293 results on '"Acha-Orbea H"'
Search Results
52. Two monoclonal rat antibodies with specificity for the beta-chain variable region V beta 6 of the murine T-cell receptor.
- Author
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Payne, J, Huber, B T, Cannon, N A, Schneider, R, Schilham, M W, Acha-Orbea, H, MacDonald, H R, and Hengartner, H
- Abstract
Two rat monoclonal antibodies (mAbs), 44-22-1 and 46-6B5, which recognize an alloreactive cytotoxic clone, 3F9, have been further tested on a panel of T hybridomas and cytotoxic T-cell clones for binding and functional activities. The mAbs recognized only those cells sharing the expression of the T-cell receptor beta-chain variable region gene V beta 6 with 3F9. All V beta 6+ cells were activated by these mAbs under cross-linking conditions and their antigen-specific activation was blocked by soluble mAb. Furthermore, depletion of 46-6B5+ normal lymph node T cells eliminated all cells expressing the epitope recognized by 44-22-1 and V beta 6 mRNA.
- Published
- 1988
- Full Text
- View/download PDF
53. Phosphorylation and dephosphorylation of spectrin from human erythrocyte ghosts under physiological conditions: autocatalysis rather than reaction with separate kinase and phosphatase.
- Author
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Imhof, B A, Acha-Orbea, H J, Libermann, T A, Reber, B F, Lanz, J H, Winterhalter, K H, and Birchmeier, W
- Abstract
The mechanism of phosphosylation and dephosphorylation of spectrin from human erythrocyte membranes has been examined under closely physiological conditions. The results support the hypothesis that spectrin is an autophosphorylating and dephosphorylating system. (i) Extraction from ghosts of up to 85% of the kinase (casein kinase) suggested to catalyze the reaction [see Fairbanks, G., Avruch, J., Dino, E. J. & Patel, V. P. (1978) J. Supramol. Struct. 9, 97--112] only slightly reduced spectrin component 2 phosphorylation and did not affect ATP-induced changes in the ghosts' shapes. (ii) A spectrin--actin complex isolated from endocytotic inside-out vesicles under hyperteonic conditions contained virtually no casein kinase activity and still exhibited a largely intact phosphorylation machinery. (iii) Photoaffinity labeling experiments indicated that spectrin component 2 fulfills the necessary prerequisite of the hypothesis--i.e., it contains its own ATP-binding site. (iv) Under various conditions, spectrin phosphorylation and dephospohrylation seem to be tightly coupled. The implications of these findings for the understanding of spectrin function and the maintenance of erythrocyte shape are discussed.
- Published
- 1980
- Full Text
- View/download PDF
54. The first external domain of the nonobese diabetic mouse class II I-A beta chain is unique.
- Author
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Acha-Orbea, H and McDevitt, H O
- Abstract
The nonobese diabetic mouse is recognized as an important animal model for human insulin-dependent diabetes mellitus. One of the components of susceptibility to this disease has been mapped to the major histocompatibility complex. In this study, full-length cDNA clones encoding the I-A alpha and beta chains from the nonobese diabetic mouse have been isolated and sequenced. They are identical to the sequences previously determined from the H-2d haplotype except for the sequence encoding the first external domain, the leader peptide, and the 5' untranslated region of the I-A beta chain molecule. Most strikingly, there are five consecutive nucleotide substitutions which lead to two radical amino acid changes in a region that is conserved between human and mouse. We suggest that the unique structure of the first external I-A beta chain domain is a major determinant in the disease susceptibility that maps to the major histocompatibility complex of the nonobese diabetic mouse.
- Published
- 1987
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55. Predominant expression of a T cell receptor V beta gene subfamily in autoimmune encephalomyelitis.
- Author
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Zamvil, S S, Mitchell, D J, Lee, N E, Moore, A C, Waldor, M K, Sakai, K, Rothbard, J B, McDevitt, H O, Steinman, L, and Acha-Orbea, H
- Abstract
TCR beta chain gene expression of individual T cell clones that share the same MHC class II restriction and similar fine specificity for the encephalitogenic NH2 terminus of the autoantigen myelin basic protein (MBP) has been examined. TCR V beta expression was examined by FACS analysis with mAbs specific for the V beta 8 subfamily of TCR beta chain genes. 14 of 18 (78%) NH2-terminal MBP-specific clones examined express a member of the TCR V beta 8 subfamily. Southern analysis was used to identify which member(s) of the TCR V beta 8 subfamily is expressed by these clones. Each of four clones examined uses V beta 8.2, though two different V beta 8.2-J beta 2 combinations were identified. Our findings indicate that there is restricted TCR V beta usage in the autoimmune T cell response to the dominant encephalitogenic NH2-terminal epitope of the MBP. The use of an mAb to the antigen-specific TCR in the prevention of T cell-mediated autoimmune disease has been investigated. Our results demonstrate that in vivo administration of a TCR V beta 8-specific mAb prevents induction of autoimmune encephalomyelitis.
- Published
- 1988
- Full Text
- View/download PDF
56. Gene rearrangement in cells with natural killer activity and expression of the β-chain of the T-cell antigen receptor.
- Author
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Yanagi, Y., Caccia, N., Kronenberg, M., Chin, B., Roder, J., Rohel, D., Kiyohara, T., Lauzon, R., Toyonaga, B., Rosenthal, K., Dennert, G., Acha-Orbea, H., Hengartner, H., Hood, L., and Mak, T. W.
- Published
- 1985
- Full Text
- View/download PDF
57. Impact of Sirtuin 2 knockout on innate immune responses
- Author
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Ciarlo, E., Lugrin, J., Dos Santos Pinheiro, I., Le Roy, D., Moulan, N., Yamamoto, H., Acha-Orbea, H., Calandra, T., Auwerx, J., and Roger, T.
58. Building a lymph node in vitro
- Author
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Siegert, S., Britschgi, M. R., Tomel, A. A., Link, A., Acha-Orbea, H., Swartz, M. A., and Sanjiv Luther
59. Lymph node stromal cells promote naive T-cell survival via an IL-7-dependent mechanism
- Author
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Vogt, T. K., Link, A., Favre, S., Britschgi, M. R., Acha-Orbea, H., Hinz, B., Cyster, J. G., and Sanjiv Luther
60. Unexpected role for lymph node stromal cells in suppressing T cell proliferation
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Sanjiv Luther, Scarpellino, L., Flavell, S., Tomei, A., Swartz, M., Nelson, P., Heikenwaelder, M., Acha-Orbea, H., Buckley, C., and Siegert, S.
61. Pseudoprolines: Targeting a cis conformation in a mimetic of the gp120 v3 loop of HIV-1
- Author
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Wittelsberger, A, Keller, M, Scarpellino, L, Patiny, L, Acha-Orbea, H, and Mutter, M
62. In vivo transformation of mouse conventional CD8{alpha}+ dendritic cells leads to progressive multisystem histiocytosis
- Author
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Steiner, Q G, Otten, L A, Hicks, M J, Kaya, G, Grosjean, F, Saeuberli, E, Lavanchy, C, Beermann, F, McClain, K L, and Acha-Orbea, H
- Abstract
Division and proliferation of dendritic cells (DCs) have been proposed to contribute to homeostasis and to prolonged antigen presentation. Whether abnormal proliferation of dendritic cells causes Langerhans cell histiocytosis (LCH) is a highly debated topic. Transgenic expression of simian virus 40 (SV40) T antigens in mature DCs allowed their transformation in vivo while maintaining their phenotype, function, and maturation capacity. The transformed cells were differentiated splenic CD8 alpha-positive conventional dendritic cells with increased Langerin expression. Their selective transformation was correlated with higher steady-state cycling compared with CD8 alpha-negative DCs in wild-type and transgenic mice. Mice developed a DC disease involving the spleen, liver, bone marrow, thymus, and mesenteric lymph node. Surprisingly, lesions displayed key immunohistologic features of Langerhans cell histiocytosis, including expression of Langerin and absence of the abnormal mitoses observed in Langerhans cell sarcomas. Our results demonstrate that a transgenic mouse model with striking similarities to aggressive forms of multisystem histiocytosis, such as the Letterer-Siwe syndrome, can be obtained by transformation of conventional DCs. These findings suggest that conventional DCs may cause some human multisystem LCH. They can reveal shared molecular pathways for human histiocytosis between humans and mice.
63. B cell response after MMTV infection: extrafollicular plasmablasts represent the main infected population and can transmit viral infection
- Author
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Ardavin, C., Martin, P., Ferrero, I., Azcoitia, I., Anjuere, F., Diggelmann, H., Luthi, F., Sanjiv Luther, and Acha-Orbea, H.
