Protocol

Molecular Diagnosis of Infectious Diseases

Volume 94 of the series Methods in Molecular Medicine™ pp 159-175

Application of Single-Cell Cultures of Mouse Splenocytes as an Assay System to Analyze the Immunomodulatory Properties of Bacterial Components

  • Ludwig DemlAffiliated withInstitute for Medical Microbiology and Hygiene, University of Regensburg
  • , Michael AignerAffiliated withInstitute for Medical Microbiology and Hygiene, University of Regensburg
  • , Alexander EckhardtAffiliated withInstitute for Medical Microbiology and Hygiene, University of Regensburg
  • , Jochen DeckerAffiliated withAccenture GmbH
  • , Norbert LehnAffiliated withInstitute for Medical Microbiology and Hygiene, University of Regensburg
  • , Wulf Schneider-BrachertAffiliated withInstitute for Medical Microbiology and Hygiene, University of Regensburg

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Abstract

Recently, various bacterial components have been suggested as initiating and modulating immune activation, thereby substantially affecting the complex and dynamic host/pathogen interactions. Herein, we present a valuable and simple methodology for determining the capacity of bacteria as well as defined bacterial structures to stimulate cellular effectors of the innate and cognate immune system. This assay format is based on the exposure of freshly prepared single-cell cultures of splenic cells derived from naive mice with the immunogen of interest. Herein, the determination of exclusive panels of cytokines by the ELISA, ELISpot, and FACS technology will serve as an indicator for the activation of defined arms of the immune system. An increased knowledge about microbial components with immunomodulatory properties will substantially contribute to a more detailed understanding of the dynamic interplay between the host and potential pathogens and, based on this knowledge, to the development of novel substances for the prevention and therapy of microbial infections.