Bordetella Pertussis Cpg Odns - The Experimental Study Of Allergic Asthma Mice Toll Signal Molecules

Allergic asthma is a chronic airway inflammation with excess express of IgE, infiltration of eosinophil and mast cells, airway hyperreactivity and remodeling. In nowadays, the incidence of asthma is rising, many patients are suffered by this disease and sometime it is fatal. Allergic asthma is an airway disease with unbalance of Thl/Th2effect. In recent research, the scientists found Th2cytokines make the main role when one get the disease, Th2cytokines like IL-4, IL-5, IL-9and IL-13would be secreted to suppress Thl cytokines. This process plays an important role in hypereosinophilia and releasing IgE by B cell. Regulating the balance of Thl/Th2is one of the accepted therapeutic of allergic asthma.Bordetella pertussis is one pathogenic microorganism which could develop whooping cough. Recently, the researchers gather the vaccines of Bordetella pertussis, bacillus diphtheriae and bacillus tetani for preventing whooping cough, diphtheria and tetanus. And also vaccine of bordetella pertussis was found to be a fierce inducer of Thl reactivity, the researchers from Korea found it could reverse the development of allergic asthma and it might relate with unmethylated CpG-ODNs which were found in bacteria.CpG-ODNs is the short name of unmethylated CpG motif containing oligodeoxynucleotides which have immuno-stimulatory activities. There are four type subsets of CpG-ODNs, group A to group D. Methylated sequences have less action than unmethylated ones. CpG-ODNs are recognized by Toll like receptor9(TLR-9), one of the members of the Toll family of innate immune receptors and activating nuclear factor kB to promote the expression of Thl cytokines and inhibiting Th2cytokines for rebuilding the balance of Thl/Th2activities.We found whole cell of Bordetella pertussis can down-regulate WBC/EOS and rebalance Thl/Th2cytokines before. Our research focus on extracting CpG-ODNs from Bordetella pertussis for preventing and therapeutic of allergic asthma. We used a biological software named Vector NTI Advance11for comparing the genome of Bordetella pertussis and three known CpG sequences which have effects for allergic diseases to find the most similar sequences, after that we found nine CpG-ODNs of bordetella pertussis for the process of anti-inflammatory cells. One sequence named seq A has the best effect. We made chronic asthma model with BABL/c mice, and gave them seq A, ODN-1018(positive) and cont ODN-1018(negative) through intragastric (ig) and intraperitoneal injection (ip) to evaluate the best administration. After executed the whole lung lavage was collected and the spleen was excised with forceps. We examined the concentration of IL-4, IFN-y and VEGF in BALF with ELISA protocol. The expression of TLR-9in spleen was examined by reverse transcription polymerase chain reaction (RT-PCR) and it is examined by westernblot in lung. Results:(1) Compare with model group, CpG-ODNs from Bordetella pertussis can downregulate eosinophil significantly (P<0.05).(2) The concentrate of IL-4in seq A and ODN-1018ip groups are lower than model group and IFN-γ in both of them are higher than model group (P<0.05), though the ig groups are not significant compared with model group (P>0.05);(3) Compared with model group, the expression of spleen TLR-9mRNA in seq A and ODN-1018ip groups are higher than model group (P<0.05), but the ig groups are not significant (P>0.05);(4) Compared with model group, the expression of lung TLR-9protein in seq A and ODN-1018ip groups are higher than model group (P>0.05), but the ig groups are not significant(P>0.05). Conclusion:CpG-ODNs from Bordetella pertussis are able to prevent airway inflammation, seqA ip group could regulate the Th1/Th2balance in BALF, but VEGF does not have significant change in both ig group and ip group. Due to the high expression of TLR-9mRNA in spleen and TLR-9protein in lung, we demonstrate the function of CpG-ODNs is indeed relating with TLR-9; We found ig groups do not have significant changes compared with model group. It is obviously that intraperitoneal injection is better than intragastric during the administration of CpG-ODNs.