Acellular tissue matrix scaffolds are much closer to tissue’s complex natural structure and biological characteristics,thus assess great advantages in cartilage engineering.We used rabbit costal cartilage to prepare ...Acellular tissue matrix scaffolds are much closer to tissue’s complex natural structure and biological characteristics,thus assess great advantages in cartilage engineering.We used rabbit costal cartilage to prepare acellular microfilaments and further 3D porous acellular cartilage scaffold via crosslinking.Poly(_L-lysine)/hyaluronic acid(PLL/HA)multilayer film was then built up onto the surface of the resulting porous scaffold.Furthermore,TGF-β3 was loaded into the PLL/HA multilayer film coated scaffold to obtain a 3D porous acellular cartilage scaffold with sustained releasing of TGF-β3 up to 60 days.The success of this project will provide a new way for the treatment of articular cartilage defects.Meanwhile,the anchoring and on-site sustained releasing of growth factors mediated by polyelectrolyte multilayered film can also provide a new method for improving the biocompatibility and the biofunctionality for other implanted biomaterials.展开更多
tThe tumor suppressor p53 is the most common mutated gene in cancer,with the R175H as the most frequent p53 missense mutant.However,there are currently no approved targeted therapies or immunotherapies against mutant ...tThe tumor suppressor p53 is the most common mutated gene in cancer,with the R175H as the most frequent p53 missense mutant.However,there are currently no approved targeted therapies or immunotherapies against mutant p53.Here,we characterized and inves-tigated a monoclonal antibody(mAb)that recognizes the mutant p53-R175H for its affinity,specificity,and activity against tumor cells in vitro.We then delivered DNA plasmids expres-sing the anti-R175H mAb or a bispecific antibody(BsAb)into mice to evaluate their therapeutic effects.Our results showed that the anti-R175H mAb specifically bound to the p53-R175H an-tigen with a high affinity and recognized the human mutant p53-R175H antigen expressed on HEK293T or MC38 cells,with no cross-reactivity with wild-type p53.In cultured cells,the anti-R175H mAb showed higher cytotoxicity than the control but did not induce antibody-dependent cellular cytotoxicity.We made a recombinant MC38 mouse cell line(MC38-p53-R175H)that overexpressed the human p53-R175H after knocking out the endogenous mutant p53 alleles.In vivo,administration of the anti-R175H mAb plasmid elicited a robust anti-tumor effect against MC38-p53-R175H in mice.The administration of the anti-R175H BsAb plasmid showed no therapeutic effects,yet potent anti-tumor activity was observed in combination with the anti-PD-1 antibody.These results indicate that targeting specific mutant epitopes using DNA-delivered mAbs or BsAbs presents a form of improved natural immunity derived from tumor-infiltrating B cells and plasma cells against intracellular tumor antigens.展开更多
基金the National Natural Science Foundation of China(No.41506091)Zhejiang Provincial Public Welfare Project(No.2017C33035)+2 种基金Wenzhou Science&Technology Bureau(Nos.Y20170091,Y20190021)Health Commission of Zhejiang Province(No.2019KY465)Key Laboratory of Orthopaedics of Zhejiang Province(No.ZJGK1806Y)。
文摘Acellular tissue matrix scaffolds are much closer to tissue’s complex natural structure and biological characteristics,thus assess great advantages in cartilage engineering.We used rabbit costal cartilage to prepare acellular microfilaments and further 3D porous acellular cartilage scaffold via crosslinking.Poly(_L-lysine)/hyaluronic acid(PLL/HA)multilayer film was then built up onto the surface of the resulting porous scaffold.Furthermore,TGF-β3 was loaded into the PLL/HA multilayer film coated scaffold to obtain a 3D porous acellular cartilage scaffold with sustained releasing of TGF-β3 up to 60 days.The success of this project will provide a new way for the treatment of articular cartilage defects.Meanwhile,the anchoring and on-site sustained releasing of growth factors mediated by polyelectrolyte multilayered film can also provide a new method for improving the biocompatibility and the biofunctionality for other implanted biomaterials.
基金supported by the Cancer Prevention and Research Institute of Texas(No.RR190043).
文摘tThe tumor suppressor p53 is the most common mutated gene in cancer,with the R175H as the most frequent p53 missense mutant.However,there are currently no approved targeted therapies or immunotherapies against mutant p53.Here,we characterized and inves-tigated a monoclonal antibody(mAb)that recognizes the mutant p53-R175H for its affinity,specificity,and activity against tumor cells in vitro.We then delivered DNA plasmids expres-sing the anti-R175H mAb or a bispecific antibody(BsAb)into mice to evaluate their therapeutic effects.Our results showed that the anti-R175H mAb specifically bound to the p53-R175H an-tigen with a high affinity and recognized the human mutant p53-R175H antigen expressed on HEK293T or MC38 cells,with no cross-reactivity with wild-type p53.In cultured cells,the anti-R175H mAb showed higher cytotoxicity than the control but did not induce antibody-dependent cellular cytotoxicity.We made a recombinant MC38 mouse cell line(MC38-p53-R175H)that overexpressed the human p53-R175H after knocking out the endogenous mutant p53 alleles.In vivo,administration of the anti-R175H mAb plasmid elicited a robust anti-tumor effect against MC38-p53-R175H in mice.The administration of the anti-R175H BsAb plasmid showed no therapeutic effects,yet potent anti-tumor activity was observed in combination with the anti-PD-1 antibody.These results indicate that targeting specific mutant epitopes using DNA-delivered mAbs or BsAbs presents a form of improved natural immunity derived from tumor-infiltrating B cells and plasma cells against intracellular tumor antigens.
