| Objective: The organism have had two sets of immune system in the long process of evolution,that is natural immunity and acquired immunity. Natural immunity is the first defense line to protect body from the pathogen infection.Mononuclear cells mainly includes monocytes and lymphocytes,both are important cell in the immune response.The newborns have immature immune system,espetially the natural immune.The propotion of Th1/Th2 cells is not balance and the immune response is skewed to Th2,and they are highly susceptible to allergic disease.Study has confirmed that there are many important antibodies and cytokines in the maternal milk,and they have very important effection on the newborns immune system.Intestinal flora also have the ability to drive the maturation of the immune system,to activate the immune cell and to induce the cytokine's production.So,it can keep the Th1/Th2 cell response in balance and induce immunologic response or tolerance.Adopting cell culture technique and enzyme-linked immumosorbent assay,we used dust mites allergen,colostrum and bifidobacteria as stimuluts to abserve the morphology and immune change of mononuclear cells in neonatal cord blood.1.To observe the morphology change and the production of the cytokines IFN-γ,IL-4,IL-10 and the ratio of IFN-γ/IL-4 after the stimulation by dust mites allergen and to discuss the characterization of the newborn's immune system and the effection of dust mites allergen on newborn's immune system.2. To observe the morphology change and the production of the cytokines IFN-γ,IL-4,IL-10 and the ratio of IFN-γ/IL-4 after the stimulation by colostrum,bifidobacteria and to discuss their effection on newborn's immune system and the effection on newborn's immune system after the stimulation by dust mites allergen.The conclusion have provide a theoretical basis for occurrence of allergic disease after breast-feeding and administration of probiotics on perinatal.Method:Cord blood was gathered from 32 normal term newborns, 15 of them has the characterization that their mothers has allergic history and positive skin prick tests on dust mites allergen, 17 of them has the characterization that their mothers dosen't has allergic history and negative skin prick tests on dust mites allergen.The first step was to sediment the RBC of cord blood by Methylcellulose,then we used Ficoll to separate cord blood mononuclear cells from the supernants.We inoculate the mononuclear cells to the 6 holes of the culture plate with 24 holes,and cultured with PBS(blank control),dust mites allergen+ PBS,colostrum,bifidobacteria,dust mites allergen + colostrum,dust mites allergen + bifidobacteria for 96 hours.We observed the morphology change and the production of the cytokines IFN-γ,IL-4,IL-10 in the supernants by ELISA.Groups were compared by Analysis of variance and t-test.Statiscal analysis was performed by using the SSPS13.0 statistical software,data is presented as mean±SD( x±s).A probability ofα= 0.05 was considered as statistically significant.Result:1After stimulation by dust mites allergen,colostrum and bifidobacteria,the mononuclear cells of newborn's cord blood tends to differentiation.2There was no statistical difference between groups in the survival percentages of mononuclear cells(P>0.05),each group>90%.3The comparison between allergic group and ono-allergic group about the production of the cytokines and the ratio of IFN-γ/IL-4The production of IFN-γhas no statistical difference between two groups(P>0.05), the production of IL-4 of allergic group is higher than ono-allergic group(P<0.05), the production of IL-10 of allergic group is less than ono-allergic group(P<0.05), the ratio of IFN-γ/IL-4 of allergic group is less than ono-allergic group(P<0.05).4The comparison of allergic group about the production of the cytokines and the ratio of IFN -γ/IL-4 after various factors'stimulation Comparied with PBS,after the stimulation of dust mites allergen, the production of IFN-γhas no statistical difference(P>0.05) , the production of IL-4 was increased(P<0.05),the production of IL-10 and the ratio of IFN-γ/IL-4 were both decreased(P<0.05).Comparied with PBS,after the stimulation of colostrum, the production of IFN-γwas increased(P<0.05), the production of IL-4 was decreased(P<0.05),the production of IL-10 has no statistical difference(P>0.05),the ratio of IFN-γ/IL-4 was increased(P<0.05).Comparied with PBS,after the stimulation of bifidobacteria, the production of IFN-γwas increased(P<0.05), the production of IL-4 was decreased(P<0.05),the production of IL-10 has no statistical difference(P>0.05),the ratio of IFN-γ/IL-4 was increased(P<0.05).Comparied with dust mites allergen,after the stimulation of dust mites allergen +colostrum, the production of IFN-γwas increased(P<0.05), the production of IL-4 was decreased(P<0.05),the production of IL-10 and the ratio of IFN-γ/IL-4 were both increased(P<0.05).Comparied with dust mites allergen,after the stimulation of dust mites allergen + bifidobacteria, the production of IFN-γhas no statistical difference(P>0.05),the production of IL-4 was decreased(P<0.05),the production of IL-10 has no statistical difference(P>0.05) ,the ratio of IFN-γ/IL-4 was increased(P<0.05). 5The comparison between ono-allergic group about the production of the cytokines and the ratio of IFN-γ/IL-4 after various factors'stimulationComparied with PBS,after the stimulation of dust mites allergen, the production of IFN-γand IL-4 both have no statistical difference(P>0.05),the production of IL-10 was decreased(P<0.05),the ratio of IFN-γ/IL-4 has no statistical difference(P>0.05).Comparied with PBS,after the stimulation of colostrum, the production of IFN-γwas increased(P<0.05), the production of IL-4 and IL-10 were both decreased(P<0.05),the ratio of IFN-γ/IL-4 was increased(P<0.05).Comparied with PBS,after the stimulation of bifidobacteria, the production of IFN-γhas no statistical difference(P>0.05), the production of IL-4 was decreased(P<0.05),the production of IL-10 and the ratio of IFN-γ/IL-4 both have no statistical difference(P>0.05).Comparied with dust mites allergen,after the stimulation of dust mites allergen +colostrum, the production of IFN-γwas increased(P<0.05), the production of IL-4 and IL-10 both hava no statistical difference (P>0.05),the ratio of IFN-γ/IL-4 was increased(P<0.05).Comparied with dust mites allergen,after the stimulation of dust mites allergen + bifidobacteria, the production of IFN-γ,IL-4,and the ratio of IFN-γ/IL-4 have no statistical difference(P>0.05). Conclusion:1After stimulation by dust mites allergen,colostrum and bifidobacteria,the of newborn's mononuclear cells of cord blood tend to differentiate,these stimulus have no effect on the cells'percentage of the survival.2Genetic factors have effection on newborn's system.The individual whose mother was allergy has higher th2 than ono-allergic one,they have more imbalanced th1/th2.Reseach data show that the individual who has family history of allergy has more imperfect immune system.3The individual who has family history of allergy has higher th2 when stimulated by dust mites,the th1 and th2 don't change significantly in the one who have no family history of allergy .It's confirmed that the susceptible ones are more easily sensitized to allergen.4The breast-milk can ajust the newborn's balance of th1 and th2, regardless of whether the mother is allergic one,and it can rectify the imbalanced th1/th2 to the normal,it can improve the newborn's immune system to a mature level.5 The bifidobacteria can ajust the newborn's balance of th1 and th2, regardless of whether the mother is allergic one,and it can rectify the imbalanced th1/th2 to the normal,it can improve the newborn's immune system to a mature level.To promote the development of the immune system of neonatal,we should be stick to breast-feeding and administrate probiotics as early as possible,breast-milk and probiotics have effect on th1/th2's balance,as same as the case of immune imbalance caused by allergen.
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