‐radioactive technique for HLA‐DR4 subtyping. A multiplex ARMS‐RFLP (Amplification Refractory Mutation System – Restriction Fragment Length Polymorphism) system allows HLA‐DR4 subtyping by analysis of the products of two multiplex ARMS reaction mixtures. For some cases, restriction enzyme digests (Hae II and/or Sac II) of the products are analysed. The technique relies on the fact that an ARMS primer with a mismatch at its 3′‐end with respect to the template will not be elongated under PCR conditions. Hence, by designing ARMS primers such that different HLA‐DR4 alleles yield PCR products of different lengths, only two reactions, each using a mixture of different ARMS primers, are sufficient to type all of the known HLA‐DR4 alleles. This system can distinguish between HLA‐DR4 ‘homozygotes’ and ‘heterozygotes’ since every HLA‐DR4 allele can be detected. The ARMS conditions were optimized using DNA from cell lines. This technique has now been used to type a panel of rheumatoid arthritis patients and controls.
PHENOTYPE DISTRIBUTION OF THE THIRD COMPONENT OF COMPLEMENT (C3) AND OF THE PROPERDIN B FACTOR (BF) IN CHILDREN WITH MUMPS MENINGITIS
Authors: Halasa J. , Kacprzak‐Beroman I. , Maǹczak M. , Schlesinger D.
Published: April 2007

The C3 and BF phenotype frequencies were studied in children with mumps meningitis. No significant differences were found between this group and other groups; children with mumps without meningitis and healthy children.
OCCURRENCE OF CAPRINE LEUCOCYTE CLASS I AND II ANTIGENS IN SAANEN GOATS AFFECTED BY CAPRINE ARTHRITIS (CAE)
Authors: Ruff G. , Regli J.G. , Lazary S.
Published: April 2007

The distribution of CLA class I and II antigens in CAE virus‐infected, diseased and healthy Saanen goats has been investigated. Three class I antigens and two class II antigens showed a statistically different frequency in the two groups.
HLA‐DR AND ‐DQB1 GENOTYPING IN A CHINESE POPULATION
Authors: Zhao Y.P. , Yang J.Q. , Ge Y. , Fan L.A. , Loiseau P. , Colombani J.
Published: April 2007

Using molecular biological methods, 58 unrelated Chinese from Shanghai were typed for HLA‐DR and DQ. The Shanghai population possesses the principal HLA‐DR and DQ characteristics of the oriental populations but with an increase of the DRB1* 12 allele.

So HLA typing of populations appears to be important not only for anthropological studies but also for transplantations and HLA associations with diseases.
A SHARED HLA‐DRB1 SEQUENCE CONFERS RA SUSCEPTIBILITY IN GREEKS
Authors: Carthy D. , Ollier W. , Papasteriades C. , Pappas H. , Thomson W.
Published: April 2007

Previous serological studies of Greek rheumatoid arthritis (RA) patients have failed to demonstrate an association with DR4. Using sequence specific oligonucleotide typing we have identified the DRB1 alleles in panels of Greek RA patients and controls.

When patient and control HLA‐DRB1 frequencies were compared, significantly higher frequencies of DRBl*0101 (23.3% vs. 7.0%, odds ratio [OR] 4.0, 95% confidence intervals [CI] 1.4‐12.0) and DRB1 *1001 (20.9% vs. 5.8%, OR 4.3,95% CI 1.3‐13.7) were found in RA patients compared with controls. No association of DRB1*04 with RA was observed (20.9% vs. 14.0% in controls) confirming earlier reports. However DRB1*04 subtyping demonstrated a small but significant increase of DRB1*0405 in patients (14.0% vs. 3.5%, OR 4.5, 95% CI 1.1‐18.9).

When the frequency of individuals carrying the shared RA susceptibility epitope was compared between patients and controls it was found to be significantly higher in RA patients (60.5% vs. 22.1%, OR 5.4, 95% CI 2.4‐12.0). We conclude that the shared epitope is significantly associated with RA in this population, but that it is predominantly accounted for by DRB 1*0101 and DRB1*1001.

