Design and Analysis of Recombinant Vaccine Against Clostridium Perfringens Type A and Type E

Öz

Objective: Clostridium perfringens is a spore-forming, anaerobic, gram positive, bacteria that exist in environment, soils, foods, and in intestinal traces of humans or animals. However, vaccines being developed for use in animals have the potential to be developed for use in humans. This study was aimed to design and analysis of multi epitope vaccine against Clostridium perfringens type A and E. Material-Method: The B cell epitopes were predicted by IEDB and MHC II epitopes were predicted by Vaxign web server. Results: Physicochemical study of vaccine showed that the designed vaccine is 58.33 kDa. The half-life of candidate vaccine was found to be greater than 100 hours in mammalian cells, greater than 20 hours in yeast, and greater than 10 hours in Escherichia coli. The instability index of vaccine was 28.41 (<40), the aliphatic index was found to be 47.51, and the vaccine is soluble in water and is considered stable. The grand average of hydropathicity of candidate vaccine is -1.283, consequently, the vaccine is a hydrophilic protein and easily soluble in water. The result obtained from of Toxinpred and Allertop revealed that the protein don’t have tixic and allergenic effect on human and animal cells. Pepcalc and Protparam analysis showed that the vaccine doesn’t have transmembrane helix in its structure, so no expression difficulties are expected in the development of the protein from recombinant dNa technology methods. The result obtained from docking analysis proved that the vaccine has maximum affinity to HLA-DRB1*0101 with the score of -660.73. Conclusion: The result of this study showed that the candidate vaccine can be stimulate HLA-DRB1*0101 and other MCHII alleles.

Anahtar Kelimeler

• Vaccine,Clostridium perfringens

In silico design and analysis of recombinant vaccine against Clostridium perfringens Type A and Type E.

Öz

Objective: Clostridium perfringens is a spore-forming, anaerobic, gram positive, bacteria that exist in environment, soils, foods, and in intestinal traces of humans or animals. However, vaccines being developed for use in animals have the potential to be developed for use in humans. This study was aimed to design and analysis of multi epitope vaccine against Clostridium perfringens type A and E. Material-Method: The B cell epitopes were predicted by IEDB and MHC II epitopes were predicted by Vaxign web server. Results: Physicochemical study of vaccine showed that the designed vaccine is 58.33 kDa. The half-life of candidate vaccine was found to be greater than 100 hours in mammalian cells, greater than 20 hours in yeast, and greater than 10 hours in Escherichia coli. The instability index of vaccine was 28.41 (<40), the aliphatic index was found to be 47.51, and the vaccine is soluble in water and is considered stable. The grand average of hydropathicity of candidate vaccine is -1.283, consequently, the vaccine is a hydrophilic protein and easily soluble in water. The result obtained from of Toxinpred and Allertop revealed that the protein don’t have tixic and allergenic effect on human and animal cells. Pepcalc and Protparam analysis showed that the vaccine doesn’t have transmembrane helix in its structure, so no expression difficulties are expected in the development of the protein from recombinant dNa technology methods. The result obtained from docking analysis proved that the vaccine has maximum affinity to HLA-DRB1*0101 with the score of -660.73. Conclusion: The result of this study showed that the candidate vaccine can be stimulate HLA-DRB1*0101 and other MCHII alleles.

Anahtar Kelimeler

• B cell,Clostridium perfringens,HLA-DRB1*0101,Type E,Type A,Vaccine

Kaynakça

1. 1. Awad, M. M., Bryant, A. E., Stevens, D. L. and Rood, J. I. (1995) Virulence studies on chromosomal α‐toxin and Θ‐toxin mutants constructed by allelic exchange provide genetic evidence for the essential role of α‐toxin in Clostridium perfringens‐mediated gas gangrene. Molecular microbiology, 15, 191-202.
2. 2. Bryant, A. E., Chen, R. Y., Nagata, Y., Wang, Y., Lee, C., Finegold, S., Guth, P. H. and Stevens, D. L. (2000) Clostridial Gas Gangrene. I. Cellular and Molecular Mechanisms of Microvascular Dysfunction Induced by Exotoxins of Clostvidium pevfvingens. The Journal of infectious diseases, 182, 799-807.
3. 3. Dar, H. A., Zaheer, T., Shehroz, M., Ullah, N., Naz, K., Muhammad, S. A., Zhang, T. and Ali, A. (2019) Immunoinformatics-Aided Design and Evaluation of a Potential Multi-Epitope Vaccine against Klebsiella Pneumoniae. Vaccines, 7, 88.
4. 4. Dikhit, M. R., Kumar, A., Das, S., Dehury, B., Rout, A. K., Jamal, F., Sahoo, G. C., Topno, R. K., Pandey, K. and Das, V. (2017) Identification of potential MHC Class-II-restricted epitopes derived from Leishmania donovani antigens by reverse vaccinology and evaluation of their CD4+ T-cell responsiveness against visceral leishmaniasis. Frontiers in immunology, 8, 1763.
5. 5. Dimitrov, I., Bangov, I., Flower, D. R. and Doytchinova, I. (2014) AllerTOP v. 2—a server for in silico prediction of allergens. Journal of molecular modeling, 20, 2278.
6. 6. Doyle, M. P. and Buchanan, R. L. (2012) Food microbiology: fundamentals and frontiers. ed. American Society for Microbiology Press.
7. 7. Fernández Miyakawa, M. E. and Uzal, F. A. (2005) Morphologic and physiologic changes induced by Clostridium perfringens type A α toxin in the intestine of sheep. American journal of veterinary research, 66, 251-255.
8. 8. Gupta, S., Kapoor, P., Chaudhary, K., Gautam, A., Kumar, R., Raghava, G. P. and Consortium, O. S. D. D. (2013) In silico approach for predicting toxicity of peptides and proteins. PloS one, 8, e73957.

