SEARCH

SEARCH BY CITATION

Keywords:

  • γδ T cells;
  • caspase recruitment domain-containing protein 9;
  • cystitis;
  • interleukin-17A

ABSTRACT

  1. Top of page
  2. ABSTRACT
  3. MATERIALS AND METHODS
  4. RESULTS
  5. DISCUSSION
  6. ACKNOWLEDGMENTS
  7. DISCLOSURE
  8. REFERENCES

Hemorrhagic cystitis often arises after cyclophosphamide (CYP) administration. As yet, however, the mechanism involved in its pathogenesis is unknown. In this study, it was found that the Fc receptor γ chain (FcRγ)- caspase recruitment domain-containing protein 9 (CARD9)-dependent pathway rather than the myeloid differentiation primary response gene 88 (MyD88)-dependent pathway is involved in the pathogenesis of acute CYP-induced cystitis in mice. Rapid and transient production of interleukin (IL)-6 and IL-1β was detected in the bladder at 4 hr, preceding IL-23 and IL-17A production and an influx of neutrophils, which reached a peak at 24 hr after injection. As assessed by weight, edema and neutrophil infiltration, cystitis was significantly attenuated in CARD9 knockout (KO) and FcRγKO mice, this attenuation being accompanied by impaired production of IL-1β, IL-6, IL-23 and IL-17A. The major source of IL-17A is the vesical γδ T cell population: IL-17AKO, CδKO and Tyk2KO mice showed little IL-17A production and reduced neutrophil infiltration in the bladder after CYP injection. These results suggest that FcRγ-CARD9-dependent production of proinflammatory cytokines such as IL-1β, IL-6, and IL-23 and the subsequent activation of IL-17A-producing γδ T cells are at least partly involved in the pathogenesis of acute CYP-induced cystitis in mice.

List of Abbreviations
ATP

adenosine triphosphate

CARD9

caspase recruitment domain-containing protein 9

CδKO

γδT cell-deficient knock out

CLR

C-type lectin receptor

CXCL

chemokine (C-X-C motif) ligand

CYP

cyclophosphamide

DAMP

damage-associated molecular pattern

E. coli

Escherichia coli

FcRγ

Fc receptor γ chain

HC

hemorrhagic cystitis

ITAM

immunorecepotor tyrosine-based activation motif

KO

knockout

MyD88

myeloid differentiation primary response gene 88

NFκB

nuclear factor of κ light polypeptide gene enhancer in B cells

PE

phycoerythrin

P2X7R

purinergic receptor

SAP130

spliceosome-associated protein 130

TCR

T cell receptor

TDM

trehalose-6,6′-dimycolate

TLR

toll-like receptor

TNF

tumor necrosis factor

Hemorrhagic cystitis is one of the adverse effects of treatment with CYP, which is widely used as both an antineoplastic agent and an immunosuppressor for autoimmune disorders such as systemic lupus erythematosus and rheumatoid arthritis [1]. Severe inflammation in the bladder occurs in both mice and rats receiving CYP [2, 3]. Hence, a CYP-induced mouse model can be used to clarify the mechanism of disease progression of HC.

Inflammatory responses are triggered by tissue-resident innate immune cells after recognition of pathogen-associated molecular patterns and DAMPs through TLRs and CLRs [4-6]. The interactions via TLRs and CLRs elicit different downstream pathways through MyD88 and CARD9, respectively, and subsequently induce various inflammatory mediators [5, 7]. It has previously been shown that pretreatment of CYP-induced cystitis with plant-derived glucose–mannose binding lectin reduces leucocyte infiltration and ameliorates tissue damage [8]. In the bladders of mouse models of CYP-induced cystitis, marked increases in cytokines such as γ–interferon, TNF-α, IL-1β and IL-6 and chemokines such as CXCL9, CXCL10 and CXCL11 reportedly occur [9-13]. Neutralization of CXCL10 reduces the severity of CYP-induced cystitis in mice [11]. However, it is still unclear what mechanism triggers the array of inflammatory responses involved in the development of acute CYP-induced cystitis.

