Effect of E. coli thioredoxin, a homologous to the adult T-cell leukemia-derived factor, on Epstein-Barr Virus-transformed B lymphocytes

In order to investigate the role of autocrine growth factors on cell growth of Epstein-Barr virus (EBV)-transformed human B lymphocytes, we have studied the effect of E. coli recombinant thioredoxin (r-thioredoxin), a homologue of adult-T-cell leukemia-derived factor (ADF), on the cell growth of various human lymphoid cell lines, either EBV-positive or -negative. It was found that, in serum-free culture conditions, exogenous r-thioredoxin increases DNA synthesis in lymphoblastoid and Burkitt's cell lines, independently of EBV genome expression; moreover, the combined effect of r-thioredoxin and interleukin-I (IL-1) or IL-6 resulted in a marked increase in DNA synthesis in lymphoblastoid cells, but not in Burkitt's cell lines. Anti r-thioredoxin monoclonal antibodies were developed and used to test the possibility of interfering with r-thioredoxin-induced cell proliferation. It was found that the growth-promoting activity of r-thioredoxin was inhibited by an anti-r-thioredoxin monoclonal antibody in a dose-dependent manner in P3HR-1 cells, a Burkitt's lymphoma cell line harboring an EBNA-2-defective virus, but not in other lymphoid cell lines, thus indicating that target cells may play an active role in antibody-mediated thioredoxin neutralization

DETECTION OF AN IDIOTOPE ON A HUMAN MONOCLONAL AUTOANTIBODY BY MONOCLONAL ANTIIDIOTYPIC ANTIBODY AND ITS RELATIONSHIP TO EPSTEIN-BARR VIRUS-INDUCED AUTOIMMUNITY

We have recently described a human IgM monoclonal antibody (mAb), reactive with both self antigens, i.e., cytoskeleton filaments and smooth muscle, and Epstein-Barr virus (EBV)-induced nuclear antigen (EBNA), produced by EBV-transformed B lymphocytes isolated from a patient with infectious mononucleosis (IM). In order to achieve higher antibody secretion in culture supernatant, the mAb-producer cells were fused with ouabain-resistant mouse myeloma cells and a stable human-mouse heterohybrid, coded HY 5488, producing up to 80 micrograms/ml IgM mAb, was isolated after 4 cloning procedures. Purified HY 5844 mAb was used to immunize mice for the production of a murine anti-idiotypic mAb, which was used to probe the expression of the idiotope of HY 5488 mAb (Id 5488) in sera of IM patients and normal controls by ELISA. It was found that Id 5488 is expressed both in IM patients and normal controls, and that Id 5488 expression is significantly higher in IM patients' sera; furthermore, in IM sera a statistically significant correlation between Id 5488 expression and anti-cytoskeleton and anti-smooth muscle autoantibodies was found. It is suggested that at least part of EBV-induced IgM autoantibodies appearing during IM are secreted by B lymphocytes programmed to the production of "natural antibodies" bearing Id 5488-like idiotopes

Potential role of the Epstein-Barr virus in systemic lupus erythematosus autoimmunity

OBJECTIVE: To investigate the possibility that Epstein-Barr virus (EBV), the agent of infectious mononucleosis (IM), may play a role in systemic lupus erythematosus (SLE). METHODS: EBV was searched for by PCR and by culture isolation in oropharyngeal lavage fluids of 15 SLE patients and, as controls, in 13 IM patients and in 28 healthy individuals with past EBV infection. Computer analysis was performed to select an antigenic domain of the virus-encoded nuclear antigen EBNA-2, in order to set up a synthetic peptide-based immunoassay. IgG antibodies to a 20-amino acid synthetic peptide derived from the selected domain of EBNA-2 (354GRGKGKSRDKQRKPGGPWRP373) were titrated in the sera of 20 SLE patients, 24 IM patients and 12 healthy subjects. RESULTS: EBV type 1 DNA was demonstrated by PCR in the oropharyngeal secretions of 8 SLE patients and the virus was isolated from 6 DNA-positive specimens. Moreover, 50% of the patients with SLE and 100% of the patients in the acute phase of IM, but none of the EBV-seropositive normal individuals, produced IgG antibodies to the EBNA-2-derived synthetic peptide. Computer analysis revealed a high degree of homology between the EBNA-2 354GRGKGKSRDKQRKPGGPWRP373 sub-sequence and the antigenic C-terminal domain 101GRGRGRGRGRGRGRGGPRR119 of the SmD1 ribonucleoprotein, a target of autoantibodies in a portion of SLE patients. CONCLUSION: We suggest the possibility that EBV may establish a persistent infection at least in a certain number of SLE patients. The antibodies elicited by the viral antigen EBNA-2 may cross-react with SmD1, thus indicating a role of EBV-specific immune responses in the outcome of SmD1 autoantibodies in SLE patients

DENSITY-DEPENDENT RESPONSIVENESS TO AUTOCRINE GROWTH-FACTORS OF EPSTEIN-BARR-VIRUS TRANSFORMED HUMAN LYMPHOCYTES-B

Analysis of the growth requirements of Epstein-Barr virus (EBV)-transformed B lymphocytes shows that interleukin 1 and thioredoxin, a disulfide reducing enzyme, are able to induce a marked increase in DNA synthesis in the early phases of in vitro culture. By contrast, interleukin 6 induces a steady increase in DNA synthesis comparable to that observed with crude conditioned supernatant. Furthermore, EBV-transformed B cells exhibit a density-dependent responsiveness to autocrine growth factors, thus suggesting that growth regulation of EBV-transformed B cells might result from the interplay between different self-stimulating soluble factors and from the competence of the cells to respond to autocrine growth factors