Evolution of the Immune Response against Recombinant Proteins (TcpA, TcpB, and FlaA) as a Candidate Subunit Cholera Vaccine

J Immunol Res. 2017:2017:2412747. doi: 10.1155/2017/2412747. Epub 2017 Jan 16.

Abstract

Vibrio cholerae is the causative agent of cholera and annually leads to death of thousands of people around the globe. Two factors in the pathogenesis of this bacterium are its pili and flagella. The main subunits of pili TcpA, TcpB, and FlaA are the constituent subunit of flagella. In this study, we studied the ability of pili and flagella subunits to stimulate immune responses in mice. After amplification of TcpA, TcpB, and FlaA genes using PCR, they were cloned in expression plasmids. After production of the above-mentioned proteins by using IPTG, the proteins were purified and then approved using immunoblot method. After injection of the purified proteins to a mice model, immune response stimulation was evaluated by measuring the levels of IgG1 and IgG2a antibody titers, IL5 and IFN-γ. Immune response stimulation against pili and flagella antigens was adequate. Given the high levels of IL5 titer and IgG1 antibody, the stimulated immune response was toward Th1. Humoral immune response stimulation is of key importance in prevention of cholera. Our immunological analysis shows the appropriate immune response in mice model after vaccination with recombinant proteins. The high level of IL5 and low level of IFN-γ show the activation of Th2 cell response.

MeSH terms

  • Animals
  • Antibodies, Bacterial / blood*
  • Antibodies, Bacterial / immunology
  • Cholera / immunology
  • Cholera Vaccines / chemistry*
  • Cholera Vaccines / immunology*
  • Disease Models, Animal
  • Fimbriae Proteins / immunology*
  • Flagella / immunology
  • Flagellin / immunology*
  • Humans
  • Immunity, Humoral
  • Immunoglobulin G / blood
  • Interferon-gamma / blood
  • Interleukin-5 / blood
  • Mice
  • Recombinant Proteins / immunology
  • Spleen / cytology
  • Th1 Cells
  • Th2 Cells / immunology
  • Vaccination
  • Vaccines, Subunit / chemistry
  • Vaccines, Subunit / immunology

Substances

  • Antibodies, Bacterial
  • Cholera Vaccines
  • Immunoglobulin G
  • Interleukin-5
  • Recombinant Proteins
  • TcpA protein, Vibrio cholerae
  • TcpB protein, Vibrio cholerae
  • Vaccines, Subunit
  • Flagellin
  • flaA protein, bacteria
  • Fimbriae Proteins
  • Interferon-gamma