Clostridium perfringens phospholipase C(plc),also calledα-toxin,is encoded by the plc gene Clostridium perfringers.The production ofα-toxin can lead to the occurrence of gas gangrene.Vaccination is considered as one...Clostridium perfringens phospholipase C(plc),also calledα-toxin,is encoded by the plc gene Clostridium perfringers.The production ofα-toxin can lead to the occurrence of gas gangrene.Vaccination is considered as one of the best solutions against Clostridium infections.In this study,an anti-Cpα-toxin monoclonal antibody(mAbs)A10E5 was successfully prepared,which had better biological reactivity.Then,the phage random 12-peptide library was used to screen mAb A10E5 protein.After four rounds of screening,three peptides with high affinity to the anti-α-toxin mAbs were screened.Two 12-peptide peptide Q and peptide E with higher inhibition rate were obtained by indirect ELISA.Two polypeptides of 500μg·mL^(-1)synthesized in vitro were mixed with30μg·mL^(-1)α-toxin at a concentration to treat Hela cells.Cell viability was determined by MTT assay.The results showed that both of the peptides significantly increased the survival rate of Hela cells compared with theα-toxin group,and the effect of peptide Q was more obvious.The chickens were immunized with phages expressing two different affinity polypeptides and then challenged.The results of chicken weight change,intestinal lesion score,bacterial count,and antibody titer in peripheral blood showed that the two phages expressing the polypeptides had a certain protective effect on the chickens compared with the PBS group,and peptide Q had better protection effect.In conclusion,the high affinity peptide with mAb A10E5 was screened in this study,and the protective effect of the plc polypeptide vaccine was verified by in vivo and in vitro experiments,which was of great significance for the comprehensive prevention and treatment of the disease.展开更多
僵人综合征(stiff-person syndrome,SPS)是一种病因未明、以波动性肌肉僵硬和痛性痉挛为核心症状的自身免疫性中枢神经系统罕见病,临床分型包括经典SPS、僵肢综合征、伴强直及肌阵挛的进行性脑脊髓炎(progressive encephalomyelitis wit...僵人综合征(stiff-person syndrome,SPS)是一种病因未明、以波动性肌肉僵硬和痛性痉挛为核心症状的自身免疫性中枢神经系统罕见病,临床分型包括经典SPS、僵肢综合征、伴强直及肌阵挛的进行性脑脊髓炎(progressive encephalomyelitis with rigidity and myoclonus,PERM)、重叠综合征[1]。展开更多
基金Supported by the National Natural Science Foundation of China(31370140,31372438)。
文摘Clostridium perfringens phospholipase C(plc),also calledα-toxin,is encoded by the plc gene Clostridium perfringers.The production ofα-toxin can lead to the occurrence of gas gangrene.Vaccination is considered as one of the best solutions against Clostridium infections.In this study,an anti-Cpα-toxin monoclonal antibody(mAbs)A10E5 was successfully prepared,which had better biological reactivity.Then,the phage random 12-peptide library was used to screen mAb A10E5 protein.After four rounds of screening,three peptides with high affinity to the anti-α-toxin mAbs were screened.Two 12-peptide peptide Q and peptide E with higher inhibition rate were obtained by indirect ELISA.Two polypeptides of 500μg·mL^(-1)synthesized in vitro were mixed with30μg·mL^(-1)α-toxin at a concentration to treat Hela cells.Cell viability was determined by MTT assay.The results showed that both of the peptides significantly increased the survival rate of Hela cells compared with theα-toxin group,and the effect of peptide Q was more obvious.The chickens were immunized with phages expressing two different affinity polypeptides and then challenged.The results of chicken weight change,intestinal lesion score,bacterial count,and antibody titer in peripheral blood showed that the two phages expressing the polypeptides had a certain protective effect on the chickens compared with the PBS group,and peptide Q had better protection effect.In conclusion,the high affinity peptide with mAb A10E5 was screened in this study,and the protective effect of the plc polypeptide vaccine was verified by in vivo and in vitro experiments,which was of great significance for the comprehensive prevention and treatment of the disease.
文摘僵人综合征(stiff-person syndrome,SPS)是一种病因未明、以波动性肌肉僵硬和痛性痉挛为核心症状的自身免疫性中枢神经系统罕见病,临床分型包括经典SPS、僵肢综合征、伴强直及肌阵挛的进行性脑脊髓炎(progressive encephalomyelitis with rigidity and myoclonus,PERM)、重叠综合征[1]。
