[Establishment of B16-HLA-MAGE-3 melanoma cell line coexpressing HLA-A 0201/K(b) and MAGE-3 and its role in tumor vaccine evaluation].
OBJECTIVE
To establish a tumor model in HLA-A2.1 transgenic mice to examine the efficacy of MAGE-3 vaccine, a cell line coexpressing HLA-A 0201/K(b) and MAGE-3 is established.


METHODS
B16-HLA-MAGE-3 melanoma was obtained by means of cotransfection of HLA-A 0201/K(b) and MAGE-3 to B16 melanoma. RT-PCR, FCM analysis and Western blot were used to detect the mRNA or protein of HLA-A 0201/K(b) or MAGE-3 expression in B16-HLA-MAGE-3. The ability of MAGE-3 antigen to be processed and presented in the B16-HLA-MAGE-3 cell line were observed by CTL activity detection and tumor challenge test.


RESULTS
Transcription and protein expression of HLA-A 0201/H-2k(b) and MAGE-3 were demonstrated in B16-HLA-MAGE-3 cells. CTL activity of splenocytes in immunized mice against B16-HLA-MAGE-3 was detected and the growth of B16-HLA-MAGE-3 in immunized mice was also inhibited.


CONCLUSIONS
MAGE-3 antigen is able to be processed and presented efficiently by B16-HLA-MAGE-3 melanoma cells and this cell can be employed to test HLA-A2 restricted epitope immunogenicity in the A2-transgenic mice.
Song
Shu-Xia
SX
Department of Animal Laboratory, Hebei Medical University, Shijiazhuang 050017, China.
Yan
Cai-Zhen
CZ
Zheng
Long
L
You
Hong-Yu
HY
Wang
Jun-Xia
JX
Liu
Fu-Ying
FY
D000818:Animals; D000951:Antigens, Neoplasm / Q000235:genetics / Q000276:immunology* / Q000378:metabolism; D015153:Blotting, Western; D019496:Cancer Vaccines / Q000235:genetics / Q000276:immunology* / Q000378:metabolism; D002460:Cell Line; D006680:HLA Antigens / Q000235:genetics / Q000276:immunology* / Q000378:metabolism; D008546:Melanoma, Experimental / Q000276:immunology* / Q000378:metabolism; D051379:Mice; D008822:Mice, Transgenic; D009363:Neoplasm Proteins / Q000235:genetics / Q000276:immunology* / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction
D000951:Antigens, Neoplasm; D019496:Cancer Vaccines; D006680:HLA Antigens; C072013:MAGEA3 protein, human; D009363:Neoplasm Proteins