Effect of cyclosporin A treatment on the production of antibody in insulin-dependent (type I) diabetic patients.

Anti-islet cell and anti-insulin antibody production was studies over a 12-mo period in 82 recently diagnosed diabetics randomly receiving either cyclosporin or placebo. Cyclosporin had only minimal effects on the production of anti-islet cell antibodies whether directed to islet cytoplasmic (immunofluorescence) or membrane (cytotoxicity assay) antigens even in patients undergoing remission. These data suggest that these antibodies do not play a major role in the pathogenesis of the disease particularly since their (irregular) presence is not predictive of the clinical response to cyclosporin. Conversely, cyclosporin completely suppressed the synthesis of antibodies elicited by exogenous insulin irrespective of the insulin doses received, and decreased the autoantibody production against thyroid antigens, indicating that cyclosporin has variable effects on antibody production against various antigens.

A combination of HLA-DQ beta Asp57-negative and HLA DQ alpha Arg52 confers susceptibility to insulin-dependent diabetes mellitus.

Family and population studies indicate that predisposition to insulin-dependent (type I) diabetes mellitus (IDDM) is polygenic. It has been shown that the absence of the aspartic acid in position 57 (Asp57) of the DQ beta chain is positively correlated to IDDM. However, Asp57-negative haplotypes do not always confer susceptibility and conversely, some Asp57-positive haplotypes seem to be disease associated. It has been suggested that other HLA class II sequences, probably belonging to the HLA DQA1 gene, confer susceptibility to IDDM. This report, based on extensive oligonucleotide dot blot hybridization of PCR-amplified DQA1 and DQB1 genes, reinforces the importance of the Asp57-negative DQ beta chain, but also introduces the possibility that a DQ alpha chain bearing an arginine in position 52 (Arg52) confers susceptibility to IDDM. A molecular model of susceptibility to IDDM is proposed. This model strongly suggests that the disease susceptibility correlates quantitatively with the expression at the cell surface of a heterodimer, composed of a DQ alpha-chain bearing an Arg52 and a DQ beta chain lacking an Asp57. In view of the respective positions of the two residues and their charge, we might anticipate that both residues DQ beta Asp57 and DQ alpha Arg52 are critical for modulation of susceptibility, presumably via viral-antigenic peptide and/or autoantigen presentation.

Dose effect of cis- and trans-encoded HLA-DQ alpha beta heterodimers in IDDM susceptibility.

Insulin-dependent diabetes mellitus (IDDM) in whites is strongly associated with particular HLA-DQ alpha beta heterodimers composed of a DQ alpha chain with an arginine at residue 52 (Arg52+) combined to a DQ beta chain lacking an aspartic acid at residue 57 (Asp57-). With the aim of confirming this association, clarifying which heterodimers account for the highest risk of IDDM and explaining the excess risk of DR3-DQw2/DR4-DQw8, 115 unrelated white IDDM patients and 108 unrelated healthy nondiabetic control subjects were studied. With polymerase chain reaction and sequence-specific oligonucleotide probes, both patients and control subjects were typed for their HLA-DQA1 and DQB1 alleles and their DQA1-DQB1 haplotype and genotype frequencies were compared. Four major findings emerged from our analysis. 1) Arg52+ DQ alpha/Asp57- DQ beta heterodimers, formed in cis and/or in trans, are strongly associated with susceptibility to IDDM; 97% of patients and 46% of control subjects had at least one such susceptibility heterodimer (relative risk [RR] 32, confidence interval [Cl] 14.25-71.86, P less than 10(-7). 2) The degree of disease susceptibility depends on the number of such DQ heterodimers that a subject can express according to his or her DQA1-DQB1 genotype. The highest RR was observed in patients with four susceptibility DQ heterodimers (RR 41, Cl 17.05-95.9). 3) Only part of the susceptibility DQ heterodimers were significantly increased in patients, conferring IDDM susceptibility of different strength. The strongest association was with the DQA1*0501-DQB1*0302 combination formed in trans position (RR 35.2, CI 12.88-96.78, P less than 10(-7).(ABSTRACT TRUNCATED AT 250 WORDS)

Inhibition of insulin release in vitro mediated by mononuclear cells from diabetic patients treated with cyclosporin A or placebo.

