Open Access Research

Heat shock protein 70 down-regulates the production of toll-like receptor-induced pro-inflammatory cytokines by a heat shock factor-1/constitutive heat shock element-binding factor-dependent mechanism

Eduardo Ferat-Osorio12, Aldair Sánchez-Anaya1, Mireille Gutiérrez-Mendoza1, Ilka Boscó-Gárate1, Isabel Wong-Baeza13, Rodolfo Pastelin-Palacios6, Gustavo Pedraza-Alva5, Laura C Bonifaz1, Pedro Cortés-Reynosa4, Eduardo Pérez-Salazar4, Lourdes Arriaga-Pizano1, Constantino López-Macías1, Yvonne Rosenstein57* and Armando Isibasi18*

Author Affiliations

1 Unidad de Investigación Médica en Inmunoquímica, Hospital de Especialidades, Centro Médico Nacional Siglo XXI, Instituto Mexicano del Seguro Social, Av. Cuauhtémoc 330, Col. Doctores, México D.F. CP 06020, México

2 Servicio de Cirugía Gastrointestinal, Hospital de Especialidades, Centro Médico Nacional Siglo XXI, Instituto Mexicano del Seguro Social, Av. Cuauhtémoc 330, Col. Doctores, México D.F. CP 06020, México

3 Departamento de Inmunología, Instituto Politécnico Nacional, Escuela Nacional de Ciencias Biológicas, México D.F., México

4 Departamento de Biología Celular, (CINVESTAV) Instituto Politécnico Nacional, Centro de Investigación y Estudios Avanzados, México D.F., México

5 Departamento de Medicina Molecular y Bioprocesos, Instituto de Biotecnología, Universidad Nacional Autónoma de México, Cuernavaca, México

6 Facultad de Química, Universidad Nacional Autónoma de México (UNAM), México D.F., México

7 Instituto de Biotecnología, Universidad Nacional Autónoma de México, Av. Universidad 2001, Col. Chamilpa, Cuernavaca Mor. 62210, México

8 Coordinación de Investigación en Salud, Piso 4 Bloque B Unidad de Congresos Centro Médico Nacional Siglo XXI, Av. Cuauhtémoc 330, Col. Doctores, México D.F. CP 06020, México

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Journal of Inflammation 2014, 11:19  doi:10.1186/1476-9255-11-19

Published: 12 July 2014

Abstract

Background

Heat shock protein 70 (Hsp70) is an intracellular chaperone protein with regulatory and cytoprotective functions. Hsp70 can also be found in the extracellular milieu, as a result of active secretion or passive release from damaged cells. The role of extracellular Hsp70 is not fully understood. Some studies report that it activates monocytes, macrophages and dendritic cells through innate immune receptors (such as Toll-like receptors, TLRs), while others report that Hsp70 is a negative regulator of the inflammatory response. In order to address this apparent inconsistency, in this study we evaluated the response of human monocytes to a highly purified recombinant Hsp70.

Methods

Human peripheral blood monocytes were stimulated with Hsp70, alone or in combination with TLR agonists. Cytokines were quantified in culture supernatants, their mRNAs were measured by RT-PCR, and the binding of transcription factors was evaluated by electrophoretic mobility shift assay (EMSA). Kruskal-Wallis test or one-way or two-way ANOVA were used to analyze the data.

Results

The addition of Hsp70 to TLR-activated monocytes down-regulated TNF-α as well as IL-6 levels. This effect was independent of a physical interaction between Hsp70 and TLR agonists; instead it resulted of changes at the TNF-α gene expression level. The decrease in TNF-α expression correlated with the binding of HSF-1 (heat shock transcription factor 1, a transcription factor activated in response to Hsp70) and CHBF (constitutive HSE-binding factor) to the TNF-α gene promoter.

Conclusion

Extracellular Hsp70 negatively regulates the production of pro-inflammatory cytokines of monocytes exposed to TLR agonists and contributes to dampen the inflammatory response.

Keywords:
Heat shock protein 70; Inflammatory response; HSF-1; CHBF; Toll-like receptors