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. 1998 Jul;72(7):5552-8.
doi: 10.1128/JVI.72.7.5552-5558.1998.

Epstein-Barr virus uses different complexes of glycoproteins gH and gL to infect B lymphocytes and epithelial cells

X Wang  1 , W J Kenyon, Q Li, J Müllberg, L M Hutt-Fletcher
Affiliations

Epstein-Barr virus uses different complexes of glycoproteins gH and gL to infect B lymphocytes and epithelial cells

X Wang et al. J Virol. 1998 Jul.

Abstract

The Epstein-Barr virus (EBV) gH-gL complex includes a third glycoprotein, gp42. gp42 binds to HLA class II on the surfaces of B lymphocytes, and this interaction is essential for infection of the B cell. We report here that, in contrast, gp42 is dispensable for infection of epithelial cell line SVKCR2. A soluble form of gp42, gp42.Fc, can, however, inhibit infection of both cell types. Soluble gp42 can interact with EBV gH and gL and can rescue the ability of virus lacking gp42 to transform B cells, suggesting that a gH-gL-gp42.Fc complex can be formed by extrinsic addition of the soluble protein. Truncated forms of gp42.Fc that retain the ability to bind HLA class II but that cannot interact with gH and gL still inhibit B-cell infection by wild-type virus but cannot inhibit infection of SVKCR2 cells or rescue the ability of recombinant gp42-negative virus to transform B cells. An analysis of wild-type virions indicates the presence of more gH and gL than gp42. To explain these results, we describe a model in which wild-type EBV virions are proposed to contain two types of gH-gL complexes, one that includes gp42 and one that does not. We further propose that these two forms of the complex have mutually exclusive abilities to mediate the infection of B cells and epithelial cells. Conversion of one to the other concurrently alters the ability of virus to infect each cell type. The model also suggests that epithelial cells may express a molecule that serves the same cofactor function for this cell type as HLA class II does for B cells and that the gH-gL complex interacts directly with this putative epithelial cofactor.

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Figures

FIG. 1
FIG. 1
Effect of MAbs to HLA class II (HB55 and ALVA 42) or to gH-gL (E1D1) on the infection of SVKCR2 cells by Akata strain virus. Cells were harvested 4 days after infection, and equal numbers were analyzed by Western blotting for expression of EBNA 1. Blots were reacted with human serum containing antibody to EBNA 1 and with goat anti-human Ig conjugated to alkaline phosphatase. Uninduced Akata cells were included on the far right to demonstrate the electrophoretic mobility of EBNA 1 from this strain of virus. +, present; −, absent.
FIG. 2
FIG. 2
Effect of gp42.Fc, Cont.Fc, and MAb E1D1 on the infection of SVKCR2 cells by Akata strain virus. Cells were harvested 4 days after infection, and equal numbers were analyzed by Western blotting for expression of EBNA 1. Blots were reacted with human serum containing antibody to EBNA 1 and with goat anti-human Ig conjugated to alkaline phosphatase. Uninduced Akata cells were included on the far right to demonstrate the electrophoretic mobility of EBNA 1 for this strain of virus. The amounts of soluble proteins added (in micrograms) are indicated. +, present; −, absent.
FIG. 3
FIG. 3
Effect of gp42.Fc and Cont.Fc on infection of SVKCR2 cells by P3HR1 strain virus. Cells were harvested 4 days after infection, and equal numbers were analyzed by Western blotting for expression of EBNA 1. Blots were reacted with human serum containing antibody to EBNA 1 and with goat anti-human Ig conjugated to alkaline phosphatase. Uninduced P3HR1 cells were included on the far right to demonstrate the electrophoretic mobility of EBNA 1 for this strain of virus. +, present; −, absent.
FIG. 4
FIG. 4
Binding of iodinated gp42.Fc to SVKCR2 cells cannot be competitively inhibited by excess unlabeled protein and shown to be specific unless cells are induced with gamma interferon to express HLA class II. The left side of the figure compares the amount of iodinated gp42.Fc bound to HLA class II-negative (HLA class II-ve) cells in the absence of any other protein (arbitrarily set at zero; bar to the extreme left) with the amount bound in the presence of a 10-fold excess of unlabeled gp42.Fc or Cont.Fc or 100 μg of MAb F-2-1. Since the data represent the averages of three experiments done with different batches of iodinated protein, they are expressed as percent changes in binding rather than as counts per minute. The right side of the figure shows the relative increase in binding of gp42.Fc to cells expressing HLA class II (HLA class II +ve) (almost 100 percent more, or twice the amount bound to HLA class II-negative cells) and shows that the addition of 100 μg of F-2-1 or a 10-fold excess of soluble gp42.Fc, but not of soluble Cont.Fc, significantly inhibited this binding. Vertical lines indicate the standard deviations of the averages of the three experiments.
FIG. 5
FIG. 5
Comparison of the abilities of wild-type Akata virus and recombinant virus lacking gp42 to infect SVKCR2 cells in the presence or absence of MAb E1D1. Cells were harvested 4 days after infection, and equal numbers were analyzed by Western blotting for expression of EBNA 1. Blots were reacted with human serum containing antibody to EBNA 1 and with goat anti-human Ig conjugated to alkaline phosphatase. Uninduced Akata cells were included on the far right to demonstrate the electrophoretic mobility of EBNA 1 for this strain of virus. +, present; −, absent.
FIG. 6
FIG. 6
SDS-PAGE analysis of proteins precipitated from Akata cells harboring wild-type (Akata) or recombinant (Akata gp42-) episomes. Cells were induced with anti-human Ig and labeled with [3H]glucosamine. Proteins were immunoprecipitated by MAb ALVA 42 to HLA class II or anti-peptide antibody to gL, or by gp42.Fc or Cont.Fc (A) or were precipitated by truncated constructs ΔN58, ΔN90, and ΔN122 (B).
FIG. 7
FIG. 7
Effect of truncated constructs ΔN58, ΔN90, and ΔN122 and gp42.Fc, Cont.Fc, and MAb E1D1 on infection of SVKCR2 cells with Akata strain virus. Cells were harvested 4 days after infection, and equal numbers were analyzed by Western blotting for expression of EBNA 1. Blots were reacted with human serum containing antibody to EBNA 1 and with goat anti-human Ig conjugated to alkaline phosphatase. Uninduced Akata cells were included on the far right to demonstrate the electrophoretic mobility of EBNA 1 for this strain of virus.
FIG. 8
FIG. 8
Effect of truncated constructs ΔN58, ΔN90, and ΔN122 and Cont.Fc on superinfection of Raji cells by P3HR1 strain virus. Cells were harvested 3 days after infection, and equal numbers were analyzed by Western blotting with a MAb for expression of EA-D.
FIG. 9
FIG. 9
SDS-PAGE analysis of proteins immunoprecipitated by MAb F-2-1 to gp42 or anti-peptide antibody to gL from virus harvested from the supernatant medium of Akata cells labeled with [3H]glucosamine and purified by centrifugation on a discontinuous dextran gradient. Prior to immunoprecipitation solubilized virus proteins were sedimented through a gradient of 5 to 25% sucrose and separated into 10 fractions. Each fraction was divided in two for immunoprecipitated. The fraction immunoprecipitated in each pair of lanes is indicated at the top (F5 [fifth from the top of the gradient] to F8 [toward the bottom of the gradient]).
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