Cysteine-tailed class I-binding peptides bind to CpG adjuvant and enhance primary CTL responses.
Immunostimulatory CpG motifs in synthetic oligonucleotides can be effective adjuvants for the priming of CTLs. We first observed that a single male-specific peptide (KCSRNRQYL) (HY2) was more efficient than another male-specific peptide (WMHHNMDLI) (HY1) at priming IFN-gamma-secreting CTLs in vivo when combined with lipid A and CpG and that it also visibly precipitated CpG. The addition of the six N-terminal residues (KCSRNR) from HY2 to HY1 yielded a peptide, KCSRNR-HY1, that both precipitated CpG and primed increased numbers of HY1-specific CTLs. We refer to this type of peptide as a primotope that includes a class I binding peptide tailed with amino acids that increase priming. Ala residues were substituted for the Arg/Lys residues (ACSANA-HY1), and these substitutions did not reduce in vivo priming potential. However, the substitution of Ala for Cys (KASRNR-HY1) resulted in the complete loss of priming, demonstrating the importance of Cys for in vivo priming when mixed with CpG. This result suggested that increased priming was based in disulfide bonding between Cys residues and internal phosphorothioate groups of synthetic CpG. The addition of Cys-bearing primotopes to radiolabeled CpG with a single thioate group resulted in the appearance of a new band that was inhibited by 1) Cys > Ala substitution and 2) reduction and alkylation of CpG. These results reveal a novel mechanism for complexing class I binding peptides and CpG adjuvant for development of new peptide-adjuvant combinations for vaccines for cancer and infectious diseases.
Wettstein
Peter J
PJ
Department of Immunology, Mayo Clinic, College of Medicine, Rochester, MN 55905, USA.
Borson
Nancy D
ND
Park
Jewn G
JG
McNallan
Kelly T
KT
Reed
Ann M
AM
D000276:Adjuvants, Immunologic / Q000737:chemistry* / Q000494:pharmacology; D000478:Alkylation; D000595:Amino Acid Sequence; D019943:Amino Acid Substitution; D000818:Animals; D017951:Antigen Presentation; D018899:CpG Islands; D003545:Cysteine; D006182:H-Y Antigen / Q000737:chemistry; D015395:Histocompatibility Antigens Class I / Q000737:chemistry / Q000276:immunology / Q000494:pharmacology*; D008297:Male; D051379:Mice; D008815:Mice, Inbred Strains; D009841:Oligonucleotides / Q000138:chemical synthesis* / Q000276:immunology / Q000494:pharmacology; D013329:Structure-Activity Relationship; D013602:T-Lymphocytes, Cytotoxic / Q000187:drug effects*; D014612:Vaccines / Q000737:chemistry* / Q000276:immunology
D000276:Adjuvants, Immunologic; D006182:H-Y Antigen; D015395:Histocompatibility Antigens Class I; D009841:Oligonucleotides; D014612:Vaccines; D003545:Cysteine