Candida Albicans Against Protective Antifungal Adaptive Immunity Cutting Edge: Nlrp10 Is Essential

Cutting Edge: Nlrp10 Is Essential for Protective Antifungal Adaptive Immunity against Candida albicans

This information is current as Sophie Joly, Stephanie C. Eisenbarth, Alicia K. Olivier, of September 28, 2021. Adam Williams, Daniel H. Kaplan, Suzanne L. Cassel, Richard A. Flavell and Fayyaz S. Sutterwala J Immunol 2012; 189:4713-4717; Prepublished online 15 October 2012;

doi: 10.4049/jimmunol.1201715 Downloaded from http://www.jimmunol.org/content/189/10/4713

Supplementary http://www.jimmunol.org/content/suppl/2012/10/15/jimmunol.120171 Material 5.DC1 http://www.jimmunol.org/ References This article cites 19 articles, 7 of which you can access for free at: http://www.jimmunol.org/content/189/10/4713.full#ref-list-1

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The Journal of Immunology is published twice each month by The American Association of Immunologists, Inc., 1451 Rockville Pike, Suite 650, Rockville, MD 20852 Copyright © 2012 by The American Association of Immunologists, Inc. All rights reserved. Print ISSN: 0022-1767 Online ISSN: 1550-6606. Cutting Edge: Nlrp10 Is Essential for Protective Antifungal Adaptive Immunity against Candida albicans x Sophie Joly,* Stephanie C. Eisenbarth,† Alicia K. Olivier,‡ Adam Williams, { ,‖ x,# Daniel H. Kaplan, Suzanne‖ L. Cassel,* Richard A. Flavell, and Fayyaz S. Sutterwala*, ,** Nucleotide-binding domain leucine-rich repeat con- Of interest, one NLR family member, Nlrp10, lacks the C- taining receptors (NLRs) are cytosolic receptors that terminal leucine-rich repeat domain and, therefore, was hy- initiate immune responses to sterile and infectious pothesized to function as a negative regulator of inflamma- insults to the host. Studies demonstrated that Nlrp3 some activation (6, 7). is critical for the control of Candida albicans infections In this study, we demonstrate that Nlrp10, unlike Nlrp6, and in the generation of antifungal Th17 responses. In Nlrp12, and Nlrc4, is required for control of a disseminated Downloaded from this article, we show that the NLR family member C. albicans infection in vivo. We also show that, in contrast Nlrp10 also plays a unique role in the control of dis- to Nlrp3, the absence of Nlrp10 in macrophages (MFs) and seminated C. albicans infection in vivo. Nlrp10-deficient dendritic cells (DCs) does not affect inflammasome activation mice had increased susceptibility to disseminated candi- in response to C. albicans or other inflammasome activators. A recent study also demonstrated that Nlrp10-deficient DCs diasis, as indicated by decreased survival and increased http://www.jimmunol.org/ have defective migration (8); in this study, we demonstrate fungal burdens. In contrast to Nlrp3, Nlrp10 deficiency that, despite normal inflammasome activation, Nlrp10-deficient did not affect innate proinflammatory cytokine pro- mice display a profound defect in the generation of Candida- duction from macrophages and dendritic cells chal- specific Th1 and Th17 responses. Thus, our results implicate lenged with C. albicans. However, Nlrp10-deficient Nlrp10 as a novel NLR involved in the generation of anti- mice displayed a profound defect in Candida-specific fungal adaptive immune responses against C. albicans through Th1 and Th17 responses. These results demonstrate a mechanism that is independent of the Nlrp3 inflammasome a novel role for Nlrp10 in the generation of adaptive and the production of IL-1b. immune responses to fungal infection. The Journal by guest on September 28, 2021 of Immunology, 2012, 189: 4713–4717. Materials and Methods Mice and bone marrow chimeras 2/2 2/2 2/2 2/2 2/2 embers of the NLRP subfamily contain a central The generation of Nlrp10 , Nlrp6 , Nlrp12 , Nlrc4 , and ASC mice was described previously (8–12). Sex- and age-matched C57BL/6 (Na- nucleotide-binding domain (NACHT), an N-termi- tional Cancer Institute) mice were used as controls. All protocols used in this M nal pyrin domain, and C-terminal leucine-rich repeats study were approved by the Institutional Animal Care and Use Committee at the University of Iowa. Bone marrow chimeras were generated, as described thought to function in ligand sensing (1). Recently, we and 2 2 (8). Reconstitution was .97% in Nlrp10 / mice reconstituted with wild- 2 2 other investigators showed that the nucleotide–binding domain type (WT) bone marrow and .82% in WT mice reconstituted with Nlrp10 / leucine-rich repeat containing receptor (NLR) family member bone marrow. Nlrp3 plays an important role in host defense against Candida albicans through triggering the assembly and activation of the In vivo infection with C. albicans Nlrp3 inflammasome (2–4). Nlrc4 also functions within the The C. albicans clinical isolate FC20 was used in this study (2). Culture con- mucosal stroma to control oral C. albicans infections (5). ditions for C. albicans yeast and hyphae were described previously (2). Mice were infected i.v. with 5 3 105 CFU C. albicans, and survival was assessed; mice However, other than Nlrp3 and Nlrc4, the role of NLR found in a moribund state for .4 h were considered terminal and euthanized. family members in fungal pathogenesis remains unknown. Kidneys were harvested at the indicated time postinfection (p.i.), and dilutions

