Study On The Combination Ability Between Recombinant BCG Mediated By Membrane Protein PEB1 Of C.Jejuni And Epithelium Cells

BACKGROUUND:The vaccine in the prevention and treatment of the human variety of epidemic plays an indelible role, among them, genetic engineering live carrier recombination vaccine were aroused many researchers' attention by its elicits cellular, humoral and even mucosa immune response. Gene recombinant Bacillus Calmette-Guerin vaccine(rBCG) is a single vaccination which can continue to target antigens induced the body of genetic engineering vaccine immune response. Construction is using genetic engineering technology to BCG gene transformation make its improve immunogenicity, which induction of one or more produced lasting defense, so as to disease prevention and cure disease. So far, a variety of rBCG which carry different preparation antigen had successfully produced by researchers, and has been used in many kinds of disease prevention such as tuberculosis [1], tumor [2] and parasitic infection [3] and so on. We research group prophase has successfully created the cell walls Der p2 -rBCG and type of cell walls PEB1-Der p2-rBCG. In this study, we use the basis of in vitro cell culture method, evaluation of combination ability between the vaccine of PEB1- Der p2-rBCG and epithelial cells.AIM:To investigate the combination ability of the PEB1-Der p2-rBCG and epithelium cells.METHODS AND RESULTS:1. Adherence experiment between cell wall type PEB1-Der p2-rBCG and epithelium CellsThe method of cell culture in vitro was used to culture the bacterial strains of common BCG, membrane form Der p2-rBCG and membrane form PEB1-Der p2- rBCG with HeLa and human intestinal epithelial cells (HIEC), HE staining and acid-fast staining were used and the adhesion of cells was detected at light microscopy. After incubated for 24 h, no matter HeLa cells or HIEC, adhesion rate of PEB1-Derp2-rBCG was obviously improved than common BCG and Der p2-rBCG, and the differences were statistically significant (P < 0.05). However, the Der p2-rBCG bacteria group was not significantly different from common BCG group (P > 0.05).2. Inhibition and blocking experiment between cell wall type PEB1-Der p2-rBCG and epithelium CellsEach group of vaccine and cells are incubated using the same methods,purified PEB1 or anti-PEB1 polyclonal antibodies were added into each group to adhesion inhibition and blocking,and were dyed at different time points,then the influence of combining ability was observed.The results showd that the differences between PEB1-Der p2-rBCG performed in the absence and presence of PEB1 protein or anti-PEB1 polyclonal antibodies were significant (P < 0.05); However, no adhesion blocking effects in the Der p2-rBCG bacteria group and the common BCG group which added PEB1 protein or PEB1 polyclonal antibodies(P > 0.05).CONCLUSIONS:Membrane Protein PEB1 of C.Jejuni increased the adhesion ability which mediated PEB1-Der p2 -rBCG and epithelium cells.