Hsp70 expression and induction as a readout for detection of immune modulatory components in food.
Stress proteins such as heat shock proteins (Hsps) are up-regulated in cells in response to various forms of stress, like thermal and oxidative stress and inflammation. Hsps prevent cellular damage and increase immunoregulation by the activation of anti-inflammatory T-cells. Decreased capacity for stress-induced Hsp expression is associated with immune disorders. Thus, therapeutic boosting Hsp expression might restore or enhance cellular stress resistance and immunoregulation. Especially food- or herb-derived phytonutrients may be attractive compounds to restore optimal Hsp expression in response to stress. In the present study, we explored three readout systems to monitor Hsp70 expression in a manner relevant for the immune system and evaluated novel Hsp co-inducers. First, intracellular staining and analysis by flow cytometry was used to detect stress and/or dietary compound induced Hsp70 expression in multiple rodent cell types efficiently. This system was used to screen a panel of food-derived extracts with potent anti-oxidant capacity. This strategy yielded the identity of several new enhancers of stress-induced Hsp70 expression, among them carvacrol, found in thyme and oregano. Second, CD4(+) T-cell hybridomas were generated that specifically recognized an immunodominant Hsp70 peptide. These hybridomas were used to show that carvacrol enhanced Hsp70 levels increased T-cell activation. Third, we generated a DNAJB1-luc-O23 reporter cell line to show that carvacrol increased the transcriptional activation of a heat shock promoter in the presence of arsenite. These assay systems are generally applicable to identify compounds that affect the Hsp level in cells of the immune system.
Wieten
Lotte
L
Department of Infectious Diseases and Immunology, Utrecht University, The Netherlands.
van der Zee
Ruurd
R
Goedemans
Renske
R
Sijtsma
Jeroen
J
Serafini
Mauro
M
Lubsen
Nicolette H
NH
van Eden
Willem
W
Broere
Femke
F
D000818:Animals; D015496:CD4-Positive T-Lymphocytes / Q000276:immunology* / Q000378:metabolism; D002460:Cell Line; D000081007:Cymenes; D005434:Flow Cytometry / Q000379:methods; D005504:Food Analysis; D050956:HSP40 Heat-Shock Proteins / Q000235:genetics / Q000378:metabolism; D018840:HSP70 Heat-Shock Proteins / Q000378:metabolism*; D007376:Interleukin-2 / Q000378:metabolism; D051379:Mice; D039821:Monoterpenes / Q000494:pharmacology; D010936:Plant Extracts / Q000494:pharmacology; D013696:Temperature
D000081007:Cymenes; C494641:Dnajb1 protein, mouse; D050956:HSP40 Heat-Shock Proteins; D018840:HSP70 Heat-Shock Proteins; D007376:Interleukin-2; D039821:Monoterpenes; D010936:Plant Extracts; C073316:carvacrol