Previous studies of UK RA patients have demonstrated a negative association of DR2 with disease and articular erosions. HLA‐DR2 variants, DRB1*1501 and *1502 are not at reduced frequency in Greek RA patients (DRB1*7507, 14.0% in patients vs. 7.0% in controls; DRB1*1502, 7.0% in patients vs. 7.0% in controls). Genes conferring RA resistance may be in linkage disequilibrium with DR2 in UK patients. This does not appear to be the case in Greek RA patients.

No association was seen between RA and HLA‐DPB1 type.
THE INDUCIBLE FORM OF NITRIC OXIDE SYNTHASE (NOS2) ISOLATED FROM MURINE MACROPHAGES MAPS NEAR THE NUDE MUTATION ON MOUSE CHROMOSOME 11
Authors: Mock B. A. , Krall M. M. , Byrd L. G. , Chin H. , Barton C. H. , Charles I. , Liew F. Y. , Blackwell J.
Published: April 2007

Nitric oxide synthase has been shown to mediate streptozocin‐induced diabetes and to act as an antimicrobial agent in murine macrophages. Using a cDNA probe for the inducible form of nitric oxide synthase (Nos2) isolated from murine macrophages we have determined that the gene maps within 1 cM of the nude mutation on mouse Chromosome 11. The position of Nos2 was also mapped relative to the markers 115, Evi 2, Cchlbl (previously unmapped), and Gfap. This map location is discussed relative to map locations for disease susceptibility loci involved in mediating cutaneous leishmaniasis (Sell) and autoimmune type‐I diabetes (Idd4).
POLYMORPHISM OF BOVINE MHC CLASS I GENES. JOINT REPORT OF THE FIFTH INTERNATIONAL BOVINE LYMPHOCYTE ANTIGEN (BoLA) WORKSHOP, INTERLAKEN, SWITZERLAND, 1 AUGUST 1992
Authors: Davies C. J. , Joosten I. , Bernoco D. , Arriens M. A. , Bester J. , Ceriotti G. , Ellis S. , Hensen E. J. , Hines H. C. , Horin P. , Kristensen B. , Lewin H. A. , Meggiolaro D. , Morgan A. L. G. , Morita M. , Nilsson Ph.R. , Oliver R. A. , Orlova A. , ØStergarrd H. , Park C. A. , Schuberth H.‐J. , Simon M. , Spooner R. L. , Stewart J. A.
Published: April 2007

The objectives of the Fifth International BoLA Workshop were to: standardize nomenclature, compare typing methods, and characterize BoLA haplotypes. The workshop was based on the distribution of blood samples (cells) from 60 selected cattle to 14 laboratories. Results for the class I (BoLA‐A) region are presented in this paper while results for the class II regions are presented in a separate report. Thirty‐six of the 50 previously established serological class I specificities were represented in the cell panel. However, only 30 specificities could be confirmed. Two specificities, A16 and A32, were upgraded from provisional, workshop (w) specificities to BoLA‐A locus specificities and three new specificities, w51(w28), w52 and w53(w28), were defined. The 39 specificities distinguished 30 class I haplotypes in the 60 animals. Class I isoelectric focusing proved to be a useful adjunct to the serology. Isoelectric focusing confirmed several serologically defined splits and detected splits of A15(A8), A18(A6) and A22(w49) that had not been detected by serology. Subsequently, serological support for splits of A15(A8) and A22(w49) was found.
INDICATIONS OF AN AUTOIMMUNE COMPONENT IN LP(a) ASSOCIATED DISORDERS
Authors: Dahlén G. H.
Published: April 2007

The exceptionally strong independent association found between Lp(a) lipoprotein [Lp(a)] levels and atherosclerotic disorders indicate that Lp(a) is a factor of considerable importance in the pathogenesis of atherosclerosis. The association between Lp(a) and diabetes, rheumatoid arthritis and renal diseases suggest that Lp(a) may be involved in immunological mechanisms.

