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Mem. Inst. Oswaldo Cruz ; 116: e200560, 2021. graf
Article in English | LILACS | ID: biblio-1154882


BACKGROUND Anisakis simplex antigens present immunomodulatory properties by the induction of tolerogenic dendritic cells (DCs) in mice. OBJECTIVES To study the capacity of DCs stimulated with A. simplex excretory-secretory (ES) or crude extract (CE) to generate Tregs. To investigate in vitro effects of antigens on the metabolic activity of splenocytes induced by LPS or CpG. METHODS Phenotypic and functional characterization of T cells co-cultured with A. simplex-pulsed DCs was performed by flow cytometry. Lymphocyte mitochondrial respiratory activity was estimated by the Alamar Blue® Assay. FINDINGS In C57BL/6J, CD4+CD25-Foxp3+ and CD8+CD25-Foxp3+ populations increased by CE-stimulated-DCs. In BALB/c, CE-stimulated-DCs caused the expansion of CD4+CD25+Foxp3+IL-10+ and CD8+CD25+Foxp3+IL-10+. IFN-γ expression raised in BALB/c CD4+CD25+ and CD4+CD25- for CE and ES, respectively. ES-stimulated-DCs increased CD4+CD25+ Foxp3+ and CD8+CD25- Foxp3+ expression in T cells. The association of ES or CE with LPS produced the increase in splenocyte activity in C57BL/6J. The association of CE with CpG decreased the proliferation caused by CpG in C57BL/6J. MAIN CONCLUSIONS A. simplex increase the frequency of Tregs, which in turn produce IL-10 and IFN-γ. The host genetic base is essential in the development of anti-Anisakis immune responses (Th2, Th1, Treg).

Animals , Mice , Anisakis , T-Lymphocytes, Regulatory , Antigens/metabolism , Bone Marrow , Dendritic Cells , Forkhead Transcription Factors , Interleukin-2 Receptor alpha Subunit , Larva , Mice, Inbred BALB C , Mice, Inbred C57BL
Article in Chinese | WPRIM | ID: wpr-849702


[Abstract] Tolerance is a state in which the human body is less responsive to changes in internal environmental status caused by drugs or other factors. Tolerance is a biological phenomenon, a natural consequence of the action of drugs or chemical factors. Altered organismal tolerance status often leads to the development of multiple autoimmune diseases. Autoimmune thyroiditis (AIT) is a particular type of autoimmune disease, and is exactly the result of an altered immune tolerance of the thyroid gland to a series of "redundant" antigen-antibody binding reactions that produce specific or non-specific inflammation leading to tissue destruction of the thyroid gland. In recent years, a variety of somatic cell therapies have been developed for the purpose of improving the body's tolerance, partially used in the clinical treatment of autoimmune diseases such as systemic lupus erythematosus, multiple sclerosis and Crohn's disease, etc. Such somatic cells that can regulate tolerance are called tolerogenic cells. The present paper will focus mainly on the specific autoimmune diseases such as AIT, discuss the promising therapeutic implications of tolerogenic cells for this group of diseases, and provide a summary of relevant studies. Hopefully, it will provide new research directions for the treatment of the disease.

Article in Chinese | WPRIM | ID: wpr-614473


OBJECTIVE To investigate the immunoregulatory role of tolerogenic dendritic cells(TolDC) in allergic airway inflammation in mouse model of allergic rhinitis(AR).METHODS A total of 24 Balb/c mice were equally and randomly divided into 4 groups with 6 mice in each group. AR group was established with oval bumin(OVA) sensitization and challenge, meanwhile the physiological saline(PBS) sensitization and challenge as the control group. AR group were treated by adoptive transfer of TolDC as treatment group. And treatment group were injected intraperitoneally with TGF-β/IL-10R neutralizing antibody as blockade group. In each group, allergic nasal symptoms score, inflammatory cell infiltration in lung, the expression of Th1/Th2-derived cytokines in the bronchoalveolar lavage fluid(BALF) and nasal lavage fluid(NALF), the expression of OVA-specific serum IgE and the expression of Treg in lung were measured.RESULTS The mouse model of OVA-induced AR was successfully developed. Compared with AR group, treatment group exhibited lower allergic nasal symptoms score, a decrease in inflammatory cell infiltration in lung, lower expression of Th1/Th2-derived cytokines in BALF and NALF and OVA-specific serum IgE, as well as up-regulation of Treg in lung, which were abolished by TGF-β/IL-10 neutralizing antibody shown in blockade group.CONCLUSION TolDC suppress airway inflammation in AR by inducing regulatory T cells through TGF-β/IL-10-dependent mechanisms.

Chinese Journal of Immunology ; (12): 633-638, 2014.
Article in Chinese | WPRIM | ID: wpr-448441


Objective:To establish the methods of isolated culture and functional identification of mice bone marrow derived tolerogenic dendritic cells (CD11b+F4/80 +TDCs) in vitro.Methods: Mice bone marrow cells were isolated and cultured to obtain iDCs with the simulation of mouse rmGM-CSF and rmIL-4.CD11b+F4/80 +TDCs were purified by fluorescence-activated cell sorting on day 6.The morphological changes of TDCs were observed with the inverted microscope dynamically .The expression of CD11b+F4/80 +TDCs were analyzed by the flow cytometry .Tolerogenic function of CD11b+F4/80 +TDCs was evaluated by the expression of MHCⅡ, CD83, IDO, TLR-2, IL-10 and TGF-β1.The expression of MHCⅡ was analyzed by the flow cytometry , and the expression of CD83, IDO and TLR-2 were analyzed by immune-histochemistry.The levels of IL-10 and TGF-β1 in the supernatant of CD11b+F4/80 +TDC were analyzed by ELISA .Meanwhile mature DCs ( mDCs) induced by LPS were used as control .Results:The fresh isolated bone marrow cells look like round and small under microscope .After two days of culture , cells became big and formed into clusters . Five or six days later, cells clusters increased, and the morphology of cells became irregular .At the same time, more dendrite ap-peared on the surface of cells .The percentage of CD11b+F4/80 +TDCs induced by rmGM-CSF and rmIL-4 was about 23%, and the purity of the purified BM CD11b+F4/80 +iDC was about 99%.Compared with mDCs, CD11b+F4/80 +TDCs expressed low levels of MHCⅡand CD83 and high levels of IDO, TLR-2, IL-10 and TGF-β1.Conclusion:CD11b+F4/80 +TDCs derived from mouse bone marrow could be induced successfully by rmGM-CSF and rmIL-4 in vitro.CD11b+F4/80 +TDCs showed tolerogenic function by the expressions of IL-10, TGF-β1, IDO and TLR-2.