PMID- 16923618
OWN - NLM
STAT- MEDLINE
DCOM- 20070504
LR  - 20080424
IS  - 1465-3249 (Print)
IS  - 1465-3249 (Linking)
VI  - 8
IP  - 4
DP  - 2006
TI  - A mutated human tumor necrosis factor-alpha improves the therapeutic index in 
      vitro and in vivo.
PG  - 415-23
AB  - BACKGROUND: Tumor necrosis factor-alpha (TNF-alpha) is a multifunctional cytokine 
      that has cytotoxic, cytostatic and immunomodulatory effects on malignant tumors. 
      However, clinical trials have revealed high systemic toxicity and this has 
      hampered its utilization as an anti-cancer agent. In this study, a human 
      TNF-alpha mutant was created and tested for its anti-tumor effects. METHODS: The 
      TNF mutant (recombinant mutated human TNF; rmhTNF) was prepared by protein 
      engineering in which amino acids Pro, Ser and Asp at positions 8, 9 and 10 of 
      TNF-alpha were substituted by Arg, Lys and Arg, and C terminal Leu157 was 
      substituted by Phe, along with deletion of the first seven N-terminal amino 
      acids. Prokaryotic expression recombinant vector pBV-mhTNF containing the PLPR 
      promotor was constructed and transformed into E. coli DH5alpha. The rmhTNF was 
      expressed in a partially soluble form in DH5alpha, purified from the supernatant 
      of cell lysate by ammonia sulfate precipitation and two sequential 
      chromatographic steps. RESULTS: The purified rmhTNF was >95% pure by SDS-PAGE 
      stained with silver and high-pressure size exclusion chromatography (SEC-HPLC). 
      Its yield was about 1.22 mg/g wet cell paste. The mutant rmhTNF exhibited an 
      approximately 50-fold increase in cytotoxicity relative to the wild-type rhTNF on 
      the mouse fibroblast cell line L929 in a standard cytotoxicity test, and at least 
      and at least 50 times higher LD50 as wild type rhTNF in mice. In vivo biological 
      activity studies carried out on tumor cell transplanted mice and nude mice also 
      showed a more effective cytotoxicity of rmhTNF than rhTNF. DISCUSSION: These 
      results suggest that rmhTNF has potential for developing an effective anti-tumor 
      reagent for some tumors.
FAU - Yan, Z
AU  - Yan Z
AD  - Biotechnology Center, School of Pharmacy of Fourth Military Medical University, 
      Shaanxi, People's Republic of China.
FAU - Zhao, N
AU  - Zhao N
FAU - Wang, Z
AU  - Wang Z
FAU - Li, B
AU  - Li B
FAU - Bao, C
AU  - Bao C
FAU - Shi, J
AU  - Shi J
FAU - Han, W
AU  - Han W
FAU - Zhang, Y
AU  - Zhang Y
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PL  - England
TA  - Cytotherapy
JT  - Cytotherapy
JID - 100895309
RN  - 0 (Antineoplastic Agents)
RN  - 0 (Recombinant Proteins)
RN  - 0 (Tumor Necrosis Factor-alpha)
SB  - IM
MH  - Animals
MH  - *Antineoplastic Agents/metabolism/therapeutic use/toxicity
MH  - Cell Line, Tumor
MH  - Dose-Response Relationship, Drug
MH  - Female
MH  - Humans
MH  - Male
MH  - Mice
MH  - Mice, Inbred BALB C
MH  - Mice, Inbred C57BL
MH  - Mice, Nude
MH  - *Mutation
MH  - Neoplasm Transplantation
MH  - *Recombinant Proteins/genetics/metabolism/therapeutic use/toxicity
MH  - *Tumor Necrosis Factor-alpha/genetics/metabolism/therapeutic use/toxicity
EDAT- 2006/08/23 09:00
MHDA- 2007/05/05 09:00
CRDT- 2006/08/23 09:00
PHST- 2006/08/23 09:00 [pubmed]
PHST- 2007/05/05 09:00 [medline]
PHST- 2006/08/23 09:00 [entrez]
AID - G249252T80382P47 [pii]
AID - 10.1080/14653240600845278 [doi]
PST - ppublish
SO  - Cytotherapy. 2006;8(4):415-23. doi: 10.1080/14653240600845278.