*Inﬂammation Program, University of Iowa Carver College of Medicine, Iowa City, IA Address correspondence and reprint requests to Dr. Fayyaz Sutterwala, University of 52242; †Department of Laboratory Medicine, Yale University School of Medicine, New Iowa, 2501 Crosspark Road, D156 MTF, Coralville, IA 52241. E-mail address: fayyaz- Haven, CT 06520; ‡Department of Pathology, University of Iowa Carver College of [email protected] x Medicine, Iowa City, IA 52242; Department of Immunobiology, Yale University { The online version of this article contains supplemental material. School of Medicine, New Haven, CT 06520; Department of Dermatology, Center ‖ for Immunology, University of Minnesota, Minneapolis, MN 55455; Department of Abbreviations used in this article: BMDC, bone marrow–derived dendritic cell; BMMF, Internal Medicine, University of Iowa Carver College of Medicine, Iowa City, IA 52242; bone marrow–derived macrophage; BUN, blood urea nitrogen; DC, dendritic cell; LVS, #Howard Hughes Medical Institute, Yale University School of Medicine, New Haven, live vaccine strain; MF, macrophage; MOI, multiplicity of infection; NLR, nucleotide- CT 06520; and **Veterans Affairs Medical Center, Iowa City, IA 52241 binding domain leucine-rich repeat containing receptor; p.i., postinfection; WT, wild-type. Received for publication June 21, 2012. Accepted for publication September 21, 2012. Copyright Ó 2012 by The American Association of Immunologists, Inc. 0022-1767/12/$16.00 This work was supported by National Institutes of Health Grants R01 AI087630 (to F.S.S.), R01 AR060744 (to D.H.K.), K08 AI067736 (to S.L.C.), and K08 AI085038 (to S.C.E.), and a scholarship from the Edward Mallinckrodt, Jr. Foundation (to F.S.S.). R.A.F. is an Investigator of the Howard Hughes Medical Institute. www.jimmunol.org/cgi/doi/10.4049/jimmunol.1201715 4714 CUTTING EDGE: NLRP10 CONTROL OF C. ALBICANS INFECTION of homogenized organs were plated and counted to determine CFU. Serum blood urea nitrogen (BUN) and creatinine levels were quantiﬁed at the Animal Fluid Analysis Core at the University of Iowa. To assess renal cytokine levels, kidneys were homogenized and resuspended in lysis buffer (50 mM Tris·HCl, 5 mM EDTA, 150 mM NaCl, 1% Triton X-100, and a protease inhibitor mixture [Roche]), and cytokine levels were measured by ELISA.

Ex vivo lymphocyte restimulation Mice were infected i.v. with a sublethal dose (5 3 104 CFU) of C. albicans. Fourteen days p.i., spleens were collected and splenocytes were cultured in 2 the presence or absence of 1 3 107 ml 1 heat-killed C. albicans for 72 h. Supernatants were collected, and IL-17 and IFN-g levels were assessed by ELISA (eBiosciences).