- Subjects
B-Lymphocytes ,Mice, Inbred BALB C ,Leukosialin ,Sialoglycoproteins ,Immunology ,Mice ,Tumor Virus Infections ,Mammary Tumor Virus, Mouse ,Antigens, CD ,Animals ,Leukocyte Common Antigens ,Immunology and Allergy ,Female ,L-Selectin ,Retroviridae Infections - Abstract
The immune response to mouse mammary tumor virus (MMTV) relies on the presentation of an MMTV-encoded superantigen by infected B cells to superantigen-specific T cells. The initial extrafollicular B cell differentiation involved the generation of B cells expressing low levels of B220. These B220low B cells corresponded to plasmablasts that expressed high levels of CD43 and syndecan-1 and were CD62 ligand− and IgD−. Viral DNA was detected nearly exclusively in these B220low B cells by PCR, and retroviral type-A particles were observed in their cytoplasm by electron microscopy. An MMTV transmission to the offspring was also achieved after transfer of B220low CD62 ligand− CD43+ plasmablasts into noninfected females. These data suggest that B220low plasmablasts, representing the bulk of infected B cells, are capable of sustaining viral replication and may be involved in the transmission of MMTV.
64. B cell response and histology of a retroviral infection in vivo
- Author
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Sanjiv Luther, Gulbranson-Judge, A., Acha-Orbea, H., Maclennan, I. C. M., Chiorazzi, N., Lahita, R. G., Pavelka, K., and Ferrarini, M.
65. Building a lymph node in vitro
- Author
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Siegert, S., Britschgi, M. R., Tomei, A. A., Acha-Orbea, H., Swartz, M. A., and Sanjiv Luther
66. Preferential infection of immature dendritic cells and B cells by mouse mammary tumor virus
- Author
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Vacheron, S., Sanjiv Luther, and Acha-Orbea, H.
67. Characterization of cloned cytotoxic lymphocytes with NK-like activity.
- Author
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Acha-Orbea, H, primary, Groscurth, P, additional, Lang, R, additional, Stitz, L, additional, and Hengartner, H, additional
- Published
- 1983
- Full Text
- View/download PDF
68. Limited heterogeneity of autoantigens and T cells in autoimmune diseases?
- Author
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Acha-Orbea, H.
- Published
- 1991
- Full Text
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69. The role of superantigens in resistance to retroviral infection
- Author
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Acha-Orbea, H.
- Published
- 1997
- Full Text
- View/download PDF
70. The 3' half of the mouse mammary tumor virus orf gene is not sufficient for its superantigen function in transgenic mice
- Author
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Lambert, J.-F., Acha-Orbea, H., Kolb, E., and Diggelmann, H.
- Published
- 1993
- Full Text
- View/download PDF
71. MHC class II transactivator is an in vivo regulator of osteoclast differentiation and bone homeostasis co-opted from adaptive immunity
- Author
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⁎, E., Mariani, E., Scolari, M., Perilli, E., Barras, E., Fazzalari, N.L., Campana, L., Otten, L., Particelli, F., Acha-Orbea, H., Baruffaldi, F., Faccio, R., Sitia, R., Reith, W., and Cenci, S.
- Published
- 2012
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72. The integrin CD11b inhibits MSU-induced NLRP3 inflammasome activation in macrophages and protects mice against MSU-induced joint inflammation.
- Author
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Ehirchiou D, Bernabei I, Pandian VD, Nasi S, Chobaz V, Castelblanco M, So A, Martinon F, Li X, Acha-Orbea H, Hugle T, Zhang L, and Busso N
- Subjects
- Animals, Mice, Male, NLR Family, Pyrin Domain-Containing 3 Protein metabolism, Macrophages metabolism, CD11b Antigen metabolism, Inflammasomes metabolism, Mice, Knockout, Uric Acid, Arthritis, Gouty chemically induced, Arthritis, Gouty metabolism, Mice, Inbred C57BL
- Abstract
Objective: In gout, monosodium urate crystals are taken up by macrophages, triggering the activation of the NLRP3 inflammasome and the maturation of IL-1β. This study aimed to investigate the role of integrin CD11b in inflammasome activation in macrophages stimulated by MSU., Methods: BMDM from WT and CD11b KO mice were stimulated in vitro with MSU crystals. Cellular supernatants were collected to assess the expression of the inflammatory cytokines by enzyme-linked immunosorbent assay and western blot methods. The role of integrin CD11b in MSU-induced gouty arthritis in vivo was investigated by intra-articular injection of MSU crystals. Real-time extracellular acidification rate and oxygen consumption rate of BMDMs were measured by Seahorse Extracellular Flux Analyzer., Results: We demonstrate that CD11b-deficient mice developed exacerbated gouty arthritis with increased recruitment of leukocytes in the joint and higher IL-1β levels in the sera. In macrophages, genetic deletion of CD11b induced a shift of macrophage metabolism from oxidative phosphorylation to glycolysis, thus decreasing the overall generation of intracellular ATP. Upon MSU stimulation, CD11b-deficient macrophages showed an exacerbated secretion of IL-1β. Treating wild-type macrophages with a CD11b agonist, LA1, inhibited MSU-induced release of IL-1β in vitro and attenuated the severity of experimental gouty arthritis. Importantly, LA1, was also effective in human cells as it inhibited MSU-induced release of IL-1β by peripheral blood mononuclear cells from healthy donors., Conclusion: Our data identified the CD11b integrin as a principal cell membrane receptor that modulates NLRP3 inflammasome activation by MSU crystal in macrophages, which could be a potential therapeutic target to treat gouty arthritis in human patients., (© 2024. The Author(s).)
- Published
- 2024
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73. A Multi-Omics Approach Reveals Features That Permit Robust and Widespread Regulation of IFN-Inducible Antiviral Effectors.
- Author
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Göczi L, Csumita M, Horváth A, Nagy G, Póliska S, Pigni M, Thelemann C, Dániel B, Mianesaz H, Varga T, Sen K, Raghav SK, Schoggins JW, Nagy L, Acha-Orbea H, Meissner F, Reith W, and Széles L
- Subjects
- Animals, Mice, Nucleotide Motifs, Promoter Regions, Genetic genetics, Response Elements genetics, Interferons metabolism, Antiviral Restriction Factors, Chromatin genetics
- Abstract
The antiviral state, an initial line of defense against viral infection, is established by a set of IFN-stimulated genes (ISGs) encoding antiviral effector proteins. The effector ISGs are transcriptionally regulated by type I IFNs mainly via activation of IFN-stimulated gene factor 3 (ISGF3). In this study, the regulatory elements of effector ISGs were characterized to determine the (epi)genetic features that enable their robust induction by type I IFNs in multiple cell types. We determined the location of regulatory elements, the DNA motifs, the occupancy of ISGF3 subunits (IRF9, STAT1, and STAT2) and other transcription factors, and the chromatin accessibility of 37 effector ISGs in murine dendritic cells. The IFN-stimulated response element (ISRE) and its tripartite version occurred most frequently in the regulatory elements of effector ISGs than in any other tested ISG subsets. Chromatin accessibility at their promoter regions was similar to most other ISGs but higher than at the promoters of inflammation-related cytokines, which were used as a reference gene set. Most effector ISGs (81.1%) had at least one ISGF3 binding region proximal to the transcription start site (TSS), and only a subset of effector ISGs (24.3%) was associated with three or more ISGF3 binding regions. The IRF9 signals were typically higher, and ISRE motifs were "stronger" (more similar to the canonical sequence) in TSS-proximal versus TSS-distal regulatory regions. Moreover, most TSS-proximal regulatory regions were accessible before stimulation in multiple cell types. Our results indicate that "strong" ISRE motifs and universally accessible promoter regions that permit robust, widespread induction are characteristic features of effector ISGs., (Copyright © 2022 by The American Association of Immunologists, Inc.)
- Published
- 2022
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- View/download PDF
74. SMRT and NCoR1 fine-tune inflammatory versus tolerogenic balance in dendritic cells by differentially regulating STAT3 signaling.
- Author
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Jha A, Ahad A, Mishra GP, Sen K, Smita S, Minz AP, Biswas VK, Tripathy A, Senapati S, Gupta B, Acha-Orbea H, and Raghav SK
- Subjects
- Animals, CD8-Positive T-Lymphocytes metabolism, Cytokines metabolism, Dendritic Cells metabolism, Interleukin-12 metabolism, Interleukin-23 metabolism, Mice, Nuclear Receptor Co-Repressor 1 genetics, Nuclear Receptor Co-Repressor 1 metabolism, Nuclear Receptor Co-Repressor 2, STAT3 Transcription Factor, TOR Serine-Threonine Kinases metabolism, Interleukin-10 metabolism, Interleukin-6 metabolism
- Abstract
Dendritic cell (DC) fine-tunes inflammatory versus tolerogenic responses to protect from immune-pathology. However, the role of co-regulators in maintaining this balance is unexplored. NCoR1-mediated repression of DC immune-tolerance has been recently reported. Here we found that depletion of NCoR1 paralog SMRT (NCoR2) enhanced cDC1 activation and expression of IL-6, IL-12 and IL-23 while concomitantly decreasing IL-10 expression/secretion. Consequently, co-cultured CD4
+ and CD8+ T-cells depicted enhanced Th1/Th17 frequency and cytotoxicity, respectively. Comparative genomic and transcriptomic analysis demonstrated differential regulation of IL-10 by SMRT and NCoR1. SMRT depletion represses mTOR-STAT3-IL10 signaling in cDC1 by down-regulating NR4A1. Besides, Nfkbia and Socs3 were down-regulated in Ncor2 ( Smrt ) depleted cDC1, supporting increased production of inflammatory cytokines. Moreover, studies in mice showed, adoptive transfer of SMRT depleted cDC1 in OVA-DTH induced footpad inflammation led to increased Th1/Th17 and reduced tumor burden after B16 melanoma injection by enhancing oncolytic CD8+ T-cell frequency, respectively. We also depicted decreased Ncor2 expression in Rheumatoid Arthritis, a Th1/Th17 disease., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2022 Jha, Ahad, Mishra, Sen, Smita, Minz, Biswas, Tripathy, Senapati, Gupta, Acha-Orbea and Raghav.)- Published
- 2022
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75. Zbtb10 transcription factor is crucial for murine cDC1 activation and cytokine secretion.
