Hossein Forghani; Mahin Jamshidi Makiani; Hossein Zarei Jaliani; Seyed Mohsen Zahraei; Seyedeh Mahdieh Namayandeh; Parisa Khani
Abstract
Background: Currently evidence indicates the resurgence of whooping cough despite high coverage of whole-cell (wP) and acellular (aP) pertussis vaccines. Objective: In this study, we investigated the cell-mediated immune response of a genetically inactivated protein containing the S1 subunit of pertussis ...
Read More
Background: Currently evidence indicates the resurgence of whooping cough despite high coverage of whole-cell (wP) and acellular (aP) pertussis vaccines. Objective: In this study, we investigated the cell-mediated immune response of a genetically inactivated protein containing the S1 subunit of pertussis toxin (PTS1) without and with the Listeriolysin O (LLO-PTS1), developed by the researchers (the authors of this study), in comparison with current wP and aP vaccines in the mice model. Methods: Thirty-six female NMRI mice aged 8 to 12 weeks (25 ± 5 g) were divided into six groups including control (n=6), and five treated groups (n=6/each). Treated groups comprising recombinant PTS1, recombinant fusion LLO-PTS1, aP, wP, and sham (phosphate-buffered saline) were injected intraperitoneally whereas the control group did not receive anything. After 60 days, the serum levels of IFN-γ, IL-4, and IL-17 cytokines (as the T-helper 1, 2, and 17 responses, respectively) were evaluated by mouse ELISA Kit. Results: Our findings showed LLO-PTS1 significantly increased IL-17 and IL-4 cytokines compared with wP and aP vaccines (superiority). IFN-γ failed to significantly increase in the LLO-PTS1 group compared to others but it was non-inferior to standard vaccines (non-inferiority). Conclusion: Our alum free mono-component monovalent recombinant fusion protein (LLO-PTS1), registered as a patent in the www.iripo.ssaa.ir, could bear the capacity to stimulate the Th-1 response similar to wP and aP vaccines (non-inferiority) in the mice model. In addition, it showed better results in Th-17 and Th-2 response (superiority). This study can be regarded as a springboard for further probes in booster pertussis vaccine development.
Iraj Pakzad; Abbas Rezaee; Mohammad Javad Rasaee; Bahman Tabbaraee; Ali Delpisheh
Volume 6, Issue 1 , March 2009, , Pages 12-21
Abstract
Background: The immunogenic Brucella abortus ribosomal protein L7/L12 is a promising candidate antigen for the development of subunit vaccines against brucellosis. Objective: This study was aimed to evaluate the protection of recombinant Human Serum Albumin (HAS)-L7/L12 fusion protein in Balb/c mice. ...
Read More
Background: The immunogenic Brucella abortus ribosomal protein L7/L12 is a promising candidate antigen for the development of subunit vaccines against brucellosis. Objective: This study was aimed to evaluate the protection of recombinant Human Serum Albumin (HAS)-L7/L12 fusion protein in Balb/c mice. Methods: The amplified L7/L12 gene was cloned in pYHSA5 vector, pYHSA5-L7/L12 construct was transformed in Saccharomyces cerevisiae and the expressed protein from supernatant was purified by affinity chromatography. Balb/c mice were immunized in five groups by tHSA-L7/L12 fusion protein (group 1), Brucella abortus S19 (group 2), HSA (group 3), recombinant L7/L12 (group 4), PBS (group 5). ELISA to detect antibody production, LTT test to assess antigen specific lymphocyte response were conducted prior to virulent B. abortus strain 544 challenge two weeks after the last injection. Bacterial counts from spleens of immunized mice were done four weeks after challenge. Results: In ELISA tests, the specific antibodies exhibited a dominance of immunoglobulin IgG1 over IgG2a. In addition, the tHSA-L7/L12 fusion protein and L7/L12 elicited a strong T-cell proliferative response upon restimulation in vitro with recombinant tHSA-L7/L12 and L7/L12, suggesting the induction of a cellular immunity response in vivo. However, there was no significant difference in proliferative response of L7/L12 and tHSA-L7/L12 fusion protein (p>0.05). The L7/L12 and tHSA-L7/L12 fusion protein vaccines could also induce significant protection against challenge with the virulent strain B. abortus 544 in Balb/c mice (p≤0.05). Conclusion: The tHSA-L7/L12 fusion protein, similar to L7/L12 has the ability to induce antigen specific lymphocyte proliferation, stimulate humoral immunity and engender protection.