[Construction, expression and targeting therapeutic of single-chain immunotoxin against hepatocellular carcinoma].
OBJECTIVE
To obtain high therapeutic effect and low toxicity single-chain immunotoxin against hepatocellular carcinoma (HCC).


METHODS
Human mutant tumor necrosis factor-alpha (mTNFalpha) was linked with the 3' end of humanized single-chain Fv against HCC (hscFv25) in pGEX4T-1 vector. The anti-HCC immunotoxin was expressed in Escherichia coli and identified by western blot. The primary tumor regression trial in nude mice bearing HCC was evaluated the targeting therapeutic value of hscFv25-mTNFalpha. The tumor tissues were stained by immunohistochemical with TNFalpha antibody.


RESULTS
The expression of single-chain immunotoxin hscFv25-mTNFalpha was 12% of total bacteria proteins. The tumor regression trials of hscFv25-mTNFalpha showed 5/5 effective. It had 2/5 completely remission and 3/5 partly remission. The therapeutic result of hscFv25-mTNFalpha was better than that of mTNFalpha (F=8.70, 0.05). The HCC tissue treated by hscFv25-mTNFalpha expressed TNFalpha positive reaction. The positive granule mainly existed in HCC cytoplasm.


CONCLUSIONS
The single-chain immunotoxin hscFv25-mTNFalpha has high therapeutic effect and low toxicity. It has potentialities for clinical application.
Zhang
Jing
J
Department of Pathology, Xijing Hospital, Fourth Military Medical University, Xi'an 710033, China.
Liu
Yan-fang
YF
Yang
Shou-jing
SJ
Qiao
Qing
Q
Cheng
Hong
H
Ma
Fu-cheng
FC
Chen
Wan-lu
WL
D000818:Animals; D007128:Immunoglobulin Fragments / Q000627:therapeutic use*; D007150:Immunohistochemistry; D000922:Immunotoxins / Q000627:therapeutic use*; D008114:Liver Neoplasms, Experimental / Q000628:therapy*; D051379:Mice; D008807:Mice, Inbred BALB C; D014409:Tumor Necrosis Factor-alpha / Q000627:therapeutic use*
D007128:Immunoglobulin Fragments; D000922:Immunotoxins; D014409:Tumor Necrosis Factor-alpha; C055966:immunoglobulin Fv