Prepared and screened a modified TNF-alpha molecule as TNF-alpha autovaccine to treat LPS induced endotoxic shock and TNF-alpha induced cachexia in mouse.
Overexpression of TNF-alpha in the body is critically involved in many diseases. A strategy to construct TNF-alpha autovaccine by introducing a T cell helper epitope to the protein has been developed and may be an alternative because it is cheaper and highly efficient. However, the induction of high level anti-TNF-alpha neutralizing autoantibodies by TNF-alpha autovaccine is depend on a proper T cell help epitope. In order to evaluate the effect of different T helper cell epitopes on the immunogenicity of mouse TNF-alpha (mTNF-alpha), three T helper cell epitopes, TT (QYIKANSKFIGITEL), HEL (NTDGSTDYGILQINSR), and PADRE (AKFVAAWTLKA), were chosen for this study. The sequence (amino acids 126-140) of mTNF-alpha was replaced with those of the T cell help epitopes, respectively. The three fusion proteins (mTNF-TT, mTNF-HEL, mTNF-PADRE) were expressed in Escherichia coli and purified with a simple strategy. The abilities of the proteins elicited TNF-alpha autoantibodies in BALB/c mice were investigated. The results showed that mTNF-PADRE is the most effective among the three modified TNF-alpha molecules. In the absence of adjuvant, the therapeutic effect of TNF-PADRE on LPS induced endotoxic shock mice and mTNF-alpha induced cachexia mice was observed. This study suggests that mTNF-PADRE may be a better candidate of mTNF-alpha autovaccine.
Wan
Yi
Y
Biotechnology Center of The Fourth Military Medical University, 17 Changle West Road, 710032 Xi'an, People's Republic of China.
Xue
Xiaochang
X
Li
Meng
M
Zhang
Xiaoyong
X
Qin
Xin
X
Zhang
Cun
C
You
Yanjie
Y
Wang
Weihua
W
Jiang
Changli
C
Wu
Shouzhen
S
Liu
Yan
Y
Zhu
Wenhua
W
Ran
Yonggang
Y
Zhang
Zhen
Z
Han
Wei
W
Zhang
Yingqi
Y
D000595:Amino Acid Sequence; D000818:Animals; D001323:Autoantibodies / Q000097:blood; D002100:Cachexia / Q000188:drug therapy*; D018984:Epitopes, T-Lymphocyte / Q000235:genetics / Q000276:immunology / Q000627:therapeutic use*; D008070:Lipopolysaccharides; D008297:Male; D051379:Mice; D008807:Mice, Inbred BALB C; D008969:Molecular Sequence Data; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000276:immunology / Q000627:therapeutic use; D012772:Shock, Septic / Q000139:chemically induced / Q000188:drug therapy*; D014409:Tumor Necrosis Factor-alpha / Q000737:chemistry / Q000235:genetics / Q000276:immunology* / Q000627:therapeutic use; D014612:Vaccines / Q000737:chemistry / Q000276:immunology / Q000627:therapeutic use*
D001323:Autoantibodies; D018984:Epitopes, T-Lymphocyte; D008070:Lipopolysaccharides; D011993:Recombinant Fusion Proteins; D014409:Tumor Necrosis Factor-alpha; D014612:Vaccines; C523999:mTNF-PADRE