[Immunoadjuvant effect of Hsp70L1 in tumor vaccine].
OBJECTIVE
To explore the immune enhancement of Hsp70L1 in the tumor cell vaccines.


METHODS
TRP2(153-243) and Hsp70L1 genes were obtained by RT-PCR from B16 cells in murine melanoma and from spleens of C57BL/6 mice and then were inserted into pcDNA3.1/V5-His eukaryotic expression vectors respectively. The recombinants of pTRP2, pHSP70L1 or pTRP2-Hsp fusion gene were obtained and transfected into B16 cells respectively. TRP2(153-243), HSP70L1 or TRP2-Hsp fusion gene-expressing B16 cells were then induced to necrosis by freezing-thawing or to apoptosis by mitomycin C. C57BL/6 mice were immunized with the necrotic or apoptotic B16 cells twice, then the live B16 tumor cells were transplanted into the immunized mice and the tumor growth was observed in some tumor-bearing mice. IFN-gamma-producing cells in splenocytes were measured by flow cytometry and the CTL activity of spleno-lymphocyte was detected by LDH release assay.


RESULTS
After the normal mice were immunized with the necrotic or apoptotic tumor vaccines modified with TRP2(153-243), Hsp70L1 or TRP2-Hsp fusion genes, CTL lysis activity and IFN-gamma production from the splenic lymphocytes were promoted in the groups of Hsp70L1 and TRP2-Hsp modified tumor vaccines (P<0.05 or P<0.01). Additionally, the tumor growth was inhibited obviously in the groups of mice immunized with necrotic tumor vaccines (P<0.05 or P<0.01). However, no marked inhibition of tumor growth was observed in the groups of mice immunized with apoptotic tumor vaccines (P>0.05).


CONCLUSIONS
Hsp70L1 remarkably improves the immunogenicity of B16 tumor vaccines, especially that of necrotic tumor vaccines.
You
Hong-yu
HY
Department of Molecular Biology & Key Lab. of Experimental Animal Sciences, Hebei Medical University, Shijiazhuang 050017, Chinaa. you_hongyu@126.com
Zhang
Kai-xia
KX
Wang
Jun-xia
JX
Zhang
Huan-ling
HL
Lian
Wei-guang
WG
Wu
Wei
W
Song
Shu-xia
SX
D000276:Adjuvants, Immunologic / Q000096:biosynthesis / Q000235:genetics / Q000494:pharmacology*; D000818:Animals; D016827:CD8 Antigens / Q000378:metabolism; D019496:Cancer Vaccines / Q000276:immunology*; D045744:Cell Line, Tumor; D049109:Cell Proliferation; D003001:Cloning, Molecular; D005822:Genetic Vectors / Q000235:genetics; D018840:HSP70 Heat-Shock Proteins / Q000096:biosynthesis / Q000235:genetics / Q000276:immunology*; D007371:Interferon-gamma / Q000378:metabolism; D019746:Intramolecular Oxidoreductases / Q000235:genetics; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D013154:Spleen / Q000166:cytology / Q000276:immunology; D013602:T-Lymphocytes, Cytotoxic / Q000276:immunology; D014162:Transfection
D000276:Adjuvants, Immunologic; D016827:CD8 Antigens; D019496:Cancer Vaccines; D018840:HSP70 Heat-Shock Proteins; D007371:Interferon-gamma; D019746:Intramolecular Oxidoreductases; C042501:dopachrome isomerase