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Heat shock protein-peptide complexes, reconstituted in vitro, elicit peptide-specific cytotoxic T lymphocyte response and tumor immunity.

Blachere NE, Li Z, Chandawarkar RY, Suto R, Jaikaria NS, Basu S, Udono H, Srivastava PK - J. Exp. Med. (1997)

Bottom Line: The immunogenicity of HSP preparations has been attributed to peptides associated with the HSPs.The studies reported here demonstrate that immunogenic HSP-peptide complexes can also be reconstituted in vitro.These observations demonstrate that HSPs are CD8+ T cell response-eliciting adjuvants.

View Article: PubMed Central - PubMed

Affiliation: Center for Immunotherapy of Cancer and Infectious Diseases, University of Connecticut School of Medicine, Farmington 06030, USA.

ABSTRACT
Heat shock protein (HSP) preparations derived from cancer cells and virus-infected cells have been shown previously to elicit cancer-specific or virus-specific immunity. The immunogenicity of HSP preparations has been attributed to peptides associated with the HSPs. The studies reported here demonstrate that immunogenic HSP-peptide complexes can also be reconstituted in vitro. The studies show that (a) complexes of hsp70 or gp96 HSP molecules with a variety of synthetic peptides can be generated in vitro; (b) the binding of HSPs with peptides is specific in that a number of other proteins tested do not bind synthetic peptides under the conditions in which gp96 molecules do; (c) HSP-peptide complexes reconstituted in vitro are immunologically active, as tested by their ability to elicit antitumor immunity and specific CD8+ cytolytic T lymphocyte response; and (d) synthetic peptides reconstituted in vitro with gp96 are capable of being taken up and re-presented by macrophage in the same manner as gp96- peptides complexes generated in vivo. These observations demonstrate that HSPs are CD8+ T cell response-eliciting adjuvants.

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Related in: MedlinePlus

Chaperoning of  peptides by HSPs is required for  generation of an effective CD8+  T cell response. gp96, hsp70, or  mouse serum albumin (MSA)  were complexed with radiolabeled VSV9 and analyzed by (A)  SDS-PAGE followed by Coomassie blue staining and autoradiography. In addition, mice  were immunized (B) with peptides complexed or simply mixed  with each of the proteins. Splenocytes of these mice were  tested for induction of CD8+ T  lymphocytes, as described in legend to Fig. 4. N1 (closed circle)  and EL4 (open circle) were used as  targets.
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Figure 5: Chaperoning of peptides by HSPs is required for generation of an effective CD8+ T cell response. gp96, hsp70, or mouse serum albumin (MSA) were complexed with radiolabeled VSV9 and analyzed by (A) SDS-PAGE followed by Coomassie blue staining and autoradiography. In addition, mice were immunized (B) with peptides complexed or simply mixed with each of the proteins. Splenocytes of these mice were tested for induction of CD8+ T lymphocytes, as described in legend to Fig. 4. N1 (closed circle) and EL4 (open circle) were used as targets.

Mentions: Reconstitution of peptides with hsp70 molecules was observed to require neither a heating and cooling cycle, nor exposure to high salt concentrations. Incubation of apparently homogeneous preparations of hsp70 with radiolabeled peptide A in sodium phosphate buffer containing 1 mM ADP and 1 mM MgCl2 at 37°C was found to be sufficient to generate SDS-stable hsp70–peptide complexes as judged by autoradiography (see Fig. 5 A).


Heat shock protein-peptide complexes, reconstituted in vitro, elicit peptide-specific cytotoxic T lymphocyte response and tumor immunity.

Blachere NE, Li Z, Chandawarkar RY, Suto R, Jaikaria NS, Basu S, Udono H, Srivastava PK - J. Exp. Med. (1997)

Chaperoning of  peptides by HSPs is required for  generation of an effective CD8+  T cell response. gp96, hsp70, or  mouse serum albumin (MSA)  were complexed with radiolabeled VSV9 and analyzed by (A)  SDS-PAGE followed by Coomassie blue staining and autoradiography. In addition, mice  were immunized (B) with peptides complexed or simply mixed  with each of the proteins. Splenocytes of these mice were  tested for induction of CD8+ T  lymphocytes, as described in legend to Fig. 4. N1 (closed circle)  and EL4 (open circle) were used as  targets.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC2199095&req=5

Figure 5: Chaperoning of peptides by HSPs is required for generation of an effective CD8+ T cell response. gp96, hsp70, or mouse serum albumin (MSA) were complexed with radiolabeled VSV9 and analyzed by (A) SDS-PAGE followed by Coomassie blue staining and autoradiography. In addition, mice were immunized (B) with peptides complexed or simply mixed with each of the proteins. Splenocytes of these mice were tested for induction of CD8+ T lymphocytes, as described in legend to Fig. 4. N1 (closed circle) and EL4 (open circle) were used as targets.
Mentions: Reconstitution of peptides with hsp70 molecules was observed to require neither a heating and cooling cycle, nor exposure to high salt concentrations. Incubation of apparently homogeneous preparations of hsp70 with radiolabeled peptide A in sodium phosphate buffer containing 1 mM ADP and 1 mM MgCl2 at 37°C was found to be sufficient to generate SDS-stable hsp70–peptide complexes as judged by autoradiography (see Fig. 5 A).

Bottom Line: The immunogenicity of HSP preparations has been attributed to peptides associated with the HSPs.The studies reported here demonstrate that immunogenic HSP-peptide complexes can also be reconstituted in vitro.These observations demonstrate that HSPs are CD8+ T cell response-eliciting adjuvants.

View Article: PubMed Central - PubMed

Affiliation: Center for Immunotherapy of Cancer and Infectious Diseases, University of Connecticut School of Medicine, Farmington 06030, USA.

ABSTRACT
Heat shock protein (HSP) preparations derived from cancer cells and virus-infected cells have been shown previously to elicit cancer-specific or virus-specific immunity. The immunogenicity of HSP preparations has been attributed to peptides associated with the HSPs. The studies reported here demonstrate that immunogenic HSP-peptide complexes can also be reconstituted in vitro. The studies show that (a) complexes of hsp70 or gp96 HSP molecules with a variety of synthetic peptides can be generated in vitro; (b) the binding of HSPs with peptides is specific in that a number of other proteins tested do not bind synthetic peptides under the conditions in which gp96 molecules do; (c) HSP-peptide complexes reconstituted in vitro are immunologically active, as tested by their ability to elicit antitumor immunity and specific CD8+ cytolytic T lymphocyte response; and (d) synthetic peptides reconstituted in vitro with gp96 are capable of being taken up and re-presented by macrophage in the same manner as gp96- peptides complexes generated in vivo. These observations demonstrate that HSPs are CD8+ T cell response-eliciting adjuvants.

Show MeSH
Related in: MedlinePlus