CD4+ T cell adoptive transfer WT mice were infected i.v. with 5 3 104 CFU C. albicans; 10 d p.i., mice were rechallenged i.v. with 1 3 106 heat-killed C. albicans. Five days later, splenic CD4+ T cells were isolated using MACS MicroBeads (Miltenyi Biotec). CD4+ T cells isolated from uninfected WT mice were used as naive controls. A total of 5 3 106 naive or immune CD4+ T cells was transferred 2 2 i.v. into Nlrp10 / mice; 24 h following the adoptive transfer mice were infected i.v. with 5 3 105 CFU C. albicans, and survival was monitored. Downloaded from

In vitro stimulation of MFs and DCs Bone marrow–derived MFs(BMMFs) and bone marrow–derived DCs (BMDCs) (13, 14) were either left unprimed or were primed with 50 ng/ml LPS (InvivoGen) for 3–4 h and then infected with C. albicans (multiplicity of infection [MOI] 10:1), Francisella tularensis live vaccine strain (LVS) (MOI 50:1), or Pseudomonas aeruginosa PAK strain (MOI 10:1) for 6 h or as in- http://www.jimmunol.org/ dicated. LPS-primed BMMFs were challenged with 50 mg/cm2 silica (Min- U-Sil-5; Pennsylvania Glass Sand), 5 mM ATP (Sigma), or 20 mM nigericin (Sigma) for 6 h. For ATP- and nigericin–treated cells, media were replaced with fresh media 30 min after stimulation. Ab pairs for ELISA were from eBiosciences, with the exception of IL-1b (R&D Systems). Results and Discussion Nlrp10-deﬁcient mice are highly susceptible to disseminated

C. albicans infection by guest on September 28, 2021 FIGURE 1. Nlrp10-deficient mice have increased susceptibility to systemic 2 2 Phylogenetic analysis of the NLR family demonstrates that C. albicans infection. Kaplan–Meier survival curves of WT (n = 17), Nlrp6 / 2 2 2 2 Nlrp6, Nlrp10, and Nlrp12 NACHT sequences are closely (n =16),Nlrp12 / (n =14),andNlrc4 / (n = 13) mice (A)orWT(n = 10) 2/2 5 related to that of Nlrp3 (1). In addition, Nlrc4, which acti- and Nlrp10 (n =10)mice(B) infected i.v. with 5 3 10 CFU of C. albicans yeast. Results are pooled from two independent experiments. p , 0.01, WT vates caspase-1 in response to cytosolic flagellin and bacterial 2 2 versus Nlrp10 / mice, log–rank test. (C) Histology of kidneys, at 9 d p.i., from 2 2 type III secretion systems (1), was also shown to play a role in WT and Nlrp10 / mice stained with H&E, Grocott’s methenamine silver controlling mucosal C. albicans infections (5). We hypothe- stain (GMS), or Masson’s trichrome stain (MT). Black arrows indicate yeast sized that, similarly to Nlrp3, these receptors might contrib- and hyphae in the GMS stain and collagen deposition (light blue) in the MT ute to the in vivo immune response against a systemic C. stain. Scale bars, 50 mM. (D) Serum creatinine and BUN levels were measured albicans infection. To assess this, we tested the susceptibility at the indicated times p.i. with 5 3 105 CFU of C. albicans yeast. Data rep- 6 of Nlrp6-, Nlrp10-, Nlrp12-, and Nlrc4-deficient mice to resent the mean SEM (n =9forday3,n = 6 for day 6, and n = 14–16 for E 2/2 