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M Colombani

Publications and source records attributed to M Colombani.

At least 19 recordsLinked to original sources

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

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

[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

Evidence for a new HLA class II determinant present on cells from HLA-DR1 and/or -DR4 individuals.

Evidence for a new HLA class II specificity is presented. It is recognized by LE serum, which reacts with most DR1 and/or DR4 individuals (r = 0.86). Its frequency in the French population is 0.33. Absorption-elution experiments showed that the serum reactivity was not due to a mixture of anti-DR1 and anti-DR4 antibodies, but to a single antibody population which could be absorbed on and eluted from both DR1(+) or DR4(+) cells. LE specificity seemed to be expressed on DR but not on DQ molecules since the serum reacted with and could be absorbed by DR+,DQw- cells; it did not react with a DR-,DQw+ mutant cell, but did react with the DR+,DQw+ parental cell. The relationship between LE specificity and MC1 and Te23 specificities remains to be determined.

Adult

Serological cross-reactivity between products of separate I regions.

A B10.S(7R) anti-B10.S(9R) serum (anti-IJEkCd) contained, as expected, antibodies specific for the I-E-subregion-encoded determinant Ia.7. However, tests on recombinant haplotypes demonstrated a series of unexpected weak extrareactions which could be interpreted to be directed against antigenic determinants encoded in the I-A subregion of the H-2 complex. The same type of extrareaction was observed in eluates from I-As, I-Ek cells coated with A.TH anti-A.TL (I-As, I-Ek anti-I-Ak, I-Ek) serum. This reactivity in serum and eluates could be interpreted as cross-reactivity between products of the I-E and I-A subregions.

Animals

Inhibition of secondary mouse mixed lymphocyte reaction by anti-Ia sera.

Secondary mixed lymphocyte reaction (MLR-II) was studied in A.TH anti A.TL and A.TL anti-A.TH combinations in which stimulation was mainly due to H-21-region differences. In both cases of MLR-II was specifically inhibited by the responder anti-stimulator Ia serum. The level of inhibition was dependent on the ratio of the amount of immune serum to the number of stimulating cells. The inhibitory activity and Ia antibodies were specifically absorbed and eluted together. The results confirm that the lymphocyte-activating determinants of the MLR-II (1) are carried by the Ia molecules and (2) are identical to the serologically defined Ia determinants. - Anti-Ia sera directed against private and public specificities of the stimulating cell induced a higher level of inhibition than anti-Ia sera directed only against public specificities, indicating that both private and public Ia specificities are involved in re-stimulation during MLR-II. - These results, in connection with others, suggest that the receptor of the proliferating T cell recognizes the same Ia determinant as the combining site of the Ia-recognizing antibody.

Animals

[Serological study of Ia antigens determined at the I-A and I-E sub-regions of the H-E complex in the H-2 complex of mice].

Immune serum B10.S (7R) anti-B10.S (9R)(anti I-JEkCd) contained as expected an anti-Ia7 antibody. A series of weaker but reproducible extra-reactions might recognize Ia3 specificity coded at the I-A subregion of the H-2 complex. Results with recombinant haplotypes confirmed this mapping. Such a reactivity could be interpreted as an interlocus cross-reaction (I-E/I-A) since the immunization was induced against an I-E subregion product. Another interpretation was possible: the immune serum would thus contain an antibody recognizing Ia7 (on the E alpha k Ia chain) and another antibody recognizing an antigenic determinant carried by the E beta k Ia chain. The latter antibody might recognize by cross-reaction as specificity carried by the A beta chain of various haplotypes (H-2b,k,q).

Animals

H-2 restriction for lymphocyte homing into lymph nodes.

Migratory patterns into lymph nodes of labeled spleen lymphocytes, injected intravenously, showed considerable differences according to the genetic identity or disparity of donor and recipient mice. The use of mice carrying different H-2 haplotypes in the same B 10 background, or, conversely, the same H-2b haplotype in different backgrounds, showed that no homing was observed in the case of complete disparity at the H-2 complex, contrasting with optimal homing in the case of H-2 identity. Homing was not influenced by disparity at the genetic background. Irradiation of recipient mice did not change the results and suggested that no immunological host-vs.-graft reaction was involved in the H-2 restriction observed. The use of donor-recipient pairs carrying recombinant H-2 haplotypes showed that a single H-2 K and/or H-2 D identity allowed almost optimal homing. The results indirectly suggested that an immunological graft-vs.-host reaction was not involved in the phenomenon but rather a positive recognition of H-2 K or H-2 D identity between donor and recipient cells.

