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Biomedical subjects

M Zouali

Publications and source records attributed to M Zouali.

At least 19 recordsLinked to original sources
PubMed1997

Identification of the B cell superantigen-binding site of HIV-1 gp120.

Previous studies showed that the gp120 envelope protein of HIV-1 is able to crosslink membrane IgM on normal human B cells and to induce their activation in a V(H)3 immunoglobulin gene-family-specific manner. Because this V(H) gene family is the largest in the human repertoire, this superantigen (SAg) property is thought to have deleterious consequences for the host, including a progressive decline of B cells with progression of the HIV-1-induced disease. Here, we have identified the sequence motifs on gp120 involved in SAg binding to normal Igs. We show that this SAg-binding activity is present in gp120s from highly divergent isolates of HIV-1 belonging to clades derived from various geographical origins, and that carbohydrate residues are not essential for its expression. The SAg-binding site is formed by protein sequences from two regions of the gp120 molecule. The core motif is a discontinuous epitope spanning the V4 variable domain and the amino-terminal region flanking the C4 constant domain. The most critical residues appear to be Leu395-Asp397 and Ile425-Gln427. Residues from the C2 constant domain (positions 252-272) also seem to play an accessory role in SAg binding of gp120 to normal human Igs. These findings are important in the design of a successful gp120-based vaccine against HIV-1.

Amino Acid Sequence↗
PubMed1997

The structure of human lupus anti-DNA antibodies.

B-cell hyperactivity and production of pathogenic autoantibodies are the main immunological events in the pathogenesis of the human autoimmune disease systemic lupus erythematosus. One approach to studying the pathogenesis of this disease and to understanding how the autoantibody response is initiated and sustained has been to analyze the variable genes expressed by antibodies that have the hallmarks of the disease-causing subset of pathogenic autoantibodies. Such studies have provided important clues. Quantitation of this repertoire revealed the presence of a specific expansion of IgG clonotypes that impart reactivity with disease-related autoantigens. The sequences of the genes encoding autoantibodies derived from these patients and expressing nephritogenic idiotopes (present in immune complexes and renal eluates of subjects with active disease) show features of molecular diversification with a high rate of replacement/silent mutations and clustering of the mutations in the hypervariable regions. These characteristics imply that a pure polyclonal B-cell activation process cannot be the only mechanism responsible for autoantibody production. More likely, an antigen drive plays a role in generation of pathogenic autoantibodies. Based on additional studies of their light chain variable region genes, we have offered a novel genetically based model whereby B cells secreting pathogenic lupus autoantibodies are blocked in their capacity to turn off their autoreactivity and to acquire a new specificity, a mechanism called receptor editing. Studies of the molecular events involved in this mechanism and of the clonal fates of B cells in vivo will be important for future research.

Amino Acids↗
PubMed1997

Polymorphism of the human immunoglobulin heavy chain locus in rheumatoid arthritis.

The genetic origin of Rheumatoid Arthritis (RA) is largely unknown. The purpose of this investigation was to assess the potential genetically determined involvement of the immunoglobulin (Ig) heavy chain variable region (VH) locus in the pathogenesis of RA. We tested the hypothesis of whether there is a genetic linkage between a structural abnormality of the VH gene complex and autoantibody hyperproduction in RA. We used restriction endonuclease generated polymorphism with human VH gene-family-specific probes to examine genomic DNA from a RA family and from unrelated RA patients from both the Tunisian and the European populations. The use of DNA samples from these ethnic origins permitted a further evaluation of the polymorphism of the human VH locus. While we found that the polymorphism of the VH locus was lower in the Tunisian population, we could not detect a restriction site polymorphism pattern restricted to RA. Together, our results do not support the involvement of major abnormalities of the Ig VH locus as a primary source in the development of RA.

Arthritis, Rheumatoid↗
PubMed1995

Selective variations in vivo of VH3 and VH1 gene family expression in peripheral B cell IgM, IgD and IgG during HIV infection.