γδ T cells are naturally occurring IL-17A-producing cells that are generated within the fetal thymus [14]. In naive mice, they are localized in the periphery, including the bladder, as long-lived effector cells [15]. IL-17A-producing γδ T cells constitutively express large amounts of IL-23R, which induce prompt IL-17A production after binding of IL-23 [16]. Recently, several lines of evidence have suggested that vesical IL-17A-producing γδ T cells are potent inflammatory cells that are directly involved in host defense and anti-tumor activity [17, 18]. In a murine model of urinary tract infection with E. coli, IL-17A-producing γδ T cells reportedly contribute to clearance of the pathogens by mediating neutrophil infiltration and macrophage activation [17]. In bladder cancer, IL-17A-producing γδ T cells contribute to the antitumor effect in the context of adjuvant therapy with bacillus Calmette–Guérin [18]. In fact, accumulation of γδ T cells has been observed in the bladder in human cystitis but not in healthy controls [19]; however, the role of vesical γδ T cells in human cystitis remain unknown.

In this study, we found that the pathogenesis of acute CYP-induced cystitis in mice depends, at least in part, on CARD9, which elicits an inflammatory cascade of IL-1β, IL-6 and IL-23-induced IL-17A production by vesical γδ T cells.

MATERIALS AND METHODS

  1. Top of page
  2. ABSTRACT
  3. MATERIALS AND METHODS
  4. RESULTS
  5. DISCUSSION
  6. ACKNOWLEDGMENTS
  7. DISCLOSURE
  8. REFERENCES

Mice

C57BL/6(WT) mice were purchased from Japan SLC (Shizuoka, Japan). IL-17AKO, CδKO, Tyk2KO, MyD88KO, FcRγKO and CARD9KO mice (B6 background) were generated as previously described [20, 21]. These mice were bred under specific pathogen-free conditions in our institute. Six- to 8-week-old female mice were used for the experiments. The study was approved by the Committee on the Ethics of Animal Experiments of the Faculty of Medicine, Kyushu University. Experiments were performed according to the Guidelines for Animal Experiment.

Induction of cystitis

Cystitis was induced by a single intraperitoneal administration of 300 mg/kg CYP (Sigma–Aldrich, St. Louis, MO, USA).

Measurement of vesical edema

Vesical edema was quantified either by increase in bladder wet weight, reported as mean ± SEM/20 g of body weight or determination of vesical vascular permeability, quantified by Evans blue dye extravasation [22].

Measurement of interleukin-6, interleukin-1β, tumor necrosis factor-α, chemokine (C-X-C motif) ligand 10, interleukin-17A and interleukin-23 in the bladder

At the indicated time after administration, the bladder was minced to yield 1–2 mm pieces, which were then incubated in a mixture of 1 mg/mL collagenase (Invitrogen, Carlsbad, CA, USA) and 20 μg/mL DNase (Sigma–Aldrich) in RPMI 1640 medium containing 10% FCS for 90 min at 37°C. After incubation, amounts of IL-6, IL-1β, TNF-α, IL-17A and IL-23 and CXCL10 in the supernatant were measured using a mouse DuoSet ELISA development system (R & D Systems, Minneapolis, MN, USA), a mouse IL-23 (p19/p40) ELISA Ready-Set-Go set (eBioscience, San Diego, CA, USA), and a CXCL10 Mouse Singleplex Bead Kit (Invitrogen), according to the manufacturers' instructions.

Flow cytometric analysis

Flow cytometric analysis was performed as previously described [20]. The antibodies used in this study were as follows. Fluorescein isothiocyanate-conjugated anti-CD4 (RM4-5), anti-CD11b (H57-797), allophycocyanin-conjugated anti-TCRγδ (GL3), peridinin chlorophyll protein (PerCP)-Cy5.5-conjugated streptavidin, PE-conjugated anti-CD3ϵ (145-2C11) and anti-mIL-17A (TC11-18H10.1) mAbs were purchased from BD Biosciences (San Diego, CA, USA). PE-conjugated anti-Gr1 (RB6-8C5) mAb was purchased from Caltag Laboratories (Burlingame, CA, USA). Allophycocyanin-conjugated F4/80(BM8), biotin-conjugated anti-major histocompatibility complex class II (M5/114.15.2), anti-F4/80 (BM8), anti-TCRβ (H57-597) and anti-B220 (RA3-6B2) mAbs were purchased from eBioscience. Stained cells were run on a fluorescent-activated cell sorting Calibur flow cytometer (BD Biosciences) after adding propidium iodide (1 μg/mL) to exclude dead cells. For intracellular staining, vesical cells were incubated for 4 hr at 37°C. Ten microgram/milliliter of brefeldin A was added for the last 3 hr of incubation. After incubation, the cells were stained with various mAbs for 30 min at 4°C. Intracellular staining was performed according to the manufacturer's instructions (BD Biosciences). Briefly, 100 μL of BD Cytofix/Cytoperm solution (BD Biosciences) was added to the cell suspensions with mild mixing. They were then placed for 20 min at 4°C. Fixed cells were washed twice with 250 μL of BD Perm/Wash solution (BD Biosciences) and then stained intracellularly with PE-conjugated anti-mIL-17A mAb for 30 min at 4°C. The data were analyzed using CellQuest software (BD Biosciences).

