This attenuated strain could also be used for developing the recombinant vaccine against other enteric pathogens. Acknowledgements This work was supported by the grant from Department of Biotechnology, Govt. of India (Project No. BT/PR14489/Med/29/207/2010). We thank Himanshu Singh Chandel for his support during the experiments. Electronic supplementary material Additional file 1: Figure S1: Evaluation of attenuation profile of mig14::aphT mutant in comparison to wild-type strain of Salmonella Typhimurium. Competitive index profile of mig-14::aphT mutant when compared against selleckchem wild-type strain.
n.s. = not significant; * = p < 0.05). Figure S2. Infection profile of mig14::aphT mutant in comparison to wild-type strain of Salmonella Typhimurium .Infection profile and systemic attenuation of mig14::aphT mutant. Bar indicates 200 μm. n.s. = not significant; * = p < 0.05). Figure S3. Flowcytometric analysis of T-cell population after Salmonella infection.
The whole cells were isolated from the mLN of the vaccinated mice. The cells were then suspended in appropriate www.selleckchem.com/products/ink128.html medium and processed for flow cytometric analysis (see materials and methods). The cells were detected by using specific conjugated antibodies against specific T-cells. Figure S4. Luminal and serum specific antibody responses in mice immunized with MT5 and MT4. Serum and gut wash from mice treated with PBS and vaccinated with MT4 and MT5 were collected, diluted to a highest dilution of 1:120 (serum) and 1:9 (gut wash). The presence of Salmonella specific IgG and secretory IgA were detected by bacterial flow cytometric (A) and Western blot (B). Each coloured line indicates data obtained from individual mice of respective group. The representative Western blot analysis of the antibody responses was done by developing the blots from the overnight cultures of MT5, MT4, SB300 (wt S. Typhimurium) and M1525 (S. Enteritidis; negative control) by using the sera and gut luminal sIgA of the
immunized mice. (PDF 434 KB) References 1. Okamura M, Lillehoj HS, Raybourne RB, Babu US, Heckert click here RA: Cell-mediated immune responses to a killed Salmonella enteritidis vaccine: lymphocyte proliferation, T-cell changes and interleukin-6 (IL-6), IL-1, IL-2, and IFN-gamma production. Comp Immunol Microbiol Infect Dis 2004,27(4):255–272.PubMedCrossRef 2. Thatte J, Rath S, Bal V: Analysis of immunization route-related variation in the immune response to heat-killed Salmonella typhimurium in mice. Infect Immun 1995,63(1):99–103.PubMed 3. Penha Filho RA, Moura BS, de Almeida AM, Montassier HJ, Barrow PA, Berchieri Junior A: Humoral and cellular immune response generated by different vaccine programs before and after Salmonella Enteritidis challenge in chickens. Vaccine 2012,30(52):7637–7643.PubMedCrossRef 4.