- Author
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Smita S, Ghosh A, Biswas VK, Ahad A, Podder S, Jha A, Sen K, Acha-Orbea H, and Raghav SK
- Subjects
- Animals, Biomarkers, Cell Differentiation genetics, Cell Differentiation immunology, Cell Line, Gene Expression Profiling, Gene Expression Regulation, Lymphocyte Activation immunology, Mice, T-Lymphocyte Subsets immunology, T-Lymphocyte Subsets metabolism, Cytokines biosynthesis, Dendritic Cells immunology, Dendritic Cells metabolism, Transcription Factors metabolism
- Abstract
Dendritic cell (DC) activation and cytokine production is tightly regulated. In this study, we found that Zbtb10 expression is activation dependent and it is essential for the immunogenic function of cDC1. Zbtb10 knockdown (KD) significantly reduced the expression of co-stimulatory genes CD80 and CD86 along with cytokines including IL-12, IL-6, and IL-10, in activated cDC1 Mutu-DC line. Consequently, the clonal expansion of CD44
+ effector T cells in co-cultured CD4+ T cells was drastically reduced owing to significantly reduced IL-2. At the same time, these CD44+ effector T cells were unable to differentiate toward Tbet+ IFNγ+ Th1 subtype. Instead, an increased frequency of Th2 cells expressing GATA3+ and IL-13+ was observed. Interestingly, in Zbtb10 KD condition the co-cultured T cells depicted increased expression of PD1 and LAG3, the T-cell anergic markers. Moreover, the global transcriptome analysis identified that Zbtb10 is pertinent for DC activation and its depletion in cDC1 completely shuts down their immune responses. Mechanistic analysis revealed that Zbtb10 KD enhanced the expression of NKRF (NF-κB repressing factor) leading to drastic suppression of NF-κB related genes. Zbtb10 KD abrogated p65 and RelB nuclear translocation, thereby controlling the activation and maturation of cDC1 and the ensuing adaptive T cell responses., (© 2021 Wiley-VCH GmbH.)- Published
- 2021
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76. IL10- and IL35-Secreting MutuDC Lines Act in Cooperation to Inhibit Memory T Cell Activation Through LAG-3 Expression.
- Author
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Koga MM, Engel A, Pigni M, Lavanchy C, Stevanin M, Laversenne V, Schneider BL, and Acha-Orbea H
- Subjects
- Animals, Arthritis, Rheumatoid immunology, Arthritis, Rheumatoid metabolism, Arthritis, Rheumatoid pathology, Arthritis, Rheumatoid therapy, Cell Communication immunology, Cell Line, Cytokines metabolism, Female, Gene Expression, Histocompatibility Antigens Class II genetics, Histocompatibility Antigens Class II immunology, Immune Tolerance, Immunomodulation, Immunotherapy methods, Lymphocyte Activation genetics, Lymphocyte Activation immunology, Mice, Mice, Knockout, Vaccines administration & dosage, Vaccines immunology, Lymphocyte Activation Gene 3 Protein, Antigens, CD genetics, Dendritic Cells immunology, Dendritic Cells metabolism, Immunologic Memory, Interleukin-10 biosynthesis, Interleukin-12 Subunit p35 biosynthesis, T-Lymphocytes immunology, T-Lymphocytes metabolism
- Abstract
Dendritic cells (DCs) are professional antigen-presenting cells involved in the initiation of immune responses. We generated a tolerogenic DC (tolDC) line that constitutively secretes interleukin-10 (IL10-DCs), expressed lower levels of co-stimulatory and MHCII molecules upon stimulation, and induced antigen-specific proliferation of T cells. Vaccination with IL10-DCs combined with another tolDC line that secretes IL-35, reduced antigen-specific local inflammation in a delayed-type hypersensitivity assay independently on regulatory T cell differentiation. In an autoimmune model of rheumatoid arthritis, vaccination with the combined tolDCs after the onset of the disease impaired disease development and promoted recovery of mice. After stable memory was established, the tolDCs promoted CD4 downregulation and induced lymphocyte activation gene 3 (LAG-3) expression in reactivated memory T cells, reducing T cell activation. Taken together, our findings indicate the benefits of combining anti-inflammatory cytokines in an antigen-specific context to treat excessive inflammation when memory is already established., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2021 Koga, Engel, Pigni, Lavanchy, Stevanin, Laversenne, Schneider and Acha-Orbea.)
- Published
- 2021
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77. NCoR1 fine-tunes type-I IFN response in cDC1 dendritic cells by directly regulating Myd88-IRF7 axis under TLR9.
- Author
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Ahad A, Smita S, Mishra GP, Biswas VK, Sen K, Gupta B, Garcin D, Acha-Orbea H, and Raghav SK
- Subjects
- Animals, Cells, Cultured, Female, Mice, Mice, Inbred C57BL, Mice, Knockout, Signal Transduction physiology, Dendritic Cells metabolism, Interferon Regulatory Factor-7 metabolism, Interferon Type I metabolism, Myeloid Differentiation Factor 88 metabolism, Nuclear Receptor Co-Repressor 1 metabolism, Toll-Like Receptor 9 metabolism
- Abstract
Plasmacytoid dendritic cells (DCs) are reported to induce robust type-I interferon (IFN) response, whereas cDC1 DCs develop moderate type-I IFN response upon TLR9 stimulation. It is very interesting to understand how this signaling under TLR9 is tightly regulated for the induction of type-I IFNs. Here, we report co-repressor protein NCoR1 as the major factor fine-tuning the signaling pathways regulating IFN-β expression under TLR9 in cDC1 DCs. We found that NCoR1 knockdown induced a robust IFN-β-mediated antiviral response upon TLR9 activation in cDC1 DCs. At the molecular level, we showed that NCoR1 directly repressed MyD88-IRF7 signaling axis in cDC1 cells. Therefore, NCoR1 depletion enhanced pIRF7 levels, IFN-β secretion, and downstream pSTAT1-pSTAT2 signaling, leading to sustained induction of IFN stimulatory genes. Integrative genomic analysis depicted strong enrichment of an antiviral gene-module in CpG-activated NCoR1 knockdown DCs upon TLR9 activation. Moreover, we confirmed our findings in primary DCs derived from splenocytes of WT and NCoR1 DC
-/- animals, which showed protection from Sendai and Vesicular Stomatitis viruses upon CpG activation. Ultimately, we identified that NCoR1-HDAC3 complex is involved in repressing the type-I IFN response in cDC1 DCs., (© 2020 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.)- Published
- 2020
- Full Text
- View/download PDF
78. Novel Autoantibody Signatures in Sera of Patients with Pancreatic Cancer, Chronic Pancreatitis and Autoimmune Pancreatitis: A Protein Microarray Profiling Approach.
- Author
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Ghassem-Zadeh S, Hufnagel K, Bauer A, Frossard JL, Yoshida M, Kutsumi H, Acha-Orbea H, Neulinger-Muñoz M, Vey J, Eckert C, Strobel O, Hoheisel JD, and Felix K
- Subjects
- Adult, Aged, Aged, 80 and over, Autoimmune Diseases diagnosis, Autoimmune Diseases immunology, Autoimmune Pancreatitis immunology, Diagnosis, Differential, Female, Humans, Immunoglobulin G blood, Male, Middle Aged, Pancreatic Neoplasms immunology, Pancreatitis, Chronic diagnosis, Pancreatitis, Chronic immunology, Patients, Pancreatic Neoplasms, Autoantibodies blood, Autoimmune Pancreatitis diagnosis, Pancreatic Neoplasms diagnosis, Protein Array Analysis methods
- Abstract
Identification of disease-associated autoantibodies is of high importance. Their assessment could complement current diagnostic modalities and assist the clinical management of patients. We aimed at developing and validating high-throughput protein microarrays able to screen patients' sera to determine disease-specific autoantibody-signatures for pancreatic cancer (PDAC), chronic pancreatitis (CP), autoimmune pancreatitis and their subtypes (AIP-1 and AIP-2). In-house manufactured microarrays were used for autoantibody-profiling of IgG-enriched preoperative sera from PDAC-, CP-, AIP-1-, AIP-2-, other gastrointestinal disease (GID) patients and healthy controls. As a top-down strategy, three different fluorescence detection-based protein-microarrays were used: large with 6400, intermediate with 345, and small with 36 full-length human recombinant proteins. Large-scale analysis revealed 89 PDAC, 98 CP and 104 AIP immunogenic antigens. Narrowing the selection to 29 autoantigens using pooled sera first and individual sera afterwards allowed a discrimination of CP and AIP from PDAC. For validation, predictive models based on the identified antigens were generated which enabled discrimination between PDAC and AIP-1 or AIP-2 yielded high AUC values of 0.940 and 0.925, respectively. A new repertoire of autoantigens was identified and their assembly as a multiplex test will provide a fast and cost-effective tool for differential diagnosis of pancreatic diseases with high clinical relevance.
