Rotavirus-specific intestinal immune response in mice assessed by enzyme-linked immunospot assay and intestinal fragment culture

Abstract

Primate rotavirus strain RRV and bovine strain WC3 or reassortants made between these animal viruses and human rotaviruses have been administered to infants as candidate vaccines. We compared RRV and WC3 in a murine model of oral infection. We determined the relative capacities of these viruses to induce a virus-specific humoral immune response by intestinal lymphocytes as tested by enzyme-linked immunospot assay, intestinal fragment culture, and enzyme-linked immunosorbent assay of intestinal contents. We found that inoculation of mice with RRV induced higher frequencies of virus-specific immunoglobulin A (IgA)-secreting cells in the lamina propria, greater quantities of virus-specific IgA in intestinal fragment cultures, and greater quantities of virus-specific IgA in intestinal secretions than did inoculation with WC3 or inactivated RRV (iRRV). The induction of an IgA response in serum was predictive of an IgA response among intestinal lymphocytes after inoculation with RRV but not WC3. In addition, large quantities of IgG, IgA, and IgM not specific for rotavirus were produced in fragment cultures from mice inoculated with RRV but not in cultures from mice inoculated with WC3 or iRRV. Possible mechanisms of RRV-induced polyclonal stimulation of intestinal B cells are discussed.

Systematic and intestinal antibody-secreting cell responses and correlates of protective immunity to human rotavirus in a gnotobiotic pig model of disease

“…Several investigations have shown that after rotavirus or coronavirus enteric infections, virus-specific IgA ASC prevailed over IgG ASC in the intestinal lamina propria (10,25,32,47). In mice inoculated with simian rotavirus, the magnitude of the intestinal IgA ASC response was about 10 times greater than the IgG ASC response (32).…”

Specific mucosal IgA immunity in turkey poults infected with turkey coronavirus

“…The culture system for the measurement of intestinal antibody was modi®ed from previously published methods for mice (Khoury et al, 1994) and human (Losonsky et al, 1999). A 0.5 cm long segment of duodenum was collected from the loop of duodenum adjacent to pancreas.…”

“…These results revealed that local intestinal mucosal IgA antibodies was stimulated and reached the plateau in a similar fashion in different sites of intestine after infection with TCV. It was demonstrated that induction of virus-speci®c IgA antibody responses in local mucosal tissue was related to replication of porcine coronavirus (VanCott et al, 1993) and rotavirus (Khoury et al, 1994). The virus-speci®c IgA antibody response seemed to decline faster in jejunum than in duodenum or ileum when the viral antigen is no longer detectable at 6 and 9 weeks PI.…”

“…Intestinal segment culture has been successfully used for the measurement of mucosal antibody production in mouse (Khoury et al, 1994) and human (Losonsky et al, 1999). Since the whole tissue sample is used, the mucosal microenvironment remains intact.…”

“…The antibody responses detected by intestinal segment cultures were positively correlated with the numbers of antibody secreting cells enumerated by enzymelinked immunospot assay (ELISPOT) (Losonsky et al, 1999). The intestinal segment culture had been used to evaluate candidate vaccines consisting of rotavirus primate strain RRV, bovine strain WC3, or reassortant viruses between these animal viruses and human rotaviruses (Khoury et al, 1994).…”