TLR2, TLR4 and the MYD88 Signaling Pathway Are Crucial for Neutrophil Migration in Acute Kidney Injury Induced by Sepsis

The aim of this study was to investigate the role of TLR2, TLR4 and MyD88 in sepsis-induced AKI. C57BL/6 TLR2(-/-), TLR4(-/-) and MyD88(-/-) male mice were subjected to sepsis by cecal ligation and puncture (CLP). Twenty four hours later, kidney tissue and blood samples were collected for analysis. the TLR2(-/-), TLR4(-/-) and MyD88(-/-) mice that were subjected to CLP had preserved renal morphology, and fewer areas of hypoxia and apoptosis compared with the wild-type C57BL/6 mice (WT). MyD88(-/-) mice were completely protected compared with the WT mice. We also observed reduced expression of proinflammatory cytokines in the kidneys of the knockout mice compared with those of the WT mice and subsequent inhibition of increased vascular permeability in the kidneys of the knockout mice. the WT mice had increased GR1(+low) cells migration compared with the knockout mice and decreased in GR1(+high) cells migration into the peritoneal cavity. the TLR2(-/-), TLR4(-/-), and MyD88(-/-) mice had lower neutrophil infiltration in the kidneys. Depletion of neutrophils in the WT mice led to protection of renal function and less inflammation in the kidneys of these mice. Innate immunity participates in polymicrobial sepsis-induced AKI, mainly through the MyD88 pathway, by leading to an increased migration of neutrophils to the kidney, increased production of proinflammatory cytokines, vascular permeability, hypoxia and apoptosis of tubular cells.Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)National Institute of Science and Technology (INCT)Universidade Federal de São Paulo, Dept Med, Disciplina Nefrol, São Paulo, BrazilUniv São Paulo, Dept Imunol, Lab Imunobiol Transplantes, São Paulo, BrazilHosp Israelita Albert Einstein, IIEP, São Paulo, BrazilUniv Fed Triangulo Mineiro, Uberaba, BrazilUniversidade Federal de São Paulo, Dept Med, Disciplina Nefrol, São Paulo, BrazilFAPESP: 07/07139-3Web of Scienc

Balance between the two kinin receptors in the progression of experimental focal and segmental glomerulosclerosis in mice

Focal and segmental glomerulosclerosis (FSGS) is one of the most important renal diseases related to end-stage renal failure. Bradykinin has been implicated in the pathogenesis of renal inflammation, whereas the role of its receptor 2 (B2RBK; also known as BDKRB2) in FSGS has not been studied. FSGS was induced in wild-type and B2RBK-knockout mice by a single intravenous injection of Adriamycin (ADM). in order to further modulate the kinin receptors, the animals were also treated with the B2RBK antagonist HOE-140 and the B1RBK antagonist DALBK. Here, we show that the blockage of B2RBK with HOE-140 protects mice from the development of FSGS, including podocyte foot process effacement and the re-establishment of slit-diaphragm-related proteins. However, B2RBK-knockout mice were not protected from FSGS. These opposite results were due to B1RBK expression. B1RBK was upregulated after the injection of ADM and this upregulation was exacerbated in B2RBK-knockout animals. Furthermore, treatment with HOE-140 downregulated the B1RBK receptor. the blockage of B1RBK in B2RBK-knockout animals promoted FSGS regression, with a less-inflammatory phenotype. These results indicate a deleterious role of both kinin receptors in an FSGS model and suggest a possible cross-talk between them in the progression of disease.Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)Universidade Federal de São Paulo, Clin & Expt Immunol Lab, Div Nephrol, BR-04023900 São Paulo, BrazilUniv São Paulo, Inst Biomed Sci 4, Dept Immunol, Lab Transplantat Immunobiol, BR-05508000 São Paulo, BrazilUniversidade Federal de São Paulo, Translat Med Div, Clin & Expt Immunol Lab, BR-04039002 São Paulo, BrazilInst Butantan, Lab Cellular Biol, BR-05503900 São Paulo, BrazilFed Univ São Paulo UNIFESP, Dept Biophys, BR-04023062 São Paulo, BrazilFed Univ São Paulo UNIFESP, Dept Microbiol Immunol & Parasitol, BR-04023062 São Paulo, BrazilINSERM, Unite Mixte Rech 699, F-75870 Paris, FranceAlbert Einstein Hosp, Inst Israelita Ensino & Pesquisa Albert Einst, Renal Transplantat Unit, BR-05521000 São Paulo, BrazilUniversidade Federal de São Paulo, Clin & Expt Immunol Lab, Div Nephrol, BR-04023900 São Paulo, BrazilUniversidade Federal de São Paulo, Translat Med Div, Clin & Expt Immunol Lab, BR-04039002 São Paulo, BrazilFed Univ São Paulo UNIFESP, Dept Biophys, BR-04023062 São Paulo, BrazilFed Univ São Paulo UNIFESP, Dept Microbiol Immunol & Parasitol, BR-04023062 São Paulo, BrazilFAPESP: 2012/05605-5FAPESP: 07/07139-3FAPESP: 12/02270-2CNPq: 140739/2008-4Web of Scienc

