Dynamic study of HBsAg and HBeAg in saliva samples from patients with hepatitis B infection: Diagnostic and epidemiological significance

Abstract

Analysis of 505 cases history of patients among men with viral hepatitis demonstrates that HBV infected patients represent 68.9% of the total and that a non-parenteral rate of transmission is the most likely means of hepatitis B infection. Saliva and serum testing for the presence of specific HBV markers (HBsAg, HBeAg and HBV DNA) at different phases of the infection process were carried out to review the diagnostic and epidemiological value of saliva samples from patients with acute viral hepatitis B. The frequency of HBsAg detection by Enzyme Immune Assay (EIA) in saliva of patients in acute period was found to correlate with the frequency of its detection in serum. In early convalescence the frequency of detection of that antigen in serum (59.5% of patients) was significantly higher than in saliva (23.8%) (P < 0.001). The frequencies of HBeAg detection by EIA in saliva samples was significantly higher than that in serum samples in both acute phase (84.3% and 28.1% of patients, respectively) and in early convalescence (56.2% and 3.1% of patients, respectively). The study of frequencies of detection of these antigens in the dynamics of the disease up to the total recovery of patients (observations were carried out for the period of 60 days and longer) showed that in most patients there was a faster disappearance HBsAg from saliva than from serum. By the end of second month this antigen was detected in saliva of only 8.3% of patients whereas in serum in the same period HBsAg was detected in 33.3% of patients. HBeAg became undetectable in blood whereas HBs-antigenemia was still pronounced, and a month after the beginning of the disease it was not found in serum specimens. In saliva, HBeAg was detected in 95.8% of patients observed directly after admission. A month after the beginning of the disease it was detected in saliva of 66.7% of patients and, by the end of observation period, in 12.5% of patients recovered from viral hepatitis. HBV DNA revealed by PCR in saliva and serum of HBV-infected patients was detected in acute period not only in serum (84.6% of cases) but also in saliva (46.2% of cases). The data illustrate the diagnostic value of saliva and point to the possible role of saliva as a source of HBV infection.

Development of a nasal vaccine for chronic hepatitis B infection that uses the ability of hepatitis B core antigen to stimulate a strong Th1 response against hepatitis B surface antigen

“…The ELISPOT assay using the P815 mastocytome cell line pulsed with the CD8 + -restricted HBs (28)(29)(30)(31)(32)(33)(34)(35)(36)(37)(38)(39) peptide indicated that nasal immunization with the combined HBs and HBc formulation activates antigen specific CD8 + cells, consistent with a strong Th1 response. This result is in line with the finding that B cells that bind viral capsids can prime cytotoxic T cells.…”

“…38 Recent reports from Bertoletti et al reveal an important degree of T-cell functional recovery after antiviral treatment with Lamivudine. 39,40 This suggests that it is possible to rescue T cells from non-responsive states. Other reports demonstrate that in patients naturally recovering from HBV chronic infection, there is a rebound in cellular immunity that correlates with viral clearance.…”

“…The cells were extensively washed with medium, resuspended in RPMI-1640 (Gibco, Scotland, UK) supplemented with 10% FCS, 2 mmol/L glutamine, 2 mmol/L piruvate, 50 mmol/L 2-mercaptoethanol and antibiotics (complete medium) and counted. Meanwhile, H-2 d mastocytome cells p815 were pulsed for 1 h at 37 ° C, 5% CO 2 in complete medium with 10 mmol/L S (28)(29)(30)(31)(32)(33)(34)(35)(36)(37)(38)(39) peptide IPQSLDSWWTSL from HBsAg. 33 After incubation p815 cells were further incubated for another 15 min with Mitomycin C (Sigma).…”

“…After washing, the cells were counted and distributed at 2 × 10 6 cells/mL in 10 mL RPMI FCS in 25 cm 2 flasks (Nalge-Nunc, Rochester, NY, USA) and stimulated with 10 mg/mL of peptide S (28)(29)(30)(31)(32)(33)(34)(35)(36)(37)(38)(39) . After culturing for 4 days in 5% CO 2 , half of the total medium was substituted and new medium containing 2 × 10 4 U/mL of IL-2 was added.…”

“…immunization with HBsAg in alum. The IFN-γ ELISPOT assay was conducted using the immunodominant HBs (28)(29)(30)(31)(32)(33)(34)(35)(36)(37)(38)(39) CTL peptide from HBsAg as previously described. Measurement of lymphoproliferative activity revealed the induction of strong proliferative responses in systemic compartments against both HBsAg and HBcAg after nasal-combined immunization.…”