key: cord-0010222-vo7lz4cz
authors: Yokochi, Takashi; Inoue, Yoshiko; Iwata, Hitoshi; Miyadai, Toshiaki; Kimura, Yoshinobu
title: Differential Expression of the Major Histocompatibility Antigen Complex (MHC) on a Series of Burkitt's Lymphoma Lines
date: 2013-11-14
journal: Microbiol Immunol
DOI: 10.1111/j.1348-0421.1987.tb01354.x
sha: 69f2fa711dd9078be28950554564cdd620c618b0
doc_id: 10222
cord_uid: vo7lz4cz

We compared the expressions of class I and class II major histocompatibility antigen complex (MHC) on the surface of Jijoye and P3HR‐1 cells of Burkitt's lymphoma sublines. Jijoye cells had a large amount of class I and class II MHC antigens, whereas these antigens were less expressed on P3HR‐1 cells. On a subline of P3HR‐1 K cells the expression of class I antigen markedly diminished and class II antigen was undetectable. On the other hand, Jijoye, P3HR‐1, and P3HR‐1 K cell lines were confirmed to be Epstein‐Barr virus (EBV) nonproducer, low producer, and high producer, respectively. The chemical activation of EBV genome by treating P3HR‐1 cells with 12‐O‐tetradecanyl phorbol‐13 acetate (TPA) and n‐butyrate resulted in inhibition of the expression of class I and II antigens, while the addition of retinoic acid, an inhibitor of virus replication, blocked the decrease in the MHC antigen expression. These findings suggested that there might be an inverse correlation between the virus production and the expression of class I and II MHC antigens.

The major histocompatibility antigen complex (MHC) is a key element in restricting immunological recognition of viral antigens (24, 25) . Virus infection often modulates the cell surface expression of class I (7, 16, 17) and class II MHC antigens (12) . Epstein-Barr virus (EBV) selectively infects and transforms human resting B cells expressing both class I and class II antigens of MHC. Usually EBVtransformed B lymphoblastoid lines and Burkitt's lymphoma lines express human leukocyte antigens (HLA) and can be killed by MHC class I-specific cytotoxic T cell lines (18) . The Burkitt's lymphoma subline of P3HR-1 cells has been obtained by cloning a high virus-producing cell from the parental Jijoye cells (6). The P3HR-1 cells express only low level of MHC class II antigen, in contrast to the parental Jijoye (19) . In this paper we present the various patterns of MHC antigen expression in sublines of a Jijoye-P3HR-1 family and discuss the relationship between the MHC antigen expression and EBV production.

Cell lines. Jijoye, P3HR-1, and P3HR-1 K lines were maintained in PRMI 1640 medium supplemented with 10% fetal calf serum and antibiotics. Jijoye and The order in the amount of class I antigen per cell was Jijoye> P3HR-1 > P3HR-1 K. Anti-class II antibody precipitated two polypeptides with molecular weight of 34,000 and 27,000 from both Jijoye and P3HR-1 cell lines, whereas no such polypeptide bands were precipitated from P3HR-1 K cells (Fig. 1 b) . In the next experiment, the quantitative analysis of their surface expression was done with the aid of a flow cytometer (Fig. 2) . Jijoye cell line was the most reactive with anticlass I and class II antibodies among three cell lines. P3HR-1 cells had a lesser amount of class I and II antigens than Jijoye cells. Especially, P3HR-1 K cells expressed the smallest amount of class I antigen, and its intensity of the peak fluorescence was about one-eighth of that of Jijoye. Class II antigen was undetectable on P3HR-1 K cells by two kinds of monoclonal antibodies which were reactive with the different antigenic determinant sites of class II molecule. On the other hand, there was no difference among these three cell lines in the expression of transferrin receptors detected by a monoclonal antibody (data not shown).

Virus Productivity of Jijoye, P3HR-1, and P3HR-1 K Cell Lines Virus-producing activity was measured by detecting the EBV-associated antigens in the cells (Table 1) . P3HR-1 K cell line contained a small percent of immunofluorescence-positive cells, while no such positive cells were detected in either P3HR-1 or Jijoye cells. After activation of the EBV genome by treatment with TPA and n-butyrate for 3 days, the frequency of fluorescence-positive cells in P3HR-1 K cell culture increased up to about 40 % and also a significant number of fluorescence-positive cells appeared in P3HR-1 line. However, no positive cells were detected in Jijoye line even after the same treatment. This finding confirmed the fact that the order in virus productivity among these cell lines is P3HR-1 K> P3HR-1 > Jijoye (9).

The results in the preceding section suggested that the virus production might be inversely correlated with the expression of MHC class I and II antigens. Therefore, we investigated the MHC antigen expression after chemical activation of EBV genome (Fig. 3, a and b) . When virus production in P3HR-1 cells was stimulated by TPA and n-butyrate treatment, the expression of MHC class I and class II antigens was significantly reduced and was less than half that of untreated control cells. The additional treatment with retinoic acid blocked this reduction of MHC expressions (data not shown). In contrast to P3HR-1 cells, no reduction of MHC antigens was found on Jijoye cells, EBV-negative BJAB Burkitt's lymphoma cells, and some EBV-transformed B lymphoblastoid cell lines even after chemical stimulation of virus genome.

In the present study we have found an inverse correlation between the expression of class I and II MHC antigens and the EBV productivity. Klein et al (9) have also reported that certain nonproducer revertant cells from P3HR-1 line again express a high level of class II antigen (9). Although virus structural proteins have been detected in only a limited number of cells even in virus producer P3HR-1 K cells (Table 1) , a significant number of virus DNA copies are synthesized in these cells (1). Further, chemical treatment markedly enhances the copy number of virus genomes per cell (4). Therefore, it seems likely that the amplification of virus genome may participate in the decrease of MHC antigen expression.

The parental Jijoye and other EBV-positive Burkitt's lymphoma lines release viruses with transforming activity (15) . When normal peripheral blood lymphocytes are transformed by these EBV (13) , class I molecules markedly become increased. On the contrary, P3HR-1 cells produce nontransforming EBV, and this variant virus is unable to immortalize normal B lymphocytes and shows cytopathic effects on EBV receptor-carrying cells (14) . The P3HR-1 EBV DNA has a deletion of 2 x 106 dalton out of the Jijoye EBV DNA and the deleted part of the DNA sequence encodes the mRNAs detected in restringently-infected, growth-transformed cells (5, 8) . Therefore, this transformation-related DNA might be involved in the regulation of MHC antigen expression. 

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