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J Colombani

Publications and source records attributed to J Colombani.

At least 37 records · Page 2Linked to original sources
PubMed1989

[HLA class II typing of peripheral blood B lymphocytes separated using monoclonal antibodies].

The different methods proposed for preparation of B lymphocyte suspensions are not always simple, rapid and reliable. Presence of monocytes in the B cell suspension makes difficult HLA-DQ typing. For these reasons we propose a method using monoclonal antibodies (McAb) and complement to lyse the non B cells. Mononuclear cells, isolated from peripheral blood by Ficoll-Hypaque, are suspended in a mixture of anti-CD2, CD3, CD11b (OKM1 recognizing monocytes and granulocytes) McAb. This method of preparation of B cell suspension is rapid and provide better typing reactions than did the suspensions prepared by depletion of sheep red blood cell rosetting cells.

Antibodies, Monoclonal↗
PubMed1989

[HLA class II typing in chronic myeloid leukemia with the complement fixation test using phytohemagglutinin-activated lymphocytes in the presence of interleukin 2 and gamma interferon].

In 23 cases of chronic myelogenous leukemia (CML) in remission, HLA class II typing was performed by complement fixation technique on phytohemagglutin (PHA) activated lymphocytes. In 10 cases satisfactory results were obtained, whereas in 13 cases the cells were weakly or non reactive, making impossible a correct determination of HLA-DR,DQ specificities. In the 13 cases a conditioned medium containing Interleukin-2 (Il-2) and gamma interferon (IFN) was added to PHA, inducing a good HLA class II reactivity and making possible a satisfactory definition of HLA-DR,DQ specificities. The complement fixation technique on lymphocytes cultured with PHA, Il-2 and gamma IFN made possible a correct HLA-DR,DQ typing in 23 consecutive cases of CML, while such a typing is often difficult or impossible when using the microlymphocytotoxicity technique on B lymphocyte targets.

Antilymphocyte Serum↗
PubMed1989

Definition of a new supertypic HLA class II determinant (LAR) associated with HLA-DR2 and -DR7.

A serum from a patient (LAR), immunized by pregnancies and blood transfusions, reacted with cells carrying HLA-DR2 and/or -DR7 specificities (titer 1:200-1:1000). Absorption-elution experiments showed that the allo-serum recognized a determinant shared by DR2 and DR7 cells. The high correlation coefficients (0.90-1) with these specificities suggested that the supertypic specificity LAR was carried by the first DR molecule encoded by DRB1 gene. LAR is another example of new supertypic specificities, reflecting structural homologies between alleles at HLA class II loci.

Epitopes↗
PubMed1989

HLA typing with monoclonal antibodies: evaluation of 356 HLA monoclonal antibodies including 181 studied during the 10th International Histocompatibility Workshop.

During the 10th International Histocompatibility Workshop (10th WS), 181 HLA MoAbs were studied using lymphocytotoxicity micro-technique (LCT) and/or enzyme immuno-assay (EIA), and their capacity to serve as typing reagents was evaluated. 129 MoAbs were tested by both techniques. Results obtained with 92 class I and 86 class II polymorphic MoAbs (10th WS) were compared to published data concerning 180 class I and 176 class II polymorphic MoAbs, listed in an HLA-MoAbs Register maintained in our laboratory. The following conclusions can be proposed: 1/HLA-A, B typing by LCT with MoAbs is possible for about 14 specificities. Some specificities are clearly recognized (HLA-A3, B8, B13, Bw4, Bw6), others are recognized as cross-reacting groups (B7+27+w22+40), others are not currently recognized by any MoAb with restricted specificity (B5, B15). Several MoAbs confirmed the existence of shared epitopes between products from a single locus (A2-A28, A25-A32), or from A and B loci (A2-B17, Bw4-A9-A32). A single HLA-Cw MoAb has been described. 2/HLA class II typing by LCT with MoAbs is more difficult than class I typing. DR2, DR3, DR4, DR5 and DR7 as well as DRw52 and DRw53 are well defined; other DR specificities are poorly or not at all defined. Particular associations (DR1+DR4, DR3+DRw6, all DR except DR7) are recognized by several MoAbs. All DQw specificities are well recognized, including new specificities defined only by MoAbs: WA (DQw4), TA10 (DQw7), 2B3 (DQw6+w8+w9). Only two HLA-DP MoAbs have been described. 3/Satisfactory results, similar to those of LCT, were obtained with EIA using lymphoid cell lines as targets. 4/Human MoAbs (12 in the Register) are satisfactory typing reagents. They could represent in the future a significant contribution to HLA typing with MoAbs.

Antibodies, Monoclonal↗
PubMed1988

Epitope mapping of HLA-B27 and HLA-B7 antigens by using intradomain recombinants.

