This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. The known roles of Heat Shock Proteins (HSPs) can be grouped into two broad categories: chaperones of antigenic peptides and stimulators of innate immunity. Both of these functions depend on HSP interaction with receptors on Antigen Presenting Cells (APC). It is not clear which HSP receptors are involved with the innate versus the adaptive component of HSP-elicited immunity. Receptors that signal but do not endocytose (i.e. are logical candidates for innate immunity) include TLR2, TLR4, CD40 and CD36. Receptors that endocytose (i.e. are logical candidates for adaptive immunity) include LOX-1 and CD91. CD91 was the first HSP receptor to be identified and its role in HSP-elicited immunity is presently the most thoroughly understood. This study aims to further assess and identify HSP receptors on antigen-presenting murine (mouse) and human Dendritic Cells (DC). This will be accomplished via two mostly independent experimental aims.
Aim 1 a will assess the role of LOX-1 and other HSP receptors relative to CD91 in the cross-presentation of HSP-chaperoned peptides.
Aim 1 b will generate monoclonal antibodies to human HSP receptors to identify new HSP receptors and better characterize already-known receptors.
Aim 1 a will utilize established mouse and human re-presentation systems to evaluate the ability of receptor ligands to inhibit the re-presentation of HSP-chaperoned antigens. Inhibition of the re-presentation system would support the involvement of that particular receptor in the cross-presentation of HSP-chaperoned antigens. The role of CD40 will be further investigated by determining the ability of a DM40-/-mouse to generate an antigen-specific CD8+ T-cell response.
Aim 1 b will utilize blood monocyte-derived immature human DCs to generate monoclonal antibodies in mice and rats. Following immunization and sera collection, the spleen cells will be fused with mouse or rat myeloma cells to create hybridomas. The resulting monoclonal anti-human DC antibodies will be tested for their ability to inhibit re-presentation of HSP-chaperoned antigen. Hybridomas that produce anti-HSP receptor antibodies will be further characterized. The results of this study have enormous potential for affecting human health. Numerous studies have shown that HSP-peptide complexes isolated from cancer or virus-infected cells can be used as vaccines against the cancer or virus concerned without eliciting detectable autoimmunity.
Showing the most recent 10 out of 638 publications
