DBL2β-PfEMP1 Recombinant Protein Dose Effect on Inducing Humoral and Cellular Immune Responses in Wistar Rats: A Study on Malaria Vaccine Development
Abstract
The DBL2β-PfEMP1 is a promising malaria vaccine candidate because of its essential role in malaria pathogenesis. This study aimed to determine the effective dose of DBL2β-PfEMP1 recombinant protein in inducing humoral and cellular immune responses by measuring immunoglobulin M (IgM), immunoglobulin G (IgG), and a cluster of differentiation 4 (CD4+) concentration in Wistar rats. The DBL2β-PfEMP1 recombinant protein was produced in Escherichia coli BL21(DE3) and purified by affinity chromatography using Ni-NTA resin. Wistar rats (Rattus norvegicus) weighing 150–200 g were randomly divided into control and three treatment groups. A control group was subcutaneously injected with 0.9% NaCl, and treatment groups were injected with 100, 150, and 200 µg of purified DBL2β-PfEMP1 recombinant protein three times at 3-week intervals. Sera were collected two weeks after each injection for immune response measurement using enzyme-linked immunosorbent assay (ELISA). Data were statistically analyzed using mixed-method ANOVA followed by a post hoc Bonferroni test. IgM and CD4+ concentrations consistently increased in all treatment groups after consecutive injections. However, IgG showed a slightly different result. The highest levels of IgM, IgG, and CD4+ were observed at a dose of 200 µg after the third injection. Statistical analysis confirmed the difference between the control and treatment groups with p=0.000, 0.034, and 0.006 for IgM, IgG, and CD4+. These findings imply that in the dose range of the study, a dose of 200 µg DBL2β-PfEMP1 recombinant protein resulted in the highest response to induce humoral and cellular immunity and could be an effective dose for malaria vaccine.
Keywords: immune response, malaria, PfEMP1, vaccine.
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