- Published
- 2020
- Full Text
- View/download PDF
79. Targeted knock-in mice expressing the oxidase-fixed form of xanthine oxidoreductase favor tumor growth.
- Author
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Kusano T, Ehirchiou D, Matsumura T, Chobaz V, Nasi S, Castelblanco M, So A, Lavanchy C, Acha-Orbea H, Nishino T, Okamoto K, and Busso N
- Subjects
- Animals, Cell Proliferation, Female, Gene Knock-In Techniques, Humans, Macrophages enzymology, Macrophages metabolism, Male, Mice, Mice, Inbred C57BL, Neoplasms genetics, Neoplasms metabolism, Neoplasms physiopathology, Reactive Oxygen Species metabolism, Xanthine Dehydrogenase metabolism, Xanthine Oxidase metabolism, Neoplasms enzymology, Xanthine Dehydrogenase genetics, Xanthine Oxidase genetics
- Abstract
Xanthine oxidoreductase has been implicated in cancer. Nonetheless, the role played by its two convertible forms, xanthine dehydrogenase (XDH) and oxidase (XO) during tumorigenesis is not understood. Here we produce XDH-stable and XO-locked knock-in (ki) mice to address this question. After tumor transfer, XO ki mice show strongly increased tumor growth compared to wild type (WT) and XDH ki mice. Hematopoietic XO expression is responsible for this effect. After macrophage depletion, tumor growth is reduced. Adoptive transfer of XO-ki macrophages in WT mice increases tumor growth. In vitro, XO ki macrophages produce higher levels of reactive oxygen species (ROS) responsible for the increased Tregs observed in the tumors. Blocking ROS in vivo slows down tumor growth. Collectively, these results indicate that the balance of XO/XDH plays an important role in immune surveillance of tumor development. Strategies that inhibit the XO form specifically may be valuable in controlling cancer growth.
- Published
- 2019
- Full Text
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80. NCoR1: Putting the Brakes on the Dendritic Cell Immune Tolerance.
- Author
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Ahad A, Stevanin M, Smita S, Mishra GP, Gupta D, Waszak S, Sarkar UA, Basak S, Gupta B, Acha-Orbea H, and Raghav SK
- Abstract
Understanding the mechanisms fine-tuning immunogenic versus tolerogenic balance in dendritic cells (DCs) is of high importance for therapeutic approaches. We found that NCoR1-mediated direct repression of the tolerogenic program in conventional DCs is essential for induction of an optimal immunogenic response. NCoR1 depletion upregulated a wide variety of tolerogenic genes in activated DCs, which consequently resulted in increased frequency of FoxP3
+ regulatory T cells. Mechanistically, NCoR1 masks the PU.1-bound super-enhancers on major tolerogenic genes after DC activation that are subsequently bound by nuclear factor-κB. NCoR1 knockdown (KD) reduced RelA nuclear translocation and activity, whereas RelB was unaffected, providing activated DCs a tolerogenic advantage. Moreover, NCoR1DC-/- mice depicted enhanced Tregs in draining lymph nodes with increased disease burden upon bacterial and parasitic infections. Besides, adoptive transfer of activated NCoR1 KD DCs in infected animals showed a similar phenotype. Collectively, our results demonstrated NCoR1 as a promising target to control DC-mediated immune tolerance., (Copyright © 2019 The Authors. Published by Elsevier Inc. All rights reserved.)- Published
- 2019
- Full Text
- View/download PDF
81. Absence of MHC-II expression by lymph node stromal cells results in autoimmunity.
- Author
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Dubrot J, Duraes FV, Harlé G, Schlaeppi A, Brighouse D, Madelon N, Göpfert C, Stokar-Regenscheit N, Acha-Orbea H, Reith W, Gannagé M, and Hugues S
- Abstract
How lymph node stromal cells (LNSCs) shape peripheral T-cell responses remains unclear. We have previously demonstrated that murine LNSCs, lymphatic endothelial cells (LECs), blood endothelial cells (BECs), and fibroblastic reticular cells (FRCs) use the IFN-γ-inducible promoter IV (pIV) of the MHC class II (MHCII) transactivator CIITA to express MHCII. Here, we show that aging mice (>1 yr old) in which MHCII is abrogated in LNSCs by the selective deletion of pIV exhibit a significant T-cell dysregulation in LNs, including defective Treg and increased effector CD4
+ and CD8+ T-cell frequencies, resulting in enhanced peripheral organ T-cell infiltration and autoantibody production. The proliferation of LN-Tregs interacting with LECs increases following MHCII up-regulation by LECs upon aging or after exposure to IFN-γ, this effect being abolished in mice in which LECs lack MHCII. Overall, our work underpins the importance of LNSCs, particularly LECs, in supporting Tregs and T-cell tolerance., Competing Interests: The authors declare that they have no conflict of interest.- Published
- 2018
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82. Importance of EMT Factor ZEB1 in cDC1 "MutuDC Line" Mediated Induction of Th1 Immune Response.
- Author
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Smita S, Ahad A, Ghosh A, Biswas VK, Koga MM, Gupta B, Acha-Orbea H, and Raghav SK
- Subjects
- Adaptive Immunity immunology, Adoptive Transfer methods, Animals, CD4-Positive T-Lymphocytes immunology, CD8-Positive T-Lymphocytes immunology, Cell Differentiation immunology, Cells, Cultured, Coculture Techniques methods, Female, HEK293 Cells, Humans, Interleukin-10 immunology, Interleukin-12 immunology, Interleukin-6 immunology, Lymphocyte Activation immunology, Mice, Mice, Inbred C57BL, T-Lymphocytes, Helper-Inducer immunology, Th2 Cells immunology, Toll-Like Receptor 9 immunology, Dendritic Cells immunology, Epithelial-Mesenchymal Transition immunology, Th1 Cells immunology, Zinc Finger E-box-Binding Homeobox 1 immunology
- Abstract
The role of Epithelial to Mesenchymal Transition (EMT) factor Zeb1 is well defined in metastasis and cancer progression but it's importance in dendritic cells (DCs) is unexplored until now. For the first time we report here that Zeb1 controls immunogenic responses of CD8α
+ conventional Type-I (cDC1) DCs. We found that ZEB1 expression increases significantly after TLR9 stimulation and its depletion impairs activation, co-stimulation and secretion of important cytokines like IL-6, IL-10 and IL-12 in cDC1 MutuDC line. We further confirmed our findings in primary cDC1 DCs derived from bone marrow. Co-culture of these Zeb1 knock down (KD) DCs with OT-II CD4+ T helper cells skewed their differentiation toward Th2 subtype. Moreover, adoptive transfer of activated Zeb1 KD DCs cleared intestinal worms in helminth infected mice by increasing Th2 responses in vivo . Integrative genomic analysis showed Zeb1 as an activator of immune response genes in cDC1 MutuDCs as compared to other pathway genes. In addition, differentially regulated genes in Zeb1 KD RNA-seq showed significant enrichment of Th2 activation pathways supporting our in vitro findings. Mechanistically, we showed that decreased IL-12 secreted by Zeb1 KD DCs is the plausible mechanism for increased Th2 differentiation. Collectively our data demonstrate that Zeb1 could be targeted in DCs to modulate T-cell mediated adaptive immune responses.- Published
- 2018
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83. Establishment and Characterization of a Functionally Competent Type 2 Conventional Dendritic Cell Line.
