Journal: Frontiers in Immunology
Article Title: Process development and preclinical evaluation of a major Plasmodium falciparum blood stage vaccine candidate, Cysteine-Rich Protective Antigen (CyRPA)
doi: 10.3389/fimmu.2022.1005332
Figure Lengend Snippet: Humoral and Cellular Immune Responses against Tag-free CyRPA Vaccine Formulated with Commonly used Adjuvants. (A) BALB/c mice (n = 6 per group) were immunized intramuscularly with tag-free CyRPA formulated with three adjuvants- Alhydrogel, AddaVax and Montanide ISA 720 and Freund’s adjuvant (CFA/IFA) as a control. Enzyme-Linked Immunosorbent Assay (ELISA) was performed to calculate antibody titers in Day 70 sera. CyRPA (AddaVax) and CyRPA (Freund’s Adjuvant) induced high antibody titers (~32,00,000 and ~25,60,000 respectively) followed by CyRPA (Montanide ISA 720) (~6,40,000) and CyRPA (Alhydrogel) (~4,20,000). ELISA cut-offs were calculated as the mean OD 490 of pre-bleed sera plus three times standard deviation. The bars and error bars show mean and standard error values of each vaccine group respectively. Individual mice titers are also shown. The P values were calculated by two-way ANOVA with Bonferroni post-hoc testing. (B) Levels of T H 1 and T H 2 responses were measured in BALB/c mice serum immunized with CyRPA+adjuvant or adjuvant alone by cytometric bead arrays (CBA) for mouse serum cytokines using flow-cytometry. CyRPA/Alhydrogel vaccine induced IL-4, IL-6, TNF-α when compared to the Alhydrogel only group while no detectable responses were observed for CyRPA/AddaVax and CyRPA/Montanide ISA 720 groups. However the control group- CyRPA/Freund’s Adjuvant observed increased levels of IFN-γ and IL-4. The pre-bleed, adjuvant and CyRPA+adjuvant plots refer to the cytokines detected in sera from groups of 6 mice before immunization, post-immunization of adjuvant and post-immunization of CyRPA formulation respectively. The bars and error bars show mean and standard error values of each group respectively. The P values were calculated by two-way ANOVA with Bonferroni post-hoc testing. * P≤0.05 ** P≤0.01, *** P≤0.001, **** P≤0.0001.
Article Snippet: AddaVax (MF-59 like squalene adjuvant) (InvivoGen), Alhydrogel (2%) (Brenntag Biosector), Montanide ISA 720 (Seppic) and complete and incomplete Freund’s adjuvants (CFA/IFA) (Sigma) was used in the mice and rabbit experiments.
Techniques: Enzyme-linked Immunosorbent Assay, Standard Deviation, Flow Cytometry
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