Lp(a) has a great tendency to aggregate and bind to glucosaminoglycans, fibrin and fibronectin and is preferentially retained in the extracellular matrix during development of atherosclerosis and is in vitro phagocytosed by macrophages, probably as small aggregates. It was previously found that the Lp(a) level is significantly related to the HLA class II genotype in male patients with early coronary artery disease. In this paper additional results of interleukin determinations in relation to HLA type and Lp(a) levels are presented and discussed. It is suggested that an autoimmune process, perhaps triggered by a concomitant intracellular infection may occur, especially in patients with inherited high Lp(a) levels in combination with certain inherited HLA class II genotypes.
IMMUNOGENETIC ANALYSIS OF SUCCESSFUL AND REJECTED BONE MARROW GRAFTS WITHIN ONE FAMILY*
Authors: Martinez O. P. , Witt C. S. , Tay G. , Christiansen F. T. , Dawkins R. L.
Published: April 2007

We report the case of an Indonesian patient who required urgent bone marrow transplantation for acute leukaemia and who received successive transplants from two siblings. The first transplant failed while the second was successful. There were some uncertainties in serological typing due to the presence of cross‐reacting HLA‐B alleles, lack of paternal typing and the use of Caucasoid sera for Indonesian patients. Distinction between the two donors was also difficult. Interestingly, the use of a new DNA technique identified the presence of differences between the patient and the first unsuccessful donor but not the second successful donor.
HLA‐DQB1‘LOW‐RESOLUTION’ TYPING BY PCR AMPLIFICATION WITH SEQUENCE‐SPECIFIC PRIMERS (PCR‐SSP)
Authors: Aldener‐Cannavá A. , Olerup O.
Published: April 2007

In the study described here primers were designed for DQB1‘low‐resolution’, i.e. generic, typing by PCR amplification with sequence‐specific primers (PCR‐SSP) considering all the currently recognized DQB 1 alleles, i.e. 0501‐0504.0601‐0609,0201,0301‐0305 and 0401‐0402. This resolution was achieved by performing eight PCR reactions per individual. The DQB1 alleles corresponding to the serological specificities DQ4, DQ5 and DQ6 were uniquely identified, whereas the DQ2. DQ7, DQS and DQ9 specificities were amplified by two primer mixes. All homozygous and heterozygous combinations of the serological series DQI to DQ9 could be distinguished. The yield of amplified products were increased compared to our previously described DQB1‘high‐resolution’ typing technique by lengthening many of the primers, modifying the PCR cycling parameters and by including glycerol in the PCR reaction mixtures. Thirty‐one cell lines and 90 donor spleen cells were investigated by the DQB1‘low‐resolution’ PCR‐SSP technique as well as by Taq1 DRB‐DQA‐DQB RFLP analysis. The concordance between PCR‐SSP typing and RFLP analysis was 100%. The cell lines and 20 of the spleen cells were typed twice with complete reproducibility. No false positive or false negative typing results were obtained. DQB1‘low‐resolution’ PCR‐SSP typing, including DNA extraction, PCR amplification, gel detection, documentation and interpretation, were performed in 2 h which renders the PCR‐SSP technique suitable also for the genotyping of cadaveric organ donors.
HLA CLASS II REGION NUCLEOTIDE SEQUENCES, 1994
Authors: Marsh S. G. E. , Bodmer J. G.
Published: April 2007

The HLA class II region sequences included in this compilation are taken from publications listed in the papers: Nomenclature for factors of the HLA system, 1994 (Bodmer et al., 1994), 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. In the following Tables, allele alignments include sequences across all exons. However, since for many alleles sequence information is incomplete, such alleles will be cut short in the Tables and the alignments compressed to conserve space.
ABSENCE OF TAP 2D IN YORUBA NIGERIANS
Authors: Awomoyi A. A. O. , Donn R. P. , Davies E. J. , Carthy D. , Thomson W. , Ollier W. E. R.
Published: April 2007

We have characterized TAP allele frequencies in a panel of 71 Yoruba Nigerians using ARMS‐PCR. With the exception that TAP 2D was absent in Nigerians, TAP 2 allele frequencies in this population were found to be similar to those in a UK white population. HLA‐DR4 also was found to be at a low frequency in Yoruba Nigerians (1.4%). This may reflect the absence of TAP 2D in Nigerians as DR4 and TAP 2D are in linkage disequilibrium in UK Caucasoids.

