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Thromb Haemost 2025; 125(03): 243-254
DOI: 10.1055/a-2369-8680
Cellular Haemostasis and Platelets

Pneumococcal Neuraminidases Increase Platelet Killing by Pneumolysin

Kristin J. Fritsch   *
1   Department of Transfusion Medicine, Institute of Transfusion Medicine, University Medicine Greifswald, Greifswald, Germany
,
Laura Krüger   *
1   Department of Transfusion Medicine, Institute of Transfusion Medicine, University Medicine Greifswald, Greifswald, Germany
2   Department of Molecular Genetics and Infection Biology, Interfaculty Institute for Genetics and Functional Genomics, University of Greifswald, Greifswald, Germany
,
Stefan Handtke
1   Department of Transfusion Medicine, Institute of Transfusion Medicine, University Medicine Greifswald, Greifswald, Germany
3   Department of Transfusion Medicine, Institute of Transfusion Medicine, University Medicine Rostock, Rostock, Germany
,
Thomas P. Kohler
2   Department of Molecular Genetics and Infection Biology, Interfaculty Institute for Genetics and Functional Genomics, University of Greifswald, Greifswald, Germany
,
Arina Ozhiganova
2   Department of Molecular Genetics and Infection Biology, Interfaculty Institute for Genetics and Functional Genomics, University of Greifswald, Greifswald, Germany
,
Kristin Jahn
2   Department of Molecular Genetics and Infection Biology, Interfaculty Institute for Genetics and Functional Genomics, University of Greifswald, Greifswald, Germany
,
Jan Wesche
1   Department of Transfusion Medicine, Institute of Transfusion Medicine, University Medicine Greifswald, Greifswald, Germany
,
Andreas Greinacher    *
1   Department of Transfusion Medicine, Institute of Transfusion Medicine, University Medicine Greifswald, Greifswald, Germany
,
Sven Hammerschmidt    *
2   Department of Molecular Genetics and Infection Biology, Interfaculty Institute for Genetics and Functional Genomics, University of Greifswald, Greifswald, Germany
› Author Affiliations Funding Support is acknowledged from the DFG (Project no. 374031971 to A.G. and Sv.H.—TRR 240 and DFG HA 3125/8-1 project no. 523973396 to Sv.H.)
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Abstract

Background Platelets prevent extravasation of capillary fluids into the pulmonary interstitial tissue by sealing gaps in inflamed endothelium. This reduces respiratory distress associated with pneumonia. Streptococcus pneumoniae is the leading cause of severe community-acquired pneumonia. Pneumococci produce pneumolysin (PLY), which forms pores in membranes of eukaryotic cells including platelets. Additionally, pneumococci express neuraminidases, which cleave sialic acid residues from eukaryotic glycoproteins. In this study, we investigated the effect of desialylation on PLY binding and pore formation on platelets.

Materials and Methods We incubated human platelets with purified neuraminidases and PLY, or nonencapsulated S. pneumoniae D39/TIGR4 and isogenic mutants deficient in PLY and/or NanA. We assessed platelet desialylation, PLY binding, and pore formation by flow cytometry. We also analyzed the inhibitory potential of therapeutic immunoglobulin G preparations (IVIG [intravenous immunoglobulin]).

Results Wild-type pneumococci cause desialylation of platelet glycoproteins by neuraminidases, which is reduced by 90 to 100% in NanA-deficient mutants. NanC, cleaving only α2,3-linked sialic acid, induced platelet desialylation. PLY binding to platelets then x2doubled (p = 0.0166) and pore formation tripled (p = 0.0373). A neuraminidase cleaving α2,3-, α2,6-, and α2,8-linked sialic acid like NanA was even more efficient. Addition of polyvalent IVIG (5 mg/mL) decreased platelet desialylation induced by NanC up to 90% (p = 0.263) and reduced pore formation >95% (p < 0.0001) when incubated with pneumococci.

Conclusion Neuraminidases are key virulence factors of pneumococci and desialylate platelet glycoproteins, thereby unmasking PLY-binding sites. This enhances binding of PLY and pore formation showing that pneumococcal neuraminidases and PLY act in concert to kill platelets. However, human polyvalent immunoglobulin G preparations are promising agents for therapeutic intervention during severe pneumococcal pneumonia.

Keywords

Streptococcus pneumoniae - neuraminidase - pneumolysin - platelets - immunoglobulins

Authors' Contribution

K.J.F. performed the experiments with purified toxins, evaluated the data, and wrote the manuscript. L.K. performed the experiments with whole bacteria, evaluated the data, and wrote the manuscript. S.H. and T.P.K. planned the experiments, evaluated the data, and edited the manuscript. K.J. and J.W. contributed to the experiments and edited the manuscript. A.G. and Sv.H. designed the project, supervised the project, evaluated the data, and edited the manuscript. All authors reviewed the final version of the manuscript.


* These authors contributed equally to the study.


Supplementary Material



Publication History

Received: 11 November 2023

Accepted: 18 July 2024

Accepted Manuscript online:
19 July 2024

Article published online:
20 August 2024

© 2024. Thieme. All rights reserved.

Georg Thieme Verlag KG
Rüdigerstraße 14, 70469 Stuttgart, Germany

 

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