Anti-beta-cell-specific cell-mediated immunity was studied over a 12-mo period in 65 recently diagnosed diabetic patients randomly receiving either cyclosporin or placebo. Anti-beta-cell cellular immunity was assessed by an in vitro test based on the inhibition of insulin release from cultured rat islet cells by patients' mononuclear cells. This beta-cell-suppressive effect disappeared in cyclosporin A-treated patients within 1 mo and did not reappear during 12 mo of follow-up. Conversely, the suppressive effect persisted unchanged in placebo-treated patients during 12 mo of follow-up. These changes were predictive neither of cyclosporin A-induced remission nor of relapses. Results of the insulin-release inhibition test were not correlated to islet cell autoantibodies or HLA phenotype.

HLA DQB1*0301 allele is involved in the susceptibility to erythema multiforme.

Erythema multiforme (EM) is an acute, episodic inflammatory disorder of the skin and mucous membranes of various etiology that could be related to immunologic hypersensitivity response. EM has been previously reported to be associated with serologically defined HLA-DRw53 and DQw3 antigens. In this report, we reevaluate the role of HLA class II alleles in EM manifestations. With use of the polymerase chain reaction, followed by sequence-specific oligonucleotide hybridization, 35 unrelated Caucasian EM patients and 80 randomly selected healthy subjects were studied, and the DRB3, DRB4, DQA1, and DQB1 alleles were analyzed. The comparison of frequencies of these alleles indicates that (i) susceptibility to EM disease is more associated with the HLA-DQ than the HLA-DR subregions and (ii) that the DQB1*0301 is the most frequent allele among EM patients. Sixty-six percent of the patients had the DQB1*0301 allele compared to 31% of the controls (RR = 4.1; p less than 0.001). An even stronger DQB1*0301 association was found in the patient group with herpes-associated EM (76%; RR = 6.5; p less than 0.001). Our data demonstrate a clear association between an HLA-DQB1 allele and susceptibility to EM.

A linkage disequilibrium map of the MHC region based on the analysis of 14 loci haplotypes in 50 French families.

A sample of 100 individuals from 50 French families of known pedigrees were typed for 14 loci of the HLA region (DPB1, DQB1, DQA1, DRB1, DRB3, 4, 5, C4B, C4A, Bf, C2, TNFa, TNFb, B, Cw, A). Linkage disequilibrium in each pair of loci was investigated by an exact test using a Markov chain algorithm. The results indicate no disequilibrium between DPB1 and the other loci, whereas the other class II genes are all significantly linked to each other. Linkage disequilibrium is also detected between some pairs of class I and class II-class I loci despite the long physical distance separating the loci (e.g. A-B, Cw-DRB1). On the other hand, some contiguous loci of the class III region are found to be in equilibrium with each other. Several hypotheses including selection, but also unequal allelic diversity at different MHC loci are discussed to explain this complex pattern of linkage disequilibrium.

Detection of islet cell antibodies by a microcytotoxicity method.

A microcytotoxicity test for antibodies against islet cells (ICA) is described. Sera from patients with insulin-dependent diabetes, their healthy first degree relatives, and normal controls, genotyped for HLA-A, -B and -DR, were tested by 4 different methods. Cytoplasmic ICA and complement fixing ICA were detected by indirect immunofluorescence with human pancreas sections, and cytotoxic complement dependent ICA and surface ICA were tested against murine beta cell suspensions. Strong correlation was found between cytotoxic and surface antibodies (P less than 10(-7). The technique described is appropriate for use in the screening of large numbers of sera.

Is the strength of single HLA antigen mismatch variable in kidney transplant survival?