a systemic infection with C. albicans. Nlrp6-, Nlrp12-, and day 9). ( )WTandNlrp10 mice (n = 10 for day 3, n = 8 for day 6, and n = 15–18 for day 9) were infected i.v. with 5 3 105 CFU of C. albicans; fungal Nlrc4-deficient mice did not show increased susceptibility to burdens in the kidney were assessed at the indicated times p.i. *p , 0.05, i.v. infection with C. albicans compared to WT mice (Fig. , 2 2 Student t test, ***p 0.001, Mann–Whitney U test. 1A). Surprisingly, Nlrp10 / mice were highly susceptible to C. albicans infection, with 100% mortality by day 16 p.i. (Fig. 1B). (Fig. 1C, Supplemental Fig. 1A). Very few yeast were detected histologically in kidney sections from WT mice in contrast to 2 2 Renal dysfunction in Nlrp10-deficient mice reflects increased fungal Nlrp10 / mice, in which C. albicans yeast and hyphae were invasion of kidneys at the late stage of infection readily observed in the renal cortex and medulla (Fig. 1C, Sepsis is the main cause of death in hematogenously dissem- Supplemental Fig. 1B). Surprisingly, despite increased C. albi- 2 2 inated candidiasis; in this model, renal dysfunction strongly cans within the renal parenchyma of Nlrp10 / mice, there correlates with increased kidney fungal burdens, as well as was no significant difference in the percentage of parenchymal 2 2 increased mortality (15). Therefore, we evaluated kidneys of MFs and neutrophil staining between WT and Nlrp10 / 2 2 WT and Nlrp10 / mice 9 d p.i. with C. albicans. Histologic mice (Supplemental Fig. 1C), suggesting a possible functional sections of kidneys revealed more severe early fibroplasia and defect in the inflammatory response observed in the absence 2 2 parenchymal loss in kidneys of Nlrp10 / mice compared of Nlrp10. 2 2 with WT mice (Fig. 1C). Kidneys from Nlrp10 / mice had Consistent with the increased renal damage observed by 2 2 significantly more collagen deposition, as shown by Masson’s histology, Nlrp10 / mice had diminished renal function at trichrome stain, than did WT mice, indicating greater damage 9 d p.i., as reflected by significantly higher serum BUN and The Journal of Immunology 4715 creatinine levels (Fig. 1D). Increased damage in Nlrp10-deficient kidneys correlated with elevated IL-1a and IL-6 levels within the kidney at day 9 p.i., although IL-1b, IL-18, IL-12p40, and IL-23 levels were unaffected by Nlrp10 deficiency (Sup- 2 2 plemental Fig. 1D). Nlrp10 / mice also had significantly higher fungal burdens in the kidney at 9 d p.i., indicating a role for Nlrp10 in controlling the replication of C. albicans in vivo (Fig. 1E). Surprisingly, during the early stages of infection, examined at days 3 and 6, there was no difference 2 2 between WT and Nlrp10 / mice with regard to renal function (Fig. 1D), fungal burdens (Fig. 1E), and kidney cytokines (data not shown). These data suggest that early innate mech- anisms required to control C. albicans replication in vivo remain intact in Nlrp10-deficient mice.