To study the HLA-B7 and HLA-B27 antigenic determinants, hybrid genes between these two alleles were constructed by in vivo recombination in Escherichia coli. After transfection of these genes into P815 (high transfection efficiency recipient) murine cells, the bindings of Bw6, HLA-B7, and HLA-B27 allele-specific mAb were studied, as well as that of human anti-HLA-B7 and anti-HLA-B27 monospecific alloantisera. Most of the HLA-B7 antigenic determinants were assigned to the first external domain of the molecule. Four different epitopic areas could be defined: the Bw6 epitope was associated with residues 82 and 83; the BB7.1 epitope to amino acids 63, 67, and 70; the MB40.2 and MB40.3 epitope to amino acid sequence 177-180, and human alloantisera identified as an epitope associated with residue 9. HLA-B27 antigenicity studied by TM-1 mAb was found to involve residues 77 and 80 in the alpha-1 domain. Results obtained with human monospecific alloantisera allowed the definition of an additional allospecific site associated with the NH2 terminal part on the alpha-1 domain of HLA-B27. Epitope mapping fits with data obtained by sequence comparisons and is discussed with reference to the crystallographic three-dimensional structure of the HLA-A2 molecule.

Amino Acid Sequence↗
PubMed1988

Erythema multiforme is associated to HLA-Aw33 and DRw53.

Erythema multiforme is an acute eruption of the skin and mucous membranes of various aetiologies. Forty-one unrelated patients were HLA typed for 53 specificities of the HLA-A, B, C, DR and DQ series. Frequencies of Aw33 and DRw53 were significantly increased: Aw33, 17.0% in patients vs 2.8% in controls (corrected p = 0.01, relative risk = 7.2); DRw53, 70.7% in patients vs 30.5% in controls (corrected p = 0.0005, relative risk = 5.5).

Adult↗
PubMed1987

Down-regulation of class I HLA antigens and of the Epstein-Barr virus-encoded latent membrane protein in Burkitt lymphoma lines.

Epstein-Barr virus (EBV)-carrying Burkitt lymphoma (BL) cells are relatively or completely resistant to the lytic effect of major histocompatibility complex class I HLA antigen-restricted cytotoxic T lymphocytes (CTLs) generated by stimulating lymphocytes of EBV-seropositive donors with the autologous EBV-transformed lymphoblastoid cell line (LCL). We previously found that EBV-negative and EBV-carrying BL lines derived from HLA-A11-positive donors were not only resistant to lysis by the HLA-A11-restricted CTL generated by stimulation with the autologous LCL, but also to HLA-A11-specific CTL derived from lymphocytes of an EBV-seronegative donor stimulated with an allogeneic LCL. Using the same and additional cell lines, we now show that the CTL resistance of the BL lines is probably due to a selective down-regulation of HLA-A11. We also show that the EBV-encoded latent membrane protein is expressed at a lower level in the EBV-carrying BL lines than in EBV-transformed LCLs. Only one of eight in vitro EBV-converted BL lines that shifted to a more LCL-like growth pattern expressed LMP at a high level. This line also reexpressed the HLA-A11 antigen that was undetectable in its EBV-negative progenitor. Our findings suggest that the typical BL cell phenotype is associated with low expression of both proteins.

Antigens, Neoplasm↗
PubMed1987

HLA-A, B and DR antigen frequencies in patients with AIDS-related persistent generalized lymphadenopathy (PGL) and thrombocytopenia.

HLA-A, B, DR phenotype frequencies were studied in 69 homosexual patients with persistent generalized lymphadenopathy (PGL), in 8 patients with LAV/HTLV III related thrombocytopenic purpura and in 21 homosexual controls. Antigen DR5 was significantly increased in PGL patient groups as compared to controls. Our findings suggest that genetic factors associated with HLA-DR5 antigen may predispose individuals infected by the LAV/HTLV III virus to the occurrence of PGL and/or thrombocytopenia.

AIDS-Related Complex↗
PubMed1987

HLA typing in heavily transfused haemophiliacs with or without antibodies against human immunodeficiency virus (HIV).

HLA-A, B, DR antigens were determined in 46 patients, heavily transfused with blood products. Patients were 44 haemophiliacs, one case of afibrinogenemia and one case of von Willebrand's disease. Thirty-three patients had HIV antibodies. No significant difference for HLA antigen frequencies was observed in haemophiliacs with or without HIV antibodies, nor between haemophiliacs and a control population.

Adolescent↗
PubMed1987

[HLA A-B and DR in dilated myocardiopathies].

HLA A, B typing was performed in 90 patients (90 men, 10 women), and HLA DR typing in 49 patients with dilated cardiomyopathy in order to test the hypothesis that a particular genetic background may influence the immune response in that disease. No significant difference in phenotype frequency of the HLA A and B antigens was observed between patients and control population. In contrast, the HLA DR4 antigen was significantly more frequent among patients (40.8% vs 23.8%, p corrected less than 0.001). The results suggest that genetic factors play a role in the pathogenesis of dilated cardiomyopathy, and that since DR4 is frequently associated with auto-immune manifestations, a modified immune response is possible in dilated cardiomyopathy.

