Epstein-Barr Virus Genome-positive Cell Research Articles

Glomerulonephritis in a patient with chronic active Epstein-Barr virus infection

Renal involvement is rare in chronic active Epstein-Barr virus (EBV) infection. We report an 11-year-old girl who had focal mesangial proliferative glomerulonephritis with cellular crescents and renal tubular atrophy with foam cells in the lumen at the time of the first admission. However, the patient was not diagnosed with chronic active EBV infection until the third admission, 18 months later, because she did not exhibit typical clinical manifestations of infectious mononucleosis, i.e., fever, lymphadenopathy, hepatomegaly, or increased atypical lymphocytes. We performed in situ hybridization of EBV in renal biopsy and renal autopsy tissue and found EBV genome-positive cells in the enlarged vascular areas surrounding the renal tubules in both specimens. The relationship between mesangial proliferative glomerulonephitis with crescents and chronic active EBV infection is unknown.

Interactions between Epstein-Barr Virus (EBV) and Human Cell Lines: Growth and EBV Induction

Differences in in vitro growth characteristics between Epstein-Barr virus (EBV) genome-negative and genome-positive lymphoblastoid cell lines and between EBV genome-negative and genome-positive epithelial cell lines were investigated. All EBV genome-positive cells were able to grow in medium containing only 0.75% fetal calf serum (FCS), whereas no EBV genome-negative cells were able to grow at this low concentration. However, there was no clear difference between the growth of the two EBV genome-negative lymphoblastoid cell lines (BJAB and Ramos) and two EBV genome-positive BJAB and Ramos sublines (BJAB/B95-8 and Ramos/B95-8) in medium containing more than 2.5% FCS. On the other hand, an EBV genome-positive clone of A2L/AH epithelial hybrid cells (cl-1) grew in both concentrations of FCS better than an EBV genome-negative clone of A2L/AH cells (cl-654). In addition, there was a difference in EBV induction between EBV genome-positive lymphoblastoid (BJAB/B95-8 and Ramos/B95-8) and epithelial (cl-1) cells after treatment with 12-O-tetradecanoylphorbol-13-acetate (TPA). Treatment of BJAB/B95-8 and Ramos/B95-8 cells with TPA induced a low percentage of EBV-induced early antigen but not viral capsid antigen, whereas treatment of cl-1 cells with TPA induced both early antigen and viral capsid antigen.

Immune response to intermediate filament-associated, Epstein-Barr virus-induced early antigen.

A murine monoclonal antibody (MAb) (15TD3) was found to recognize a unique filamentous structure in Epstein-Barr virus (EBV)-producing lymphoblastoid cell lines. By immunofluorescent morphology, in comparison with a control MAb to vimentin, the 15TD3 filamentous structure was judged to be associated with intermediate filaments of the cytoskeleton. Expression of the 15TD3 antigen and vimentin was induced simultaneously in some EBV genome-positive cell lines either by EBV superinfection or by 12-0-tetradecanoyl-1-phorbol-13-acetate (TPA) and n-butyrate treatment. The 15TD3 antigen was considered to be a restricted component of the EBV-induced early antigen (EA) complex. The 15TD3 antigen was expressed only in EBV genome-activated cells after either spontaneous EBV genome activation, EBV superinfection, or TPA and n-butyrate treatment. The expression of 15TD3 antigen paralleled the induction of EA in several models of induction of EBV antigens, and was detected only in EA+ cells which were stained with anti-EA+ human sera. The reactivity of 15TD3 MAb was blocked with anti-EA+ human serum, but not with anti-EA- serum. The synthesis of 15TD3 antigen was not inhibited with phosphonoacetic acid, was resistant to acetone fixation, and was sensitive to ethanol (or methanol) fixation. Human lymphoblastoid cells from patients with acute infectious mononucleosis were cloned for the production of antibodies which detected EBV-specific or -nonspecific epitopes on filamentous structures. Two human MAb were defined by two-color immunofluorescence to react to the 15TD3 determinants on intermediate filaments of EBV+ cells. This study supports the following views: that EBV genome activation induces a structure associated with intermediate filaments, and that antibodies against both the EBV-specific, intermediate filament-associated epitope and native intermediate filament epitopes are produced by some EBV-transformed lymphoblastoid cell lines from patients with infectious mononucleosis.

