Regulation of the neutralizing anti-hepatitis B surface (HBs) antibody response in vitro in HBs vaccine recipients and patients with acute or chronic hepatitis B virus (HBV) infection

Abstract

SUMMARYAntibodies directed to the HBs antigen indicate viral clearance and the development of life-long immunity in patients that recovered from HBV infection. In HBs antigen vaccine recipients anti-HBs antibodies provide protective immunity. However, little is known about the regulation of this HBsspecific antibody response. The existence of anti-HBs-secreting B cells was demonstrated using the highly sensitive ELISPOT technique compared with conventional ELISA in serum and cell culture supernatants. In the peripheral blood of patients with acute self-limited hepatitis B, HBs-specific B cells were demonstrated with a high frequency despite undetectable anti-HBs serum antibodies. HBVimmunized patients that had recovered from infection and vaccine recipients had significantly lower frequencies, whereas chronic HBV carriers and negative controls showed no anti-HBs-secreting B cells. Coculture experiments of isolated B and T cells revealed that the anti-HBs antibody response was restricted to the presence of T helper cells, but not to identical HLA class II molecules. Allogeneic T cells derived from vaccine recipients or chronic HBV carriers stimulated the HBs-specific B cell response in HBs vaccine recipients. Otherwise, isolated T helper cells could never provide sufficient help to induce the HBs-specific B cell response in chronic HBV carriers. Furthermore, peripheral blood mononuclear cells (PBMC) of six out of 10 vaccine recipients, one out of five HBV-immunized patients, but of no chronic HBV carrier showed a proliferative response to different HBs antigen preparations. This study demonstrated a high frequency of circulating anti-HBs-producing B cells in the early phase of acute HBV infection, but a lower frequency of HBs-specific B cells years after resolution of HBV infection. In chronic HBV carriers, however, deficient HBs-specific T and B cell responses were observed.

Variants of two major T cell epitopes within the hepatitis B surface antigen are not recognized by specific T helper cells of vaccinated individuals

“…Cells from 12 donors recognized 2 epitopes ( Table 2). The most vigorous T-cell responses were elicited by peptide P4 (mean SI of positive cells 27.2, range 7-55) followed by peptide P1 (mean SI of positive cells 11.6, range [7][8][9][10][11][12][13][14][15][16][17]. Experiments with PBMC subpopulations depleted of CD4 ϩ or CD8 ϩ cells showed that all T-cell responses were mediated by CD4 ϩ T-helper cells (data not shown).…”

Development of antibody to hepatitis B surface antigen after liver transplantation for chronic hepatitis B

“…A defective neutralizing anti-HBs response may be related to a HBsAg-specific T-helper cell and B cell defect. 15 This immune response is expected to be worse in liver transplant recipients because of the immunosuppressive medications. Therefore, the spontaneous development of anti-HBs is most unexpected and has not been reported previously in other series of patients receiving lamivudine prophylaxis 5,7 or in whom HBIg prophylaxis was discontinued after long-term administration.…”

Cellular and humoral immune responses induced by intradermal or intramuscular vaccination with the major hepatitis B surface antigen

“…The frequencies of anti-HBs secreting B cells within peripheral blood mononuclear cells (PBMC) were determined using an anti-HBs-specific ELISPOT technique as described previously. 18 In brief, 96-well plates with high-protein binding membranes (MAIP S4510; Millipore, Eschborn, Germany) were coated overnight with recombinant HBsAg (5 mg/well) or tetanus toxoid (Chiron Behring, 2 mg/well) as irrelevant control antigen. After washing with sterile distilled water, the plates were preincubated with RPMI 1640 ϩ 3% bovine serum albumin (BSA) for 1 hour before cells were added in triplicates in 2 different concentrations (2.5 ϫ 10 5 , 1.25 ϫ 10 5 per well) and incubated overnight in a humidified atmosphere at 37°C and 5% CO 2 .…”