Heat shock proteins (HSPs) of the hsp70, hsp90 and calreticulin families are abundant soluble intracellular proteins that elicit powerful immunological responses. These responses include components of both the innate and adaptive aspects of T cell responses as well as its down regulation. The ability of HSPs to chaperone peptides and engage internalizing receptors on antigen presenting cells are two key properties that allow them to elicit immune responses. A key role for HSPs in cross- priming of antigens during T cell responses was also envisaged and demonstrated in 2005. In this application we aim to (i) investigate the role of the HSP receptor, CD91, in cross-priming in the murine system, (ii) examine the mechanism of immunogenicity of alpha2-macroglobulin (?2M), a CD91 ligand, complexed either endogenously and artificially to various peptides, and (iii) test the role of serum levels of ?2M on the ability of mice to mount immune responses.
Aim 1 : To investigate the role of the HSP receptor CD91 in cross-priming of T cell responses. a. Determine the ability of RAP (a competitive inhibitor of CD91) -expressing cells to cross present antigens and mount effective T cell responses; b. Creation of a CD91 conditional knock-out mouse and the characterization of immunological responses in this mouse.
Aim 2 : To investigate the mechanism of ?2M-mediated immunogenicity and its application to immunotherapy of cancer and infectious disease. a. Examine the mechanism of co-stimulatory activation by ?2M; b. Examination of the intracellular trafficking and processing of ?2M-peptide complexes. c. Test the complexes of tumor derived-peptides with ?2M in prophylaxis and therapy of cancers; d. Examination of endogenously formed ?2M-peptide complexes as immunogens.
Aim 3 : To investigate the role of blodd ?2M levels on immune system. a. Test the effects of blood ?2M levels on the ability of mice to elicit T cell responses after tumor inoculation; b. Test the effects of blood ?2M levels on the ability of mice to elicit T cell responses after immunization with HSP-peptide complexes.

Public Health Relevance

/RELEVANCE We have identified a role for heat shock proteins in cross-priming, an event that is central to the initiation of adaptive immune responses against cancer and infectious disease. In this proposal we aim to examine the HSP receptor(s) involved and how we may exploit this receptor by channeling other ligands, such as ?2-macroglobulin through this pathway. This will allow one to generate novel therapeutic agents against cancer and infectious disease by targeting cell surface receptors.

Agency
National Institute of Health (NIH)
Institute
National Institute of Allergy and Infectious Diseases (NIAID)
Type
Research Project (R01)
Project #
5R01AI079057-05
Application #
8488395
Study Section
Transplantation, Tolerance, and Tumor Immunology (TTT)
Program Officer
Gondre-Lewis, Timothy A
Project Start
2009-07-01
Project End
2014-06-30
Budget Start
2013-07-01
Budget End
2014-06-30
Support Year
5
Fiscal Year
2013
Total Cost
$308,784
Indirect Cost
$101,492
Name
University of Pittsburgh
Department
Microbiology/Immun/Virology
Type
Schools of Medicine
DUNS #
004514360
City
Pittsburgh
State
PA
Country
United States
Zip Code
15213
Zhou, Yu Jerry; Messmer, Michelle Nicole; Binder, Robert Julian (2014) Establishment of tumor-associated immunity requires interaction of heat shock proteins with CD91. Cancer Immunol Res 2:217-28
Messmer, Michelle Nicole; Pasmowitz, Joshua; Kropp, Laura Elizabeth et al. (2013) Identification of the cellular sentinels for native immunogenic heat shock proteins in vivo. J Immunol 191:4456-65
Pawaria, Sudesh; Binder, Robert J (2011) CD91-dependent programming of T-helper cell responses following heat shock protein immunization. Nat Commun 2:521
Pawaria, Sudesh; Messmer, Michelle Nicole; Zhou, Yu Jerry et al. (2011) A role for the heat shock protein-CD91 axis in the initiation of immune responses to tumors. Immunol Res 50:255-60
Kropp, Laura E; Garg, Manish; Binder, Robert J (2010) Ovalbumin-derived precursor peptides are transferred sequentially from gp96 and calreticulin to MHC class I in the endoplasmic reticulum. J Immunol 184:5619-27