Th-cytokine Profile Evaluation In Mice Model With Either Immunization Of Natural Echinococcus-antigens/BCG Or Second Infection Of Alveolar Echinococcosise

Objective: To investigate the cytokine profiles in mice models with either immunizationby use of natural Echinococcus-antigens and BCG or second alveolar echinococcosisinfection after immunization. To be insight in the immune-regulation mechanisms of hostprotective and immune-pathological inflammation processes to anti-invading pathogen(alveolar echinococcosis) through T-help cells and their cytokines.Methods:(1) Establishment of immuniztion with the Echinococcus-antigens/BCG andalveolar echinococcosis infectious mice models: to immunize mice by use of natural antigensfrom cystic/alveolar echinococcosis (CE/AE) and commercial BCG with0.2mgsubcutaneous injection in first and boost twice with0.1mg/per month, after immunization,half of them were challenged by alveolar echinococcosis by ip-injection dose of2000protoscolexes. Every group has20mouse with half females and males; but10normal micecontrols and10no-immunization infectous controls with five-males and females,respectively.(2) Mice model process and sample collections: The samples of blood andtissues from portion of liver/spleen were collected before mice-euthanasia by anesthetization.Serum separated from clot and stored at-80C, tissues immediately put into RNALater frommice body. Storage were at-80C after4C over night balance. After death, carefully checkingabdominal cavity and counting lesion numbers and measuring its zise; the cytokine detctionswere carried out by use of flow-fluorescence for serum-cytokines and by use of suspension microarray for tissue-cytokines.(3) Analysis the cytokine-data: using SPSS to comparison ofcytokine profiles between various group and set up the significant difference at95%intervalsResults:(1) There was73%(102/140) mouse were successfully set-up as designedmice-model.(2) The immune response of mice in different models, the Th1cytokines ofIFN-γ, IL-1were significately increased in CE antigen-immunized to comparing with PBScontrol-group. The Th2cytokines of IL-10, IL-13in AE-antigen immunized group wassignificantly increased when compared with the PBS control group. The Th2cytokines ofIL-5, IL-6, IL-13in BCG immunization group were decreased when compared with thecontrol group though significant value was closed to be meanful. However, the Th2cytokinesof IL-1, IL-12, IFN-γ in BCG immunization group were significantly increased whencompared with the PBS control group. The Th17cytokines of IL-17, IL-21, IL-22in AEantigen immunization group and in challenged group after AE-antigen immunization wassignificant difference when compared with infection control and normal control groups.Conclusion:(1) The flow fluorescence and suspension microarray technology has variedadvantages such as high-flux, less volume of samples necessary, and multiple indicators(multicriteria). Therefore, it can be used in the cytokine network study effectly.(2) There is akind of complex cytokine network regulation in the infection of hydatid, in which CE antigeninduced Th1cell response advantage, but the preferential immune response can not prohibitthe alveclar echinococcosis development. AE antigen induced Th2type cellular immuneresponse, but the levels of Th2cytokines decreased after BCG immunization. The Th1immune response was induced by BCG immunization. Th1cytokines showed in decreasedprofile in the AE immunized group and the challenged group after AE-immunization, thismay reflect immune-suppression induced by AE-antigen stimulations.(3) Echinococcusmultilocularis affected the proliferation and differentiation of memory Th17cell, andcytokines of Th17may play a role in chronic inflammation processes of hydatid disease.However, this pathway still remains largely unclear how Th17cell at play in hydatid disease courses, and it needs further study to identify the effect of specific protective or immunepathological functions in intermediate hosts.