The survival rate among 458 cadaver kidney grafts with "full-house" four HLA-A,B antigens detected in the donor, three being identical to those of the recipient and only one mismatched, was studied specifically in relation to antigen incompatibility. At the A locus, the A3 incompatibility was associated with a lower transplant survival (44% at 2 years) than all of the others, and particularly more than the A11 (80% at 2 years) (P less than 0.003 but without significance after correction multiplying by the number of tested alleles). At the B locus, there was no significant difference in survival rate among the alleles. These results are only preliminary and need confirmation based on longer series permitting possible cross-reactions which exit between donor and recipient antigens to be taken into consideration.

Selective reactivity of sera from alloimmunized sheep and cattle against human T and leukemia cells.

Human B and T lymphocytes from a panel of healthy individuals were tested against serial dilutions of 68 mare, 81 cow, 7 sow, and 87 ewe sera. All the animals had been alloimmunized by pregnancies and/or blood transfusions. Weak correlations with HLA-A, B, C, and DR specificities were found in 20 sera. Twelve other sera, 9 from ewes and 3 from cows, had a strong reactivity against T lymphocytes but weak or no reactivity against B cells, spleen null cells, granulocytes, and platelets, suggesting a non-major histocompatibility complex (MHC) cross-reactivity. They were cytotoxic for most of the cells of malignant proliferative origin tested thus far, including T acute lymphoblastic leukemia (T ALL), common ALL (cALL), acute myeloblastic leukemia (AML), and Sezary cells, but were negative with B lymphoblastoid cell lines and cells from patients with B chronic lymphocytic leukemia (CLL) and chronic myelocytic leukemia (CML). The hypothesis that humans and certain other mammals share a common determinant on T-lineage cells and some malignant cells is advanced.

Islet-cell antibodies (ICA-CFICA) and HLA genotypes in 107 IDD patients and their first-degree relatives.

The prevalence of ICA and CFICA in relation to HLA-DR genotypes was analyzed in 107 insulin-dependent diabetic (IDD) patients with a duration of IDD ranging from onset to 30 years, and 247 nondiabetic first-degree relatives. In 46 patients tested at onset of IDD, the prevalence of ICA and of CFICA was 61 and 50%, respectively; with the longer duration of IDD, the prevalence of CFICA decreased more rapidly than that of ICA. No significant association was observed between ICA and any HLA allele in patients tested from onset till 2 years after onset. However, a higher prevalence of ICA was found in DR3 positive patients with a duration of IDD of more than 2 years (p less than 0.02). Among the healthy relatives, the prevalence of ICA was 7% in parents and 13% in siblings; one out of 101 siblings had CFICA. No association was found between ICA and any HLA marker. The presence of ICA in sibs was independent of the number of haplotypes they shared with the IDD proband: among the 13 ICA-positive healthy sibs, one shared 2 haplotypes, nine shared 1 haplotype and three shared no haplotype with the proband, which is not different from the random distribution.

HLA-G gene polymorphism segregation within CEPH reference families.

HLA-G, a nonclassical HLA class I antigen, presents tissue-restricted expression on human trophoblasts and may play an important role in immune tolerance of mother-versus-fetus. In this work we have demonstrated extensive HLA-G genomic polymorphism within three CEPH reference families, by PCR-SSCP analysis and direct sequencing. Among six unrelated parents we assigned eight HLA-G alleles, seven of which are new. We observed the segregation of HLA-G alleles of heterozygous parents among their offspring that matched the segregation of the HLA class I haplotypes. Only one of the mutations observed was found to be nonsynonymous indicating low polymorphism of the HLA-G molecule.

No evidence of HTLV-I infection in French patients with multiple sclerosis using the polymerase chain reaction.

