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Transmission of Anaplasma phagocytophilum from endothelial cells to peripheral granulocytes in vitro under shear flow conditions

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Abstract

Anaplasma phagocytophilum (Ap) is a tick-borne pathogen, which can cause granulocytic anaplasmosis in humans and animals. In vivo this obligate intracellular pathogen is primarily located in circulating mature granulocytes, but it also infects endothelial cells. In order to study the interaction between Ap-infected endothelial cells and human granulocytes under conditions similar to those found naturally in the infected host, an in vitro model that mimics physiological flow conditions in the microvasculature was established. Cell-to-cell interactions were then visualized by microscopy, which showed that granulocytes adhered strongly to Ap-infected endothelial cells at a shear stress of 0.5 dyne/cm2. In addition, Ap-transmission assays under flow conditions showed that the bacteria transferred from infected endothelial cells to circulating granulocytes and were able to establish infection in constantly moving granulocytes. Cell surface analysis showed that Ap induced up-regulation of the cell adhesion molecules ICAM-1 and VCAM-1 on infected endothelial cells in a dose-dependent manner. Furthermore, IL-8 secretion by endothelial cells indicated that the presence of Ap induced a pro-inflammatory response. In summary, the results of this study suggest that endothelial cells of the microvasculature (1) provide an excellent site for Ap dissemination to peripheral blood granulocytes under flow conditions and therefore may play a crucial role in the development of persistent infection, and (2) are stimulated by Ap to express surface molecules and cytokines that may lead to inflammatory responses at the site of the infection.

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Acknowledgments

The authors thank Anke Schiller and Stephanie Hiereth for their assistance in cell and bacterial culture; Dr. Silke Kadlez-Gebhardt, Dr. Winfried Kapfhammer and Dr. Vollert Lüthje for collecting blood samples from our donors; and Dr. Sonja Härtle for her useful discussion about flow cytometric data analysis. The authors thank Dr. Evelyn Overzier and Dr. Anna Rettinger for their careful reading of our manuscript. J.W. is a research fellow of the China Scholarship Council (CSC).

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Authors and Affiliations

  1. Institute for Infectious Diseases and Zoonoses, Department of Veterinary Sciences, Faculty of Veterinary Medicine, LMU Munich, Veterinärstr. 13, 80539, Munich, Germany

    Jinyong Wang, Viktor Dyachenko & Reinhard K. Straubinger

  2. Department of Entomology, University of Minnesota, St. Paul, MN, 55108, USA

    Ulrike G. Munderloh

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  1. Jinyong Wang
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  2. Viktor Dyachenko
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Correspondence to Reinhard K. Straubinger.

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Wang, J., Dyachenko, V., Munderloh, U.G. et al. Transmission of Anaplasma phagocytophilum from endothelial cells to peripheral granulocytes in vitro under shear flow conditions. Med Microbiol Immunol 204, 593–603 (2015). https://doi.org/10.1007/s00430-015-0387-0

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  • DOI: https://doi.org/10.1007/s00430-015-0387-0

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