Induction of mucosal and systemic humoral immune responses in murine system by intranasal immunization with peptide antigens of P. vivax and CpG oligodeoxynucleotide (ODN) in microparticle delivery

In the present study we have investigated the immunomodulatory effects of two adjuvants, CpG 1826 (two copies of CpG motifs) and CpG 2006 (three copies of CpG motifs) to the five peptide antigens of Plasmodium vivax derived from circumsporozoite protein (CSP), merozoite surface protein-1 (MSP1#1, MSP1#23), apical membrane antigen-1 (AMA-1) and gametocyte surface antigen (Pvs24) in alum and microparticle formulations, using intramuscular and intranasal routes of immunization. Alum formulation without CpG ODN generated low serum IgG and IgA antibody titers and the predominant IgG isotypes were IgG1 but with the addition of CpG ODN (1826 or 2006), the antibody titers were increased by four fold with the predominance of IgG2a/2b isotypes. The SIgA peak titers in lung and intestinal washes were significantly increased with the intranasal mode of administration. Specific activity measurement was done to calculate for the accurate amounts of total serum IgG, IgA and SIgA in washes and showed direct correlation between antibody titer and its concentration. High titer anti-Pvs24 antibodies have significant inhibitory effects on parasite development in the mosquito midgut when tested in membrane feeding assays. The immunofluorescence results show that the peptide specific antisera reacted with the air-dried parasite antigens isolated from P. vivax patients. The present study demonstrates that intranasal route of immunization appears to be an alternate mode of inducing protective immunity in P. vivax malaria.