- Author
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Pigni M, Ashok D, Stevanin M, and Acha-Orbea H
- Subjects
- Animals, Mice, Mice, Knockout, Spleen cytology, Spleen immunology, Adaptive Immunity, Cell Line, Dendritic Cells cytology, Dendritic Cells immunology
- Abstract
Dendritic cells (DCs) are the most potent antigen presenting cells and possess an incomparable ability to activate and instruct T cells, which makes them one of the cornerstones in the regulation of the cross-talk between innate and adaptive immunity. Therefore, a deep understanding of DC biology lays the foundations to describe and to harness the mechanisms that regulate the development of the adaptive response, with clear implications in a vast array of fields such as the study of autoimmune diseases and the development of new vaccines. However, the great difficulty to obtain large quantities of viable non-activated DCs for experimentation have considerably hindered the progress of DC research. Several strategies have been proposed to overcome these limitations by promoting an increase of DC abundance in vivo , by inducing DC development from DC progenitors in vitro and by generating stable DC lines. In the past years, we have described a method to derive immortalized stable DC lines, named MutuDCs, from the spleens of Mushi1 mice, a transgenic mouse strain that express the simian virus 40 Large T-oncogene in the DCs. The comparison of these DC lines with the vast variety of DC subsets described in vivo has shown that all the MutuDC lines that we have generated so far have phenotypic and functional features of type 1 conventional DCs (cDC1s). With the purpose of deriving DC lines with characteristics of type 2 conventional DCs (cDC2s), we bred a new Batf3
-/- Mushi1 murine line in which the development of the cDC1 subset is severely defective. The new MutuDC line that we generated from Batf3-/- Mushi1 mice was phenotypically and functionally characterized in this work. Our results demonstrated that all the tested characteristics of this new cell line, including the expression of subset-determining transcription factors, the profile of cytokine production and the ability to present antigens, are comparable with the features of splenic CD4- cDC2s. Therefore, we concluded that our new cell line, that we named CD4- MutuDC2 line, represents a valuable model for the CD4- cDC2 subset.- Published
- 2018
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84. Sirtuin 2 Deficiency Increases Bacterial Phagocytosis by Macrophages and Protects from Chronic Staphylococcal Infection.
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Ciarlo E, Heinonen T, Théroude C, Herderschee J, Mombelli M, Lugrin J, Pfefferlé M, Tyrrell B, Lensch S, Acha-Orbea H, Le Roy D, Auwerx J, and Roger T
- Abstract
Sirtuin 2 (SIRT2) is one of the seven members of the family of NAD
+ -dependent histone deacetylases. Sirtuins target histones and non-histone proteins according to their subcellular localization, influencing various biological processes. SIRT2 resides mainly in the cytoplasm and regulates cytoskeleton dynamics, cell cycle, and metabolic pathways. As such, SIRT2 has been implicated in the pathogenesis of neurodegenerative, metabolic, oncologic, and chronic inflammatory disorders. This motivated the development of SIRT2-directed therapies for clinical purposes. However, the impact of SIRT2 on antimicrobial host defense is largely unknown. Here, we address this question using SIRT2 knockout mice. We show that SIRT2 is the most highly expressed sirtuin in myeloid cells, especially macrophages. SIRT2 deficiency does not affect immune cell development and marginally impacts on intracellular signaling and cytokine production by splenocytes and macrophages. However, SIRT2 deficiency enhances bacterial phagocytosis by macrophages. In line with these observations, in preclinical models, SIRT2 deficiency increases survival of mice with chronic staphylococcal infection, while having no effect on the course of toxic shock syndrome toxin-1, LPS or TNF-induced shock, fulminant Escherichia coli peritonitis, sub-lethal Klebsiella pneumoniae pneumonia, and chronic candidiasis. Altogether, these data support the safety profile of SIRT2 inhibitors under clinical development in terms of susceptibility to infections.- Published
- 2017
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85. Sirtuin 3 deficiency does not alter host defenses against bacterial and fungal infections.
- Author
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Ciarlo E, Heinonen T, Lugrin J, Acha-Orbea H, Le Roy D, Auwerx J, and Roger T
- Subjects
- Animals, Biomarkers, Dendritic Cells immunology, Dendritic Cells metabolism, Disease Resistance genetics, Humans, Immunophenotyping, Macrophages immunology, Macrophages metabolism, Mice, Mice, Knockout, Thymocytes immunology, Thymocytes metabolism, Bacterial Infections genetics, Host-Pathogen Interactions genetics, Mycoses genetics, Sirtuin 3 deficiency
- Abstract
Sirtuin 3 (SIRT3) is the main mitochondrial deacetylase. SIRT3 regulates cell metabolism and redox homeostasis, and protects from aging and age-associated pathologies. SIRT3 may drive both oncogenic and tumor-suppressive effects. SIRT3 deficiency has been reported to promote chronic inflammation-related disorders, but whether SIRT3 impacts on innate immune responses and host defenses against infections remains essentially unknown. This aspect is of primary importance considering the great interest in developing SIRT3-targeted therapies. Using SIRT3 knockout mice, we show that SIRT3 deficiency does not affect immune cell development and microbial ligand-induced proliferation and cytokine production by splenocytes, macrophages and dendritic cells. Going well along with these observations, SIRT3 deficiency has no major impact on cytokine production, bacterial burden and survival of mice subjected to endotoxemia, Escherichia coli peritonitis, Klebsiella pneumoniae pneumonia, listeriosis and candidiasis of diverse severity. These data suggest that SIRT3 is not critical to fight infections and support the safety of SIRT3-directed therapies based on SIRT3 activators or inhibitors for treating metabolic, oncologic and neurodegenerative diseases without putting patients at risk of infection.
- Published
- 2017
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86. Distinct pathophysiological cytokine profiles for discrimination between autoimmune pancreatitis, chronic pancreatitis, and pancreatic ductal adenocarcinoma.
- Author
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Ghassem-Zadeh S, Gaida MM, Szanyi S, Acha-Orbea H, Frossard JL, Hinz U, Hackert T, Strobel O, and Felix K
- Subjects
- Adenocarcinoma blood, Adenocarcinoma physiopathology, Adult, Autoimmune Diseases blood, Autoimmune Diseases physiopathology, Carcinoma, Pancreatic Ductal physiopathology, Diagnosis, Differential, Female, Humans, Male, Middle Aged, Pancreatic Neoplasms blood, Pancreatitis, Chronic physiopathology, ROC Curve, Pancreatic Neoplasms, Adenocarcinoma diagnosis, Autoimmune Diseases diagnosis, Carcinoma, Pancreatic Ductal blood, Carcinoma, Pancreatic Ductal diagnosis, Cytokines blood, Pancreatic Neoplasms diagnosis, Pancreatitis, Chronic blood, Pancreatitis, Chronic diagnosis
- Abstract
Background: Discriminating between autoimmune pancreatitis (AIP), chronic pancreatitis (CP), and pancreatic ductal adenocarcinoma (PDAC) can be challenging. In this retrospective study, levels of serum and tissue cytokines were analyzed as part of the clinical strategy for the preoperative differentiation between AIP and PDAC. The identification of differential cytokine profiles may help to prevent unnecessary surgical resection and allow optimal treatment of these pathologies., Methods: To compare the cytokine profiles of AIP, CP, and PDAC patients, serum and pancreatic tissue homogenates were subjected to multiplex analysis of 17 inflammatory mediators. In total, serum from 73 patients, composed of 29 AIP (14 AIP-1 and 15 AIP-2), 17 CP, and 27 PDAC, and pancreatic tissue from 36 patients, including 12 AIP (six AIP-1 and six AIP-2), 12 CP, and 12 PDAC, were analyzed., Results: Comparing AIP and PDAC patients' serum, significantly higher concentrations were found in AIP for interleukins IL-1β, IL-7, IL-13, and granulocyte colony-stimulating factor (G-CSF). G-CSF also allowed discrimination of AIP from CP. Furthermore, once AIP was divided into subtypes, significantly higher serum levels for IL-7 and G-CSF were measured in both subtypes of AIP and in AIP-2 for IL-1β when compared to PDAC. G-CSF and TNF-α were also significantly differentially expressed in tissue homogenates between AIP-2 and PDAC., Conclusions: The cytokines IL-1β, IL-7, and G-CSF can be routinely measured in patients' serum, providing an elegant and non-invasive approach for differential diagnosis. G-CSF is a good candidate to supplement the currently known serum markers in predictive tests for AIP and represents a basis for a combined blood test to differentiate AIP and particularly AIP-2 from PDAC, enhancing the possibility of appropriate treatment.
- Published
- 2017
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87. CD11b regulates the Treg/Th17 balance in murine arthritis via IL-6.
- Author
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Stevanin M, Busso N, Chobaz V, Pigni M, Ghassem-Zadeh S, Zhang L, Acha-Orbea H, and Ehirchiou D
- Subjects
- Adoptive Transfer, Animals, Antibodies, Blocking pharmacology, CD11b Antigen genetics, Cell Differentiation, Cells, Cultured, Collagen Type II immunology, Lymphocyte Activation, Mice, Mice, Inbred C57BL, Mice, Knockout, Receptors, Interleukin-6 immunology, Arthritis, Experimental immunology, CD11b Antigen metabolism, Cartilage immunology, Dendritic Cells immunology, Interleukin-6 metabolism, T-Lymphocytes, Regulatory immunology, Th17 Cells immunology
- Abstract
Th17 cells are often associated with autoimmunity and been shown to be increased in CD11b
-/- mice. Here, we examined the role of CD11b in murine collagen-induced arthritis (CIA). C57BL/6 and CD11b-/- resistant mice were immunized with type II collagen. CD11b-/- mice developed arthritis with early onset, high incidence, and sustained severity compared with C57BL/6 mice. We observed a marked leukocyte infiltration, and histological examinations of the arthritic paws from CD11b-/- mice revealed that the cartilage was destroyed in association with strong lymphocytic infiltration. The CD11b deficiency led to enhanced Th17-cell differentiation. CD11b-/- dendritic cells (DCs) induced much stronger IL-6 production and hence Th17-cell differentiation than wild-type DCs. Treatment of CD11b-/- mice after establishment of the Treg/Th17 balance with an anti-IL-6 receptor mAb significantly suppressed the induction of Th17 cells and reduced arthritis severity. Finally, the severe phenotype of arthritis in CD11b-/- mice was rescued by adoptive transfer of CD11b+ DCs. Taken together, our results indicate that the resistance to CIA in C57BL/6 mice is regulated by CD11b via suppression of IL-6 production leading to reduced Th17-cell differentiation. Therefore, CD11b may represent a susceptibility factor for autoimmunity and could be a target for future therapy., (© 2017 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.)- Published
- 2017
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88. Interleukin-35-Producing CD8α + Dendritic Cells Acquire a Tolerogenic State and Regulate T Cell Function.
