Intranasal immunization with liposome-encapsulated plasmid DNA encoding influenza virus hemagglutinin elicits mucosal, cellular and humoral immune responses

Abstract

Influenza viral infections are a significant global public health concern due to the morbidity and mortality associated with acute respiratory disease, associated secondary complications and pandemic threat. Currently, the most effective preventative measure is an annual intramuscular (i.m.) injection of a trivalent vaccine. Intramuscular immunization induces strong systemic humoral responses but not mucosal immune responses which are important in the first line of defense against influenza. A plasmid encoding influenza A/PR/8/34 (H1N1) hemagglutinin (HA; pCI-HA10) was evaluated with respect to the mucosal, cellular and humoral immune responses generated and to its efficacy in protection against a challenge with a lethal dose of influenza. BALB/c mice were immunized with pCI-HA10 DNA or liposome-encapsulated pCI-HA10 by either an intranasal (i.n.) or i.m. route. Sera and bronchoalveolar lavage (BAL) fluid were collected at various times and evaluated for HA-specific IgG and IgA antibodies and T cells, isolated from draining lymph nodes and spleens, were analyzed for their proliferative ability. Immunized mice were challenged with a lethal dose (5LD(50)) of influenza and monitored for survival. Intranasal immunization with liposome-encapsulated pCI-HA10 stimulated both IgG and IgA humoral responses and increased IgA titers in BAL fluid, whereas immunization with naked pCI-HA10 had no effect on the antibody response. Intramuscular immunization with both naked and liposome-encapsulated pCI-HA10 elevated serum IgG levels, but had no effect on IgA levels in either the serum or BAL fluid. Both i.n. and i.m. administration of HA vaccine (naked and liposome-encapsulated) elicited T cell proliferative responses. These results suggest that i.n. administration of liposome-encapsulated HA-encoding DNA is effective at eliciting mucosal, cellular and humoral immune responses. Mice immunized i.n. were able to withstand a lethal challenge of influenza virus, confirming that the immune responses mediated by the vaccine were protective.