CD4(+)CD69(+) T cells and CD4(+)CD25(+)FoxP3(+) Treg cells imbalance in peripheral blood, spleen and peritoneal lavage from pristane-induced systemic lupus erythematosus (SLE) mice

Author(s):Show less -

Peixoto, Tatiana Vasconcelos ^[1] ; Carrasco, Solange ^[1] ; Ciccone Botte, Domingos Alexandre ^[1] ; Catanozi, Sergio ^[2] ; Parra, Edwin Roger ^[3] ; Lima, Thais Martins ^[4] ; Ugriumov, Natasha ^[1] ; Soriano, Francisco Garcia ^[4] ; Verissimo de Mello, Suzana Beatriz ^[1] ; Rodrigues, Caio Manzano ^[5] ; Goldenstein-Schainberg, Claudia ^[6]

Affiliation

^[1] Univ Sao Paulo, Lab Imunol Celular LIM 17, Fac Med, Sao Paulo, SP - Brazil

^[2] Univ Sao Paulo, Fac Med, Lab Lipides LIM 10, Sao Paulo, SP - Brazil

^[3] Univ Sao Paulo, Fac Med, Dept Patol Clin, Sao Paulo - Brazil

^[4] Univ Sao Paulo, Fac Med, Lab Emergencias Clin LIM 51, Sao Paulo, SP - Brazil

^[5] Univ Estadual Paulista Julio de Mesquita Filho Un, FMB, Botucatu, SP - Brazil

^[6] Univ Sao Paulo, Fac Med, Hosp Clin, Lab Imunol Celular LIM 17, Sao Paulo, SP - Brazil

• Document type: Journal article
• Source: ADVANCES IN RHEUMATOLOGY; v. 59, JUL 24 2019.
• Web of Science Citations: 0

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 (R) 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-gamma (p = 0.017) levels, whereas reduced Treg cells in the blood were negatively correlated with TNF alpha levels (p = 0.043) and positively correlated with TGF beta 1 (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. (AU)

• FAPESP's process: 13/19292-1 - Analysis of activator and regulatory molecules of CD4+ T cells and CD4+CD25+ t reg cells in mice with Systemic Lupus Erythematosus (SLE) pristane-induced
• Grantee: Tatiana Vasconcelos Peixoto
• Support Opportunities: Scholarships in Brazil - Doctorate (Direct)