Counteracting breast-cancer induced immune suppression by reactivating lymph node-resident conventional dendritic cells (LNR-cDC)

Breast cancer (BrC)-derived factors inhibit proper development and activation of dendritic cells (DC), which in turn promote the expansion of regulatory T cells (Tregs), thus contributing to tumor progression and spread. In two separate BrC sentinel lymph node (SLN) cohorts, one from the EACRI in Po...

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Published in:	Journal for immunotherapy of cancer Vol. 3; no. Suppl 2; p. P276
Main Authors:	van de Ven, Rieneke, van Pul, Kim, Aliabadi, Shaghayegh, van den Tol, Petrousjka, te Velde, Lisette A, Rutgers, Emiel JTh, Vuylsteke, Ronald JCLM, Stockmann, Hein BAC, Dubay, Christopher, Harrington, Chris, Fox, Bernard A, de Gruijl, Tanja D
Format:	Journal Article
Language:	English
Published:	London BioMed Central 04.11.2015 BioMed Central Ltd BMJ Publishing Group LTD
Subjects:	Antigens Breast cancer Complications and side effects Health aspects Immunology Immunosuppression Immunotherapy Lymph nodes Lymphatic system Lymphocytes Medicine Medicine & Public Health Metastasis Oncology Poster Presentation Prevention Sentinel Lymph Node Treg Frequency Efficient Antigen Immune Suppression Dendritic Cell
ISSN:	2051-1426, 2051-1426
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Abstract	Breast cancer (BrC)-derived factors inhibit proper development and activation of dendritic cells (DC), which in turn promote the expansion of regulatory T cells (Tregs), thus contributing to tumor progression and spread. In two separate BrC sentinel lymph node (SLN) cohorts, one from the EACRI in Portland and one from the VUmc in Amsterdam, we found metastasis-related immune suppression of CD11c+CD14-lymph node-resident conventional DC (LNR-cDC).Genome-wide microarray analyses on FACS sorted migratory CD1a+ cDC, LNR-cDC and CD14+ LNR-cDC isolated from breast-draining LN were performed. A focused analysis of c-type lectin receptors (CLR) and transcription factor genes associated with Ag uptake and cross-presentation, showed LNR-cDC to express relatively high transcript levels of Interferon-regulatory factor 8 (IRF8), chemokine receptor 1 (XCR1), BDCA3/thrombomodulin, and, remarkably, of Langerin, CLEC7A and CLEC4A. These are interesting observations, since these are all factors previously linked to efficient antigen cross-presentation and cross-priming of CD8+ T cells.The EACRI cohort (n=38) included LN from ductal carcinoma in situ (DCIS), tumor-negative SLN (SLN-) and metastasis-positive SLN (SLN+), analyzed with a 12-parameter flow cytometry panel. The VUmc cohort included healthy, prophylactically removed breast LN (HLN, n=16), SLN- and SLN+ (n=40). In both cohorts, there was significantly reduced activation of LNR-cDC, but not of migratory cDC, in SLN-versus DCIS/HLN with further reductions in SLN+. Notably, reduction in activation of LNR-cDC went hand-in-hand with increased Treg frequencies.We next explored the possibility to counteract this BrC-related suppression in SLN ex vivo. The combined activity of CpG-B and the JAK2/STAT3 inhibitor (STAT3i) AG490 resulted in increased activation of LNR-plasmacytoid DC and LNR-cDC. It also promoted production of IFNγ by CD8+ T cells, reduced production of IL-4 by CD4+ T cells, and decreased rates of suppressive Tregs, which were slightly elevated by CpG-B alone. Finally, the combination of CpG-B and STAT3i increased tumor-specific effector T cell responses as measured by IFNγ ELISPOT assay after in vitro restimulation against a pool of overlapping 15-mer peptides, covering the sequence of the BrC–associated antigen Mammaglobin-A.Combined, our data show that BrC-mediated suppression in SLN is primarily directed against the LNR-cDC subset (with particular cross-presenting characteristics and abilities), rather than CD1a+ cDC subsets that migrate to the SLN from local tissues. Stimulation with CpG-B and STAT3i could reactivate these suppressed LNR-cDC ex vivo, thereby restoring type-1 mediated anti-tumor immunity. In view of increasing evidence that immune-regulated pathways influence response to (neo)adjuvant chemotherapy, we anticipate clinical benefit of combination therapy with this immune-stimulatory cocktail.
