Dendritic Cell Sphingosine 1-Phosphate Receptor-3 Regulates Th1–Th2 Polarity in Kidney Ischemia–Reperfusion Injury

Abstract

Dendritic cells (DCs) are central to innate and adaptive immunity of early kidney IRI and strategies to alter DC function may provide new therapeutic opportunities. Sphingosine 1-phosphate (S1P) modulates immunity through binding to its receptors (S1P1-5) and protection from kidney IRI occurs in S1P3-deficient mice. Through a series of experiments we determined that this protective effect was due in part to differences between S1P3-sufficient and deficient DCs. Mice lacking S1P3 on bone marrow cells were protected from IRI and S1P3-deficient DCs displayed an immature phenotype. WT but not S1P3-deficient DCs injected into mice depleted of DCs prior to kidney IR reconstituted injury. Adoptive transfer (i.e, i.v. injection) of glycolipid (antigen)-loaded WT but not S1P3-deficient DCs into WT mice exacerbated IRI, suggesting that WT but not S1P3-deficient DCs activated NKT cells. Whereas WT DC transfers activated the Th1-IFN-γ pathway, S1P3-deficient DCs activated the Th2-IL4 pathway and an IL4 blocking antibody reversed protection from IRI supporting the concept that IL-4 mediates the protective effect of S1P3-deficient DCs. Administration of S1P3-deficient DCs 7 days prior to or 3 h after IRI protected mice from IRI and suggests their potential use in cell-based therapy. We conclude that absence of DC S1P3 prevents DC maturation and promotes a Th2-IL4 response. These findings highlight the importance of DC S1P3 in modulating NKT cell function and IRI and support development of selective S1P3 antagonists for tolerizing DCs for cell based therapy or for systemic administration for the prevention and treatment of IRI and autoimmune diseases.

Sphingosine 1-Phosphate Receptor 3–Deficient Dendritic Cells Modulate Splenic Responses to Ischemia-Reperfusion Injury

“…2/2 BMDCs Have an Immature Phenotype BMDCs were generated from wild-type (WT) or S1pr3 2/2 mice as described previously 10 and treated with LPS for 24 hours. Compared with C57BL/6 BMDCs from WT mice, S1pr3…”

“…Data from our laboratory 10,11 and the laboratories of others 12,13 show that modulation of sphingosine 1-phosphate receptors (S1PRs) significantly influences AKI development and progression in animal models. These receptors belong to a family of five G proteincoupled receptors (sphingosine 1-phosphate 1 [S1P1], S1P2, S1P3, S1P4, and S1P5) and modulate diverse physiologic responses, including cellular growth and proliferation, angiogenesis, apoptosis, and lymphocyte trafficking.…”

“…[24][25][26] S1pr3 2/2 mice are protected from renal IRI through a mechanism involving BMDCs and their immune modulatory function. 10 However, the phenotype of S1pr3 2/2 BMDCs and their site of action in vivo (intrarenal versus extrarenal) remain unclear.…”

“…For example, activation of S1P3 protects hearts from IRI, 18,19 but S1P3-deficient mice are protected from kidney IRI. 10,20 Use of pharmacologic agents has limitations because of offtarget and other associated adverse side effects. Cell-based therapeutic approaches have advantages; transferred cells are capable of sensing diverse signals, moving to specific sites in the body, making decisions, and executing complex responses.…”

Sphingosine 1-Phosphate Receptor-1 Enhances Mitochondrial Function and Reduces Cisplatin-Induced Tubule Injury

“…For histology, kidneys were fixed overnight in 0.2% sodium periodate/1.4% DL-lysine/4% paraformaldehyde in 0.1 M phosphate buffer, pH 7.4 (4% PLP), and embedded in paraffin. Kidney sections were prepared for hematoxylin and eosin staining as previously described 47 and viewed by light microscopy (Carl Zeiss AxioSkop) under 3200 magnification. Photographs were taken and brightness/ contrast adjustment was made with a SPOT RT camera (software version 3.3; Spot Imaging Solutions/Diagnostic Instruments Inc., Sterling Heights, MI).…”