9. 9. Guruprasad, K., Reddy, B. B. and Pandit, M. W. (1990) Correlation between stability of a protein and its dipeptide composition: a novel approach for predicting in vivo stability of a protein from its primary sequence. Protein Engineering, Design and Selection, 4, 155-161.
10. 10. Jain, A., Tripathi, P., Shrotriya, A., Chaudhary, R. and Singh, A. (2015) In silico analysis and modeling of putative T cell epitopes for vaccine design of Toscana virus. 3 Biotech, 5, 497-503.
11. 11. Jenssen, H. and Aspmo, S. I. (2008), in Peptide-based drug design, Springer, pp. 177-186.
12. 12. Khan, A., Junaid, M., Kaushik, A. C., Ali, A., Ali, S. S., Mehmood, A. and Wei, D.-Q. (2018) Computational identification, characterization and validation of potential antigenic peptide vaccines from hrHPVs E6 proteins using immunoinformatics and computational systems biology approaches. PloS one, 13, e0196484.
13. 13. Li, J., Adams, V., Bannam, T. L., Miyamoto, K., Garcia, J. P., Uzal, F. A., Rood, J. I. and McClane, B. A. (2013) Toxin plasmids of Clostridium perfringens. Microbiol. Mol. Biol. Rev., 77, 208-233.
14. 14. McClane, B., Uzal, F., Miyakawa, M., Lyerly, D. and Wilkins, T. (2006) The Enterotoxic 644 Clostridia.
15. 15. Meza, B., Ascencio, F., Sierra-Beltrán, A. P., Torres, J. and Angulo, C. (2017) A novel design of a multi-antigenic, multistage and multi-epitope vaccine against Helicobacter pylori: an in silico approach. Infection, Genetics and Evolution, 49, 309-317.
16. 16. Nagahama, M., Yamaguchi, A., Hagiyama, T., Ohkubo, N., Kobayashi, K. and Sakurai, J. (2004) Binding and internalization of Clostridium perfringens iota-toxin in lipid rafts. Infection and immunity, 72, 3267-3275.
17. 17. Naylor, C. E., Eaton, J. T., Howells, A., Justin, N., Moss, D. S., Titball, R. W. and Basak, A. K. (1998) Structure of the key toxin in gas gangrene. Nature Structural & Molecular Biology, 5, 738.
18. 18. Redondo, L. M., Carrasco, J. M. D., Redondo, E. A., Delgado, F. and Miyakawa, M. E. F. (2015) Clostridium perfringens type E virulence traits involved in gut colonization. PloS one, 10, e0121305.
19. 19. Richard, J. F., Mainguy, G., Gibert, M., Marvaud, J. C., Stiles, B. G. and Popoff, M. R. (2002) Transcytosis of iota‐toxin across polarized CaCo‐2 cells. Molecular microbiology, 43, 907-917.
20. 20. Sakurai, J., Nagahama, M. and Oda, M. (2004) Clostridium perfringens alpha-toxin: characterization and mode of action. Journal of biochemistry, 136, 569-574.
21. 21. Shahsavani, N., Sheikhha, M. H., Yousefi, H. and Sefid, F. (2018) In silico Homology Modeling and Epitope Prediction of NadA as a Potential Vaccine Candidate in Neisseria meningitidis. International journal of molecular and cellular medicine, 7, 53.
22. 22. Shey, R. A., Ghogomu, S. M., Esoh, K. K., Nebangwa, N. D., Shintouo, C. M., Nongley, N. F., Asa, B. F., Ngale, F. N., Vanhamme, L. and Souopgui, J. (2019) In-silico design of a multi-epitope vaccine candidate against onchocerciasis and related filarial diseases. Scientific reports, 9, 4409.
23. 23. Songer, J. G. (1998) Clostridial diseases of small ruminants.
24. 24. http://hex.loria.fr/manual800/hex_manual.pdf.
25. 25. http://www.rcsb.org/structure/5V4N).
26. 26. https://en.wikipedia.org/wiki/Diphtheria
27. 27. https://www.cdc.gov/diphtheria/about/index.html
28. 28. https://www.cdc.gov/diphtheria/about/symptoms.html
29. 29. https://www.medicalnewstoday.com/articles/159534.php
30. 30. https://www.ncbi.nlm.nih.gov/protein/4AE1_B.