The most frequent TAP 1 allele in Yoruba Nigerians was TAP 1A (49%). However, this value will be an underestimate as TAP1 alleles could not be unequivocally assigned in 41 % of subjects using the ARMS‐PCR methodology.
STUDY OF HLA CLASS I/II AND T LYMPHOCYTE SUBSETS IN KUWAITI VITILIGO PATIENTS
Authors: AL‐Fouzan A. , Al‐Arbash M. , Fouad F. , Kaaba S.A. , Mousa M.A. , Al‐Harbi S.A.
Published: April 2007

HLA polymorphisms of class I and class II MHC were investigated in 40 Kuwaiti vitiligo patients and in 40 controls using microcytotoxicity assay. HLA‐B21, Cw6 and DR53 were increased significantly in patients compared to controls (P= 0.00001, 0.00001 and P= 0.0053 respectively) while HLA‐A19, DR52, were significantly decreased (P= 0.00236,0.05, respectively). Total T‐cells, T4 and T8 were measured as CD2, CD4 and CD8 respectively by flow cytometry. Vitiligo patients showed significant increase in CD4 compared to controls (P= 0.03). Our findings suggest that HLA‐B21 and Cw6 and DR53, are susceptible genes of vitiligo, while A19 and DR52 are protective genes in the Kuwaiti population.
DELETION OF THE 3 ′ SPLICE SITE OF THE LEADER‐VARIABLE REGION INTRON OF IMMUNOGLOBULIN HEAVY CHAIN GENES INDUCES A DIRECT SPLICING OF LEADER TO CONSTANT REGION, RESULTING IN THE PRODUCTION OF TRUNCATED μ‐CHAINS
Authors: Komori T. , Sugiyama H.
Published: April 2007

An Abelson virus‐transformed immature B cell line, AT8‐1‐12‐5‐2, produced truncated μ‐chains. Sequencing analysis of the VHDJH complex on the expressed H‐chain allele revealed the deletion of 75 nucleotides that involved leader‐variable region intron and the 5’ end of the variable region, which resulted in the loss of the 3′ splice site of leader‐variable region intron. Sequence studies of a leader‐ and CH1 ‐containing cDNA clone showed that leader region was directly spliced to the CH1 exon, resulting in the production of the truncated μ‐chains without variable portion. Our results demonstrated for the first time that the only loss of the 3′ splice site of leader‐variable region intron could induce an aberrant splicing between leader and constant region.
AT LEAST FOUR MHC CLASS I GENES ARE TRANSCRIBED IN THE HORSE: PHYLOGENETIC ANALYSIS SUGGESTS AN UNUSUAL EVOLUTIONARY HISTORY FOR THE MHC IN THIS SPECIES
Authors: Ellis S. A. , Martin A. J. , Holmes E. C. , Morrison W. I.
Published: April 2007

Nineteen horse MHC class I specificities have been serologically identified previously at a single locus (ELA‐A), and two other specificities appear to be coded at other loci. Biochemical studies indicate that there are at least two expressed loci. In order to establish the number of transcribed horse MHC class I genes, we made a cDNA library from a heterozygous animal (ELA‐A3/A7), and screened for positive clones using a bovine class I probe. More than 200 class I clones were isolated in this way, and so far seven unique full length sequences have been identified. All of the sequences are predicted to code for surface expressed, functional molecules. The number of different sequences identified demonstrate that at least four genes are transcribed, although variations in transmembrane length (which is generally conserved in class I loci) suggest that five genes could be represented. Evolutionary analysis of these sequences (and two additional sequences known to represent different horse class I loci) reveals no firm relationships, such that the division between the different loci cannot be discerned. These results suggest an unusual evolutionary history for the horse MHC, the precise nature of which may be revealed only following further cross‐species comparisons.
STRONG ASSOCIATION BETWEEN THE RESPONDER STATUS OF THE FCγ II RECEPTOR and RECURRENT SPONTANEOUS ABORTION
Authors: Lorenz A. , Blasczyk R. , Kuhn U. , Venjakob U. , Mendonca M. , Grosse‐Wilde H.
Published: April 2007

