# Anti-Coxsackievirus B4 Activity of Serum and Saliva from Mice Exposed to the Virus via the Mucosal Route

## TL;DR

This study shows that mice exposed to Coxsackievirus B4 through the mouth or nose develop both blood and saliva antibodies that can neutralize the virus.

## Contribution

The study demonstrates that mucosal exposure to CVB4 induces both systemic and mucosal immune responses in mice.

## Key findings

- Mice exposed to CVB4 via the mucosal route developed serum and saliva neutralizing activity.
- Oral or intranasal inoculation induced systemic IgG and mucosal IgA responses.
- Serum from these mice showed in vitro anti-CVB4 enhancing activity.

## Abstract

Coxsackieviruses B are single-stranded RNA viruses belonging to the Enterovirus genus and are associated with various clinical outcomes, ranging from acute infections to chronic diseases, such as type 1 diabetes (T1D). It was previously shown that inoculation of Swiss albino mice with CVB4 by the intraperitoneal route induced both anti-CVB4 neutralizing and enhancing activities of serum. This study aimed to investigate the humoral immune response of mice inoculated with CVB4 by the mucosal route. Mice were inoculated orally or intranasally with CVB4, and the anti-CVB4 neutralizing activity of serum and saliva was assessed by a cell culture neutralization assay. Anti-enterovirus (EV) IgG and IgA antibodies were detected in serum and saliva, respectively, by ELISA. The serum-dependent enhancement of CVB4 infection in cultures of murine splenocytes was evaluated by detecting intracellular viral RNA using RT-qPCR. At day 45 post-inoculation, an anti-CVB4 neutralizing activity, the extent of which depends on the amount of inoculated infectious particles, was detected in the serum of mice exposed orally or intranasally. An increase in anti-CVB4 neutralizing activity was observed in the saliva of mice inoculated orally or intranasally during the follow-up. Oral or intranasal inoculation of CVB4 induced a systemic IgG and mucosal IgA response. In addition, serum from these mice harbored an anti-CVB4 enhancing activity in vitro. These data indicate that Swiss albino mice exposed to CVB4 via the mucosal route constitute a potentially useful model for testing strategies to promote the production of protective mucosal and systemic anti-CVB4 antibodies and for verifying whether or not enhanced antibodies are produced.