Animals

Reactivity of anti-HLA sera against mouse lymphocytes.

The reactivity of anti-HLA hyper-immune sera was tested by the microlymphocytotoxicity technique (LCT) against a panel of mouse lymphocytes. Three levels of reactivity were observed: negative (immune sera titre 1:20 or less), weak (titre 1:40 to 1:80 with a percentage of dead cells less than 50%) or strong (titre 1:60 or more with 100% killing). Thirteen normal human sera were non-reactive. Eleven out of 60 hyper-immune sera were strongly reactive. Tests using congenic lines showed that the reactivity was controlled at the H-2 complex. One serum (CODRON) was studied in detail. When tested against a panel of strains carrying 10 different H-2 haplotypes, it reacted strongly against lymphocytes H-2d, ja, k, p and r; and did not react, or at least only weakly, against lymphocytes H-2b, f, q, s and v. Tests using mouse strains carrying the recombinant H-2 haplotypes h4, i5, i, y2, g, g2 and t1, suggested that the observed reactivity was directed against structures controlled at the K end of the H-2 complex (H-2.47?). Absorption-elution experiments with human and murine lymphocytes and platelets confirmed that the structures recognised by serum CODRON were determined at the major histocompatibility complex.

Animals

Separation of anti-Ia (I-region associated antigens) from anti-H-2 antibodies in complex sera, by absorption on blood platelets. description of three new Ia specificities.

H-2 antigens are expressed in substantial amounts of murine blood platelets (for H-2 antigenic content 1 lymphocyte approximately 50 platelets) whereas Ia antigens are probably not expressed at all (minimal Ia antigenic content more than 35 times lower than for H-2). This property of blood platelets makes them very useful for the selective absorption of anti-H-2 antibodies from complex sera and for the preparation of specific anti-Ia antibodies from such sera. In 20 sera produced against the complete H-2 complex, 12 sera contained anti-Ia antibodies beside the expected anti-H-2 antibodies. In two sera, separation of the anti-Ia antibodies was easily obtained by absorption of the anti-H--2 antibodies on platelets. The analysis of one serum (C3H.Q X B10.D2) anti-C3H [(q X d) anti-k] showed that, in addition to be expected anti-H--2.23 and anti-Ia.2 antibodies, it contained at least three other Ia antibodies, separable by absorption on lymphocytes, which recognized three antigens--Ia.17, determined by the haplotypes k, f, s, r, j; Ia.18, determined by the haplotypes k, f, s; and Ia.19 determined by the haplotypes k and r. The genes are located in the I--A and/or I--B subregions of the H--2 complex.

Absorption

A molecular defect in thrombasthenic platelets.

An IgG antibody found in the serum of a thrombasthenic patient reacted in complement fixation with platelets from 350 normal individuals but was nonreactive with platelets from eight other thrombasthenic patients. ADP-induced aggregation of normal platelets was inhibited by the patient's antibody. Family studies using the quantitative complement fixation test showed that healthy heterozygotes were easily distinguishable from normal or thrombasthenic individuals since their platelets had an intermediate amount of the reactive antigen. Indirect immunoprecipitation tests using this serum and soluble membrane antigens labeled with iodine-125 that had been extracted from normal platelets by the detergent Nonidet P-40 gave a single radioactive peak at 120,000 mol wt in sodium dodecyl sulfate polyacrylamide gel electrophoresis. A similar estimate of the molecular weight was obtained from Sephadex G-200 filtration of the soluble antigens extracted from normal platelets by spontaneous release or chaotropic agents and tested in complement fixation with the patient's serum. These findings strongly suggest that the molecule recognized by this antibody is absent or structurally modified in thrombasthenia cases and that it may be involved in platelet aggregation.

Adenosine Diphosphate