Several models exist for the aetiology and therapy of recurrent spontaneous abortion (RSA). They are based in part on the assumption that an increased materno‐fetal histocompatibility results in an insufficient maternal immunological recognition of the fetus, and thus renders the maternal immune system unable to sustain the pregnancy. The involvement of the FCγII receptor (FcγIIR) in RSA was suggested, since FcγIIR‐blocking antibodies, present in normal pregnancies, could not be found in patients suffering from RSA.

The FcγIIR is known to be functionally and structurally dimorphic, which results in a responder, respectively non‐responder pattern. We used an in vitro proliferation assay to distinguish between FCγIIR responder and non‐responder phenotypes. In 29 RSA couples we found 97% (28/29) of RSA patients and 100% (29/29) of their partners to be responders, whereas 50 fertile couples and a further 100 unrelated controls revealed distributions of responder vs. non‐responder of 73% vs. 27% and 70% vs. 30%, respectively. These differences (RSA vs. controls) are highly significant (P≤ 0.0001). Our results suggest further that the FcγIIR dimorphism might be involved in the pathogenesis of RSA.
DETERMINATION OF DQB1 ALLELES USING PCR AMPLIFICATION AND ALLELE‐SPECIFIC PRIMERS
Authors: Lepage V. , Ivanova R. , Loste M.N. , Mallet C. , Douay C. , Naoumova E. , Charron D.
Published: April 2007

Molecular genotyping of HLA class II genes is commonly carried out using polymerase chain reaction (PCR) in combination with sequence‐specific oligotyping (PCR‐SSO) or a combination of the PCR and restriction fragment length polymorphism methods (PCR‐RFLP). However, the identification of the DQB1 type by PCR‐SSO and PCR‐RFLP is very time‐consuming which is disadvantageous for the typing of cadaveric organ donors. We have developed a DQB1 typing method using PCR in combination with allele‐specific amplification (PCR‐ASA), which allows the identification of the 17 most frequent alleles in one step using seven amplification mixtures. PCR allele‐specific amplification HLA‐DQB1 typing is easy to perform, and the results are easy to interpret in routine clinical practice. The PCR‐ASA method is therefore better suited to DQB1 typing for organ transplantation than other methods.
THE EFFECT OF TNF*B GENE POLYMORPHISM ON TNF‐α AND ‐β SECRETION LEVELS IN PATIENTS WITH INSULIN‐DEPENDENT DIABETES MELLITUS AND HEALTHY CONTROLS
Authors: Whichelow C. E. , Hitman G. A. , Raafat I. , Bottazzo G. F. , Sachs J. A.
Published: April 2007

TNF‐α and ‐β have been implicated in the development of HLA‐associated autoimmune diseases. It has been suggested that inter‐individual differences in the secretion levels of these cytokines may contribute to the predisposition of certain individuals to the development of diseases such as insulin‐dependent diabetes mellitus (IDDM). We have investigated whether a diallelic TNF*B polymorphism detected using the enzyme Ncol influences the TNF‐α and/or ‐β secretory capacity of peripheral blood mononuclear cells (PBMC) from PHA stimulated healthy individuals and IDDM patients.

We have shown that the level of TNF‐β secreted correlates with the TNF*B genotype in healthy individuals: those with the TNF B*2 allele secreted significantly higher levels of TNF‐β (P= 0.025) than those with the TNF*B1 allele. In IDDM patients, the reverse situation was observed, with those patients with the TNF*B1 allele secreting higher levels of TNF‐β than those with the TNF*B2 allele. No correlation was found between TNF‐α levels and TNF*B genotype. Furthermore, when IDDM patients and controls were matched for TNF*B genotype, the IDDM patients with the TNF*B2 allele secreted significantly lower levels of TNF‐β than controls with this allele.

