Goldberg J B, Hatano K, Meluleni G S, Pier G B
Channing Laboratory, Department of Medicine, Brigham and Women's Hospital, Harvard Medical School, Boston, MA 02115-5899.
Proc Natl Acad Sci U S A. 1992 Nov 15;89(22):10716-20. doi: 10.1073/pnas.89.22.10716.
As a step toward developing recombinant oral vaccines, we have explored the feasibility of expression of O polysaccharide antigens from Pseudomonas aeruginosa by Escherichia coli. We cloned in E. coli HB101 a 26.2-kilobase DNA fragment from P. aeruginosa strain PA103 that specifies the production of the O polysaccharide of Fisher immunotype 2 (IT-2) strains. The recombinant organism incorporated the P. aeruginosa IT-2 O polysaccharide onto the core of the E. coli lipopolysaccharide (LPS). Transfer of the recombinant plasmid to three LPS-rough strains of P. aeruginosa resulted in synthesis of IT-2 O antigen, and two of these transconjugant strains also synthesized a second O polysaccharide, presumably representing expression of a repressed, or an incomplete, set of genes for an endogenous O polysaccharide. Rabbits injected with the purified recombinant LPS made antibody specific for P. aeruginosa IT-2 O side chains, as did mice fed the recombinant E. coli strain. Expression of P. aeruginosa O antigens by enteric bacteria makes it possible to study these recombinant strains as oral vaccines to prevent P. aeruginosa infections.
作为开发重组口服疫苗的第一步，我们探索了大肠杆菌表达铜绿假单胞菌O多糖抗原的可行性。我们在大肠杆菌HB101中克隆了来自铜绿假单胞菌菌株PA103的一个26.2千碱基的DNA片段，该片段决定了费舍尔免疫型2（IT-2）菌株O多糖的产生。重组生物体将铜绿假单胞菌IT-2 O多糖整合到大肠杆菌脂多糖（LPS）的核心上。将重组质粒转移到三株铜绿假单胞菌LPS粗糙型菌株中，导致IT-2 O抗原的合成，其中两株转接合子菌株还合成了第二种O多糖，推测这代表了一组被抑制或不完整的内源性O多糖基因的表达。注射纯化重组LPS的兔子产生了针对铜绿假单胞菌IT-2 O侧链的特异性抗体，喂食重组大肠杆菌菌株的小鼠也是如此。肠道细菌表达铜绿假单胞菌O抗原使得将这些重组菌株作为口服疫苗来预防铜绿假单胞菌感染成为可能。