Statistical analyses

All values are reported as mean ± SEM. Statistical significance was calculated by one-way ANOVA with Dunnett as a post-hoc test using GraphPad Prism software. Differences with P values of <0.05 were considered statistically significant.

RESULTS

  1. Top of page
  2. ABSTRACT
  3. MATERIALS AND METHODS
  4. RESULTS
  5. DISCUSSION
  6. ACKNOWLEDGMENTS
  7. DISCLOSURE
  8. REFERENCES

Genetic ablation of caspase recruitment domain-containing protein 9 or Fc receptor γ chain, but not of myeloid differentiation primary response gene 88, attenuate the severity of acute cyclophosphamide-induced cystitis in mice

Consistent with a previous finding [1], we observed infiltration of inflammatory cells in the bladder but not in other tissues such as the kidney, liver and spleen (data not shown) in mice with acute CYP-induced cystitis. To determine whether the clinical manifestations of acute CYP-induced cystitis are associated with signaling through TLRs or CLRs, we examined pathological changes in the bladders of MyD88KO and CARD9KO mice after administration of CYP. As shown in Figure 1a,b, compared with CARD9KO mice, in WT and MyD88KO mice we found significant increases in bladder weight and thickened mucosal areas, which correlate with progression of disease [3], as early as 4 hr after administration of CYP (Fig. 1a,b). The bladder weights of WT and MyD88KO mice gradually increased until 24 hr after administration (Fig. 1a). CARD9 is a downstream molecule of CLRs, which are classified into two groups based on whether their cytoplasmic tails do or do not have an ITAM. Some CLRs without an ITAM interact with ITAM-containing FcRγ, which delivers their signaling intracellularly [23, 24]. We found that acute CYP-induced cystitis was less severe in FcRγKO mice and CARD9KO mice than in WT and MyD88KO mice (Fig. 1a,b). There was less severe inflammation in the bladders of CARD9KO and FcRγKO mice than in those of WT and MyD88KO mice as evidenced by vesical permeability (quantified by Evans blue dye extravasation) (Fig. 1c).

image

Figure 1. The FcRγ-CARD9 rather than the MyD88 pathway is involved in development of acute CYP-induced cystitis. (a) Changes in bladder weight after CYP administration. Bladder weight in WT, CARD9KO, FcRγKO and MyD88KO mice was assessed at 0, 4, 12 and 24 hr after intraperitoneal injection of 300 mg/kg of CYP. Data are shown as the mean ± SEM of five mice for each group. *P < 0.05, ***P < 0.001. Data are representative of three independent experiments. (b) Histochemical analysis of bladders taken from mice 4 hr after CYP injection (original magnification, 40 ×). (c) Vesical vascular permeability in the bladders of WT, CARD9 KO, FcRγKO and MyD88KO mice 4 hr after CYP administration was quantified by leakage of Evans blue. Data are shown as the mean ± SEM of five mice for each group. *P < 0.05. Data are representative of three independent experiments.

Download figure to PowerPoint

The interleukin-23-interleukin-17A axis is significantly impaired in the absence of caspase recruitment domain-containing protein 9-mediated signaling

We next investigated which inflammatory mediators are involved in acute CYP-induced cystitis. In agreement with previous studies [9-11, 13], we found that production of IL-6, IL-1β and CXCL10 is rapidly induced in mice with acute CYP-induced cystitis. Production of IL-6 and IL-1β peaked 4 hr after onset of the disease whereas CXCL10 production gradually increased with development of clinical evidence of CYP-induced cystitis (Figs 1a, 2a). Recently, CARD9-dependent signaling was found to be involved in production of IL-17A [25, 26]. We found that IL-17A is produced in the inflamed bladder and rapidly increases for the next 24 hr (Fig. 2a). In addition, IL-23, a well-known inducer of IL-17A production [27], is simultaneously produced (Fig. 2a). IL-6 and IL-1β were also partially decreased in the absence of either FcRγ or CARD9 (Fig. 2b). Of note, IL-23 production was particularly dependent on FcRγ-CARD9 signaling (Fig. 2b). The lack of IL-23 production impaired IL-17A production and neutrophil infiltration in FcRγKO and CARD9KO mice (Fig. 2b,c).

