Identification of peptide mimotopes of gp96 using single-chain antibody library.
Heat shock proteins such as gp96 are immunogenic and are widely used as vaccines in immunotherapy of cancers. The present study focuses on the use of peptide mimotopes as immunotherapeutic vaccines for prostate cancer. To this end, we developed a 15-mer gp96 peptide mimotope specifically reactive to MAT-LyLu gp96-peptide complex using combinatorial single-chain antibody and peptide phage display library. The immunogenicity of the synthesized gp96 mimotope was analyzed initially in normal BALB/c mice in combination with various adjuvants such as complete Freund's adjuvant (CFA), aluminum salts (ALUM), granulocyte-macrophage colony-stimulating factor (GM-CSF), and liposome, of which CFA served as a positive control. The antibody response was determined and found that the gp96 mimotope with ALUM showed a significant increase in antibody titer, followed by GM-CSF and liposomes. Further, the T cell (CD4(+) and CD8(+)) populations from splenocytes, as well as IgG isotypes, interleukin-4, and interleukin-5 of gp96 mimotope with ALUM-immunized animals, were analyzed. The results suggest that the gp96 mimotope may elicit a potent and effective antitumor antibody response. Further, the study identifies ALUM and GM-CSF as adjuvant options to drive an appropriate protective immune response as these adjuvants have prior use in humans.
Shanmugam
Arulkumaran
A
Department of Microbiology and Immunology, New York Medical College, Valhalla, NY 10595, USA.
Suriano
Robert
R
Goswami
Neha
N
Chaudhuri
Devyani
D
Ashok
Badithe T
BT
Rajoria
Shilpi
S
George
Andrea L
AL
Mittelman
Abraham
A
Tiwari
Raj K
RK
D000818:Animals; D002460:Cell Line; D004797:Enzyme-Linked Immunosorbent Assay; D000939:Epitopes / Q000097:blood* / Q000737:chemistry; D008297:Male; D008562:Membrane Glycoproteins / Q000276:immunology; D051379:Mice; D008807:Mice, Inbred BALB C; D018716:Molecular Mimicry; D019151:Peptide Library; D010455:Peptides / Q000276:immunology; D011471:Prostatic Neoplasms / Q000276:immunology / Q000517:prevention & control; D051381:Rats; D057127:Single-Chain Antibodies / Q000097:blood*; D014612:Vaccines / Q000276:immunology
D000939:Epitopes; D008562:Membrane Glycoproteins; D019151:Peptide Library; D010455:Peptides; D057127:Single-Chain Antibodies; D014612:Vaccines; C053387:endoplasmin