Morphological Effect Of Autologous Hsp70

Published Date: 02 Nov 2017

P.K. GAUTAM,1 S. KUMAR,1 P. DEEPAK,1., and A. ACHARYA1*

1Department of zoology, Faculty of Science, Banaras Hindu University,

Varanasi-221005, U.P., India

*Mailing Address: Prof. Arbind Acharya

Immunology Laboratory, Department of Zoology,

Faculty of Science, Banaras Hindu University

Tel: +91 542 2307149 (Office), 6702518 (Lab)

Fax: +91 542 2368174

E-mail: [email protected],

Keywords: lamellipodia; Macrophages; Tumor associated macrophages; Hsp70; Morphology

Introduction

Inflammation is well known to play an important role in initiating and promoting cancer, although, it might be participated to both specific and nonspecific tumor rejection [1-3]. Epidemiological studies have been reported that chronological inflammation predisposed to different cancer, including T cells leukemia is a prototype. Inflammation recruits different type of immune cells to the tumor milieu. Tumor microenvironment suffer from, hypoxia, nutrient deprivation, acidosis, abrent stroma, and buildup metabolic waste product and surrounded by different non malignant such as endothelial cells, fibroblast and various immune cells including macrophages (Mɸ) which are leading immune shoulder of tumor regression. Tumor cell produces different immunosuppressive molecules that promote tumor growth by releasing tumor growth factor such as IL-10, TGF-β, IL-4, to tumor cells survival releasing M-CSF, VEGF and many other molecules that MDF, PGE2 that deactivate or suppress immune cell function including Mɸ. Mɸ are specialized myeloid derived phagocytic cell that recruit to tumor milieu due to influence of different chemoattractant produces by tumor cells such as CSF-1, CCL-1, CCL-2, CCL-3, CCL-4, CCL-5, VEGF, MIP-1 which induces the migration of macrophages and other leukocytes at the site of tumor growth [4,5]. Mɸ releases several immunomodulatory molecules such as ROIs, RNIs, IL-6, TNF-α that kill tumor cell, IL-10 inhibit angiogenesis, produces several other molecules that inhibit MMPs, VEGF and IL-10 production by tumor cells that assist tumor growth, angiogenesis, metastasis and invasion. Due to high plastic phenotype and their functional polarization is determined by cytokines, tumor derived factor with in local microenvironment transform M1 polarized phenotype of Mɸ into M2 phenotype Mɸ in advanced stage of tumor growth termed TAMs or Myeloid- derived suppressor cells (MDSC) that suppresses adaptive anti-tumor immune responses[6]. MDSC produces several factor such as IL-4. Il-10, for M2 differentiation and growth, IL-1, 6, 8, bFGF, CSF, TGF-β for angeogenesis, IL-23 Invasion and MMPs for metastasis of tumor cells. MDSC also shows suppressed expression of antigen, phagocytosis and immune cell activation of T cell response.

During inflammation different type of anti-inflammatory molecules and MHC restricted molecules produces by different cells including Heat Shock Proteins (HSPs) is expressed nearly in all prokaryotic and eukaryotic cells. It is phyllogenetically highly conserved molecule performing important function in folding, refolding and translocation of protein molecules [7]. HSP is expressed inducibly as well as constitutively upon stimulation by different cell damaging stimuli and protects the cell from the subsequent damage. There are different types of HSPs and among all Hsp70 is highly conserved [8] and immunogenic capable for inducing antibody production and T cell activation. The Hsp70 plays an important role in antigen presentation and cross presentation of tumor antigen [9,10]. It stimulates macrophages which are the most abundant cell population at the tumor site. Now a day, it is emerging as biochemical regulators of cell growth, apoptosis, protein homeostasis and cellular targets of peptides.

Numerous studies have demonstrated that Hsp70-peptides derived from a tumor cell can elicit cancer-specific immunity against the same tumor by virtue of their ability to bind tumor-specific peptides [11]. It has been observed that Hsp70 preparations derived from normal tissues do not elicit tumor immunity, but the immunity elicited by Hsp70-peptides complex has been shown only due to antigenic peptides associated with it. Further studies indicate that immunity against tumor elicited by immunization with HSP peptide complexes, including Hsp70 and gp96 family is mediated by CD8+ T lymphocytes, and its mechanism involves MHC-I class molecule restricted response which is required to channel the peptides into class I presentation pathway[12].

The surfaces of macrophages and monocytes are covered in ruffles and micrivilli, and small surface blebs which helps macrophage locomotion, adherence to endothelial cell, endocytosis of antigens. It has been reported that ruffling structures of cell membrane of macrophage extend after activation with Biologic Response Modifiers (BRM), which includes LPS, lipoprotein, cytokines, PMA, PGE2, nucleic acids etc [13] .Recently, it has been shown that heat shock proteins (HSPs) are able to activate macrophages to induce the production of specific and non-specific effectors molecules and extend surface blebs of macrophage [14-16].

In the present study, an investigate have shown that how Hsp70 was isolated and purified from DL cells isolated from tumor bearing mice and their effect on pseudopodial expression in normal and tumor associated macrophages has been studied.