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Streptococcal pyrogenic exotoxin B causes mitochondria damage to polymorphonuclear cells preventing phagocytosis of Group A streptococcus

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Abstract

The streptococcal pyrogenic exotoxin B (SpeB) is known to be involved in group A streptococcus (GAS) survival in blood, but the detailed mechanism is not clear. For clarification of this issue, speB isogenic mutants of strains M6 and M49 were constructed by using an integrational plasmid and confirmed by Southern blot analysis. The resistance to phagocytosis of wild-type strains and their speB isogenic mutants was analyzed. The results demonstrated a five-fold increase in phagocytosis of speB mutants compared to that of wild-type strains in whole blood, but no significant difference in plasma. To further clarify whether this effect is due to a functional SpeB protein, recombinant SpeB (r-SpeB) and a SpeB mutant protein lacking proteinase activity (r-C192S) were purified and incubated with a speB mutant in whole blood. The results showed a two- to threefold increase in resistance to phagocytosis when the M6 speB mutant was incubated with r-SpeB, but not with r-C192S. Incubation with the wild-type strain, speB mutant, or the r-SpeB protein did not affect the total cell number of polymorphonuclear (PMN) cells in whole blood under laboratory conditions. However, the PMN cells’ mitochondria showed decreasing dehydrogenase activity and loss of membrane potential after r-SpeB treatment. These data indicate that SpeB could cause the mitochondria damage to the PMN cells, preventing immune clearance at an early infectious stage.

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Acknowledgements

We thank Chiou-Feng Lin and Peng-Ju Chien for providing many helpful discussions and technical supports with mitochondria integrity measurement. This work was partly supported by grants NSC-90-2320-B-006-088 and NSC-91-2314-B-006-088 from the National Science Council, and NHRI-EX91-9027SP from National Health Research Institutes, Taiwan.

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Authors and Affiliations

  1. Institute of Basic Medical Sciences, College of Medicine, National Cheng Kung University, Tainan, Taiwan

    Chuan Chiang-Ni & Chih-Hung Wang

  2. Department of Biochemistry, College of Medicine, National Cheng Kung University, Tainan, Taiwan

    Woei-Jer Chuang

  3. Department of Microbiology and Immunology, College of Medicine, National Cheng Kung University, Tainan, Taiwan

    Yee-Shin Lin

  4. Department of Pediatrics, College of Medicine, National Cheng Kung University, Tainan, Taiwan

    Ching-Chuan Liu

  5. Department of Medical Laboratory Science & Biotechnology, College of Medicine, National Cheng Kung University, No. 1 Ta-Hsueh Road, Tainan, Taiwan, 70101

    Jiunn-Jong Wu

  6. Department of Laboratory Science& Biotechnology, College of Medicine, Tsu Chi University, Hualien, Taiwan

    Pei-Jane Tsai

  7. Department of Biochemistry, College of Medicine, Tsu Chi University, Hualien, Taiwan

    Ming-T. Lin

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  1. Chuan Chiang-Ni
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Correspondence to Jiunn-Jong Wu.

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Chiang-Ni, C., Wang, CH., Tsai, PJ. et al. Streptococcal pyrogenic exotoxin B causes mitochondria damage to polymorphonuclear cells preventing phagocytosis of Group A streptococcus. Med Microbiol Immunol 195, 55–63 (2006). https://doi.org/10.1007/s00430-005-0001-y

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  • DOI: https://doi.org/10.1007/s00430-005-0001-y

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