Nlrp10 functions within the hematopoietic compartment to control disseminated infection with C. albicans

To better understand the biological function of Nlrp10, we Downloaded from examined the tissue distribution of Nlrp10 in WT mice. Consistent with previous reports, we found high expression of Nlrp10 mRNA in the heart (7); in addition, Nlrp10 was highly expressed in the tongue, testis, and spleen (Supplemental Fig. 1E). Within the hematopoietic compartment, Nlrp10 was + + expressed in MFs, DCs, CD4 T cells, CD19 B cells, and http://www.jimmunol.org/ neutrophils but minimally in CD8+ T cells (Supplemental Fig. 1F). Stimulation of MFs and DCs with live C. albicans in vitro resulted in a reduction in Nlrp10 expression in these cells (Supplemental Fig. 1G). In contrast, LPS and heat-killed C. albicans did not signiﬁcantly alter Nlrp10 mRNA expression (Supplemental Fig. 1G). Given that Nlrp10 is expressed in both hematopoietic cells

and stromal cells, we wanted to determine whether the in- by guest on September 28, 2021 2 2 creased susceptibility of Nlrp10 / mice to disseminated candidiasis was the result of a loss of Nlrp10 in the hematopoietic FIGURE 2. Nlrp10 in hematopoietic cells is required for control of a systemic C. albicans infection. (A) Kaplan–Meier survival curves of bone marrow compartment. To do this, we generated bone marrow chimeric chimeras (donor/recipient) infected i.v. with 5 3 105 CFU of C. albicans mice in which Nlrp10 deficiency was restricted to either the yeast. Results are pooled from two independent experiments (n = 14–15). 2 2 hematopoietic or nonhematopoietic compartment. WT mice There was no significant (ns) difference between WT/Nlrp10 / and 2 2 2 2 2 2 2 2 that received Nlrp10 / bone marrow were susceptible to dis- WT/WT mice or between Nlrp10 / /WT and Nlrp10 / /Nlrp10 / 2 2 2 2 seminated candidiasis and recapitulated the phenotype observed mice. *p , 0.01, WT/Nlrp10 / versus Nlrp10 / /WT mice, log–rank 2 2 2/2 in Nlrp10-deficient mice (Fig. 2A). Conversely, Nlrp10 / test. (B) Unprimed and LPS-primed BMMFs from WT and Nlrp10 mice mice that received WT bone marrow did not have signifi- were stimulated or not for 6 h with C. albicans yeast (MOI 10:1), and IL-1b secretion was quantified by ELISA. (C) LPS-primed BMMFs from WT, 2 2 2 2 cantly increased mortality compared with WT mice that re- Nlrp10 / , and ASC / mice were challenged for 6 h with C. albicans (MOI ceived WT bone marrow (Fig. 2A). These results suggest that 10:1) or P. aeruginosa (MOI 10:1) or for 9 h with F. tularensis LVS (MOI 2/2 the increased susceptibility of Nlrp10 mice to C. albicans 50:1); IL-1b secretion was quantified by ELISA. (D) LPS-primed BMMFs 2 2 2 2 infection was primarily due to a deficiency in Nlrp10 within from WT, Nlrp10 / , and ASC / mice were challenged with silica (50 mg/ the hematopoietic compartment. A recent study by Lautz et al. cm2) or nigericin (20 mM) for 6 h, and IL-1b secretion was quantified by E 2/2 (16) demonstrated that Nlrp10 contributes to proinflammatory ELISA. ( ) BMMFs from WT and Nlrp10 mice were stimulated for 6 h a cytokine release by epithelial cells and dermal fibroblasts in with 50 ng/ml LPS, and IL-12 p40, TNF- , and IL-6 levels were assessed by ELISA. Determinations were performed in triplicate and expressed as the response to infection with Shigella flexneri,whichmayex- mean 6 SEM; results are representative of three independent experiments. plain the reduction, although not significant, in the survival 2 2 of Nlrp10 / mice that received WT bone marrow. Nlrp3 agonists silica and nigericin induced similar levels of Nlrp10 deficiency does not affect inflammasome activation IL-1b secretion from Nlrp10-deficient BMMFs compared Recent studies using in vitro overexpression of Nlrp10, as with WT BMMFs (Fig. 2D). In addition, P. aeruginosa and well as Nlrp10 transgenic mice, suggested that Nlrp10 could F. tularensis LVS, activators of the Nlrc4 and AIM2 inflam- inhibit the activation of Nlrp3 and Nlrc4 inflammasomes and masomes, respectively (1), induced comparable levels of IL- 2 2 suppress NF-kB activation (6, 7). We examined the ability of 1b secretion from Nlrp10 / and WT BMMFs (Fig. 2C). As Nlrp10-deficient MFs to secrete IL-1b in response to specific expected, IL-1b secretion in response to C. albicans, P. aer- inflammasome agonists. LPS-primed BMMFs from WT and uginosa, and F. tularensis LVS was dependent on the presence 2 2 Nlrp10 / mice secreted comparable levels of IL-1b when of the inflammasome adaptor molecule ASC (Fig. 2C). Similar challenged with C. albicans yeast (Fig. 2B, 2C). Similarly, the to our findings with BMMFs, BMDCs from WT and 4716 CUTTING EDGE: NLRP10 CONTROL OF C. ALBICANS INFECTION

2 2 Nlrp10 / mice secreted comparable levels of IL-1b when challenged with C. albicans yeast (Supplemental Fig. 2A). In 2 2 addition, both unprimed and LPS-primed WT and Nlrp10 / BMMFs failed to secrete IL-1b in response to C. albicans hyphae (Supplemental Fig. 2B). C. albicans hyphae were ca- pable of inducing the secretion of IL-1b from LPS-primed WT BMDCs, although again this was similar to the levels of IL-1b 2 2 secreted from LPS-primed Nlrp10 / BMDCs (Supplemental Fig. 2C). These data indicate that a deficiency in Nlrp10 in MFs or DCs does not affect the activation of Nlrp3, Nlrc4, and AIM2 inflammasomes. Consistent with a recent study by Eisenbarth et al. (8), in response to stimulation with the TLR4 agonist LPS, the production of IL-6, TNF-a,andIL-12 p40 was unaffected by Nlrp10 deficiency in both BMMFs and BMDCs (Fig. 2E, Supplemental Fig. 2D), suggesting that Nlrp10 also does not suppress NF-kB activation in these cells.

