Determination of the role for CD21 during Epstein-Barr virus infection of B-lymphoblastoid cells

Abstract

Epstein-Barr virus (EBV), a herpesvirus with oncogenic potential, is camouflaged with glycoprotein 350/220, which mimics the human ligand C3dg and thereby binds to and exploits complement receptor type 2 (CR2; CD21), the EBV receptor. It has not been possible to determine the role of CR2 during postbinding events of viral infection because all B lymphocytes express endogenous CR2, precluding an informative study of receptor mutants. We have overcome this obstacle through creation of a novel experimental system based on molecular dissection of the ligand-binding domains of human CR2 and murine CR2. Our results demonstrate first, that two discontinuous amino acid substitutions within the ligand-binding domain of murine CR2 render it capable of mediating EBV infection of human B-lymphoblastoid cells, and second, that the specific role of CR2 during EBV infection is to capture virions at the cell surface, after which cofactors not associated with CR2 mediate postbinding events. These are the first studies to be described in which a cell that is normally susceptible to viral infection can be manipulated so as to direct entry of virions via recombinant or endogenous receptors.

High‐potentiality preliminary selection criteria and transformation time‐dependent factors analysis for establishing Epstein–Barr virus transformed human lymphoblastoid cell lines

“…These data not only show that the previously mentioned status of the lymphocytes (blood transportation times of less than 48 h, more than 10 6 cells/ml, and cell viabilities exceeding 90%) could be the preliminary selection criteria for EBV transformation performance of isolated B lymphocytes but they also imply that this criteria is helpful specially for cryopreserved lymphocytes as starting material. Molecular mechanisms beneath the proposed selection criteria might be associated with CD21 (the CR2 complement receptor), which has been demonstrated to mediate EBV binding to and infection of B lymphocytes (Martin et al . 1994).…”

Epstein–Barr Virus Susceptibility in Activated PI3Kδ Syndrome (APDS) Immunodeficiency

“…Although a neutralizing antibody response against several viral proteins such as gp350, a particularly immunogenic EBV protein, is detectable in these patients ( 47 ), the peak of this antibody response occurs after disappearance of IM symptoms and clearance of the virus, and this delay has been attributed to B-cell dysfunction in acutely infected patients ( 46 ). Several vaccination strategies have focused on the gp350 protein ( 49 – 51 ) since it acts as a major mediator for entry of EBV into B cells through its interaction with CD21 ( 52 ). Interestingly, vaccination using recombinant gp350 in phase-I and-II trials correlated with a gp350-specific antibody response and showed a protective effect in IM development but not in asymptomatic EBV infections ( 50 , 51 ).…”

The biology of Epstein–Barr virus in post‐transplant lymphoproliferative disease

“…EBV infects B cells through an interaction between gp350/220, a viral envelope glycoprotein, and CD21, a component of the complement receptor ( 1). Following attachment, the virion is endocytosed and fuses with the vesicle membrane, releasing the nucleocapsid into the cytoplasm.…”