Antibodies against PbGAPDH epitope peptides bind to sporozoite surface-GAPDH and inhibit sporozoite liver infection. Antibodies against PbGAPDH epitope.

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Antibodies against PbGAPDH epitope peptides bind to sporozoite surface-GAPDH and inhibit sporozoite liver infection. Antibodies against PbGAPDH epitope peptides bind to sporozoite surface-GAPDH and inhibit sporozoite liver infection. Mice were immunized with the KLH-conjugated epitope peptides G6-1-20, G6-41-60, or with KLH only as a negative control (c.f., Fig 5). (A) Immunofluorescence assays verified that the anti-PbGAPDH epitope peptide antibodies bind to the surface of non-permeabilized P berghei sporozoites. (B) Anti-PbGAPDH epitope antibodies specifically recognize parasite GAPDH, not mGAPDH. Commercial anti-mouse GAPDH antibody (α-MsGAPDH) served as a positive control to stain mGAPDH from a HepG2 (a human hepatoma cell line) lysate, from a Raw cell (a mouse macrophage-like cell line) lysate, and recombinant PbGAPDH protein. Anti-actin antibody was used as a loading control. Red arrows indicate the band of P berghei GAPDH, actin, and mouse GAPDH, respectively. (C) Comparison of the immunogenicity of two PbGAPDH epitope peptides, G6-1-20 and G6-41-60. Immunization with G6-41-60 produced more antibody than G6-1-20 immunization. Data pooled from three independent experiments. Number of assayed mice denoted in parenthesis at the bottom of the panel. (D) Immunization with epitope peptides protects mice from infection. Immunized mice were challenged by the bites of two infected A stephensi mosquitos. Prevalence of mouse infection was determined by thin-blood smears and Giemsa stain until day 12 post-infection. Infect./Chal.: number infected over number of challenged mice. Ppd: average prepatent days. Data pooled from three independent experiments with Freund's adjuvant and two independent experiments with AddaVax adjuvant which showed similar results as Freund's adjuvant. P-values (*P < 0.05; **P < 0.01) were calculated using the one-way ANOVA test (C) or Mann–Whitney U tests (D). Error bars in (C) indicate SD. Sung-Jae Cha et al. LSA 2018;1:e201800111 © 2018 Cha et al.