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V Pascual

Publications and source records attributed to V Pascual.

49 records · Page 3Linked to original sources
PubMed1992

The human VH repertoire: a restricted set of VH genes may be the target of immune regulation.

During the development of the immune system, a restricted set of VH gene segments provides the bulk of the immunoglobulin heavy chain repertoire. Most of these VH genes have been found later in life encoding autospecificities either in normals or in patients with autoimmune diseases. Additionally, there is considerable evidence that the fetal/neonatal B-cell repertoire is autoreactive and idiotypically connected. In the course of sequencing the heavy chain of a panel of human autoantibodies mainly derived from patients with autoimmune diseases, we found that one of the VH families, and more specifically one single VH gene contributes to a large extent to the adult autoimmune repertoire in restricted as well as unrestricted responses. This VH gene segment is not particularly overexpressed in the fetus. Since the only common element to these autoreactive responses is the region encoded by the VH gene itself, these observations may provide an important insight into B-cell regulation.

Adult↗
PubMed1992

IgM rheumatoid factors in patients with rheumatoid arthritis derive from a diverse array of germline immunoglobulin genes and display little evidence of somatic variation.

Rheumatoid factors (RF) are present in the plasma of patients with rheumatoid arthritis (RA) although the site of synthesis of most of these antibodies is within the synovium. This report primary concerns RF of the IgM isotype. While a few of the RF derive from patients with systemic lupus erythematosus or from normal individuals, the remaining derive from the inflamed synovial tissue of patients with RA. Two RF are encoded by members of the VH1 gene family, 8 from the VH3 family and 2 from the VH4 family. Two polyreactive antibodies derive from the VH3 family and 2 come from the VH4 family. This distribution is not fundamentally different from the distributions seen in a large array of autoantibodies and antibodies to external antigens. Similarly, the light chains derive from most of the known kappa and lambda VL families. It is hard to escape the preliminary conclusion that gene segments from virtually any light chain variable region can contribute to RF or polyreactive antibody structures. Most IgM RF and polyreactive antibodies are direct copies of germline genes in one of their polypeptide chains or at most are 2 nucleotides away in one of their chains from a known germline gene.

Antibody Formation↗
PubMed1991

Nucleotide sequence analysis of the V regions of two IgM cold agglutinins. Evidence that the VH4-21 gene segment is responsible for the major cross-reactive idiotype.

Cold agglutinins are human autoantibodies, usually of the IgM class, which agglutinate RBC at low temperature. The major subset recognizes the I/i carbohydrate Ag, and many of these antibodies bear cross-reacting idiotypic determinants. An anti-idiotypic mAb that is specific for one of the idiotopes largely confined to cold agglutinins has been used to identify and monitor tumor cells that secrete these molecules in two patients. The tumor cells were immortalized with EBV and the idiotope-positive lines used to investigate the utilization of the VH and VL genes by these antibodies. Nucleotide sequence analysis of the two cold agglutinins (FS-1 and FS-2) revealed the utilization of a single common gene segment, VH4-21. Serologic analysis documented that only human antibodies utilizing the VH4-21 gene segment were reactive in the idiotope assay, other VHIV antibodies as well as a panel of antibodies derived from other VH families being negative. The DH, JH, VK, and JK gene segments of FS-1 and FS-2 were structurally distinct. These data suggest that the structural basis for the cross-reactive idiotope as well as cold agglutinin activity is the VH4-21 gene segment. A nucleotide change in H chain CDR1 of both cold agglutinins results in the substitution of an aspartic acid residue for glycine at position 31, suggesting that this amino acid might be critical to recognition of the red cell Ag.

Aged↗
PubMed1991

Human monoclonal antibodies against blood group antigens preferentially express a VH4-21 variable region gene-associated epitope.

An anti-idiotypic antibody has been raised which recognizes human immunoglobulins with cold agglutinin activity of anti-I/i specificity. The pattern of reactivity of the antibody indicates that the structural basis for the epitope is located in the VH4-21 gene segment of the VHIV family, which is preferentially utilized by these cold reactive antibodies. Using this antibody, epitope expression was investigated in a panel of 72 human monoclonal allo-antibodies specific for human blood group antigens, as compared with a control panel of 39 randomly selected human monoclonal IgM antibodies of unknown specificities. The anti-blood group panel included 44 IgM and 28 IgG monoclonal antibodies against a variety of blood group antigens including the A antigen, Rh C, c, D, E, e, G antigens, and the Kidd antigens Jka and Jkb. The epitope was expressed by 64% (28/44) of the IgM anti-blood group antibodies and by 21% (6/28) of the IgG antibodies, but by only 7.7% (3/39) of the control IgM antibodies. These data indicate that the human alloimmune response to blood group antigens is biased in the use of VH gene families, with a preference for the VH4-21 gene segment of the VHIV family, or closely related gene segments. The fact that this mirrors the findings for the autoimmune cold agglutinins suggests a link in immunoglobulin gene usage between antibodies against structurally diverse antigens on the red cell surface.

Antibodies, Monoclonal↗
PubMed1990

Nucleotide sequence of a human monoclonal anti-idiotypic antibody specific for a rabies virus-neutralizing monoclonal idiotypic antibody reveals extensive somatic variability suggestive of an antigen-driven immune response.