The polymerase chain reaction (PCR), using three primer pairs in the pol, tax, and env regions of the HTLV-I genome, was unable to detect HTLV-I in the blood samples of 54 caucasian subjects with multiple sclerosis who were seronegative for HTLV-I/II. Seventeen HTLV-I/II seropositive (by ELISA and Western blot) subjects used as positive controls were positive with the three primer pairs. The PCR was negative in 47 healthy HTLV-I/II seronegative (by ELISA) subjects at low risk of HTLV-I infection used as negative controls. These results suggest that there is no association between the occurrence of HTLV-I sequences and the development of multiple sclerosis.

Mitochondrial DNA polymorphism in the French population.

One hundred unrelated individuals of French origin were screened for mtDNA variation as restriction fragment length polymorphisms (RFLPs) with the restriction enzymes HpaI, BamHI, HaeII, MspI, AvaII and HincII. Twenty enzyme morphs were detected, four of which (AvaII-37Fr, -38Fr, HincII-18Fr and -19Fr) are new. Of the 17 mitotypes detected, five are new and they were named 1-19Fr, 6-18Fr, 100Fr-2 (2-1-2-4-1-2), 101Fr-2 (2-1-1-1-38Fr-2) and 102Fr-2 (2-1-1-4-37Fr-2). All new morphs and mitotypes derive from those already known due to a single nucleotide substitution. The French population was compared with other European, Mediterranean and Caucasian populations. Calculation of the genetic distances showed close genetic affinity with European-Mediterranean populations and especially with Calabrians, Majorcans and northern Italians (at negative values).

Mitochondrial DNA polymorphisms of a west Algerian population (Oran region).

Mitochondrial DNA codes for enzymes involved in the cellular energetic pathway. The polymorphism of this genome has been extensively analyzed for disease associations, but can also be used to characterize anthropological distances between populations. This study presents the results of mitochondrial DNA (mtDNA) sequence variation for a population sample of 50 unrelated individuals originating from western Algeria. The samples were studied with the recently developed long PCR technique followed by RFLP analysis using six restriction endonucleases: HpaI, BamHI, HaeII, MspI, AvaII and HincII. One new morph for HpaI (named HpaI-9Alg) was detected, and was found to be derived from the combination of the already known morphs 3 and 4. mtDNA restriction endonuclease fragment patterns were analyzed for potential site gain or loss and classified into 18 mtDNA types by the sequence-comparison method. Three mtDNA types (97Alg; 2-1-7-1-1, 98Alg; 2-1-1-8-37 and 99Alg; 9Alg-1-1-1-3) were detected for the first time. Another mtDNA marker--the presence of the 9 bp deletion in the COII/tRNA(Lys) region--was also studied in the Algerian sample. No deletions were observed. Our results indicate that the Algerians are genetically related to the Israeli-Arab population, with certain characteristics found in southern Europeans and others found in sub-Saharan Africans.

Familial versus sporadic longevity and MHC markers.

We have studied the polymorphism of HLA-A, B, in 2 elderly populations (> or = 90 years) compared to a control series of 429 healthy unrelated individuals less advanced in age. The aged population issued from the CHRONOS cohort consisted of 336 centenarians without familial history of longevity, and 102 nonagenarians index cases randomly selected from families. Almost all individuals (310) were previously typed for HLA-DRB1. The increased allelic frequency of HLA DR11 was observed in familial nonagenarians (18.3%) compared to controls (10%) (p < .001) and to sporadic centenarians (11.8%). Concerning HLA Class I alleles, only rare alleles (A30, B70) remain significantly different from the controls after correction of the p value. No distortion of the Mendelian sharing of haplotypes was observed among sibling pairs of familial nonagenarians. A protective effect of the HLA-DR11 molecule itself, presenting adequately immunogenic-infectious peptides, is probable rather than genes in disequilibrium. Our study strongly supports the heterogeneity of longevity, the association of HLA-DR11 in its familial form in aged populations.

Cigarette smoking-related persistent polyclonal B lymphocytosis. A premalignant state.

A case of cigarette smoking-related polyclonal B lymphocytosis, with binucleated cells, was observed. The condition was persistent for two years. We indicated the abnormal ultrastructural features and suggest that this unexplained syndrome could be a premalignant state.