We have analyzed the expression of VH gene families in IgM, IgD and IgG of peripheral blood B cells from a group of HIV-infected patients. CD19+CD20+ cells were purified and anchored reverse transcriptase-polymerase chain reaction products were hybridized with VH gene family probes. IgM, IgD and IgG that expressed a VH3 gene family segment, were decreased in patients with low CD4 counts and to a greater extend in patients with AIDS symptoms (up to 85% for IgG) compared to adult healthy donors. This was correlated with elevated levels of IgM and IgG encoded by a VH1 gene family segment (around 60% for IgG). These results confirm and extend previous work that has detected the VH3 gene family under-representation in HIV infection. Here, we show that, in vivo, this phenomenon actually affects the different B cell populations of the peripheral blood: IgM+ or IgG+ B cells and also IgM+IgD+ naive B cells. In the course of HIV infection, this results in their gradual depletion. Data presented here strengthen the hypothesis that a B-cell superantigen exists in HIV infection. These pronounced variations of the normally most-expressed VH gene family may be related to B cell abnormalities detected in HIV-infected patients.

Adult↗
PubMed1995

Analysis of human VH gene repertoire expression in peripheral CD19+ B cells.

Using CD19 B-cell selection and polymerase chain reaction-amplified cDNA libraries, we analyzed the peripheral immunoglobulin heavy chain variable repertoire of three healthy adult donors. Here we report that most of the CD19+ circulating B cells expressed unmutated VH-D-JH rearrangements. By specific VH family hybridization, we show that VH gene family utilization in the periphery roughly corresponds to the complexity of these families in the germline and appears to be relatively constant among the analyzed subjects. However, sequence data of clones picked at random from one IgM cDNA library reveals that in spite of this "random" utilization, the VH gene expression in naive circulating B cells is highly biased towards the expression of a limited set of VH genes. As previously reported by others, this restricted mechanism is also found for the D and JH segments.

Adult↗
PubMed1995

Variable region light chain genes encoding human antibodies to HIV-1.

In previous work, it was found that the heavy chain variable gene (VH) repertoire of human antibodies to HIV is markedly skewed and that the gp120 molecule is a ligand for VH3 gene products. Here, we have analysed the light chain (L-chain) variable region genes (VL) expressed by a panel of human monoclonal antibodies derived from an immunized volunteer, an AIDS patient and seropositive asymptomatic donors, and specific for HIV-1 p25, gp41 and gp120 proteins. We found that, in contrast to VH gene-family use, the VL repertoire does not exhibit a family-bias. We noticed however, a tendency to the use of VL genes that map to the downstream portion of the kappa locus. The VL genes expressed have mutated at lower rates than the corresponding VH genes and show no clustering of the replacement mutations in the hypervariable regions. We also found that the third hypervariable regions (CDR3) of the L-chains have undergone a marked diversification, with addition of untemplated nucleotides, frequent truncation at the 3' end of the VLs and somatic mutation. These molecular events result in a length heterogeneity of the CDR3s and an apparently positive selection of specific highly reactive amino acids. We conclude that the specificity of, at least some of the anti-HIV antibodies, is dictated by the L-chain CDR3 regions which bear the imprints of antigenic selection.

Amino Acid Sequence↗
PubMed1995

B-cell superantigens: implications for selection of the human antibody repertoire.

For several decades, B-cell interactions with antigens were thought to occur only through a clonal activation mechanism, in which the hypervariable regions of the immunoglobulin receptor are exclusively involved in ligand binding. However, an additional mode of interaction can occur, whereby molecules, termed B-cell superantigens, can bind human B cells bearing immunoglobulin receptors of a given variable (V)-gene family. This mechanism requires contributions from regions outside the conventional hypervariable loops and results in a B-cell response of increased magnitude. Here, Moncef Zouali reviews recent in vitro and in vivo observations on human B-cell superantigens in the context of the current consensus of B-cell development, and discusses the implications of these novel concepts with respect to pathogenesis.

Antibody Diversity↗
PubMed1995

Analysis of variable region genes encoding anti-Sm and anti-cardiolipin antibodies from a systemic lupus erythematosus patient.

We have analysed the heavy and light chain variable region genes of two monoclonal antibodies, specific for the Sm antigen (RSP1; IgG kappa) and for cardiolipin (RSP4; IgM lambda), derived from a patient with active systemic lupus erythematosus (SLE). We have established that the variable region genes of the RSP1 autoantibody are somatic mutants of two germ line genes from the VH4 and V kappa 1 gene families. RSP4 antibody uses gene segments closely related to a VH3 gene member and to a V lambda 1 gene. The presence and distribution of the somatic mutations on both monoclonal autoantibodies are compatible with an antigen-driven immune process. These data suggest that in SLE a common antigenic stimulus may govern the autoantibody response against a wide spectrum of unrelated antigens, including native DNA, cardiolipin or Sm antigens, and provide further evidence that disease-associated autoantibodies are generated through antigen-selected somatic mutations.

