Zhang Lulu, Pan Chao, Feng Erling, Hua Xiaoting, Yu Yunsong, Wang Hengliang, Zhu Li
State Key Laboratory of Pathogen and Biosecurity, Beijing Institute of Biotechnology, Beijing 100071, China.
Key Laboratory of Microbial Technology and Bioinformatics in Zhejiang, Sir Run Run Shaw Hospital of College of Medicine of Zhejiang University, Hangzhou 310016, Zhejiang, China.
Sheng Wu Gong Cheng Xue Bao. 2020 Sep 25;36(9):1899-1907. doi: 10.13345/j.cjb.200014.
The main purpose of this research is to synthesize and evaluate a new glycoconjugate vaccine against Klebsiella pneumonia (Kp). First, the gene (waaL) responsible for the expression of O antigen ligase was deleted to block the synthesis of bacterial LPS. Then the vector that encodes a glycosyltransferase (PglL) was transferred into the mutant. The enzyme PglL could catalyze the transfer of OPS units to recombinant cholera toxin B subunit (rCTB) to form glycoproteins in vivo. The protective effects of the glycoproteins were studied by the mice models with acute bacteremia that were induced by intraperitoneal injection of wildtype Kp bacteria. The results were as followings: A Kp waaL mutant was obtained and the rCTB protein could be successfully glycosylated in the mutant. The vaccine can stimulate a high antibody titer in the mice sera with or without adjuvant. It can also protect mice from the lethal dose injection of Kp. The survival rate of vaccine candidate groups could reach 75%. The glycoconjugate vaccine candidate prepared by this biosynthetic method is expected to become a novel effective vaccine against Klebsiella pneumoniae.
本研究的主要目的是合成并评估一种针对肺炎克雷伯菌(Kp)的新型糖缀合物疫苗。首先,删除负责O抗原连接酶表达的基因(waaL)以阻断细菌LPS的合成。然后将编码糖基转移酶(PglL)的载体转入突变体。酶PglL可催化OPS单位转移至重组霍乱毒素B亚基(rCTB),从而在体内形成糖蛋白。通过腹腔注射野生型Kp细菌诱导急性菌血症的小鼠模型研究了糖蛋白的保护作用。结果如下:获得了Kp waaL突变体,且rCTB蛋白可在该突变体中成功糖基化。该疫苗在有或无佐剂的情况下均可刺激小鼠血清产生高抗体滴度。它还可保护小鼠免受致死剂量的Kp注射。候选疫苗组的存活率可达75%。通过这种生物合成方法制备的候选糖缀合物疫苗有望成为一种新型有效的抗肺炎克雷伯菌疫苗。