image

Figure 2. The FcRγ-CARD9 pathway is important for production of IL-23 and IL-17A in acute CYP-induced cystitis. (a) Production of various inflammatory mediators by vesical cells was assessed at 0, 4, 12 and 24 hr after CYP administration. Data are shown as the mean ± SEM of three to five mice for each group. Data are representative of three independent experiments. (b) Production of IL-6 (4 hr), IL-1β (4 hr), IL-17A (24 hr) and IL-23 (24 hr) by vesical cells of WT, CARD9KO, FcRγKO and MyD88KO mice was assessed after CYP administration. Data are shown as the mean ± SEM of five mice for each group. *P < 0.05, **P < 0.01, ***P < 0.001. Data are representative of three independent experiments. (c) The number of Gr1 + CD11b + F4/80– neutrophils that had infiltrated the bladders 4 hr after CYP administration was calculated by flow cytometry. Data are shown as the mean ± SEM of three to five mice for each group. *P < 0.05, **P < 0.01. Data are representative of three independent experiments.

Download figure to PowerPoint

Vesical γδ T cells are the major producer of interleukin-17A in acute cyclophosphamide-induced cystitis

Interleukin-17A-producing cells have been identified in various lymphoid cell populations [28, 29]. In mice with acute CYP-induced cystitis, we observed IL-17A-producing cells, but not CD4-positive αβ T cells, in the γδ TCR-positive fraction (Fig. 3a). IL-17A production in the bladders of γδ T cell-deficient mice (CδKO) with acute CYP-induced cystitis was significantly decreased (Fig. 3b). We have previously reported that Tyk2-mediated signaling is indispensable for IL-23-induced IL-17A production by γδ T cells [30]. IL-17A production by γδ T cells was abrogated in Tyk2KO mice (Fig. 3b). In Cδ KO mice and IL-17AKO, neutrophil infiltration was decreased 24 hr after CYP administration (Fig. 3c,d). These results indicate that IL-17A was produced by γδ T cells, which may, at least in part, accelerate the pathogenesis of acute CYP-induced cystitis.

image

Figure 3. γδ T cells are major IL-17A producers in acute CYP-induced cystitis. (a) Identification of IL-17A-producing cells in vesical cells after administration of CYP. Twelve hours after intraperitoneal injection of 300 mg/kg CYP, vesical cells were cultured in the presence of brefeldin A for 4 hr. Flow cytometric analysis was performed without stimulation after gating on major histocompatibility complex class II-negative CD3-positive cells. The numbers in the upper right quadrants indicate the percentage of IL-17A+ cells in γδ TCR+ cells (left) or CD4+ cells (right). (b) CδKO and Tyk2KO mice with acute CYP-induced cystitis have significantly reduced IL-17A production. IL-17A production by vesical cells of WT, CδKO or Tyk2KO mice was measured 24 hr after CYP administration. Data are shown as the mean ± SEM of five mice for each group. **P < 0.01, ***P < 0.001. Data are representative of three independent experiments. (c) Cδ KO and IL-17AKO mice with acute CYP-induced cystitis have significantly reduced neutrophil infiltration. Flow cytometric analysis was performed after gating on CD11b positive. The numbers in the upper left quadrants indicate the percentage within the gate. The number of Gr1 + CD11b + F4/80– neutrophils that had infiltrated the bladders 24 hr after CYP administration was calculated by flow cytometry. Data are shown as the mean ± SEM of five mice for each group. *P < 0.05, ***P < 0.001. Data are representative of three independent experiments. (d) Histochemical analysis was performed on bladders taken from mice 24 hr after CYP injection (original magnification, 40 ×).

Download figure to PowerPoint

DISCUSSION

  1. Top of page
  2. ABSTRACT
  3. MATERIALS AND METHODS
  4. RESULTS
  5. DISCUSSION
  6. ACKNOWLEDGMENTS
  7. DISCLOSURE
  8. REFERENCES

Systemic administration of CYP induces chemical cystitis in both mice and rats and is used as an experimental model of HC [2, 3]. While the precise cause of HC is not known, a number of researchers have suggested that non-infectious urinary bladder inflammation is caused by innate immune responses involving pro-inflammatory cytokines and cell infiltration [9-13]. In the present study, we found that FcRγKO mice and CARD9KO mice are less susceptible to acute CYP-induced cystitis than WT mice. To our knowledge, this is the first evidence that the FcRγ-CARD9 pathway, which is an important CLR signal pathway, is involved at an early stage of acute CYP-induced cystitis.

