Identification of an Epstein-Barr virus nuclear antigen by fluoroimmunoelectrophoresis and radioimmunoelectrophoresis

Abstract

A 65,000-dalton (65K) antigen found in Raji cells by fluoroimmunoelectrophoresis and radioimmunoelectrophoresis has been identified as an Epstein-Barr virus nuclear antigen (EBNA). This identification is based on the following evidence. The 65K antigen is detected in Raji cells but not in three Epstein-Barr virus (-) human B cell lines. It is not detected with EBNA (-) sera. The 65K antigen is found predominantly in the nucleus and co-elutes with EBNA during partial purification by DNA-Sepharose and Blue Dextran-Sepharose chromatography. Finally, the partially purified 65K antigen is an effective absorbant of EBNA antibody as measured in an anticomplement immunofluorescence assay. Antigens with molecular weights of 72, 70, and 73K have been detected in B95-8, P3HR-1, and Namalwa cells, respectively. These antigens are the likely homologues of the 65K Raji EBNA. In addition, an Epstein-Barr virus-associated, 81K DNA-binding antigen has been detected in both B95-8 and Raji cells.

Characterization of a second Epstein‐Barr virus‐determined nuclear antigen associated with the BamHI WYH region of EBV DNA

“…In contrast, we found sera which were negative both for EBNA-2 in ACIF and for 86 kd on immunoblots. Our finding that both EBNA-1 and 86 kd are dsDNA-binding is in agreement with the findings of Strnad et al, (1981). and the presence of both EBNA-1 and 86 kd in the nuclear fractions of cells confirms the results of Hennessy and Kieff (1983).…”

“…A recent study has identified an 82 kd polypeptide present in the nuclear fraction of EBV-carrying cell lines, but not in EBV-negative lines (Hennessy and Kieff, 1983). Previously, Strnad et al, (1981) who first detected EBNA-1 on immunoblots also observed an 81 kd polypeptide detected in 2/4 EBV-positive cell lines by 2/5 EBV-positive antisera. It was suggested that this polypeptide might be EBV-specific.…”

Lymphoblastoid cell lines and burkitt‐lymphoma‐derivee cell lines differ in the expression of a second epstein‐barr virus encoded nuclear antigen

“…In contrast, only 10 of 27 Burkitt lymphoma lines expressed EBNA-2. Four of the lines were EBNA-2negative when analyzed by immunoblotting (Dambaugh et al, 1984;Dillner et al, 1985a,b;Strnad et al, 1981). Two of these (Daudi and P3HR-1) release or can be induced to release non-transforming virus with a completely deleted EBNA-2 region.…”

“…Antibodies to one of the 3 possible reading frames in the third internal repeat array in LT-2 conclusively identified the LT-2 gene product as a 70-92-kDa nuclear polypeptide, designated EBNA-1 (Dillner et al, 1984;Heller et al, 1981). Recently, several reports have described a second EBV-associated nuclear antigen, EBNA-2, detected by sera from EBV-seropositive donors by immunoblotting (Dillner et al, 19856;Hennessy and Kieff, 1983;Sculley et al, 1984;Strnad et al, 1981) and by immunofluorescence (Dillner et al,198%). This has also been shown by the production of antisera to a bacterial fusion protein containing part of the LT-1 region (Hennessy and Kieff, 1985).…”

Identical igm antibodies recognizing a glycine‐alanine epitope are induced during acute infection with epstein‐barr virus and cytomegalovirus

“…A primary infection is confirmed serologically by the initial detection of antibody to the EBV-specific viral capsid antigen (VCA) followed by the later appearance (usually after 1-6 mo) of antibodies to the EBV encoded nuclear antigen (EBNA) (2); for a review, see de The (3) and Henle and Henle (4). EBNA-I , the first nuclear antigen to be recognized, has been identified as a 69,000-85,000 protein by the immunoblotting technique (5)(6)(7). The size of the EBNA-I protein is correlated with the variation of the length of the IR-3 region of EBV-DNA (6).…”