[HLA and susceptibility to malignancies]. / HLA et susceptibilité aux maladies malignes.

From the large amount of data pertaining to a possible relationship between MHC and susceptibility to malignancies in humans, some significant findings emerge. 1. In population studies, HLA-A2 in ALL and A1 in Hodgkin's disease are observed significantly increased in frequency in comparison to the normal controls. Some significant associations with an HLA phenotype are also observed in several cancers. 2. HLA genotyping of familial cases of Hodgkin's disease, with multiplex affected sibs, lead to the conclusions that an excess of HLA identical pairs are observed among the patients. This could indicate a linkage between the susceptibility gene and the HLA region. Another linkage might exist with familial malignant melanoma fitting with a dominant mode of transmission of the trait. These facts strongly support the role of the MHC among the polyfactorial and polygenic determinism of some malignancies. The mechanisms are discussed.

Bone marrow transplantation in 65 patients with severe aplastic anemia.

This articles summarizes the experience of the Hospital Saint-Louis Bone Marrow Transplantation Team of bone marrow transplantation in severe aplastic anemia. Sixty-five consecutive patients have been transplanted with an HLA identical sibling marrow. Various conditioning regimen have been used. Conditioning regimen using high dose Cyclophosphamide alone or associated with Procarbazine and anti-thymocyte globulin gave a high number of bone marrow graft rejection. Therefore, a conditioning regimen using Cyclophosphamide and total body irradiation with lung shielding has been used for the last three years. This regimen suppressed bone marrow graft rejection. The main problems remain graft versus host disease and intercurrent infections. Despite these complications, 50 per cent of the patients become long term survivors and are apparently cured of their initial disease.

[Epidemiology of juvenile insulin-dependent diabetes in Algeria (Wilaya of Oran)]. / Epidemiologie du diabète insulinodépendant juvénile en Algérie (Wilaya d'Oran).

In order to precise the prevalence and incidence of IDDM in a well characterized population, we have ascertained between 1979 and 1988 all juvenile diabetic patients living in the department of Oran (Wilaya of Oran), Algeria, aged less than 20 years at the date of Dec. 31th 1988 who had onset of the disease before 15 years old. Cross comparisons of the data obtained both by a IDDM registry and a questionnaire sent to the practitioners show an excellent correlation. A total of 173 families was studied, including 155 Simplex Families (SF), and 18 Multiplex Families (MF). The prevalence was 0.27 p. 1000 among children aged less than 15 years. The annual incidence (mean = 4.4 p. 100,000) increased regularly from 1.6 p. 100,000 in 1981 to 8.1 p. 100,000 in 1988. The prevalence of IDDM among siblings was 3.8%. The birth order of index cases (first affected siblings) was lower in MF (3.0 +/- 0.4) than both in index cases of SF (4.0 +/- 0.3; p less than 0.01) and in secondary affected siblings of MF (4.8 +/- 0.05; p less than 0.001). The frequency of 2nd degree consanguineous marriages (between first cousins) was greater among the IDDM families compared with control families: 25% vs 11% (RR = 2.9; p less than 0.001).

[Interaction between HLA and Gm for susceptibility to insulin-dependent diabetes]. / Interaction entre HLA et Gm pour la susceptibilité au diabète insulino-dépendant.

It is now well established that HLA system is involved in the susceptibility to Type 1 diabetes mellitus. In this study we look for a possible effect of the Gm system. A first study (cases-controls) suggests that among individuals who had HLA-DR3 but not HLA-DR4 or HLA-DR4 without HLA-DR3, there is a possible effect of the phenotype Gm3,23,5 or Gm3,-5 in the susceptibility to IDDM. Moreover, we have tested by the sibpair method whether HLA and Gm are transmitted independently from IDDM: an unaffected sib, sharing the same HLA haplotypes than an affected individual, seems to be more often phenotypically different at the Gm loci.