See LPS information below..LPS not found with borrelia.

Grant-funded Project Nr. 274/1999/C/P�F
Final Report
Project title: Production of cytokines and expresion of heat shock proteines during the stress due to different pyrogens.
Research leader: RNDr. Stanislav Vyb�ral, CSc.
Co-researcher: Mgr. Martin Couf; Mgr. Ludmila Nov�kov�
Period of project: 1999-2001
Overall grant: 275 000 CZK

Project Results
The method for isolation of lymphocytes, their cultivation, determination of cytokines and heat shock proteins including of specialized software for quantification of HSPs concentrations after immunodetection have been established. It was found that activation of PBMC by LPS increase production of IL-1beta and TNF-alpha significantly, while production of IL-6 increased to a smaller extent. Delayed increase in production of IL-10 was also observed. Infection of PBMC by live Borrelia increases production of IL-1beta, TNF-alpha, IL-6 and IL-10 in the same way as LPS, but in contrast to LPS, Borrelia increases production of IL-12 and IFN-gamma in the later phase of the experiment. Data indicate that infection by different pyrogens activates the cells of the immune cascade in a different way. Infection with Borrelia activates not only the early steps of the immune response (macrophages and monocytes) but also the late phase of the immune cascade probably due to activation of NK and T-cells. Data further indicate that infection by Borrelia has a more extensive effect on immune cells than LPS infection. The dominant role of IL-1beta and TNF-alpha in induction of fever has been confirmed in experiments in vivo on an animal model. The significant pyrogenic effect of Borrelia with all signs of the typical one phase fever has been observed. Concentrations of HSPs (HSP 70, HSC 70, HSP 90, HSP 27) in PBMC isolated from normal human subjects were measured. HSC 70 was found to be the most prominent HSP in PBMC in contrast to HSP 27. In order to find out whether or not the bacterial or viral infections can influence HSP concentrations. The PBMC from humans suffering from EB viral or streptococal angina were also analyzed. No significant differences in HSPs concentration were discovered between control and infected subjects mainly due to a considerable variability. Under in vitro conditions LPS stimulated cytokine production in PBMC an increase in HSPs concentration was also observed, during the later phase of stimulation. This contrasts with cytokines production which occur immediately after stimulation. Thus, the increased production of cytokines is not probably directly related to HSPs expression.