Protective effects of a novel trimerized sTNFRII on acute liver injury

Luo, Mansheng; Liu, Dan; Zhang, Lingmin; Huang, Shigao; Yang, Wenjuan; Zhang, Lin; Cui, Tong; Ma, Honghui; Wang, Zhen; Sun, Qian; Xiong, Chunhui; Zhu, Jian-Hong; Li, Jinlong; Wu, Xiaobing; Jin, Liqin; Hu, Zhiming; Gao, Jimin

doi:10.1016/j.intimp.2012.03.013

Cited by 8 publications

(3 citation statements)

References 27 publications

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“…Neutralizing activity of sTNFRII-gAD was significantly higher than that of sTNFRII-Fc ( p <0.05) ( Fig. 9 ), which is consistent with our previous observation showing that sTNFRII-gAD was able to attenuate D-galactosamine/LPS-induced acute liver injury resulted from excessive TNFα more efficaciously than sTNFRII-Fc [10] .…”

Section: Resultssupporting

confidence: 92%

“…Utilizing the intrinsic trimerization property of gAD, we have developed a novel TNFα antagonist, the trimerized fusion protein named sTNFRII-gAD, which was composed of sTNFRII and gAD. We had shown that sTNFRII-gAD was superior to sTNFRII-Fc as a TNFα antagonist, highlighting the potential of sTNFRII-gAD for the treatment of excessive TNFα-associated diseases [10] . However, our previously reported sTNFRII-gAD expression system yielded less than 10 mg/L of sTNFRII-gAD.…”

Section: Introductionmentioning

confidence: 99%

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Efficient Production of sTNFRII-gAD Fusion Protein in Large Quantity by Use of the Modified CHO-S Cell Expression System

et al. 2014

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TNFα is one of the initial and important mediators to activate downstream signaling pathways by binding to trimerized TNFα receptors (TNFR), and thus is an ideal drug target for cancer therapy. Taking advantage of intrinsic homotimerization of the globular domain of adiponectin (gAD), we have developed a novel TNFα antagonist, the trimerized fusion protein named sTNFRII-gAD. However, our previously-used CHO expression system yielded less than 10 mg/L of sTNFRII-gAD. To produce large quantities of sTNFRII-gAD efficiently, we used a modified CHO-S cell expression system, which is based on a pMH3 vector with non-coding GC-rich DNA fragments for high-level gene expression. We obtained stable clones that produced 75 mg/L of sTNFRII-gAD in the 96-well plate, adapted the clones to 40 ml suspension serum-free batch culture, then optimized the culturing conditions to scale up the fed-batch culture in a 3 L shake-flask and finally in a 5 L AP30 bioreactor. We achieved a final yield of 52 mg/L of sTNFRII-gAD. The trimerized sTNFRII-gAD exhibited the higher affinity to TNFα with a dissociation constant (Kd) of 5.63 nM than the dimerized sTNFRII-Fc with a Kd of 13.4 nM, and further displayed the higher TNFα-neutralizing activity than sTNFRII-Fc (p<0.05) in a L929 cytotoxicity assay. Therefore, the strategy employed in this study may provide an efficient avenue for large-scale production of other recombinant proteins by use of the modified CHO-S cell expression system.

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Section: Resultssupporting

confidence: 92%

Section: Introductionmentioning

confidence: 99%

Efficient Production of sTNFRII-gAD Fusion Protein in Large Quantity by Use of the Modified CHO-S Cell Expression System

et al. 2014

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“…To this end, we developed a novel sTNFRII by fusing with gAD, which tends to trimerize spontaneously. In line with our hypothesis, the fusion protein sTNFRII-gAD was proven to efficaciously attenuate D-galactosamine/lipopolysaccharide-induced liver injury in which TNFa played a pivotal role (Luo et al, 2012).…”

Section: Discussionsupporting

confidence: 76%

Preventive Effect of a Novel Recombinant sTNFRII on Collagen-Induced Arthritis

Luo

Zhao

Gao

et al. 2015

Immunological Investigations

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We developed a novel trimeric sTNFRII fusion protein, named sTNFRII-gAD, which exhibited a higher in vitro antagonistic efficacy for TNFα in comparison with sTNFRII-Fc. This study aimed to investigate the arthritic protection of sTNFRII-gAD in a rat collagen-induced arthritis (CIA). The rats were injected intradermally with collagen type II at days 0 and 7. Three days after the second injection (day 10), the rats were intraperitoneally given sTNFRII-gAD or sTNFRII-Fc, or PBS. Effects of treatments were examined with respect of CIA incidence, severity and pathological changes. Serum TNFα, IL-17A and regulatory T cell (Treg) in periphery were determined at days 10 and 16, respectively. Our results showed that sTNFRIIgAD significantly reduced CIA incidence and severity (p < 0.05); meanwhile it led to a dramatic improvement in cartilage and bone damage. Moreover, the increase in serum anti-CII and IL-17A, and the reduction in Treg population were inhibited (p < 0.05) by sTNFRII-gAD or sTNFRII-Fc. Serum TNFα was found to be accumulated in the groups treated with sTNFRII-gAD or sTNFRII-Fc compared with the group treated with PBS (p < 0.05). It is noteworthy that sTNFRII-gAD displayed a better efficacy than sTNFRII-Fc in CIA incidence, pathological changes in cartilage and the elevation of anti-CII antibody, indicating that sTNFRII-gAD is potentially a more efficacious anti-TNFα agent for rheumatoid arthritis.

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