Caspase recruitment domain-containing protein 9 is an adaptor protein that has an essential role in NF-κB activation and pro-inflammatory cytokine and chemokine production in myeloid cells through CLRs [31]. Several CLRs, such as Mincle (also known as clec4e) and CLEC9a (also known as DNGR-1) have been reported as recognizing self-antigens derived from damaged cells as DAMPs [24, 32]. Mincle associates with ITAM-containing adaptor molecular FcRγ chain and recognizes TDM and spliceosome-associated protein 130 (SAP130), which is released from necrotic cells [24]. TDM is an abundant mycobacterial cell wall glycolipid. Acrolein, a toxic CYP-derived metabolite, triggers HC by damaging the layer of proteoglycan and glycoprotein that covers the epithelium of the bladder in humans and mice [1, 33]. CLEC9a recognizes an unidentified ligand on necrotic cells and contains cytoplasmic ITAM-like motif. In this study, FcRγ which has ITAM, was responsible for proinflammatory cytokine production. Taken together, we speculate that DAMPs such as SAP130 may be released from acrolein-induced damaged apoptotic bladder cells after CYP administration and thence involved in the pathogenesis of acute CYP-induced cystitis.

Recently, it was reported that p2X7 receptor, a purinergic receptor, is involved in inflammatory and nociceptive changes in CYP-induced cystitis in mice [34]. P2X7R is activated extracellularly in an ATP-dependent manner when ATP is released from dying cells, resulting in K+ efflux as a danger signal. K+ efflux activates NLRP3 inflammasome, which contain NALP3, the adapter ASC and pro-caspase-1, to result in caspase-1 activation. Activated caspase-1 converts pro-IL-1β to IL-1β. These findings suggest that extracellular ATP from damaged bladder cells may contribute to IL-1β production in acute CYP-induced cystitis. Indeed, we found that IL-1β production is significantly decreased in CARD9KO and FcRγKO mice after CYP administration. These findings suggest that activation of caspase-1 by ATP may convert pro-IL-1β produced via a FcRγ-CLR pathway to IL-1β, which would induce edema.

A notable finding of the present study is that the MyD88 pathway is not involved in the pathogenesis of acute CYP-induced cystitis. In a previous study, administration of acrolein caused more severe edema in the bladders of TLR4-deficient mice than in those of TLR4-sufficient mice [33] and acrolein inhibited lipopolysaccharide-induced homodimerization of TLR4, which resulted in down-regulation of NF-κB and interferon regulatory factor 3 activation [35]. In addition, TLR4 activates two distinct signaling pathways: the “MyD88-dependent” and “TIR-domain-containing adapter-inducing interferon-β-dependent” pathways [36]. S100A8/9 is known as DAMPs and interacts with TLR4. Strong expression of S100A8/9 has been detected in the bladder and kidney in acute urinary tract infection. However, S100A8/9 does not contribute to an effective host response against E. coli in the urinary tract [37]. In this regard, further study is necessary to elucidate the role of the TLR4 pathway in acute CYP-induced cystitis.

Vesical interstitial infiltration of neutrophils is one of the earliest events induced by CYP administration. IL-17A enhances neutrophil infiltration by promoting IL-6, granulocyte colony-stimulating factor and CXCL8 production [29]. In this study, we found that vesical γδ T cells in the neutrophil infiltrations in the bladder after CYP administration are involved in IL-17A production. Recent report suggests that innate lymphoid cells have C-type lectin receptors and produce IL-17A. Therefore it is possible that IL-17A produced by innate lymphoid cells is also involved in acute CYP-induced cystitis [38]. However, our data showing that IL-17A production is almost completely abolished in TCRδKO mice indicate that γδ T cells are the main source of IL-17A production in acute CYP-induced cystitis. IL-23, which induces IL-17A production, was produced in the bladder 24 hr after CYP administration in a FcRγ-CARD9 dependent manner but not in a MyD88-dependent manner, as were IL-1β and IL-6. Interestingly, we found that IL-17A is not involved in vesical edema. The bladder weights of IL-17AKO mice were not significantly different from those of WT mice after CYP administration (data not shown). These data suggest that edema and neutrophil infiltration are induced by different cytokines.

