Role of the capsid protein VP4 in the plasma-dependent enhancement of the Coxsackievirus B4E2-infection of human peripheral blood cells.

It has been previously shown that antibodies contained in human plasma directed towards the Coxsackievirus B4 (CVB4)E2 capsid protein VP4 can enhance the CVB4E2-induced production of IFN-alpha by peripheral blood mononuclear cells (PBMC). The aim of this study was to produce a VP4 fusion protein to investigate the role of the internal capsid protein VP4 and anti-VP4 antibodies in the plasma-dependent enhancement of CVB4E2 infection of PBMC. A fusion protein MBPVP4 containing the VP4 insert of CVB4E2 and a control fusion protein MBP-beta-gal-alpha, were produced in Escherichia coli K12 TB1. The CVB4E2 infection of PBMC was quantified by using a real time PCR method amplifying CVB4E2-negative strand RNA. IFN-alpha concentrations in culture supernatants were assayed by DELFIA. MBPVP4 but not MBP-beta-gal-alpha, preincubated with plasma inhibited the plasma-dependent enhancement of CVB4E2-induced production of IFN-alpha by PBMC. Human plasma samples, antibodies contained in plasma eluted from MBPVP4-coated plates, but not from MBP-beta-gal-alpha-coated plates, incubated with CVB4E2 enhanced the infection of PBMC and the production of IFN-alpha by infected cells. Together our results show that VP4 and anti-VP4 antibodies play a role in the plasma-dependent enhancement of CVB4E2 infection of PBMC.

Human antibodies isolated from plasma by affinity chromatography increase the coxsackievirus B4-induced synthesis of interferon-alpha by human peripheral blood mononuclear cells in vitro.

Coxsackievirus B4 (CVB4) can be found in circulating blood of patients; however, the interaction of CVB4 with peripheral blood mononuclear cells (PBMCs) is poorly understood. CVB4 induced low levels of IFN-alpha synthesis in PBMCs from healthy donors. In contrast, preincubation of infectious CVB4 with plasma from these donors containing anti-CVB4 antibodies strongly enhanced the synthesis of IFN-alpha. IgG obtained from plasma by chromatography formed immune complexes with CVB4 and increased significantly the CVB4-induced production of IFN-alpha by PBMCs. These antibodies did not have a neutralizing effect on CVB4 infection of Hep-2 cells. The role of CVB and adenovirus receptor (CAR), FcgammaRII and FcgammaRIII in the increased synthesis of IFN-alpha induced by CVB4 preincubated with IgG was shown by inhibition with specific antibodies. The major interferon-alpha-producing cells in response to CVB4-IgG complexes were CD14(+) cells and monocyte-enriched PBMCs. With the latter, detection of IFN-alpha by immunostaining was positive whereas in monocyte-depleted PBMCs it was not. This study shows that CVB4-induced synthesis of IFN-alpha by PBMCs can be enhanced by an antibody-dependent mechanism through interactions between the virus, non-neutralizing antivirus antibodies, FcgammaRII and III and CAR.