- Author
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Haller S, Duval A, Migliorini R, Stevanin M, Mack V, and Acha-Orbea H
- Abstract
Dendritic cells (DCs) play a central role in shaping immunogenic as well as tolerogenic adaptive immune responses and thereby dictate the outcome of adaptive immunity. Here, we report the generation of a CD8α
+ DC line constitutively secreting the tolerogenic cytokine interleukin (IL)-35. IL-35 secretion led to impaired CD4+ and CD8+ T lymphocyte proliferation and interfered with their function in vitro and also in vivo . IL-35 was furthermore found to induce a tolerogenic phenotype on CD8α+ DCs, characterized by the upregulation of CD11b, downregulation of MHC class II, a reduced costimulatory potential as well as production of the immunomodulatory molecule IL-10. Vaccination of mice with IL-35-expressing DCs promoted tumor growth and reduced the severity of autoimmune encephalitis not only in a preventive but also after induction of encephalitogenic T cells. The reduction in experimental autoimmune encephalitis severity was significantly more pronounced when antigen-pulsed IL-35+ DCs were used. These findings suggest a new, indirect effector mechanism by which IL-35-responding antigen-presenting cells contribute to immune tolerance. Furthermore, IL-35-transfected DCs may be a promising approach for immunotherapy in the context of autoimmune diseases.- Published
- 2017
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89. Role of proapoptotic BH3-only proteins in Listeria monocytogenes infection.
- Author
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Margaroli C, Oberle S, Lavanchy C, Scherer S, Rosa M, Strasser A, Pellegrini M, Zehn D, Acha-Orbea H, and Ehirchiou D
- Subjects
- Animals, BH3 Interacting Domain Death Agonist Protein deficiency, Bcl-2-Like Protein 11 deficiency, Cytokines genetics, Cytokines metabolism, Disease Models, Animal, Disease Resistance genetics, Disease Resistance immunology, Disease Susceptibility, Extracellular Traps immunology, Female, Gene Expression, Listeriosis mortality, Listeriosis pathology, Male, Mice, Mice, Knockout, Neutrophil Infiltration genetics, Neutrophil Infiltration immunology, Neutrophils immunology, Neutrophils metabolism, Proto-Oncogene Proteins c-bcl-2 deficiency, Reactive Oxygen Species metabolism, Spleen immunology, Spleen metabolism, Spleen pathology, Survival Rate, Apoptosis, Listeria monocytogenes, Listeriosis etiology, Listeriosis metabolism, Mitochondrial Proteins metabolism
- Abstract
The ability of pathogens to influence host cell survival is a crucial virulence factor. Listeria monocytogenes (Lm) infection is known to be associated with severe apoptosis of hepatocytes and spleen cells. This impairs host defense mechanisms and thereby facilitates the spread of intracellular pathogens. The general mechanisms of apoptosis elicited by Lm infection are understood, however, the roles of BH3-only proteins during primary Lm infection have not been examined. To explore the roles of BH3-only proteins in Lm-induced apoptosis, we studied Listeria infections in mice deficient in Bim, Bid, Noxa or double deficient in BimBid or BimNoxa. We found that BimNoxa double knockout mice were highly resistant to high-dose challenge with Listeria. Decreased bacterial burden and decreased host cell apoptosis were found in the spleens of these mice. The ability of the BH3-deficient mice to clear bacterial infection more efficiently than WT was correlated with increased concentrations of ROS, neutrophil extracellular DNA trap release and downregulation of TNF-α. Our data show a novel pathway of infection-induced apoptosis that enhances our understanding of the mechanism by which BH3-only proteins control apoptotic host cell death during Listeria infection., (© 2016 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.)
- Published
- 2016
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90. Derivation and Utilization of Functional CD8(+) Dendritic Cell Lines.
- Author
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Pigni M, Ashok D, and Acha-Orbea H
- Subjects
- Animals, Cell Transformation, Neoplastic genetics, Dendritic Cells immunology, Disease Models, Animal, HEK293 Cells, Humans, Mice, Mice, Transgenic, Promoter Regions, Genetic, Tumor Cells, Cultured, Antigens, Polyomavirus Transforming genetics, CD11c Antigen genetics, CD8 Antigens metabolism, Cell Culture Techniques methods, Cell Transformation, Neoplastic pathology, Dendritic Cells pathology
- Abstract
It is notoriously difficult to obtain large quantities of non-activated dendritic cells ex vivo. For this reason, we produced and characterized a mouse model expressing the large T oncogene under the CD11c promoter (Mushi mice), in which CD8α(+) dendritic cells transform after 4 months. We derived a variety of stable cell lines from these primary lines. These cell lines reproducibly share with freshly isolated dendritic cells most surface markers, mRNA and protein expression, and all tested biological functions. Cell lines can be derived from various strains and knockout mice and can be easily transduced with lentiviruses. In this article, we describe the derivation, culture, and lentiviral transduction of these dendritic cell lines.
- Published
- 2016
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91. Dendritic Cells Cause Bone Lesions in a New Mouse Model of Histiocytosis.
- Author
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Grosjean F, Nasi S, Schneider P, Chobaz V, Liu A, Mordasini V, Moullec K, Vezzoni P, Lavanchy C, Busso N, Acha-Orbea H, and Ehirchiou D
- Subjects
- Animals, Bone Density Conservation Agents therapeutic use, Bone and Bones drug effects, Cell Line, Cell Transdifferentiation, Diphosphonates therapeutic use, Disease Models, Animal, Histiocytosis, Langerhans-Cell complications, Humans, Mice, Mice, Inbred C57BL, Mice, Transgenic, Osteoclasts pathology, Osteolysis complications, Osteolysis prevention & control, Osteoprotegerin therapeutic use, Bone and Bones pathology, Dendritic Cells pathology, Histiocytosis, Langerhans-Cell pathology, Langerhans Cells pathology, Osteolysis pathology
- Abstract
Langerhans cell histiocytosis (LCH) is a rare disease caused by the clonal accumulation of dendritic Langerhans cells, which is often accompanied by osteolytic lesions. It has been reported that osteoclast-like cells play a major role in the pathogenic bone destruction seen in patients with LCH and these cells are postulated to originate from the fusion of DCs. However, due to the lack of reliable animal models the pathogenesis of LCH is still poorly understood. In this study, we have established a mouse model of histiocytosis- recapitulating human disease for osteolytic lesions seen in LCH patients. At 12 weeks after birth, severe bone lesions were observed in our multisystem histiocytosis (Mushi) model, when CD8α conventional dendritic cells (DCs) are transformed (MuTuDC) and accumulate. Most importantly, our study demonstrates that bone loss in LCH can be accounted for the transdifferentiation of MuTuDCs into functional osteoclasts both in vivo and in vitro. Moreover, we have shown that injected MuTuDCs reverse the osteopetrotic phenotype of oc/oc mice in vivo. In conclusion, our results support a crucial role of DCs in bone lesions in histiocytosis patients. Furthermore, our new model of LCH based on adoptive transfer of MuTuDC lines, leading to bone lesions within 1-2 weeks, will be an important tool for investigating the pathophysiology of this disease and ultimately for evaluating the potential of anti-resorptive drugs for the treatment of bone lesions.
- Published
- 2015
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92. TLR3-Mediated CD8+ Dendritic Cell Activation Is Coupled with Establishment of a Cell-Intrinsic Antiviral State.
