Antigenicity assessment of the Theileria equi merozoite antigen (EMA-2) expressed in Pichia pastoris in mice and horses

Exp Parasitol. 2019 Nov:206:107757. doi: 10.1016/j.exppara.2019.107757. Epub 2019 Sep 5.

Abstract

Equine theileriosis is a severe equine disease caused by the protozoan Theileria equi, which is prevalent in tropical and subtropical areas. In this study, a recombinant equi merozoite antigen-2 (rEMA-2) of T. equi was used as an immunogen. Two groups of 10 mice each were divided into control and vaccinated groups. Sixty mares seronegative for theileriosis were divided in two groups, one vaccinated and another group as a control animal. Mice and mares of the vaccinated groups were inoculated with 150 μL of the vaccine containing 50 μg of rEMA-2 and 2 mL of the vaccine containing 200 μg of rEMA-2, respectively, at days 0 and 21. The immunogenicity of rEMA-2 was evaluated by ELISA and fluorescent antibody test (IFAT) using serum from vaccinated mice, mares and antigenicity in naturally infected horse. At every point throughout the ELISA study, there were significant differences between the vaccinated and control groups (p < 0.05). The vaccine induced 3- and 4-fold IgG increases in mice at the 14th and 28th day, respectively, compared to the control group. The horses' IgG dynamics showed a significant (p < 0.05) increase in the total IgG titer as early as day 7, which increased until day 28 at which time a more significant (p < 0.001) IgG titer was observed. In evaluating the isotypes, we observed a trend similar to that of total IgG, where IgG(T) (IgG3-5) were significantly (p < 0.05) more elevated than the other isotypes analyzed, followed by IgGb (IgG4-7) and IgGa (IgG1). Positive fluorescence was detected by IFAT, suggesting that the protein is immunogenic and conserves some epitopes identical to the native T. equi antigens present in the equine blood smear. Thus, our results suggest that rEMA-2 can be a promising vaccinal antigen.

Keywords: EMA-2; Equids; Theileriosis; Vaccine.

MeSH terms

  • Analysis of Variance
  • Animals
  • Antibodies, Protozoan / blood
  • Antibodies, Protozoan / immunology
  • Antigens, Protozoan / biosynthesis
  • Antigens, Protozoan / genetics
  • Antigens, Protozoan / immunology*
  • Enzyme-Linked Immunosorbent Assay
  • Female
  • Fluorescent Antibody Technique
  • Horses
  • Immunoglobulin G / blood
  • Immunoglobulin G / immunology
  • Merozoites / immunology
  • Mice
  • Mice, Inbred BALB C
  • Pichia / immunology*
  • Polymerase Chain Reaction
  • Random Allocation
  • Recombinant Proteins / immunology
  • Theileria / immunology*

Substances

  • Antibodies, Protozoan
  • Antigens, Protozoan
  • Immunoglobulin G
  • Recombinant Proteins