Mesenchymal stromal cells modulate gut inflammation in experimental colitis

Inflammatory bowel diseases (IBDs) affect millions of people worldwide and their frequencies in developed countries have increased since the twentieth century. In this context, there is an intensive search for therapies that modulate inflammation and provide tissue regeneration in IBDs. Recently, the immunomodulatory activity of adipose tissue-derived mesenchymal stromal cells (ADMSCs) has been demonstrated to play an important role on several immune cells in different conditions of inflammatory and autoimmune diseases. In this study, we explored the immunomodulatory potential of ADMSC in a classical model of DSS-induced colitis. First, we found that treatment of mice with ADMSC ameliorated the severity of DSS-induced colitis, reducing colitis pathological score and preventing colon shortening. Moreover, a prominent reduction of pro-inflammatory cytokines levels (i.e., IFN-gamma, TNF-alpha, IL-6 and MCP-1) was observed in the colon of animals treated with ADMSC. We also observed a significant reduction in the frequencies of macrophages (F4/80(+)CD11b(+)) and dendritic cells (CD11c(+)CD103(+)) in the intestinal lamina propria of ADMSC-treated mice. Finally, we detected the up-regulation of immunoregulatory-associated molecules in intestine of mice treated with ADMSCs (i.e., elevated arginase-1 and IL-10). Thus, this present study demonstrated that ADMSC modulates the overall gut inflammation (cell activation and recruitment) in experimental colitis, providing support to the further development of new strategies in the treatment of intestinal diseases.FAPESPCAPESCNPqUniv Sao Paulo, Inst Biomed Sci 4, Dept Immunol, Sao Paulo, BrazilUniv Fed Sao Paulo, Nephrol Div, Dept Med, Sao Paulo, BrazilUniv Fed Alagoas, Inst Biol Sci & Hlth, Alagoas, BrazilUniv Sao Paulo, Sch Med, LIM 16, Sao Paulo, BrazilAv Prof Lineu Prestes 1730 Lab 238 Cidade Univ, BR-05508000 Sao Paulo, SP, BrazilUniv Fed Sao Paulo, Nephrol Div, Dept Med, Sao Paulo, BrazilFAPESP: 2012/02270-2, 2012/16794-3, 2013/25327-2Web of Scienc

Role of Leptin in Skin Graft Survival and Tolerance.

Univ Sao Paulo, Inst Biomed Sci, Dept Immunol, Sao Paulo, BrazilState Univ Campinas UNICAMP, Div Nephrol, Campinas, SP, BrazilUniv Fed Sao Paulo, Dept Biophys, Sao Paulo, BrazilUniv Fed Sao Paulo, Div Nephrol, Sao Paulo, BrazilUniv Fed Sao Paulo, Dept Immunol, Sao Paulo, BrazilUniv Fed Sao Paulo, Dept Biophys, Sao Paulo, BrazilUniv Fed Sao Paulo, Div Nephrol, Sao Paulo, BrazilUniv Fed Sao Paulo, Dept Immunol, Sao Paulo, BrazilWeb of Scienc

SHEDs modulate the cytokine profile in co-culture with iDCs and mDCs.

<p>Graphics indicate soluble factors levels (pg/mL; detection by CBA technique) present in the culture moDCs after seven days of differentiation (co-cultured with or without SHEDs, in a 1∶10 ratio) stimulated (to generate mDC) or not (to maintained iDC) with LPS (during the last two days of culture). Comparisons were done using t-test between iDC and iDC+SHED and between mDC and mDC+SHED; *p≤0.05 and **p≤0.01; n = 4.</p

MoDCs previously stimulated by SHEDs are deficient to induce the proliferation of allogeneic T cells.

<p>Immature and mature moDCs co-cultured with SHEDs were isolated by anti-HLA-DR magnetic beads. MoDCs isolated were co-cultured with 1×10⁵ non-adherent cells CFSE stained (1∶10 ratio). After five days, T cells were stained with anti-CD4 and anti-CD8 antibodies, and CFSE dilution was determined by flow cytometry analysis. This analysis was previously performed within FSC and SSC gate, followed by CD4 or CD8 gate for T cells type of interest. PHA (Phytohemagglutinin A) was used as a polyclonal-positive stimulus. At least 20,000 events were acquired. iDC – without SHED contact; iDC+SHED – iDC previously co-cultured with SHED; mDC – without SHED contact; mDC+SHED – mDC previously co-cultured with SHED. Comparisons were done using t-test between iDC and iDC+SHED and between mDC and mDC+SHED co-cultured with T cells; *p≤0.05 and **p≤0.01 for the groups that showed significant differences; n = 3.</p

MoDCs pre-stimulated with SHEDs induce immune modulation on cytokines production in co-culture with T cells.

<p>Graphics indicate soluble factors levels (pg/mL; detection by CBA technique) present in moDCs culture previously treated with SHEDs (in a 1∶10 ratio; without T cells); and in the co-culture of allogeneic T cells with moDCs (previously co-cultured with or without SHEDs; both cultures in a 1∶10 ratio); both stimulated or not with LPS. Comparisons were done using t-test between iDC and iDC+SHED and between mDC and mDC+SHED; *p≤0.05 and **p≤0.01 for the groups that showed significant differences; n = 4.</p