Re-evaluation of anti-HBc non-reactive serum samples from patients with persistent hepatitis B infection by immune precipitation with labelled HBV core antigen

Abstract

Background: Core antigen (HBcAg) is the most immunogenic component of hepatitis B virus (HBV) and is believed to induce virtually always antibodies (anti-HBc) in immunocompetent infected persons. However, some chronically infected persons do not develop detectable anti-HBc.

Objective: A more sensitive assay for anti-HBc was to be developed and used to re-evaluate a cohort of chronically HBV infected persons without detectable anti-HBc.

Study design: Among 3309 serum samples which had been tested by commercially available (microparticle) enzyme immune assay (M/EIA) 34 samples from 22 patients were identified having reacted positive for HBsAg and negative for anti-HBc. Nine of these patients had immunosuppression or HIV coinfection, 13 patients were immunocompetent, 5 of them were perinatally infected. Anti-HBc was re-tested for in an immune precipitation (IP) assay using (32)P-labelled recombinant HBcAg as reagent and anti-human-IgG-coated magnetic beads as separation system for immunecomplexes containing HBcAg. Specificity was controlled for by competition with unlabelled HBcAg.

Results: 27 serum samples from the 22 patients could be retested. IP was positive in 7 MEIA negative sera, unspecific positive in 4 and negative in 16. Using 5 anti-HBe positive control sera, we found IP to be 1.8-fold (1.3-2.9) more sensitive than MEIA, but IP was 6.5-fold (5.8-7.4) more sensitive with 4 anti-HBe negative, anti-HBc positive sera.

Conclusion: IP allowed specific detection of anti-HBc in about 25% of MEIA negative chronic HBV patients. The majority of these seem to produce no or very little anti-HBc, however.