Molecular adjuvant effects of a conformationally biased agonist of human C5a anaphylatoxin.
A conformationally biased decapeptide agonist of human C5a anaphylatoxin (YSFKPMPLaR) was used as a molecular adjuvant in stimulating Ab responses against peptide epitopes derived from human MUC1 glycoprotein and the human mu and kappa opioid receptors. C57BL6 mice were immunized with the MUC1 epitope (YKQGGFLGL); the C5a agonist (YSFKPMPLaR); YSFKPMPLaR and YKQGGFLGL together, but unconjugated; a C5a-active, MUC1 epitope construct (YKQGGFLGLYSFKPMPLaR); and a C5a-inactive, reversed moiety construct (YSFKPMPLaRYKQGGFLGL). High Ab titers specific for the MUC1 epitope were observed only in mice immunized with the C5a-active epitope construct. Similar results were obtained in BALB/c mice immunized with the C5a-active, MUC1 epitope construct. Abs from the sera of the C57BL6 mice were predominately of the IgG2a, IgG2b, and IgM isotypes and were reactive against human recombinant MUC1 and MUC1 expressed by the Panc-1 M1F.15 pancreatic cell line. When compared with the corresponding KLH-epitope conjugates in C57BL6 mice, the epitope-C5a agonist constructs produced titers of specific IgG Abs of isotypes distinct from those generated by the keyhole limpet hemocyanin-epitope conjugates. Rabbits immunized with a mu opioid receptor epitope-C5a agonist construct (GDLSDPCGNRTNLGGRDSLYSFKPMPLaR) or a kappa opioid receptor epitope-C5a agonist construct (FPGWAEPDSNGSEDAQLYSFKPMPLaR) generated high titer, epitope-specific Ab responses. Ab titers generated in response to the opioid epitope-C5a agonist constructs were comparable to those generated by the opioid KLH-epitope conjugates. The results of this study are discussed in terms of possible mechanisms by which the conformationally biased C5a agonist serves as a molecular adjuvant.
Tempero
R M
RM
Eppley Institute for Research in Cancer and Allied Diseases, University of Nebraska Medical Center, Omaha 68198, USA.
Hollingsworth
M A
MA
Burdick
M D
MD
Finch
A M
AM
Taylor
S M
SM
Vogen
S M
SM
Morgan
E L
EL
Sanderson
S D
SD
D000276:Adjuvants, Immunologic / Q000737:chemistry; D000595:Amino Acid Sequence; D000706:Anaphylatoxins / Q000737:chemistry*; D000818:Animals; D015936:Complement C5a / Q000819:agonists*; D018604:Epitope Mapping; D006801:Humans; D007158:Immunologic Techniques; D051379:Mice; D008807:Mice, Inbred BALB C; D008810:Mice, Inbred C57BL; D008969:Molecular Sequence Data; D018396:Mucin-1 / Q000276:immunology*; D010455:Peptides / Q000737:chemistry; D011487:Protein Conformation; D011817:Rabbits; D017473:Receptors, Opioid, kappa / Q000276:immunology; D017450:Receptors, Opioid, mu / Q000276:immunology
D000276:Adjuvants, Immunologic; D000706:Anaphylatoxins; D018396:Mucin-1; D010455:Peptides; D017473:Receptors, Opioid, kappa; D017450:Receptors, Opioid, mu; D015936:Complement C5a