Internalization and killing of Candida is an indispensable Downloaded from function of MFs in the control of candidal infections. Nlrp10- deficient BMMFs and BMDCs did not display any defect in their ability to phagocytose C. albicans compared with WT BMMFs and BMDCs (Supplemental Fig. 2E). Growth of FIGURE 3. Nlrp10-deficient mice fail to mount a protective T cell response 2 2 C. albicans within MFs was also comparable between WT against C. albicans.(A) WT and Nlrp10 / mice were infected with a sub- 2/2 3 4 and Nlrp10 BMMFs (Supplemental Fig. 2F). Similarly, lethal dose (5 10 CFU) of C. albicans yeast. Fourteen days p.i., splenocytes http://www.jimmunol.org/ we did not observe defects in the phagocytosis or growth of were restimulated with heat-killed C. albicans; 72 h later, IL-17 and IFN-g 2/2 secretion into the supernatant was assessed by ELISA. Data are pooled from C. albicans within Nlrp10 thioglycollate–elicited peritoneal three independent experiments (n = 6–8) and expressed as the mean 6 SEM. neutrophils (Supplemental Fig. 2E, 2F). Taken together, these (B) A total of 5 3 106 CD4+ T cells from either naive or immune WT mice 2 2 data suggest that phagocytosis, intracellular killing, and the was adoptively transferred into Nlrp10 / mice, followed by i.v. infection generation of proinflammatory cytokines by MFsandDCs with 5 3 105 CFU of C. albicans yeast; survival was monitored. Data are pooled from two independent experiments (n = 13–15). *p , 0.05, Student t remain intact in the absence of Nlrp10. 2 2 test, **p , 0.01, Nlrp10 / mice that received naive CD4+ T cell or PBS + Nlrp10 is necessary for generating Candida-specific Th1 and Th17 versus immune CD4 T cells, log-rank test. by guest on September 28, 2021 responses 2 2 Adaptive immune responses play a crucial role in host defense C. albicans. Nlrp10 / mice that received CD4+ T cells from against C. albicans. Th17 responses, in particular, are important naive WT mice succumbed to C. albicans infection at a sim- 2 2 for control of C. albicans infections through the recruitment ilar rate to Nlrp10 / control mice (Fig. 3B). However, 2 2 of neutrophils to the infection site. As such, mice deficient in Nlrp10 / mice that received CD4+ T cells from WT mice the cytokine receptor IL-17RA have increased susceptibility that had previously been challenged with a sublethal dose of to both disseminated and mucosal candidiasis (17, 18). Given C. albicans and boosted with heat-killed C. albicans displayed our findings that Nlrp10 deficiency had little effect on MF significantly improved survival in response to a lethal C. albicans 2 2 and DC production of proinflammatory cytokines, we next challenge compared with Nlrp10 / control mice (Fig. 3B). examined whether the generation of CD4+ Th cell responses Hence, taken together, these data suggest that Nlrp10 is required 2 2 to C. albicans remained intact in Nlrp10 / mice. WT and for the generation of protective antifungal adaptive immune 2 2 Nlrp10 / mice were infected i.v. with a sublethal dose of responses in vivo. C. albicans; Th1 and Th17 responses were evaluated 15 d p.i. The defect in the generation of specific Th cell responses 2 2 by measuring IFN-g and IL-17 release, respectively, from in Nlrp10 / mice was not restricted to C. albicans; immu- 2 2 splenocytes restimulated with heat-killed C. albicans for 72 h. nization of Nlrp10 / mice with Ag in the presence of a As expected, WT mice displayed a mixed Th1 and Th17 re- number of adjuvants, including LPS, aluminum hydroxide, sponse to infection with C. albicans,asevidencedbythese- and CFA, was also shown to result in defective adaptive im- cretion of IFN-g and IL-17 by restimulated splenocytes (Fig. mune responses (8). In addition, Nlrp10-deficient DCs were 2 2 3A). Surprisingly, Nlrp10 / mice displayed a profound shown to have an intrinsic defect in their ability to emigrate defect in the generation of Candida-specific Th1 and Th17 from a site of inflammation, resulting in a lack of Ag trans- responses (Fig. 3A), which indicates that Nlrp10 is required for port to the draining lymph node and explaining the lack of 2 2 driving appropriate adaptive immune responses to C. albicans priming of naive CD4+ T cells in Nlrp10 / mice (8). Hence, in vivo. Nlrp3 and Nlrp10 play distinct roles in shaping adaptive To determine whether the inability to generate appropriate immune responses against fungal pathogens. Although Nlrp3 adaptive immune responses was the cause of the increased inflammasome-driven IL-1b production drives Th17 differ- 2 2 mortality of Nlrp10 / mice following C. albicans infection, entiation during a C. albicans infection (19), the role of Nlrp10 we adoptively transferred naive and C. albicans–immune WT in the generation of specific Th cell responses is likely to be at 2 2 CD4+ TcellsintoNlrp10 / mice. Following CD4+ T cell the level of appropriate DC migration and Ag presentation to 2 2 adoptive transfer, Nlrp10 / mice were infected i.v. with naive CD4+ T cells. The Journal of Immunology 4717

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