- Author
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Széles L, Meissner F, Dunand-Sauthier I, Thelemann C, Hersch M, Singovski S, Haller S, Gobet F, Fuertes Marraco SA, Mann M, Garcin D, Acha-Orbea H, and Reith W
- Subjects
- Animals, Cross-Priming immunology, Humans, Interferon-beta genetics, Lymphocyte Activation immunology, Mice, Mice, Inbred C57BL, Mice, Knockout, Picornaviridae Infections immunology, Picornaviridae Infections virology, Poly I-C immunology, Receptor, Interferon alpha-beta immunology, Rhinovirus immunology, Sendai virus immunology, Vesicular stomatitis Indiana virus immunology, CD8-Positive T-Lymphocytes immunology, Dendritic Cells immunology, Interferon-beta immunology, Toll-Like Receptor 3 immunology
- Abstract
Because of their unique capacity to cross-present Ags to CD8(+) T cells, mouse lymphoid tissue-resident CD8(+) dendritic cells (DCs) and their migratory counterparts are critical for priming antiviral T cell responses. High expression of the dsRNA sensor TLR3 is a distinctive feature of these cross-presenting DC subsets. TLR3 engagement in CD8(+) DCs promotes cross-presentation and the acquisition of effector functions required for driving antiviral T cell responses. In this study, we performed a comprehensive analysis of the TLR3-induced antiviral program and cell-autonomous immunity in CD8(+) DC lines and primary CD8(+) DCs. We found that TLR3-ligand polyinosinic-polycytidylic acid and human rhinovirus infection induced a potent antiviral protection against Sendai and vesicular stomatitis virus in a TLR3 and type I IFN receptor-dependent manner. Polyinosinic-polycytidylic acid-induced antiviral genes were identified by mass spectrometry-based proteomics and transcriptomics in the CD8(+) DC line. Nanostring nCounter experiments confirmed that these antiviral genes were induced by TLR3 engagement in primary CD8(+) DCs, and indicated that many are secondary TLR3-response genes requiring autocrine IFN-β stimulation. TLR3-activation thus establishes a type I IFN-dependent antiviral program in a DC subtype playing crucial roles in priming adaptive antiviral immune responses. This mechanism is likely to shield the priming of antiviral responses against inhibition or abrogation by the viral infection. It could be particularly relevant for viruses detected mainly by TLR3, which may not trigger type I IFN production by DCs that lack TLR3, such as plasmacytoid DCs or CD8(-) DCs., (Copyright © 2015 by The American Association of Immunologists, Inc.)
- Published
- 2015
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93. Maintaining dendritic cell viability in culture.
- Author
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Vremec D, Hansen J, Strasser A, Acha-Orbea H, Zhan Y, O'Keeffe M, and Shortman K
- Subjects
- Animals, Apoptosis drug effects, Apoptosis genetics, Cell Survival drug effects, Cells, Cultured, Cytokines pharmacology, Dendritic Cells drug effects, Humans, Mice, Inbred C57BL, Signal Transduction drug effects, Signal Transduction genetics, Cell Culture Techniques methods, Dendritic Cells cytology
- Abstract
When mouse dendritic cells (DCs) are isolated from tissues, purified and placed in a nutritive culture they die more rapidly than would be expected from their normal turnover in vivo. This can distort culture assays of DC function. We therefore tested several approaches to prolonging DC survival in culture. Of several cytokines tested granulocyte-macrophage colony stimulating factor was most effective at preserving the viability of conventional DCs (cDCs) but was ineffective for plasmacytoid DCs (pDCs). Surprisingly, Fms-like tyrosine kinase 3 ligand, crucial for DC development, produced only a marginal improvement in DC survival in culture, and interleukin-3, reported to prevent apoptosis of human pDCs, produced only a minor improvement in survival of mouse DCs. Genetic manipulation of cell death pathways was also tested, to avoid activation effects exerted by cytokine signalling. The isolation of DCs from mice overexpressing Bcl-2 was especially effective in maintaining pDC viability but gave a lesser improvement in cDC viability. DCs isolated from Bim(-/-)Noxa(-/-) mice also showed improved culture survival, but in this case with pDCs showing the least improvement., (Crown Copyright © 2014. Published by Elsevier Ltd. All rights reserved.)
- Published
- 2015
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94. Beta-catenin signaling drives differentiation and proinflammatory function of IRF8-dependent dendritic cells.
- Author
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Cohen SB, Smith NL, McDougal C, Pepper M, Shah S, Yap GS, Acha-Orbea H, Jiang A, Clausen BE, Rudd BD, and Denkers EY
- Subjects
- Animals, Antigens, CD immunology, Bridged Bicyclo Compounds, Heterocyclic pharmacology, CD11c Antigen immunology, CD8 Antigens immunology, CD8-Positive T-Lymphocytes immunology, Cell Differentiation immunology, Enzyme Activation, Female, Integrin alpha Chains immunology, Interferon Regulatory Factors immunology, Interleukin-12 biosynthesis, Interleukin-12 metabolism, Mice, Mice, Inbred C57BL, Mice, Knockout, Parasite Load, Promoter Regions, Genetic, Pyrimidinones pharmacology, Receptors, Cell Surface genetics, Signal Transduction immunology, Spleen cytology, Spleen immunology, Th1 Cells immunology, Toxoplasma immunology, Toxoplasmosis immunology, Vaccinia immunology, Vaccinia virus immunology, beta Catenin antagonists & inhibitors, beta Catenin biosynthesis, Dendritic Cells cytology, Dendritic Cells immunology, Inflammation immunology, Interferon Regulatory Factors genetics, beta Catenin immunology
- Abstract
Beta-catenin signaling has recently been tied to the emergence of tolerogenic dendritic cells (DCs). In this article, we demonstrate a novel role for beta-catenin in directing DC subset development through IFN regulatory factor 8 (IRF8) activation. We found that splenic DC precursors express beta-catenin, and DCs from mice with CD11c-specific constitutive beta-catenin activation upregulated IRF8 through targeting of the Irf8 promoter, leading to in vivo expansion of IRF8-dependent CD8a+, plasmacytoid, and CD103+ CD11b2 DCs. beta-catenin–stabilized CD8a+ DCs secreted elevated IL-12 upon in vitro microbial stimulation, and pharmacological beta-catenin inhibition blocked this response in wild-type cells. Upon infections with Toxoplasma gondii and vaccinia virus, mice with stabilized DC beta-catenin displayed abnormally high Th1 and CD8+ T lymphocyte responses, respectively. Collectively, these results reveal a novel and unexpected function for beta-catenin in programming DC differentiation toward subsets that orchestrate proinflammatory immunity to infection.
- Published
- 2015
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95. Timing is everything: dendritic cell subsets in murine Leishmania infection.
- Author
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Ashok D and Acha-Orbea H
- Subjects
- Animals, Antigen Presentation immunology, Leishmania immunology, Mice, Th1 Cells immunology, Dendritic Cells immunology, Leishmaniasis immunology
- Abstract
Mouse models of Leishmania major infection have shown that a predominant CD4(+) T helper type 1 cell (Th1) response leads to protection, while T helper type 2 cell (Th2) predominance confers susceptibility. Dendritic cells (DCs) are antigen-presenting cells that orchestrate the T cell response. The immune response to L. major involves direct antigen presentation by migrating DCs or transfer of antigens to resident DCs to prime T cells. In this review, we discuss the timing and consequences of antigen presentation by DC subsets and how this affects Leishmania susceptibility. We propose a model where dermal DCs and Langerhans cells play a role early in infection, followed by inflammatory monocyte-derived DC and lymph node (LN)-resident DCs at later time points of infection to establish the resistant Th1 response., (Copyright © 2014 Elsevier Ltd. All rights reserved.)
- Published
- 2014
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96. Activated group 3 innate lymphoid cells promote T-cell-mediated immune responses.
- Author
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von Burg N, Chappaz S, Baerenwaldt A, Horvath E, Bose Dasgupta S, Ashok D, Pieters J, Tacchini-Cottier F, Rolink A, Acha-Orbea H, and Finke D
- Subjects
- Animals, Antigen Presentation immunology, B-Lymphocytes immunology, CD4-Positive T-Lymphocytes drug effects, CD4-Positive T-Lymphocytes metabolism, Cell Proliferation drug effects, Cells, Cultured, Cytokines immunology, Cytokines metabolism, Histocompatibility Antigens Class II immunology, Immunity, Cellular immunology, Interleukin-1beta pharmacology, Lymphocyte Activation drug effects, Lymphocytes drug effects, Lymphocytes metabolism, Mice, Mice, Inbred C57BL, CD4-Positive T-Lymphocytes immunology, Interleukin-1beta immunology, Lymphocyte Activation immunology, Lymphocytes immunology, Signal Transduction immunology
- Abstract
Group 3 innate lymphoid cells (ILC3s) have emerged as important cellular players in tissue repair and innate immunity. Whether these cells meaningfully regulate adaptive immune responses upon activation has yet to be explored. Here we show that upon IL-1β stimulation, peripheral ILC3s become activated, secrete cytokines, up-regulate surface MHC class II molecules, and express costimulatory molecules. ILC3s can take up latex beads, process protein antigen, and consequently prime CD4(+) T-cell responses in vitro. The cognate interaction of ILC3s and CD4(+) T cells leads to T-cell proliferation both in vitro and in vivo, whereas its disruption impairs specific T-cell and T-dependent B-cell responses in vivo. In addition, the ILC3-CD4(+) T-cell interaction is bidirectional and leads to the activation of ILC3s. Taken together, our data reveal a novel activation-dependent function of peripheral ILC3s in eliciting cognate CD4(+) T-cell immune responses.
- Published
- 2014
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97. Extensive remodeling of DC function by rapid maturation-induced transcriptional silencing.
