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Adv Rheumatol ; 59: 30, 2019. tab, graf
Article de Anglais | LILACS | ID: biblio-1088600

RÉSUMÉ

Abstract Background: Adaptive immune cells, including CD4+CD69+ and CD4+CD25+FoxP3+ regulatory T (Treg) cells, are important for maintaining immunological tolerance. In human systemic lupus erythematosus (SLE), CD4+CD25+FoxP3+ Treg cells are reduced, whereas CD69 expression is increased, resulting in a homeostatic immune imbalance that may intensify autoreactive T cell activity. To analyze the mechanisms implicated in autotolerance failure, we evaluated CD4+CD69+ and CD4+CD25+FoxP3+ T cells and interleukin profiles in a pristane-induced SLE experimental model. Methods: For lupus induction, 26 female Balb/c mice received a single intraperitoneal 0.5 ml dose of pristane, and 16 mice received the same dose of saline. Blood and spleen samples were collected from euthanized mice 90 and 120 days after pristane or saline inoculation. Mononuclear cells from peripheral blood (PBMC), peritoneal lavage (PL) and splenocytes were obtained by erythrocyte lysis and cryopreserved for further evaluation by flow cytometry using the GuavaEasyCyte TM HT. After thawing, cells were washed and stained with monoclonal antibodies against CD3, CD4, CD8, CD25, CD28, CD69, FoxP3, CD14 and Ly6C (BD Pharmingen TM). Interleukins were quantified using Multiplex® MAP. The Mann-Whitney test and the Pearson coefficient were used for statistical analysis, and p < 0.05 considered significant. Results: Compared with the controls, SLE-induced animals presented increased numbers of CD4+CD69+ T cells in the blood on T90 and T120 (p = 0.022 and p = 0.008) and in the spleen on T120 (p = 0.049), but there were decreased numbers in the PL (p = 0.049) on T120. The percentage of Treg was lower in blood (p < 0.005 and p < 0.012) on T90 and T120, in spleen (p = 0.043) on T120 and in PL (p = 0.001) on T90. Increased numbers of CD4+ CD69+ T cells in the PL were positively associated with high IL-2 (p = 0.486) and IFN-γ (p = 0.017) levels, whereas reduced Treg cells in the blood were negatively correlated with TNFα levels (p = 0.043) and positively correlated with TGFβ1 (p = 0.038). Conclusion: Increased numbers of CD4+CD69+ T cells and reduced numbers of CD4+CD25+FoxP3+ Treg cells with an altered interleukin profile suggests loss of autotolerance in pristane-induced lupus mice, which is similar to human lupus. Therefore, this model is useful in evaluating mechanisms of cellular activation, peripheral tolerance and homeostatic immune imbalance involved in human SLE.


Sujet(s)
Animaux , Femelle , Souris , Rate/cytologie , Lavage péritonéal , Lymphocytes T CD4+/cytologie , Lymphocytes T régulateurs/cytologie , Lupus érythémateux disséminé/immunologie , Rate/immunologie , Terpènes , Lymphocytes T CD4+/immunologie , Antigènes Ly/analyse , Antigènes Ly/immunologie , Antigènes de différenciation des lymphocytes T/analyse , Antigènes de différenciation des lymphocytes T/immunologie , Antigènes CD/analyse , Antigènes CD/immunologie , Sous-populations de lymphocytes T/cytologie , Sous-populations de lymphocytes T/immunologie , Lymphocytes T régulateurs/immunologie , Antigène CD28/analyse , Antigène CD28/immunologie , Numération des lymphocytes , Antigènes CD14/analyse , Antigènes CD14/immunologie , Lectines de type C/analyse , Lectines de type C/immunologie , Facteurs de transcription Forkhead/analyse , Facteurs de transcription Forkhead/immunologie , Sous-unité alpha du récepteur à l'interleukine-2/analyse , Sous-unité alpha du récepteur à l'interleukine-2/immunologie , Immunosuppresseurs , Lupus érythémateux disséminé/sang , Lupus érythémateux disséminé/induit chimiquement , Souris de lignée BALB C
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