Antibodies against glutathione S-transferase T1 in non-solid organ transplanted patients.

Background: This article describes the presence of antibodies against glutathione S-transferase T1 (GSTT1) in a group of patients who never received a solid organ graft. These antibodies have been previously detected in liver and kidney transplant subjects with donor-recipient mismatch for this enzyme at the genetic level. In liver-grafted subjects, the appearance of these antibodies correlated with de novo immune hepatitis.

Anti-centromere antibodies in patients with systemic lupus erythematosus.

Background: Anti-centromere autoantibodies (ACA) are frequently detected in systemic sclerosis (SScl), especially in the calcinosis, Raynaud phenomenon, esophageal dysmotility, sclerodactyly, telangiectasia (CREST) syndrome, in which a prevalence of 55% has been reported. The presence of ACA in systemic lupus erythematosus (SLE) is so rare that its detection can raise serious doubts about the validity of the diagnosis.

Objective: To determine the frequency of ACA positive subjects from a wide monocentric cohort of SLE patients and analyse the clinical and biological characteristics of this group.

A human monoclonal antibody reacting against HLA-DQ1-, DQ4-, and a subset of DQ7-bearing cells.

Human mAb 2A2 recognizes an epitope present in the HLA-DQ1 + 4 specifications and also on several DQ7-positive cells. We have investigated the extra reactions of this monoclonal reagent on a wider panel of DQ1-, DQ4-negative/DQ7-positive B-cell lines. The results obtained support the existence of two subtypes of the HLA-DQ7 specificity on the basis of their reactivity with human mAb 2A2; the DQ7/2A2-positive variant has been found in 12 of 29 BCLs positive for the DR11 antigen, and in four of eight BCLs bearing DR4-DQ7 haplotypes.

Application of the MAILA technique to the study of human anti-HLA monoclonal antibody specificity.

The monoclonal antibody-specific immobilization of lymphocyte antigens (MAILA) assay was developed to detect antibodies present in human alloantisera against antigens of different major histocompatibility complex loci, particularly of class II specificity. The MAILA assay has been used in our laboratory to the determination of the type of HLA molecule recognized by human monoclonal antibodies 91C2 (anti-A2 + 28), 34F11 (anti-DQ1), and 2A2 (anti-DQ1 + 4 + short DQ7), using well characterized monomorphic as well as polymorphic murine monoclonals for the specific immobilization of HLA molecules. Results obtained show that the MAILA assay is also a valuable tool for the determination of specific human MHC locus products recognized by human monoclonal antibodies.

A human monoclonal autoantibody to a nucleolar structure.

Peripheral blood lymphocytes from a scleroderma patient (CDC) were isolated, transformed with Epstein-Barr virus and fused to the heteromyeloma SHM-D33. Supernatants from cultures were screened for autoantibody production against nucleoprotamine by ELISA. Positive wells were cloned by limiting dilution.

Production of heterohybridomas secreting autoreactive and polyreactive human monoclonal antibodies.

Peripheral blood lymphocytes from two polytransfused renal dialysis patients were transformed by Epstein-Barr virus, fused to a heteromyeloma and cloned. Eight human monoclonal antibodies from the resulting clones were tested for their binding to a variety of antigens by ELISA, indirect immunofluorescence and immunoblotting. Antigens tested included B-cell lines, T and B lymphocytes, red blood cells, chronic lymphocytic leukaemic B cells, IgG, ssDNA, dsDNA, histones, nucleoprotamine, sperm nuclei, thymus and spleen extracts, MOLT4 cell lysates, affinity purified autoantigens, tetanus toxoid, bacterial lipopolysaccharide, insulin, and a tissue section screen.

Cloning of human heterohybridoma cell lines using chronic lymphocytic leukemia B cells as a feeder layer.

B cells from the peripheral blood of chronic lymphocytic leukemia patients were isolated by gradient density centrifugation and used without irradiation as a feeder layer in the cloning of human heterohybridoma cell lines by limiting dilution. Cloning efficiencies were high with all the cell lines tested. These feeder leukemia B cells could also be successfully used after having been stored in liquid nitrogen.

Characterization of three human monoclonal antibodies specific for polymorphic class I and class II HLA antigens.

Three human monoclonal antibodies were derived from a single polytransfused patient awaiting renal transplantation. In microcitotoxicity assays, the patient's serum displayed strong positive reactions against greater than 90% of a panel of cells representing the known HLA specificities. The donor's peripheral blood lymphocytes were infected with Epstein-Barr virus, cloned, and supernatants of the virus transformed cultures were screened for the presence of IgG antilymphocyte reactivity utilizing an enzyme-linked immunosorbent assay method.