On analysis of IDDM‐susceptible extended HLA haplotypes in the homozygous groups, 4/7 IDDM patients with the TNF*B2 allele were Bw62‐DR4 compared with 0/16 matched controls. Thus, the extended haplotype Bw62‐DR4‐TNF*B2/2 rather than IDDM per se is almost certainly responsible for the depressed TNF‐β secretion found in the IDDM‐TNF*B2 homozygous cohort.
GENOTYPE‐RELATED FINGERPRINTS FROM HLA‐DPB1 EXON 2 LOW‐STRINGENCY PCR
Authors: Porfirio B. , Chiarelli I. , Rombolà G. , Mattiuz P.L.
Published: April 2007

Techniques based on the formation of heteroduplexes between relevant heterologous amplified HLA loci have proved to be simple and cost‐effective screening methods for the detection of DNA sequence diversity. However, the banding patterns produced may not be as complex as required. We used the original procedure of Pena et al. (Proceedings of the National Academy of Sciences USA 91, 1946–1949, 1994) to generate fingerprints from a specific, polymorphic PCR product. HLA‐DPB1 exon 2 was amplified, recovered from agarose gel, and used as a template for subsequent low‐stringency (30°C) amplification cycles (LS‐PCR) in the presence of a single primer. The LS‐PCR products were run on 8% PAGE and silver‐stained. In total, 22 subjects were characterized by this method. The issues of the reproducibility and specificity of the patterns obtained were addressed by comparing fingerprints from individuals with the same genotype. The results showed that LS‐PCR was robust. A further step was the evaluation of the diversity that can be generated, i.e. the sensitivity of the method. Genotype‐related fingerprints were produced, and differences as small as a single nucleotide in heterozygous samples could be detected. We then demonstrated the usefulness of LS‐PCR in the evaluation of donor/recipient pairs. We believe that LS‐PCR may be a valuable adjunct to the battery of tests aimed at the verification of HLA matching before unrelated bone marrow transplantation. We suggest that it could be used to speed up the search process when several candidate donors are retrieved from registries before embarking on SSOP typing or sequencing.
ANALYSIS OF HLA HAPLOTYPES IN FAMILIES WITH TYPE 1 DIABETES MELLITUS IN LA RÉUNION ISLAND
Authors: Feugeas J. P. , Tortosa P. , Dulay S. , Augustin‐Pascalis I. , Charron D. , Krishnamoorthy R. , Caillens H. , Montchamp‐Moreau C.
Published: April 2007

To analyse HLA and insulin‐dependent diabetes mellitus (IDDM) association in the ethnically mixed population of La Réunion island, we carried out a family study on 70 diabetic subjects. HLA‐DQA1, ‐DQB1 and ‐DRB1 typing was performed by polymerase chain reaction‐restriction fragment length polymorphism (PCR‐RFLP), completed by PCR‐sequence‐specific oligonucleotide (SSO) and PCR‐sequence‐specific priming (SSP). Haplotype‐relative risks (HRR) were determined with the non‐transmitted parental haplotypes as controls, and relative risks (RR) were calculated with a classical case‐control study. The most significant risks were found for the cis and trans combinations between DQA1*03 or *0501 (Arg52+) and DQB1*02 or *0302 (Asp57−) alleles, suggesting a direct role for the HLA‐DQ heterodimer in IDDM susceptibility. Interestingly, due to the mixed origin of the population, the trans‐encoded DQ molecules in the (DR3)‐DQA1*0501‐DQB1*02/(DR4)‐DQA1*03‐DQB 1*0302 subjects were also found cis‐encoded in patients with the (DR7 or 9)‐DQA1*03‐DQB1*02 haplotype and in a patient with the rare (DR 11)‐DQA1*0501‐DQB 1*0302 haplotype. A relative predispositional effect (RPE) analysis gave significant haplotype‐IDDM+ associations in the following order: (DR3)‐DQA1*0501‐DQB1*02>(DR4)‐DQA1*03‐DQB1*0302>(DR9)‐DQA1*03‐DQB*02>(DR7)‐DQA1*03‐DQB1*02>(DR2)‐DQA1*01‐DQB1*0502. No protective effect remained significant once the susceptible haplotypes were removed. A stratification study showed a stronger influence of the DQ genes than DRB1 alleles within the DR7 haplotypes. On the other hand, IDDM subjects with only one susceptible haplotype had inherited this haplotype more often from their father than from their mother. This paternal effect could be related to the greater risk of IDDM in offspring of diabetic fathers than the risk in offspring of diabetic mothers.
FREQUENCY OF COMPLEMENT THIRD COMPONENT (C3) AND PROPERDIN FACTOR (BF) PHENOTYPES IN PATIENTS WITH VARIOUS CLINICAL MANIFESTATIONS OF HBV INFECTION
Authors: Kacprzak‐Bergman I. , Hacasa J.
Published: April 2007

