Profile Of Murine Innate Lymphoid Cell Subsets In Lung Under The Hypothermia Condition And Protection Of Yu Ping Feng Powder

Objective The lung is the vital organ for gas exchange.Immune cells in lung,especially the innate immune cells can response to challenges sensitively from the external environment(such as temperature,dust pollution and pathogens),in order to maintain immune system function.When the responses out of control,immune system would be at confusion,result in many diseases.Low temperature environment can easily lead to a variety of respiratory diseases,which is closely related to the immune disorders of the lung.Innate lymphoid cells(ILCs)were discovered recently,distributed in lung mucosal barrier widely.ILCs play an important role in maintaining balance of lung micro-environment and fighting against challenges of external environment.The sensitivity and responses of ILCs to low temperature are unclear.The famous traditional Chinese medicine Yupingfeng powder(YPF)can be uesd to strengthen the exterior of body.It is useful in prevention and treatment of respiratory diseases.Our objective is to observe responses of ILCs in lung under the low temperature environment,and explore the immune protection effects of YPF targeting ILCs.Methods1.Preparation of traditional Chinese medicine YPF: YPF consist of Saposhnikovia divaricata,Astragalus membranaceus and bighead atractylodes rhizome.YPF was prepared by the method of water-extraction and alcohol-precipitation.2.Establishment mouse model of low temperature and intervention method of YPF:Mouse was placed in low temperature device(10±1?)to establish low temperature model;the model group were treated by drinking YPF for drug intervention.3.Blood cell analyzer to detect the components of mouse peripheral blood cell.4.Flow cytometry was used to detect subsets and absolutely number T lymphocytes and ILCs in spleen and lung of mice.5.Transcription levels of functional effectors of ILC subsets were detected by RT-PCR.6.HE staining was used to observe the changes of lung tissue in mice.7.Transcription levels of cytokines and chemokines(IL-12,CXCL-10,CCL-20 and IL-25)was detected by RT-PCR.Results1.The responses of lymphocytes in peripheral blood and spleen of mice under low temperature.Under the low temperature environment,WBC in peripheral blood decreased(P< 0.01),lymphocyte count decreased(P< 0.01),eosinophils and mononuclear cell count decreased(P< 0.01).As for the spleen,proportion and absolutely number of ILCs decreased significantly(P< 0.05).The immune system was suppressed under low temperature,and ILCs were more sensitivity.2.Responses of ILCs in lung of mice induced by low temperature.Under the low temperature environment,absolutely number of ILCs decreased(P< 0.01),with the proportion and cell number of ILC1 decreased(P< 0.05)and proportions of ILC2 and ILC3 increased(P< 0.05).Expression of IFN-? decreased(P< 0.05),but IL-5 and IL-13 increased(P< 0.01).ILC1 was in disadvantage and ILC2 became advantage.The expression of cytokines and chemokines which can improve differentiation and recruitment of ILC subsets,including IL-12,CXCL-10,CCL-20,IL-25,IL-33 and CXCL-2 were increased(P< 0.05).3.Effects of YPF on immune function in lung of mice under low temperature After treatment of YPF,absolutely numbers of T cells and ILCs increased in lung of hypothermia mice(P< 0.05),with proportion of ILC1 increased(P< 0.01),proportion of ILC2 decreased(P< 0.05)and cell numbers of ILC1 and ILC3 increased(P< 0.05).Expression of IFN-? upregulated(P< 0.05),whereas IL-5 and IL-13downregulated(P< 0.01).Expressions of differentiation relative factors of ILC subsets,including IL-12,CCL-20 and IL-25,were decreased(P< 0.05).Conclusions1.The low temperature environment depressed whole immunity function in mice,and ILCs were more sensitivity under this influence.2.The low temperature environment resulted in imbalance of ILCs in lung.ILC1 was at a disadvantage,and ILC2 was became advantage.Advantage of ILC2 improved eosinophil infiltration in lung,and could increase the risk of pulmonary allergic diseases.3.YPF provided protection of mice lung immunity under low temperature.ILCs were up-regulated in lung.Inferiority of ILC1 improved,and balance of ILC subsets and T cells recovered.The incidence of allergic lung disease reduced under low temperatures.