We previously described the isolation and characterization of a human monoclonal anti-idiotypic antibody (Ab2) isolated from EBV-transformed human PBL after immunization with rabies vaccine. The present study concerns the molecular characteristics of the Ab2 and the germ-line elements that gave rise to it. The H chain of this antibody derives from the small VHV family of human V region gene segments. Parallel studies on the germ-line VHV gene isolated from the same individual revealed that the expressed molecule contains 19 nucleotide differences in the VH gene segment. The D segment of Ab2 could have arisen by a D to D fusion; the J segment is a JH6. Extensive somatic variation evident in the H chain variable region of this naturally arising monoclonal anti-idiotypic antibody suggests that this Ab2, the product of a CD5+ B cells, was the consequence of an Ag-driven immune response.

Amino Acid Sequence↗
PubMed1990

The complete nucleotide sequences of the heavy chain variable regions of six monospecific rheumatoid factors derived from Epstein-Barr virus-transformed B cells isolated from the synovial tissue of patients with rheumatoid arthritis. Further evidence that some autoantibodies are unmutated copies of germ line genes.

Structural studies of human monoclonal rheumatoid factors (RF) derived from patients with monoclonal gammapathies have revealed a restriction in the usage of heavy and light chain variable regions. These studies have suggested that germline genes with little if any somatic mutation can generate RF specificity. However, there is no information presently available regarding the primary structure and genetic origin of RF in rheumatoid arthritis. In this study, we have isolated and sequenced the VH regions of six monoclonal RF derived from the synovial membranes of two patients with rheumatoid arthritis and one with the juvenile polyarticular form of the disease. We found the same VH families as previously reported among monoclonal paraproteins with RF activity. However, our sample was diverse regarding the VH, DH, and JH gene segments used. Among VHI RF there was conservation in the length of CDRIII as well as restriction in the amino acid generated at the V-D junction, as opposed to VHIII RF and non-RF VHI molecules that are highly heterogeneous in these two aspects. We also found that different JH gene segments may contribute to RF specificity. The VH, DH, and JH elements of one RF in our study all had clearly identifiable germline counterparts. This RF displays a nearly germline configuration throughout its entire heavy chain and represents another example of an autoantibody encoded by one of the VH gene segments from the preimmune fetal repertoire.

Arthritis, Rheumatoid↗
PubMed1990

Heavy chain variable region gene utilization in human antibodies.

We have sequenced by the polymerase chain reaction the heavy chain variable regions of over thirty human antibodies of defined specificity. The distribution of VH families in this group of antibodies is disproportionately skewed toward the recently discovered VH gene families. Within a family, there is also a disproportional use of individual gene segments. Numerous explanations for these imbalances are discussed.

Autoantibodies↗
PubMed1978

Sensitivity of ribosomal bound N-acetylphenylalanyl-tRNA to hydrolysis by a spectific hydrolase.

Binding of N-acetylphenylalanyl-tRNA, either enzymatically or non-enzymatically, to yeast 80-S ribosomes renders the substrate resistant to the hydrolytic action of a specific hydrolase present in yeast. In contrast, N-acetylphenylalanyl-tRNA bound to the 40-S ribosomal subunit is sensitive to enzymatic hydrolysis. The presence of the hydrolase in the aminoacyl-tRNA binding factor preparations greatly interferes with the formation of complexes between N-acetylphenylalanyl-tRNA and the small ribosomal subunit.

Hydrolases↗
PubMedDate not supplied

Anomalous creatine kinase isoenzymes in serum from patients with rectal carcinoma.

Sera from eight patients with biopsy-proven rectal carcinoma were assayed for creatine kinase (CK). Two of the patients had enzyme activity: one with the BB and the other with an MB isoenzyme. We think that our results may be of interest because the detection of anomalous isoenzymes of CK in serum ma serve as a diagnostic marker or as an indicator of the effectiveness of treatment.

Carcinoma↗
PubMedDate not supplied

Probing of the rheumatoid factor (RF) V gene repertoire in rheumatoid arthritis (RA) by hybridoma clones.

Twenty human monoclonal antibodies with rheumatoid factor (RF) specificity were produced from fusions using B lymphocytes derived from the synovial tissue of two patients with rheumatoid arthritis (RA) and one with the polyarticular form of juvenile rheumatoid arthritis (JRA) (1). All the 20 monoclonal antibodies were IgM. Fourteen of these were classical RFs with specificity restricted to IgG, and included 12 kappa and 2 lambda proteins. When the fine specificity for IgG Fc determinants were investigated most of them showed the Ga specificity. In addition, 5 lambda and 1 kappa monoclonal RF antibodies showed polyreactivity and also reacted with various other antigens than IgG (1). The 14 monoreactive RFs were further studied for the expression of RF-related cross-reactive idiotypes (CRI) and variable heavy (VH) and light chain (VL) subgroups. Only four of the twelve kappa RFs expressed the V kappa III subgroup. Three of them belong to the V kappa IIIb sub-subgroup and expressed the CRI 17.109. One of these 3 clones in addition expressed the VH I associated CRI G6. Five other monoreactive RFs expressed either or both of the VH III associated CRI B6 and D12 (2). Using staphylococcal protein A (SPA) binding as well as Northern blotting techniques (2), studies indicated that 10 out of the 12 RFs studied expressed the VH III regions and 2 expressed the VH I region. These data, both for the heavy and light chains, indicated a different V gene usage by the RF derived from RA patients than by the RF M-components derived from patients with mixed cryoglobulinemia and Waldenström's macroglobulinemia but without RA.(ABSTRACT TRUNCATED AT 250 WORDS)

Antibodies, Monoclonal↗