The Study Of Lactobacillus Acidophilus S-layer Protein Inhibiting Virus Infection

Mechanism of antiviral activity of probiotics has been understood that the majority ofscholars. especially Flora Lactobacillus acidophilus has obvious anti-pathogenic microorganismsnature. This study Lactobacillus acidophilus the degree of participation in the antiviral activity andthe role of the principle of the S-layer protein (S-layer protein), primary intestinal epithelial cellsfor research probiotics platform Lactobacillus acidophilus the antiviral effect and the S-layerprotein level of participation in the antiviral activity to do research to provide a theoretical basisfor follow-up study of the probiotic.First, the S-layer protein of Lactobacillus acidophilus gene and protein extracts identification,experimental results show that the size of the S-layer protein gene of bacterial presence1200bp,using SDS-PAGE analysis using the the neutral the LiCl extract bacterial S-layer protein referredThe protein size45ku and higher purity of the protein, consistent with literature comparison.Purified by further extraction of the protein purification method for protein apex cut after thepreparation of the anti-S-layer protein rabbit polyclonal serum antibodies detected by the Elisamethod, the serum antibody titer of1.0x105, it is possible to meet the experimental follow-uprequirements. Ji Chang epithelial cells in primary culture, the choice to be hatched in20-day-oldSPF chicken embryos, using a scraping method intestinal villus and crypt units collagenase Ⅰand Ⅳ joint digested into pre-paved with collagen96-well cell culture plate, in accordance withcertain cell seeding density, two chicken embryo can be covered with a96-well cell plates andco-cultured with fibroblasts IEC culture.S-layer protein in order to evaluate the antiviral Lactobacillus acidophilus in the degree ofanti-viral, and set the following experimental groups:1Prevention experiment is the first contactof the different components of the microbial cells IEC, full role of co-incubated for1h make it,using the wash buffer the virus incubated1h after adding to the other ingredients, and then washaway the virus ingredients, added to the cells continue to sustain liquid cultured cells, until thevirus control group until cytopathic effect (CPE) in to stop cell culture, MTT cell viability;2treatment experiments were first adding the virus, after adding bacterial ingredients concrete stepsas above;3mixed sense of experiments is to continue to foster bacterial component and virus for1h with three co-incubated1h, added to the cells after wash away the ingredients added to the cellsto maintain liquid until the virus control group until the cytopathic effect (CPE) to stop cell culture,cell viability MTT assay. The experimental results show that the S-layer protein of the virus inhibitory strongestbacterial followed off followed by S-layer protein bacterial metabolites inhibit virus lowest rate.The reason why the S-layer protein is higher than the virus inhibition rate of the Lactobacillusacidophilus, the protein concentration within the range in the IEC can withstand greater than the Sprotein content of the surface of bacteria Lactobacillus acidophilus than bacteria, and therefore thevalue of the suppression rate obvious role of body; off compared to the S-layer protein bacterialwhole cell inhibition rate of the former than the latter, we can see from the S-layer protein plays asignificant role in cell numbers for the effects of the virus; metabolites from the rate of viralsuppression see, the secondary metabolites of the microbial cells still have certain anti-viralproperties. Inhibition of bacterial viruses can be seen by comparison with the other ingredients, theresults of the work of many hands play, the S-layer protein played an essential role in the antiviralactivity of Lactobacillus acidophilus.The sense of four kinds for prevention of bacterial component and mixed for experiment twoviral suppression rates are higher than in the therapy experiments described when the virusinfected cells, probiotic ingredients can not play a significant inhibition of virus proliferation.Prevention experiment with mixed sense of experiments compared different components vary. ForS-layer protein plus protein after virus protein in order to prevent it from infected cells andvirus-infected cells and then added to the S protein little apparent effect. This means that the Sprotein can only play a role of anti-viral infection early in the viral infection, the S-layer proteinwith the prevention of infection effect; When the virus-infected cells, the S-layer protein of thevirus infection of cells but not treatment effect, we can see that the S protein play antiviral activityperiod before the virus from entering cells. Bacterial virus inhibition rate the mixed sense for thegroup and prevention group, the rate of viral suppression of bacterial treatment of the experimentalgroup was significantly higher than that of the S protein group, off the S bacteria group andmetabolite group, indicating that the bacteria a comprehensive anti-virus ability of each ingredient,and a protective effect to the virus of the IEC.To further study the role of the relationship between the S-layer protein and virus followingexperiments. Experiment1draw indirect ELISA the adhesion of the virus and the S-layer proteinand to select bovine serum albumin as a negative control; Experiment2Evaluation of bovineserum albumin with or without viral suppression method with the S-layer protein of the virusinhibition assay. Experimental results show that the S-layer protein and albumin can be adheredIBV and NDV, but only the S protein of the virus has a significant inhibitory albumin notsignificantly inhibit viral properties, does affect the adhesion of the S-layer proteins on the virusand impede the virus infection of intestinal epithelial cells, and having the ability to inhibit IBVand NDV infection IEC.In summary, the inhibition of bacterial virus is the results of the work of many hands to play,but the S-layer protein played a significant role in the antiviral activity of Lactobacillus acidophilus. The experimental results of further study of the role in the relationship between theS-layer protein and virus the the S protein adhesion virus played a effect of inhibition of viralinfection, the S protein-virus complex hinder a viral infection of the IEC. For the S-layer protein,there are a lot of work to be done, this study only the probiotic Lactobacillus acidophilus in S-layerprotein in the chicken primary intestinal epithelial cells in antiviral research, due to the probioticS-layer protein diversity and the platform’s multi-selective cells, for a more comprehensiveunderstanding of the S-layer protein need more in-depth research.