Effect of route of inoculation on innate and adaptive immune responses to porcine epidemic diarrhea virus infection in suckling pigs

The effects of route of administration on systemic and gut mucosal immune responses induced by porcine epidemic diarrhea virus (PEDV) infection in suckling pigs were investigated. Twenty-four conventional 5-day-old suckling piglets were randomly divided into four groups and were inoculated orally, intranasally (I.N.), intramuscularly (I.M.) with PEDV or DMEM (mock). Pigs were monitored daily for clinical signs and fecal viral load. Blood samples were collected at 7, 14, 21 days post infection (dpi) and subjected for the analyses of serum antibody production, T cell and natural killer (NK) cell frequencies, NK cytotoxicity and serum cytokine levels. Oral inoculation led to higher levels of PEDV-specific IgA antibodies in both serum and gut mucosal sites than did other routes of inoculation. Intranasal inoculation elicited significantly higher titers of virus-specific IgG antibodies in serum. PEDV-infected pigs regardless of inoculation routes had significantly lower NK cell frequencies than those of the control pigs at 14 dpi. The orally inoculated pigs had significantly higher CD3+CD8+ T cell frequencies as compared to I.N. or I.M. inoculated pigs at 14 dpi, while there was no significant difference among orally, I.N. or I.M. inoculated pigs and control pigs in CD3+CD4+ T cell frequencies in peripheral blood. PEDV-infected and control pigs had low, but detectable NK cell activities at 14 and 21 dpi, however, NK cell activities were barely detectable at 7 dpi whether the pigs were infected or not. Serum IL-10 levels were induced drastically in orally infected pigs at 7 dpi and then gradually declined. Serum IL-12 levels followed a similar pattern while the fold-change was much lower. In conclusion, oral inoculation may generate more comprehensive immune responses.