Adult↗
PubMed1994

Human lupus anti-DNA autoantibodies undergo essentially primary V kappa gene rearrangements.

We have recently characterized the heavy chain variable region (VH) genes expressed by a panel of human anti-DNA antibodies derived from four patients with systemic lupus erythematosus and expressing an idiotypic marker representative of a subset of pathogenic autoantibodies. Here, we have cloned and sequenced the kappa chain variable region genes (V kappa) of the clones whose VH genes had been previously analysed. All the V kappa genes utilized map to the 280 kb portion of the 3' end of the locus, suggesting that they represent essentially the products of primary rearrangements. This proximal clustering of the V kappa genes used contrasts with the broad distribution of immunization-induced human antibody V kappa genes over 1400 kb of the locus. In addition, lupus autoantibodies show no tendency to express the downstream junctional (J kappa) exons--another indication of infrequent secondary variable gene assembly. Since successive rearrangements may extinguish high-affinity recognition of self antigens, we propose that this bias in V kappa and J kappa expression reflects a low rate of secondary light chain rearrangements among lupus autoantibodies. We also postulate that the corrective mechanism capable of editing potentially aggressive, self-reactive antibodies in these patients may be deficient--a deficit that could be genetically determined and/or somatically acquired.

Amino Acid Sequence↗
PubMed1994

Somatic diversification in the heavy chain variable region genes expressed by human autoantibodies bearing a lupus-associated nephritogenic anti-DNA idiotype.

Monoclonal anti-DNA antibodies bearing a lupus nephritis-associated idiotype were derived from five patients with systemic lupus erythematosus (SLE). Genes encoding their heavy (H)-chain variable (VH) regions were cloned and sequenced. When compared with their closest VH germ-line gene relatives, these sequences exhibit a number of silent (S) and replacement (R) substitutions. The ratios of R/S mutations were much higher in the complementarity-determining regions (CDRs) of the antibodies than in the framework regions. Molecular amplification of genomic VH genes and Southern hybridization with somatic CDR2-specific oligonucleotide probes showed that the configuration of the VH genes corresponding to VH sequences in the nephritogenic antibodies is not present in the patient's own germ-line DNA, implying that the B-cell clones underwent somatic mutation in vivo. These findings, together with the characteristics of the diversity and junctional gene elements utilized to form the antibody, indicate that these autoantibodies have been driven through somatic selection processes reminiscent of those that govern antibody responses triggered by exogenous stimuli.

Amino Acid Sequence↗
PubMed1994

Anti-idiotypic antibodies to the third variable domain of gp120 induce an anti-HIV-1 antibody response in mice.

We tested the potential of anti-idiotypic antibodies to function as molecular mimics of a neutralizing epitope of the human immunodeficiency virus (HIV). Three monoclonal antibodies to the third variable domain (V3) of gp120 from the HIV-1LAI isolate were raised in a BALB/c mouse. They bound gp120LAI with high affinity (K congruent to 10(-11) M), immunoprecipitated viral gp120LAI, recognized HIVLAI-infected cells by immunofluorescence and neutralized HIVLAI infection in vitro. Mice and rabbits were immunized against each one of these antibodies, and one monoclonal and two polyclonal anti-idiotypic reagents were selected for further study. The three anti-idiotypes recognized binding site-related idiotopes that were not present on other V3-specific mouse sera or monoclonal antibodies. Groups of mice and rabbits were immunized against these anti-idiotypes presented in a homopolymerized form, conjugated to a protein carrier, or in an uncoupled form. In the absence of antigen, mice that received polyclonal anti-idiotypes mounted an antibody response to gp120LAI, but not to V3-deleted gp120LAI recombinant proteins. Unlike the three nominal mAbs, the induced antibodies also reacted with gp120 from the divergent HIV strain SF2. The isotypic distribution of the anti-idiotype-induced antibodies and their temporal evolution resemble the restricted pattern and the rapid rise of the antibody response obtained after administration of recombinant gp120LAI to mice. These data thus present evidence that anti-idiotypes raised against HIV-specific antibodies may substitute viral antigens for induction of a humoral immune response to HIV. They also suggest that the HIV epitope variability may be, in part, overcome with the use of anti-idiotypic reagents.

Animals↗