Flow cytometry is a rapid and reliable method for evaluating heat shock protein 70 expression in human monocytes

Bachelet, Maria; Mariéthoz, Ewa; Banzet, Nathalie; Souil, Évelyne; Pinot, Françoise; Polla, C Z; Durand, Philippe; Bouchaert, Isabelle; Polla, B. S.

doi:10.1379/1466-1268(1998)003<0168:fciara>2.3.co;2

Cited by 57 publications

(35 citation statements)

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“…Flow cytometry was used for the detection of Hsp70 and Hsp27 in lymphocytes (Xiao et al 2003;Bachelet et al 1998). Each fixed lymphocyte sample was divided into two to detect Hsp70 and Hsp27 separately as follows.…”

Section: Detection Of Hsp70 and Hsp27 Levels In Lymphocytesmentioning

confidence: 99%

COPD and levels of Hsp70 (HSPA1A) and Hsp27 (HSPB1) in plasma and lymphocytes among coal workers: a case-control study

Cui

Xing

Liu

et al. 2015

Cell Stress and Chaperones

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This case-control study aimed to investigate whether the levels of Hsp70 (HSPA1A) and Hsp27 (HSPB1) in plasma and lymphocytes were associated with the risk of chronic obstructive pulmonary disease (COPD) among coal workers. A total of 76 COPD cases and 48 age-matched healthy controls from a group of coal workers were included. The case group consisted of 35 COPD patients whose condition was complicated with coal workers' pneumoconiosis (CWP) and 41 COPD patients without CWP. Heat shock proteins (Hsps) in plasma and lymphocytes were detected by ELISA and flow cytometry, respectively. Multiple logistic regression models were applied to estimate the association between Hsp levels and COPD risk. Our results showed that plasma Hsp70 and lymphocyte Hsp27 levels were significantly higher and plasma Hsp27 levels were significantly lower in COPD cases than in controls (p<0.01). No significant differences in lymphocyte Hsp70 levels were found between COPD cases and the matched subjects. Higher plasma Hsp70 levels (odds ratio (OR)=13.8, 95 % confidence interval (CI)=5.7-33.5) and lower plasma Hsp27 levels (OR=4.6, 95 % CI= 2.0-10.5) were significantly associated with an increased risk of COPD after adjusting for confounders. Higher lymphocyte Hsp27 levels were only associated with an increased risk of COPD with CWP (OR=6.6, 95 % CI=2.0-22.1) but not with an increased risk of COPD without CWP (OR=3.0, 95 % CI= 0.9-8.9). Additionally, there were strong joint effects of different Hsps on COPD risk. These results showed that higher levels of plasma Hsp70 and lower levels of plasma Hsp27 might be associated with an increased risk of COPD among coal workers. They may have the potential to serve as monitoring markers for COPD in coal workers.

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Section: Detection Of Hsp70 and Hsp27 Levels In Lymphocytesmentioning

confidence: 99%

COPD and levels of Hsp70 (HSPA1A) and Hsp27 (HSPB1) in plasma and lymphocytes among coal workers: a case-control study

Cui

Xing

Liu

et al. 2015

Cell Stress and Chaperones

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“…Earlier studies that addressed compound induced manipulation of the heat shock response also measured Hsp70 expression (Yan et al 2004;Brundel et al 2006;Kato et al 1998), enabling easy comparison of previously identified compounds with new ones. First, we addressed whether flow cytometry could be used to study stress induced Hsp70 in several mouse cell types in a simple and rapid manner (Bachelet et al 1998). We analyzed Hsp70 expression in RAW264.7 macrophages, m-IC c12 intestinal crypt cells, primary bone marrow-derived dendritic cells (BMDC) and spleen cells.…”

Section: Analysis Of Hsp70 Expression By Flow Cytometrymentioning

confidence: 99%

Hsp70 expression and induction as a readout for detection of immune modulatory components in food

Wieten

Zee

Goedemans

et al. 2010

Cell Stress and Chaperones

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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 Tcells. 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 stressinduced 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.

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“…1, B and C). Flow cytometric analysis of intracellular HSPs is a rapid, sensitive, quantitative, and more-reproducible method than Western blotting and is particularly suited for determining protein levels in individual cells (50,51). Moreover, heat-treated tumor cells displayed a characteristic phenotype with higher levels of TAAs (CEA and MUC1) and HLA-ABC (HLA-A2 and/or A24) (Fig.…”

Section: Characteristic Phenotype Of Ok/hs-fcsmentioning

confidence: 99%

Synergistic Induction of Antigen-Specific CTL by Fusions of TLR-Stimulated Dendritic Cells and Heat-Stressed Tumor Cells

Koido

Hara

Homma

et al. 2007

The Journal of Immunology

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Dendritic cell (DC)/tumor cell fusion cells (FCs) can induce potent CTL responses. The therapeutic efficacy of a vaccine requires the improved immunogenicity of both DCs and tumor cells. The DCs stimulated with the TLR agonist penicillin-killed Streptococcus pyogenes (OK-432; OK-DCs) showed higher expression levels of MHC class I and II, CD80, CD86, CD83, IL-12, and heat shock proteins (HSPs) than did immature DCs. Moreover, heat-treated autologous tumor cells displayed a characteristic phenotype with increased expression of HSPs, carcinoembryonic Ag (CEA), MUC1, and MHC class I (HLA-A2 and/or A24). In this study, we have created four types of FC preparation by alternating fusion cell partners: 1) immature DCs fused with unheated tumor cells; 2) immature DCs fused with heat-treated tumor cells; 3) OK-DCs fused with unheated tumor cells; and 4) OK-DCs fused with heat-treated tumor cells. Although OK-DCs fused with unheated tumor cells efficiently enhanced CTL induction, OK-DCs fused with heat-treated tumor cells were most active, as demonstrated by: 1) up-regulation of multiple HSPs, MHC class I and II, CEA, CD80, CD86, CD83, and IL-12; 2) activation of CD4+ and CD8+ T cells able to produce IFN- γ at higher levels; 3) efficient induction of CTL activity specific for CEA or MUC1 or both against autologous tumor; and 4) superior abilities to induce CD107+IFN-γ+CD8+ T cells and CD154+ IFN-γ+CD4+ T cells. These results strongly suggest that synergism between OK-DCs and heat-treated tumor cells enhances the immunogenicity of FCs and provides a promising means of inducing therapeutic antitumor immunity.

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Flow cytometry is a rapid and reliable method for evaluating heat shock protein 70 expression in human monocytes

Cited by 57 publications

References 0 publications

COPD and levels of Hsp70 (HSPA1A) and Hsp27 (HSPB1) in plasma and lymphocytes among coal workers: a case-control study

COPD and levels of Hsp70 (HSPA1A) and Hsp27 (HSPB1) in plasma and lymphocytes among coal workers: a case-control study

Hsp70 expression and induction as a readout for detection of immune modulatory components in food

Synergistic Induction of Antigen-Specific CTL by Fusions of TLR-Stimulated Dendritic Cells and Heat-Stressed Tumor Cells

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