Application of Single-Cell Cultures of Mouse Splenocytes as an Assay System to Analyze the Immunomodulatory Properties of Bacterial Components
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.
- 14.Shimoyama, T. and Crabtree, J. E. (1998) Bacterial factors and immune pathogenesis in Helicobacter pylori infection. Gut 43(suppl 1), S2–5.Google Scholar
- 19.Sambrook, J., Fritsch, E. F., and Maniatis, T. (1989) Molecular Cloning: A Laboratory Manual. New York, Cold Spring Harbor Laboratory.Google Scholar
- 20.Deml, L. and Wagner, R. (1998) Stable transfected Drosophila Schneider-2 cells as a novel tool to produce recombinant antigens for diagnostic, therapeutic and preventive purposes, in U Reischl (ed), Methods in Molecular Medicine. Molecular Diagnosis of Infectious Diseases. Humana, Totowa, NJ, pp. 185–200.Google Scholar
- 21.White, B. A. (1993) Methods in Molecular Biology: PCR Protocols, Current Methods and Applications. Humana, Totowa, NJ.Google Scholar