Enhanced immunogenicity of human papillomavirus 16 L1 genetic vaccines fused to an ER-targeting secretory signal peptide and RANTES.
To increase the potency of human papillomavirus (HPV) DNA vaccines, we constructed a series of HPV16 L1 vaccines genetically fused with a secretion signal and/or immune cell-recruiting RANTES. The DNA vaccines encoding secretory HPV L1 were constructed by inserting HPV L1 gene into a vector with an ER-targeting secretory signal sequence. The expression plasmid encoding secretory HPV L1 (pER/L1) was fused with cDNA of RANTES, generating pER/L1/R. For comparison, HPV L1 genes were cloned into pVAX1 vector with no signal sequence (pL1), and further linked to the N-terminus (pL1/R) or C-terminus of RANTES (pR/L1). The secretion of L1 proteins was observed in the pER/L1, pER/L1/R, and pR/L1-transfected cells, except the pL1/R-transfected group. Cytoplasmic localization of L1 protein was observed in the cells transfected with pL1/R, but not with pER/L1/R at 48 h after transfection. In mice, RANTES-fused vaccines more effectively elicited the levels of HPV16 L1-specific IgG and IgG2a antibodies than pL1. Of RANTES-fused vaccines, pER/L1/R encoding the secreted fusion protein induced the highest humoral and CD8(+) T-cell-stimulating responses. These results suggest that the immunogenicity of HPV L1 DNA vaccines could be enhanced by genetic fusion to a chemokine and secretory signal peptide sequences.
Kim
S J
SJ
Comprehensive Gynecologic Cancer Center, Pundang CHA General Hospital, Sungnam, Kyonggi-do, South Korea.
Lee
C
C
Lee
S Y
SY
Kim
I
I
Park
J S
JS
Sasagawa
T
T
Ko
J J
JJ
Park
S E
SE
Oh
Y-K
YK
D000914:Antibodies, Viral / Q000096:biosynthesis; D019677:Artificial Gene Fusion / Q000379:methods; D018414:CD8-Positive T-Lymphocytes / Q000276:immunology; D036022:Capsid Proteins; D018946:Chemokine CCL5 / Q000235:genetics / Q000276:immunology*; D004721:Endoplasmic Reticulum / Q000378:metabolism; D006801:Humans; D007111:Immunity, Cellular; D007074:Immunoglobulin G / Q000096:biosynthesis; D009856:Oncogene Proteins, Viral / Q000235:genetics / Q000276:immunology*; D027383:Papillomaviridae / Q000276:immunology*; D053918:Papillomavirus Vaccines; D021382:Protein Sorting Signals / Q000235:genetics; D021381:Protein Transport; D014162:Transfection; D019444:Vaccines, DNA / Q000276:immunology*; D014765:Viral Vaccines / Q000276:immunology*
D000914:Antibodies, Viral; D036022:Capsid Proteins; D018946:Chemokine CCL5; D007074:Immunoglobulin G; D009856:Oncogene Proteins, Viral; D053918:Papillomavirus Vaccines; D021382:Protein Sorting Signals; D019444:Vaccines, DNA; D014765:Viral Vaccines; C063317:L1 protein, Human papillomavirus type 16