HTLV-III infection of EBV-genome-positive B-lymphoid cells with or without detectable T4 antigens.

The infection of a number of new and established B-cell lines by human T-cell lymphotropic virus III (HTLV-III) was investigated. The B lymphocytes differed in their expression of T4 antigens detected by specific monoclonal antibodies (MAbs) and the presence of Epstein Barr virus (EBV)-DNA or antigens. The presence of the EBV genome was the only requirement for infection of B-lymphocytes by HTLV-III, although its presence did not ensure infection. Two EBV genome and T4 antigen-positive B-cell lines, lacking EBV early antigens (EA) and viral capsid antigens (VCA), could be productively infected with no induction of known EBV antigens. Two other EBV genome-positive cell lines, lacking T4, EA, and VCA could also be infected. Another genome-positive cell line (P3HR-I) that was EBV-EA, VCA-positive and produced non-transforming EBV, could also be infected by HTLV-III. However, 3 EBV genome- and T4 antigen-negative B-cell lines could only be infected with HTLV-III after successful conversion to an EBV-genome-positive state by pre-infection with EBV. Five other EBV-genome-positive B-cell lines lacking T4 antigens were not infectible with HTLV-III even after super-infection with EBV. Incomplete inhibition of the HTLV-III infection of a T4-positive (LDV-7) and a T4-negative (Craig) was obtained by preadsorption with specific MAb to T4 (OKT4A and Leu 3A). From these observations, it is not clear whether the presence of T4 antigen on the cell surface is needed for the infection of B lymphoblastoid cells; however, successful infection does depend upon the presence of the EBV genome. The mechanism of interaction of HTLV-III and EBV-infected B-cell lines permitting this infection is not fully understood. Although the clinical implications of these observations remain to be determined, it is possible that infection of EBV-positive B-cells may contribute to aberrant humoral responses and/or increased frequency of B-cell malignancies observed in HTLV-III-infected individuals.

Identification of multiple Epstein-Barr virus-induced nuclear antigens with sera from patients with rheumatoid arthritis.

By means of the protein immunoblot technique, the Epstein-Barr virus (EBV) nuclear antigen (EBNA) could be identified in a variety of EBV-transformed cell lines with anti-EBNA-positive sera from normal donors. The molecular weight of EBNA expressed in each of the cell lines varied between 70,000 and 75,000 and was dependent upon the strain of infecting virus. In contrast, 15 of 21 sera from patients with rheumatoid arthritis identified antigens in addition to EBNA. The most prominent of these antigens had molecular weights of 110,000 to 115,000 and 92,000. All of the EBV genome-positive cell lines except for QIMR-GOR and cell lines containing the P3HR-1 virus expressed these antigens. The antigens were not present in the EBV genome-negative Ramos and BJAB cell lines, nor were they identified with EBV seronegative sera, indicating that they were EBV related. There was no direct correlation between the presence of antibodies in sera to EBNA, viral capsid antigen or early antigen, and reaction with the 92,000-molecular-weight antigen in immunoblots, indicating that this antigen was distinct from previously described EBV-related antigens.

Enhancement of outgrowth of EB virus-transformed cells from normal human peripheral blood by a tumor promoter, TPA.

Effect of 12-0-tetradecanoyl-phorbol-13-acetate (TPA) on the establishment of lymphoblastoid cell lines (LCL) following spontaneous tranformation of B cells by Epstein-Barr virus (EBV) from peripheral lymphocytes of healthy EBV-seropositive adults was examined. In the lymphocyte culture with TPA (0.5 ng/ml), the frequency of establishment of LCL increased and the time required for appearance of transformation decreased. A larger number of EBV-positive cells in the peripheral blood were detected in the assay by co-cultures with cord blood lymphocytes in the presence of TPA than in the absence of it. The enhancement of LCL establishment by TPA was largely abolished in the culture with EBV neutralizing antibody-containing human umbilical cord serum. This suggests that the enhancement by TPA may be caused mostly by transformation of coresident B lymphocytes infected with active virus induced from the EBV genome-positive cells by TPA.