Author/Authors :
Malaei, Fatemeh Biology Research Center - Faculty of Science - Imam Hussein University , Saadati, Mojtaba Biology Research Center - Faculty of Science - Imam Hussein University , Honari, Hussein Biology Research Center - Faculty of Science - Imam Hussein University , Nazarian, Shahram Biology Research Center - Faculty of Science - Imam Hussein University , Hesaraki, Mahdi Department of Stem Cell and Developmental Biology - Cell Science Research Center - Royan Institute for Stem Cell Biology and Technology - ACECR - Tehran , Ahdi, Ali Mohammad Department of Genetics - Faculty of Science - Shahrekord University - Shahrekord , Sadraeian, Mohammad Pharmaceutical Sciences Research Center - Faculty of Pharmacy - Shiraz University of Medical Sciences - Shiraz
Abstract :
Background: Recombinant vaccine technology is one of the most developed means in
controlling infectious diseases. However, an effective vaccine against Shigella is still
missing. Objective: To evaluate recombinant IpaC protein of Shigella as a vaccine
candidate. Methods: In this study we cloned IpaC gene into an expression vector in
prokaryotic system. The protein expression was evaluated by SDS-PAGE and Western-
Blotting analysis. The recombinant protein was purified using Ni–NTA affinity
chromatography. Guinea pigs were immunized with the recombinant protein and the
level of immunogenicity was examined by ELISA and Western blotting of IpaC.
Challenge test was done through the intraoculary injection of Shigella dysenteriae
(6×108 CFU/eye) and after 48 hours was scored for keratoconjunctivitis. Results: The
results showed a remarkable level of immunogenicity in terms of antibody response and
protection against keratoconjunctivitis in tested animals. The recombinant IpaC protein
provided a protective system against Shigella dysenteriae type I during the challenge
test. Conclusion: The results showed the potential of using recombinant IpaC in
preparation of vaccine in perspective studies.
Keywords :
Expression , Shigella dysenteriae , Recombinant Vaccine , IpaC Gene