Four groups of children were tested for the distribution of C3 and BF phenotypes: HBV‐infected patients with Gianotti‐Crosti syndrome, with CAH and with glomerulonephritis, and healthy children. The frequency of the phenotype C3F was significantly higher in children with Gianotti‐Crosti syndrome in comparison with healthy children. The frequency of the phenotype BFS was significantly higher in patients with glomerulonephritis than in individuals with CAH. The difference between the frequency of the BFS phenotype in glomerulonephritis patients and that in healthy subjects neared significance. We suggest that the carriers of these phenotypes are characterized by susceptibility to some immune complex diseases associated with HBV infection.
CHARACTERIZATION OF A SEVENTH FAMILY OF IMMUNOGLOBULIN HEAVY CHAIN VH GENE SEGMENTS IN THE CHANNEL CATFISH, ICTALURUS PUNCTATUS
Authors: Ventura‐Holman T. , Ghaffari S.H. , Lobb C.J.
Published: April 2007

A VH gene segment that could not be assigned to any of the six known VH gene families of the channel catfish was identified in a genomic clone containing VH gene segments. This gene segment (designated VH7.1) exhibited the structural features characteristic of vertebrate VH genes, specifically potential upstream regulatory sequences, a leader sequence split by an intron, a reading frame that could be readily divided into framework and complementarity determining regions, and a 3′ recombination signal sequence. Two regions of nucleotide deletions coupled with degeneracy in the nonamer sequence indicate that this VH gene segment is a pseudogene. Genomic DN A restricted with different enzymes and hybridized under stringent conditions with probes derived from VH7.1 showed that 8‐10 bands were present in Southern blots. Reverse transcriptase PCR approaches were used to determine if any of these related sequences were expressed. Sequence analysis of cloned PCR products indicates that different VH gene segments exhibiting >80% similarity to germline VH7.1 are expressed. Multiple sequence alignments showed that the expressed cVH7a cDNA sequence shared less than 60% nucleotide similarity with representative cDNA sequences from the other known catfish VH gene families. These combined results thus fulfil the criteria for the definition of a new family of catfish VH gene segments. This newly defined, small VH family is designated VH7.
COMBINED PCR‐HETERODUPLEX AND PCR‐SSCP ANALYSIS FOR MATCHING OF HLA‐A, ‐B AND ‐C ALLOTYPES IN MARROW TRANSPLANTATION
Authors: Pursall M.C. , Clay T. M. , Bidwell J.L.
Published: April 2007

We describe a method for rapid matching of HLA‐A, ‐B and ‐C allotypes using simultaneous polymerase chain reaction‐single strand conformation polymorphism (PCR‐SSCP) and heteroduplex analysis. Electrophoresis is performed at ambient temperature without requirements for buffer cooling. SSCP and heteroduplexes are revealed as discrete spatially separated band clusters. Using HLA‐A, ‐B and ‐C locus‐specific PCR primers, matching for alleles at these loci can be performed in 5h. We tested 17 serologically matched patient‐unrelated donor pairs and found considerable microheterogeneity at the DNA level. We propose that this technology has several advantages over conventional low‐resolution typing methods and represents a potentially valuable screening method in unrelated donor selection.