Although IL-23 production was particularly dependent on the FcRγ-CARD9 pathway, production of IL-17A was not completely stopped. γδ T cells were able to produce IL-17A not only in the standard manner with IL-23, but also via TCR signaling. These results suggest that γδ T cells may recognize an unknown ligand in the bladder and produce IL-17A upon TCR stimulation. At present, the specificity of γδ T cells remains unknown. Therefore, it will be of interest to ascertain whether γδ T cells in the murine bladder recognize any such unique antigen.

In conclusion, we found that the CARD9-dependent pathway, which induces IL-17A production by γδ T cells, contributes at least in part to the pathogenesis of acute CYP-induced cystitis. In clear contrast, we have previously reported that the TLR2-MyD88 pathway induces IL-17A production by γδ T cells, which are involved in host defense against extracellular pathogens such as E. coli and Candida albicans [20, 39]. These findings clearly indicate the importance of innate immune cells, which regulate not only host defense but also autoimmune disorders by activating distinct signaling pathways. These responses are primarily regulated by ligand–receptor interactions. In this regard, further studies of the molecular mechanisms responsible for the initiation of HC are required. Nevertheless, the FcRγ-CARD9 pathway could be a novel target for the management of acute CYP- induced cystitis.

ACKNOWLEDGMENTS

  1. Top of page
  2. ABSTRACT
  3. MATERIALS AND METHODS
  4. RESULTS
  5. DISCUSSION
  6. ACKNOWLEDGMENTS
  7. DISCLOSURE
  8. REFERENCES

We thank K. Akasaki, M. Ohkubo, M. Kijima and A. Yano for their excellent technical assistance. This work was supported by a Grant-in-Aid for Scientific Research from the Japan Society for the Promotion of Science (K.S. and Y.Y.), by the Kaibara Morikazu Medical Science Promotion Foundation (K.S.), by Takeda Science Foundation, and in part by Grants for Excellent Graduate Schools, MEXT, Japan.

DISCLOSURE

  1. Top of page
  2. ABSTRACT
  3. MATERIALS AND METHODS
  4. RESULTS
  5. DISCUSSION
  6. ACKNOWLEDGMENTS
  7. DISCLOSURE
  8. REFERENCES

None of the authors has any conflict of interest associated with this study.