Adolescent↗
PubMed1987

In vitro proliferation of human lymphocytes measured by an enzyme immunoassay using an anti-5-bromo-2-deoxyuridine monoclonal antibody.

A previously described, non radioactive method for the measure of in vitro mouse lymphocyte proliferation was applied to human lymphocyte proliferation assays. It involved incorporation into DNA, during cell multiplication, of 5-bromo-2-deoxyuridine (BUdR), a thymidine analogue. BUdR-DNA was then assayed by a sandwich enzyme immunoassay (BUdR-EIA) using an anti-BUdR monoclonal antibody (McAb 76-7). BUdR-DNA from crude cell extracts was first immobilized on microtitration plates coated with McAb 76-7. In a second step BUdR-DNA was reacted again with McAb 76-7 conjugated to horse radish peroxydase. The quantity of peroxydase in microtitration wells was then measured by the coloration of o-phenylenediamine (492 nm). Titration curves obtained with dilutions of crude extracts were compared to the curve obtained with a purified BUdR-DNA reference solution. Results were expressed as equivalent ng BUdR-DNA/ml. BUdR-EIA was compared to 3H-thymidine incorporating assay for the measure of lymphocyte proliferation induced by PHA mitogen, candidine and tuberculine antigens and mixed lymphocyte culture. Excellent correlation between both assays was observed for each experiments (r = 0.953 to 0.999). Overall correlation coefficient for the 5 experiments was 0.785, indicating greater variation of BUdR than 3H-thymidine incorporation, according to the mitogen or antigen used and the culture conditions. This could be due to that fact that BUdR-EIA measured only BUdR incorporated into DNA, while 3H-thymidine incorporation assay measured 3H-thymidine both incorporated into DNA, and stocked into the cell before DNA incorporation. BUdR-EIA would thus reflect cell proliferation more exactly than 3H-thymidine incorporation assay. The sensitivities of both techniques were comparable.(ABSTRACT TRUNCATED AT 250 WORDS)

Antibodies, Monoclonal↗
PubMed1986

Correlation between an HLA-DQ alpha length polymorphism of messenger RNA and serologically defined specificities (DQw1, DRw53, DR3+5).

mRNAs for the two chains of the HLA-DQ molecule were analyzed, in particular the DQ alpha mRNA whose polymorphism had previously been suggested (Schenning et al. 1984). Northern blot transfers of the mRNA of 12 LCLs and of B lymphocytes from a healthy donor were carried out. We report that a length polymorphism of DQ alpha mRNA exists, and we show that it can be correlated with serologically defined specificities (DQw1, DRw53, DR3+5). This correlation could be explained by a linkage disequilibrium, as these specificities are considered to be different from those carried by the DQ molecule (except for the DQw1 specificity).

DNA Restriction Enzymes↗
PubMed1986

[Study of the HLA-DQ system by the complement fixation test on lymphocytes stimulated by phytohemagglutinin. Existence of HLA-DQX allele(s)].

The complement fixation microtechnique against PHA blasts has been used to study HLA-DQw1, 2, 3 specificities with sera from multiple transfused patients and/or from multiparous women. Several sera (6 or 7) have been used to define each DQ specificity. The sera have been chosen because of their reactivity with cells from HLA-DR 1, 2 or w6 donors (for DQw1), DR3 or 7 donors (for DQw2,) DR4 or 5 donors (for DQw3). Correlation coefficients between DQ and DR specificities were from 0.56 to 0.91. Correlation coefficients between sera were from 0.51 to 0.92 in each cluster of sera. The segregation of DQw1, 2, 3 specificities has been studied in 46 families with 234 children. This study showed haplotypes lacking DQw1, 2, 3 specificities. The segregation of such 11 DQX haplotypes has been observed in 38 children from 8 families; 5 children were DQX/DQX homozygotes. Up to now, no serological reagent defining the specificity (or specificities) corresponding to DQX has been found. No preferential association was observed between DQX and DR specificities. The gene frequencies observed in 170 haplotypes in these 46 families were as follows: DQw1: 0.400; DQw2: 0.252; DQw3: 0.282; DQX: 0.065. Detecting DQ specificities seems easier by CF on PHA blasts than by lymphocytotoxicity microtechnique against B lymphocytes and monocytes from pheripheral blood. This suggests that PHA blasts express larger quantities of DQ molecules than B lymphocytes and monocytes. The results confirm that complement fixation microtechnique against PHA blasts is efficient for HLA-DQw typing.

Alleles↗
PubMed1986

[HLA typing in patients with chronic adenopathies in a population at risk for AIDS].

HLA-A B and DR typing were performed in 77 patients with AIDS related complex (ARC)--69 lymphadenopathy associated syndrome and 8 thrombocytopenic purpura LAV/HTLV III related--and 21 symptom free homosexual males. A significant increase in the frequency of HLA DR5 antigen was observed in patients with ARC mainly in purpura thrombocytopenic patients. We suggest that increase of HLA DR5 antigen support the view that DR5 antigen could be one of the factors necessary at the spreading out clinical symptoms.

AIDS-Related Complex↗