- Author
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Seguín-Estévez Q, Dunand-Sauthier I, Lemeille S, Iseli C, Ibberson M, Ioannidis V, Schmid CD, Rousseau P, Barras E, Geinoz A, Xenarios I, Acha-Orbea H, and Reith W
- Subjects
- Animals, Humans, Mice, Proto-Oncogene Proteins metabolism, Trans-Activators metabolism, Dendritic Cells metabolism, Gene Silencing, Nuclear Proteins genetics, Trans-Activators genetics, Transcription, Genetic
- Abstract
The activation, or maturation, of dendritic cells (DCs) is crucial for the initiation of adaptive T-cell mediated immune responses. Research on the molecular mechanisms implicated in DC maturation has focused primarily on inducible gene-expression events promoting the acquisition of new functions, such as cytokine production and enhanced T-cell-stimulatory capacity. In contrast, mechanisms that modulate DC function by inducing widespread gene-silencing remain poorly understood. Yet the termination of key functions is known to be critical for the function of activated DCs. Genome-wide analysis of activation-induced histone deacetylation, combined with genome-wide quantification of activation-induced silencing of nascent transcription, led us to identify a novel inducible transcriptional-repression pathway that makes major contributions to the DC-maturation process. This silencing response is a rapid primary event distinct from repression mechanisms known to operate at later stages of DC maturation. The repressed genes function in pivotal processes--including antigen-presentation, extracellular signal detection, intracellular signal transduction and lipid-mediator biosynthesis--underscoring the central contribution of the silencing mechanism to rapid reshaping of DC function. Interestingly, promoters of the repressed genes exhibit a surprisingly high frequency of PU.1-occupied sites, suggesting a novel role for this lineage-specific transcription factor in marking genes poised for inducible repression., (© The Author(s) 2014. Published by Oxford University Press on behalf of Nucleic Acids Research.)
- Published
- 2014
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98. Large T Antigen-Specific Cytotoxic T Cells Protect Against Dendritic Cell Tumors through Perforin-Mediated Mechanisms Independent of CD4 T Cell Help.
- Author
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Duval A, Fuertes Marraco SA, Schwitter D, Leuenberger L, and Acha-Orbea H
- Abstract
Our newly generated murine tumor dendritic cell (MuTuDC) lines, generated from tumors developing in transgenic mice expressing the simian virus 40 large T antigen (SV40LgT) and GFP under the DC specific promoter CD11c, reproduce the phenotypic and functional properties of splenic wild type CD8α(+) conventional DCs. They have an immature phenotype with low co-stimulation molecule expression (CD40, CD70, CD80, and CD86) that is upregulated after activation with toll-like receptor ligands. We observed that after transfer into syngeneic C57BL/6 mice, MuTuDC lines were quickly rejected. Tumors grew efficiently in large T transgene-tolerant mice. To investigate the immune response toward the large T antigen that leads to rejection of the MuTuDC lines, they were genetically engineered by lentiviral transduction to express luciferase and tested for the induction of DC tumors after adoptive transfer in various gene deficient recipient mice. Here, we document that the MuTuDC line was rejected in C57BL/6 mice by a CD4 T cell help-independent, perforin-mediated CD8 T cell response to the SV40LgT without pre-activation or co-injection of adjuvants. Using depleting anti-CD8β antibodies, we were able to induce efficient tumor growth in C57BL/6 mice. These results are important for researchers who want to use the MuTuDC lines for in vivo studies.
- Published
- 2014
- Full Text
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99. Cross-presenting dendritic cells are required for control of Leishmania major infection.
- Author
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Ashok D, Schuster S, Ronet C, Rosa M, Mack V, Lavanchy C, Marraco SF, Fasel N, Murphy KM, Tacchini-Cottier F, and Acha-Orbea H
- Subjects
- Animals, Antibodies, Protozoan blood, Antibodies, Protozoan genetics, Antibodies, Protozoan immunology, Basic-Leucine Zipper Transcription Factors biosynthesis, Basic-Leucine Zipper Transcription Factors genetics, CD8-Positive T-Lymphocytes immunology, CD8-Positive T-Lymphocytes metabolism, CD8-Positive T-Lymphocytes pathology, Dendritic Cells metabolism, Dendritic Cells pathology, Immunoglobulin E blood, Immunoglobulin E genetics, Immunoglobulin E immunology, Interferon-gamma, Leishmania major metabolism, Leishmaniasis, Cutaneous blood, Leishmaniasis, Cutaneous genetics, Leishmaniasis, Cutaneous pathology, Mice, Mice, Inbred BALB C, Mice, Knockout, Repressor Proteins biosynthesis, Repressor Proteins genetics, Th2 Cells immunology, Th2 Cells metabolism, Th2 Cells pathology, Antigen Presentation, Basic-Leucine Zipper Transcription Factors immunology, Cross-Priming immunology, Dendritic Cells immunology, Leishmania major immunology, Leishmaniasis, Cutaneous immunology, Repressor Proteins immunology
- Abstract
Leishmania major infection induces self-healing cutaneous lesions in C57BL/6 mice. Both IL-12 and IFN-γ are essential for the control of infection. We infected Jun dimerization protein p21SNFT (Batf3(-/-) ) mice (C57BL/6 background) that lack the major IL-12 producing and cross-presenting CD8α(+) and CD103(+) DC subsets. Batf3(-/-) mice displayed enhanced susceptibility with larger lesions and higher parasite burden. Additionally, cells from draining lymph nodes of infected Batf3(-/-) mice secreted less IFN-γ, but more Th2- and Th17-type cytokines, mirrored by increased serum IgE and Leishmania-specific immunoglobulin 1 (Th2 indicating). Importantly, CD8α(+) DCs isolated from lymph nodes of L. major-infected mice induced significantly more IFN-γ secretion by L. major-stimulated immune T cells than CD103(+) DCs. We next developed CD11c-diptheria toxin receptor: Batf3(-/-) mixed bone marrow chimeras to determine when the DCs are important for the control of infection. Mice depleted of Batf-3-dependent DCs from day 17 or wild-type mice depleted of cross-presenting DCs from 17-19 days after infection maintained significantly larger lesions similar to mice whose Batf-3-dependent DCs were depleted from the onset of infection. Thus, we have identified a crucial role for Batf-3-dependent DCs in protection against L. major., (© 2014 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.)
- Published
- 2014
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100. Interferon-γ induces expression of MHC class II on intestinal epithelial cells and protects mice from colitis.
- Author
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Thelemann C, Eren RO, Coutaz M, Brasseit J, Bouzourene H, Rosa M, Duval A, Lavanchy C, Mack V, Mueller C, Reith W, and Acha-Orbea H
- Subjects
- Animals, Antibodies, Monoclonal pharmacology, Chemokines metabolism, Colitis pathology, Colon drug effects, Colon immunology, Colon pathology, Enterocytes drug effects, Enterocytes pathology, Forkhead Transcription Factors metabolism, Helicobacter drug effects, Helicobacter physiology, Helicobacter Infections immunology, Immunity, Innate drug effects, Inflammation Mediators metabolism, Keratin-14 genetics, Lymphocyte Count, Mice, Inbred C57BL, Mice, Transgenic, Nuclear Proteins genetics, Promoter Regions, Genetic genetics, Receptors, Interleukin-10 immunology, Th1 Cells drug effects, Th1 Cells immunology, Trans-Activators genetics, Up-Regulation drug effects, Colitis immunology, Colitis prevention & control, Enterocytes metabolism, Interferon-gamma metabolism, Nuclear Proteins metabolism, Trans-Activators metabolism
- Abstract
Immune responses against intestinal microbiota contribute to the pathogenesis of inflammatory bowel diseases (IBD) and involve CD4(+) T cells, which are activated by major histocompatibility complex class II (MHCII) molecules on antigen-presenting cells (APCs). However, it is largely unexplored how inflammation-induced MHCII expression by intestinal epithelial cells (IEC) affects CD4(+) T cell-mediated immunity or tolerance induction in vivo. Here, we investigated how epithelial MHCII expression is induced and how a deficiency in inducible epithelial MHCII expression alters susceptibility to colitis and the outcome of colon-specific immune responses. Colitis was induced in mice that lacked inducible expression of MHCII molecules on all nonhematopoietic cells, or specifically on IECs, by continuous infection with Helicobacter hepaticus and administration of interleukin (IL)-10 receptor-blocking antibodies (anti-IL10R mAb). To assess the role of interferon (IFN)-γ in inducing epithelial MHCII expression, the T cell adoptive transfer model of colitis was used. Abrogation of MHCII expression by nonhematopoietic cells or IECs induces colitis associated with increased colonic frequencies of innate immune cells and expression of proinflammatory cytokines. CD4(+) T-helper type (Th)1 cells - but not group 3 innate lymphoid cells (ILCs) or Th17 cells - are elevated, resulting in an unfavourably altered ratio between CD4(+) T cells and forkhead box P3 (FoxP3)(+) regulatory T (Treg) cells. IFN-γ produced mainly by CD4(+) T cells is required to upregulate MHCII expression by IECs. These results suggest that, in addition to its proinflammatory roles, IFN-γ exerts a critical anti-inflammatory function in the intestine which protects against colitis by inducing MHCII expression on IECs. This may explain the failure of anti-IFN-γ treatment to induce remission in IBD patients, despite the association of elevated IFN-γ and IBD.
- Published
- 2014
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