REFERENCES

  1. Top of page
  2. ABSTRACT
  3. MATERIALS AND METHODS
  4. RESULTS
  5. DISCUSSION
  6. ACKNOWLEDGMENTS
  7. DISCLOSURE
  8. REFERENCES
  • 1
    Cox P.J. (1979) Cyclophosphamide cystitis—identification of acrolein as the causative agent. Biochem Pharmacol 28: 20459.
  • 2
    Fraiser L., Kehrer J.P. (1992) Murine strain differences in metabolism and bladder toxicity of cyclophosphamide. Toxicology 75: 25772.
  • 3
    Gray K.J., Engelmann U.H., Johnson E.H., Fishman I.J. (1986) Evaluation of misoprostol cytoprotection of the bladder with cyclophosphamide (Cytoxan) therapy. J Urol 136: 497500.
  • 4
    Piccinini A.M., Midwood K.S. (2010) DAMPening inflammation by modulating TLR signalling. Mediators Inflamm 2010: 21 Article ID 672395.
  • 5
    Takeda K., Kaisho T., Akira S. (2003) Toll-like receptors. Annu Rev Immunol 21: 33576.
  • 6
    Gringhuis S.I., Wevers B.A., Kaptein T.M., Van Capel T.M., Theelen B., Boekhout T., De Jong E.C., Geijtenbeek T.B. (2011) Selective C-Rel activation via Malt1 controls anti-fungal T(H)-17 immunity by dectin-1 and dectin-2. PLoS Pathog 7: e1001259.
  • 7
    Hara H., Ishihara C., Takeuchi A., Imanishi T., Xue L., Morris S.W., Inui M., Takai T., Shibuya A., Saijo S., Iwakura Y., Ohno N., Koseki H., Yoshida H., Penninger J.M., Saito T. (2007) The adaptor protein CARD9 is essential for the activation of myeloid cells through ITAM-associated and Toll-like receptors. Nat Immunol 8: 61929.
  • 8
    Assreuy A.M., Martins G.J., Moreira M.E., Brito G.A., Cavada B.S., Ribeiro R.A., Flores C.A. (1999) Prevention of cyclophosphamide-induced hemorrhagic cystitis by glucose-mannose binding plant lectins. J Urol 161: 198893.
  • 9
    Gomes T.N., Santos C.C., Souza-Filho M.V., Cunha F.Q., Ribeiro R.A. (1995 Participation of TNF-alpha and IL-1 in the pathogenesis of cyclophosphamide-induced hemorrhagic cystitis. Braz J Biol Res 28: 11038.
  • 10
    Nishii H., Nomura M., Fujimoto N., Matsumoto T. (2006) Up-regulation of interleukin-6 gene expression in cyclophosphamide-induced cystitis in mice: an in situ hybridization histochemical study. Int J Urol 13: 133943.
  • 11
    Sakthivel S.K., Singh U.P., Singh S., Taub D.D., Novakovic K.R., Lillard J.W., Jr. (2008) CXCL10 blockade protects mice from cyclophosphamide-induced cystitis. J Immune Based Ther Vaccines 6: 6.
  • 12
    Chuang Y.C., Tyagi P., Huang H.Y., Yoshimura N., Wu M., Kaufman J., Chancellor M.B. (2011) Intravesical immune suppression by liposomal tacrolimus in cyclophosphamide-induced inflammatory cystitis. Nueurorol Urodyn 30: 4217.
  • 13
    Malley S.E., Vizzard M.A. (2002) Changes in urinary bladder cytokine mrna and protein after cyclophosphamide-induced cystitis. Physiol Genomics 9(1): 513.
  • 14
    Shibata K., Yamada H., Nakamura R., Sun X., Itsumi M., Yoshikai Y. (2008) Identification of CD25+ gamma delta T cells as fetal thymus-derived naturally occurring IL-17 producers. J Immunol 181: 59407.
  • 15
    Haas J.D., Ravens S., Duber S., Sandrock I., Oberdorfer L., Kashani E., Chennupati V., Fohse L., Naumann R., Weiss S., Krueger A., Forster R., Prinz, I. (2012) Development of interleukin-17-producing gammadelta T cells is restricted to a functional embryonic wave. Immunity 37: 4859.
  • 16
    Riol-Blanco L., Lazarevic V., Awasthi A., Mitsdoerffer M., Wilson B.S., Croxford A., Waisman A., Kuchroo V.K., Glimcher L.H., Oukka M. (2010) IL-23 receptor regulates unconventional IL-17-producing T cells that control bacterial infections. J Immunol 184: 171020.
  • 17
    Sivick K.E., Schaller M.A., Smith S.N., Mobley H.L. (2010) The innate immune response to uropathogenic Escherichia coli involves IL-17A in a murine model of urinary tract infection. J Immunol 184: 206575.
  • 18
    Takeuchi A., Dejima T., Yamada H., Shibata K., Nakamura R., Eto M., Nakatani T., Naito S., Yoshikai Y. (2011) IL-17 production by gammadelta T cells is important for the antitumor effect of Mycobacterium bovis bacillus Calmette-Guerin treatment against bladder cancer. Eur J Immunol 41: 24651.
  • 19
    Christmas T.J. (1994) Lymphocyte sub-populations in the bladder wall in normal bladder, bacterial cystitis and interstitial cystitis. Br J Urol 73: 50815.
  • 20
    Dejima T., Shibata K., Yamada H., Hara H., Iwakura Y., Naito S., Yoshikai Y. (2011) Protective role of naturally occurring interleukin-17A-producing gammadelta T cells in the lung at the early stage of systemic candidiasis in mice. Infect Immun 79: 450310.
