Enhancement of cytotoxic T lymphocyte activity by dendritic cells loaded with Tat-protein transduction domain-fused hepatitis B virus core antigen.
The protein transduction domain (PTD) of human immunodeficiency virus-1-Tat protein has a unique potency to penetrate the cellular membranes. To synthesize the sequence of Tat-PTD47-57 and hepatitis B virus core antigen (HBcAg), we spliced these sequences and linked a fusion gene into the pMAL-c2x vector. The fusion proteins were purified by affinity chromatography and pulsed with bone marrow-derived dendritic cells (DCs), and the transduction of recombinant protein was detected by immunofluorescence antibody assay. Results showed that recombinant PTD-HBcAg could penetrate into DC cytoplasm while recombinant HBcAg was detected on the surface of cells. The percentage of DC surface molecules, such as CD80, CD86 and major histocompatibility complex II, and production of cytokine (IL-12p70) induced by recombinant PTD-HBcAg were significantly higher than those induced by recombinant HBcAg or tumor necrosis factor-alpha. DCs treated with PTD-HBcAg induced T cells to differentiate into specific cytotoxic T lymphocytes (CTLs) and enhanced the CTL killing response. In conclusion, the expressed and purified PTD-HBcAg fusion protein could penetrate into cells through the plasma membrane, promote DC maturation, and enhance T cells response to generate HBcAg-specific CTLs efficiently.
Chen
Xiaohua
X
Department of Infectious Disease, Shanghai No. 6 People's Hospital, College of Medicine, Shanghai Jiaotong University, Shanghai, China.
Yu
Yongsheng
Y
Pan
Qingchun
Q
Tang
Zhenghao
Z
Han
Jinchao
J
Zang
Guoqing
G
D000818:Animals; D001483:Base Sequence; D002846:Chromatography, Affinity; D017931:DNA Primers; D003713:Dendritic Cells / Q000276:immunology*; D015696:Gene Products, tat / Q000276:immunology*; D006512:Hepatitis B Core Antigens / Q000276:immunology*; D006801:Humans; D007150:Immunohistochemistry; D018664:Interleukin-12 / Q000096:biosynthesis; D007959:Lymphocyte Culture Test, Mixed; D051379:Mice; D008807:Mice, Inbred BALB C; D013602:T-Lymphocytes, Cytotoxic / Q000276:immunology*
D017931:DNA Primers; D015696:Gene Products, tat; D006512:Hepatitis B Core Antigens; D018664:Interleukin-12