title: A Tobacco-Derived Thymosin β4 Concatemer Promotes Cell Proliferation and Wound Healing in Mice.
abstract: Thymosin β4 (Tβ4) is a peptide that is known to play important roles in protection, regeneration, and remodeling of injured tissues in humans, and that shows great promise in a range of clinical applications. However, current strategies to Tβ4 are insufficient to meet growing demand and have a number of limitations. In this current study we investigated whether expression of recombinant Tβ4 in plants, specifically in tobacco (Nicotiana tabacum) leaves, represents an effective approach. To address this question, a 168 bp Tβ4 gene optimized for tobacco codon usage bias was constitutively expressed in tobacco as a 4-unit repeat concatemer, fused to a polyhistidine tag. Quantitative polymerase chain reaction and Western blot analyses were used to verify 4×Tβ4 expression in 14 transgenic tobacco lines and enzyme-linked immunosorbent assay analysis indicated 4×Tβ4 protein concentrations as high as 3 μg/g of fresh weight in the leaves. We observed that direct administration of tobacco-derived Tβ4 was more effective than Tβ4 either obtained commercially or derived from expression in Escherichia coli at promoting splenocyte proliferation in vitro and wound healing in mice through an endothelial migration assay. This study provides new insights into the development of plant-derived therapeutic proteins and their application by direct administration.
authors: Joint Tomato Research Institute, School of Agriculture and Biology, Shanghai Jiao Tong University, Shanghai 200240, China; Plant Biotechnology Research Center, School of Agriculture and Biology, Shanghai Jiao Tong University, Shanghai 200240, China.
authors: Rylosona
authors: R
authors: Janarthini
authors: Joint Tomato Research Institute, School of Agriculture and Biology, Shanghai Jiao Tong University, Shanghai 200240, China; Plant Biotechnology Research Center, School of Agriculture and Biology, Shanghai Jiao Tong University, Shanghai 200240, China.
authors: Xiaolei
authors: X
authors: Wang
authors: Joint Tomato Research Institute, School of Agriculture and Biology, Shanghai Jiao Tong University, Shanghai 200240, China; Plant Biotechnology Research Center, School of Agriculture and Biology, Shanghai Jiao Tong University, Shanghai 200240, China.
authors: Lulu
authors: L
authors: Chen
authors: Joint Tomato Research Institute, School of Agriculture and Biology, Shanghai Jiao Tong University, Shanghai 200240, China; Plant Biotechnology Research Center, School of Agriculture and Biology, Shanghai Jiao Tong University, Shanghai 200240, China.
authors: Lei
authors: L
authors: Gao
authors: Joint Tomato Research Institute, School of Agriculture and Biology, Shanghai Jiao Tong University, Shanghai 200240, China; Plant Biotechnology Research Center, School of Agriculture and Biology, Shanghai Jiao Tong University, Shanghai 200240, China.
authors: Lingxia
authors: 0000-0001-5144-2413
authors: L
authors: Zhao
mesh_terms: D000818:Animals; D049109:Cell Proliferation / Q000187:drug effects*; D004926:Escherichia coli; D018506:Gene Expression Regulation, Plant / Q000187:drug effects; D006801:Humans; D051379:Mice; D011994:Recombinant Proteins / Q000008:administration & dosage / Q000096:biosynthesis* / Q000235:genetics; D013154:Spleen / Q000187:drug effects / Q000473:pathology; D013947:Thymosin / Q000008:administration & dosage / Q000096:biosynthesis* / Q000235:genetics; D014026:Nicotiana / Q000737:chemistry / Q000235:genetics; D014945:Wound Healing / Q000187:drug effects*; D000099201:Thymosin beta(4)
chemical_list: D011994:Recombinant Proteins; D013947:Thymosin; D000099201:Thymosin beta(4)