  • 21
    Park S.Y., Ueda S., Ohno H., Hamano Y., Tanaka M., Shiratori T., Yamazaki T., Arase H., Arase N., Karasawa A., Sato S., Ledermann B., Kondo Y., Okumura K., Ra C., Saito T. (1998) Resistance of Fc receptor-deficient mice to fatal glomerulonephritis. J Clin Invest 102: 122938.
  • 22
    Ribeiro R.A., Freitas H.C., Campos M.C., Santos C.C., Figueiredo F.C., Brito G.A., Cunha F.Q. (2002) Tumor necrosis factor-alpha and interleukin-1beta mediate the production of nitric oxide involved in the pathogenesis of ifosfamide induced hemorrhagic cystitis in mice. J Urol 167: 222934.
  • 23
    Osorio F., Reis E Sousa C. (2011) Myeloid C-type lectin receptors in pathogen recognition and host defense. Immunity 34: 65164.
  • 24
    Yamasaki S., Ishikawa E., Sakuma M., Hara H., Ogata K., Saito T. (2008) Mincle is an ITAM-coupled activating receptor that senses damaged cells. Nat Immunol 9: 117988.
  • 25
    Glocker E.O., Hennigs A., Nabavi M., Schaffer A.A., Woellner C., Salzer U., Pfeifer D., Veelken H., Warnatz K., Tahami F., Jamal S., Manguiat A., Rezaei N., Amirzargar A.A., Plebani A., Hannesschlager N., Gross O., Ruland J., Grimbacher B. (2009) A homozygous CARD9 mutation in a family with susceptibility to fungal infections. N Eng J Med 361: 172735.
  • 26
    Leibundgut-Landmann S., Gross O., Robinson M.J., Osorio F., Slack E.C., Tsoni S.V., Schweighoffer E., Tybulewicz V., Brown G.D., Ruland J., Reis E Sousa, C. (2007) Syk- and CARD9-dependent coupling of innate immunity to the induction of T helper cells that produce interleukin 17. Nat Immunol 8: 6308.
  • 27
    Aggarwal S., Ghilardi N., Xie M.H., De Sauvage F.J., Gurney A.L. (2003) Interleukin-23 promotes a distinct CD4 T cell activation state characterized by the production of interleukin-17. J Biol Chem 278: 19104.
  • 28
    Harrington L.E., Hatton R.D., Mangan P.R., Turner H., Murphy T.L., Murphy K.M., Weaver C.T. (2005) Interleukin 17-producing CD4+ effector T cells develop via a lineage distinct from the T helper type 1 and 2 lineages. Nat Immunol 6: 112332.
  • 29
    Cua D.J., Tato C.M. (2010) Innate IL-17-producing cells: the sentinels of the immune system. Nat Rev Immunol 10: 47989.
  • 30
    Nakamura R., Shibata K., Yamada H., Shimoda K., Nakayama K., Yoshikai Y. (2008) Tyk2-signaling plays an important role in host defense against Escherichia coli through IL-23-induced IL-17 production by gammadelta T cells. J Immunol 181: 20715.
  • 31
    Hara H., Saito T. (2009) CARD9 versus CARMA1 in innate and adaptive immunity. Trends Immunol 30: 23442.
  • 32
    Sancho D., Joffre O.P., Keller A.M., Rogers N.C., Martinez D., Hernanz-Falcon P., Rosewell I., Reis E Sousa, C. (2009) Identification of a dendritic cell receptor that couples sensing of necrosis to immunity. Nature 458: 899903.
  • 33
    Bjorling D.E., Elkahwaji J.E., Bushman W., Janda L.M., Boldon K., Hopkins W.J., Wang Z.Y. (2007) Acute acrolein-induced cystitis in mice. BJU Int 99: 15239.
  • 34
    Martins J.P., Silva R.B., Coutinho-Silva R., Takiya C.M., Battastini A.M., Morrone F.B., Campos M.M. (2012) The role of P2 × 7 purinergic receptors in inflammatory and nociceptive changes accompanying cyclophosphamide-induced haemorrhagic cystitis in mice. Br J Pharmacol 165: 18396.
  • 35
    Lee J.S., Lee J.Y., Lee M.Y., Hwang D.H., Youn H.S. (2008) Acrolein with an alpha, beta-unsaturated carbonyl group inhibits LPS-induced homodimerization of toll-like receptor 4. Mol Cells 25: 2537.
  • 36
    Kawai T., Akira S. (2011) Toll-like receptors and their crosstalk with other innate receptors in infection and immunity. Immunity 34: 63750.
  • 37
    Dessing M.C., Butter L.M., Teske G.J., Claessen N., Van Der Loos C.M., Vogl T., Roth J., Van Der Poll T., Florquin S., Leemans J.C. (2010) S100A8/A9 is not involved in host defense against murine urinary tract infection. PLoS ONE 5: e13394.
  • 38
    Sutton C.E., Mielke L.A., Mills K.H. (2012) Il-17-producing gammadelta T cells and innate lymphoid cells. Eur J Immunol 42: 222131.
  • 39
    Shibata K., Yamada H., Hara H., Kishihara K., Yoshikai Y. (2007) Resident Vdelta1+ gammadelta T cells control early infiltration of neutrophils after Escherichia coli infection via IL-17 production. J Immunol 178: 446672.