Psp140: an immunodominant antigen in the supernatant of Streptococcus pneumoniae culture
Background and Objectives: Streptococcus pneumoniae causes many lethal infections. Due to its reduced sensitivity to commonly used antibiotics, development of new strategies against pneumococcal infections seems to be necessary. We aimed to investigate immunodominant antigens in S. pneumoniae culture supernatant in order to develop novel targets for pneumococcal vaccines.
Materials and Methods: In this study S. pneumoniae ATCC49619 was sub-cultured into BHI broth from overnight culture at 37°C for 4 h. The supernatant proteins were precipitated using acetone precipitation method. A rabbit was intramuscularly immunized with alum adjuvant and 100 μg pneumococcal supernatant proteins, 6 times at 14 days' intervals to produce hyperimmune serum. ELISA assay was performed to determine the antibody level response to pneumococcal secretory proteins. Then dot blot applied for rapid evaluation of hyperimmune serum reactivity to pneumococcus supernatant proteins. The western blot was also used to determine the interaction of supernatant proteins with immunogenic rabbit's hyperimmune-serum.
Results: According to the western blot analysis, the immunodominant protein had 140KDa molecular weight and designated as pneumococcal secretory protein140 (Psp140).
Conclusion: The Psp140 protein in the supernatant of S. pneumoniae culture is an immunodominant protein and it is likely related to pneumococcal secretory protein or surface exposed protein which released into culture supernatant during bacterial growth.
2. Farajzadeh Sheikh A, Ahmadi K, Nikakhlagh S. Detection of Streptococcus pneumoniae and Moraxella catarrhalis in patients with paranasal chronic sinusitis by polymerase chain reaction method. J Chin Med Assoc 2016;79:440-444.
3. Lu CY, Ting YT, Huang LM. Severe Streptococcus pneumoniae 19A pneumonia with empyema in children vaccinated with pneumococcal conjugate vaccines.J Formos Med Assoc 2015;114:783-784.
4. Castillo EM, Rickman LS, Brodine SK, Ledbetter EK, Kelly C. Streptococcus pneumoniae: bacteremia in an era of penicillin resistance. Am J Infect Control 2000;28:239-243.
5. Black RE, Cousens S, Johnson HL, Lawn JE, Rudan I, Bassani DG, et al. Global, regional, and national causes of child mortality in 2008: a systematic analysis. Lancet 2010;375:1969-1987.
6. Weiser JN, Ferreira DM, Paton JC. Streptococcus pneumoniae: transmission, colonization and invasion. Nat Rev Microbiol 2018;16:355-367.
7. Henriques-Normark B, Tuomanen EI. The pneumococcus: epidemiology, microbiology, and pathogenesis. Cold Spring Harb Perspect Med 2013;3(7):a010215.
8. Cherazard R, Epstein M, Doan TL, Salim T, Bharti S, Smith MA. Antimicrobial resistant Streptococcus pneumoniae: prevalence, mechanisms, and clinical implications. Am J Ther 2017;24(3):e361-e369.
9. Ludwig E, Meszner Z, Hungarian Society of Infectious Disease and Clinical Microbiology, Vaccinology Section. [Prevention of Streptococcus pneumoniae (pneumococcal) infections in adults]. Orv Hetil 2014;155:1996-2004.
10. Feldman C, Anderson R. Review: current and new generation pneumococcal vaccines. J Infect 2014;69:309-325.
11. Choi CW, Lee YG, Kwon SO, Kim HY, Lee JC, Chung YH, et al. Analysis of Streptococcus pneumoniae secreted antigens by immuno-proteomic approach. Diagn Microbiol Infect Dis 2012;72:318-327.
12. Pancotto L, De Angelis G, Bizzarri E, Barocchi MA, Del Giudice G, Moschioni M, et al. Expression of the Streptococcus pneumoniae pilus-1 undergoes on and off switching during colonization in mice. Sci Rep 2013;3:2040.
13. Kazemian H, Afshar D, Garcia E, Pourmand MR, Jeddi-Tehrani M, Aminharati F, et al. CbpM and CbpG of Streptococcus pneumoniae elicit a high protection in mice challenged with a serotype 19F Pneumococcus. Iran J Allergy Asthma Immunol 2018;17:574-585.
14. Zysk G, Bongaerts RJ, ten Thoren E, Bethe G, Hakenbeck R, Heinz HP. Detection of 23 immunogenic pneumococcal proteins using convalescent-phase serum. Infect Immun 2000;68:3740-3743.
15. De Paolis F, Beghetto E, Spadoni A, Montagnani F, Felici F, Oggioni MR, et al. Identification of a human immunodominant B-cell epitope within the immunoglobulin A1 protease of Streptococcus pneumoniae. BMC Microbiol 2007;7:113.
16. Xu G, Kiefel MJ, Wilson JC, Andrew PW, Oggioni MR, Taylor GL. Three Streptococcus pneumoniae sialidases: three different products. J Am Chem Soc 2011;133:1718-1721.
17. Janapatla RP, Chen CL, Hsu MH, Liao WT, Chiu CH. Immunization with pneumococcal neuraminidases NanA, NanB and NanC to generate neutralizing antibodies and to increase survival in mice. J Med Microbiol 2018;67:709-723.
18. Yother J, White JM. Novel surface attachment mechanism of the Streptococcus pneumoniae protein PspA. J Bacteriol 1994;176:2976-2985.
19. Bongaerts RJ, Heinz HP, Hadding U, Zysk G. Antigenicity, expression, and molecular characterization of surface-located pullulanase of Streptococcus pneumoniae. Infect Immun 2000;68:7141-7143.
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