AbstractList	Breast cancer (BrC)-derived factors inhibit proper development and activation of dendritic cells (DC), which in turn promote the expansion of regulatory T cells (Tregs), thus contributing to tumor progression and spread. In two separate BrC sentinel lymph node (SLN) cohorts, one from the EACRI in Portland and one from the VUmc in Amsterdam, we found metastasis-related immune suppression of CD11c+CD14-lymph node-resident conventional DC (LNR-cDC).Genome-wide microarray analyses on FACS sorted migratory CD1a+ cDC, LNR-cDC and CD14+ LNR-cDC isolated from breast-draining LN were performed. A focused analysis of c-type lectin receptors (CLR) and transcription factor genes associated with Ag uptake and cross-presentation, showed LNR-cDC to express relatively high transcript levels of Interferon-regulatory factor 8 (IRF8), chemokine receptor 1 (XCR1), BDCA3/thrombomodulin, and, remarkably, of Langerin, CLEC7A and CLEC4A. These are interesting observations, since these are all factors previously linked to efficient antigen cross-presentation and cross-priming of CD8+ T cells.The EACRI cohort (n=38) included LN from ductal carcinoma in situ (DCIS), tumor-negative SLN (SLN-) and metastasis-positive SLN (SLN+), analyzed with a 12-parameter flow cytometry panel. The VUmc cohort included healthy, prophylactically removed breast LN (HLN, n=16), SLN- and SLN+ (n=40). In both cohorts, there was significantly reduced activation of LNR-cDC, but not of migratory cDC, in SLN-versus DCIS/HLN with further reductions in SLN+. Notably, reduction in activation of LNR-cDC went hand-in-hand with increased Treg frequencies.We next explored the possibility to counteract this BrC-related suppression in SLN ex vivo. The combined activity of CpG-B and the JAK2/STAT3 inhibitor (STAT3i) AG490 resulted in increased activation of LNR-plasmacytoid DC and LNR-cDC. It also promoted production of IFNγ by CD8+ T cells, reduced production of IL-4 by CD4+ T cells, and decreased rates of suppressive Tregs, which were slightly elevated by CpG-B alone. Finally, the combination of CpG-B and STAT3i increased tumor-specific effector T cell responses as measured by IFNγ ELISPOT assay after in vitro restimulation against a pool of overlapping 15-mer peptides, covering the sequence of the BrC–associated antigen Mammaglobin-A.Combined, our data show that BrC-mediated suppression in SLN is primarily directed against the LNR-cDC subset (with particular cross-presenting characteristics and abilities), rather than CD1a+ cDC subsets that migrate to the SLN from local tissues. Stimulation with CpG-B and STAT3i could reactivate these suppressed LNR-cDC ex vivo, thereby restoring type-1 mediated anti-tumor immunity. In view of increasing evidence that immune-regulated pathways influence response to (neo)adjuvant chemotherapy, we anticipate clinical benefit of combination therapy with this immune-stimulatory cocktail.
ArticleNumber	P276
Audience	Academic
Author	Aliabadi, Shaghayegh Vuylsteke, Ronald JCLM van de Ven, Rieneke van Pul, Kim Dubay, Christopher Harrington, Chris van den Tol, Petrousjka Fox, Bernard A te Velde, Lisette A Stockmann, Hein BAC de Gruijl, Tanja D Rutgers, Emiel JTh
AuthorAffiliation	4 Spaarne Gasthuis, Haarlem, Netherlands 3 Antoni van Leeuwenhoek Hospital, Netherlands Cancer Institute, Amsterdam, Netherlands 1 VU University medical center, Cancer Center Amsterdam, Amsterdam, Netherlands 6 Earle A. Chiles Research Institute, Portland, OR, USA 5 Oregon Health and Sciences University (OHSU), Portland, OR, USA 2 Robert W. Franz Cancer Research center at the Earle A. Chiles Research Institute (EACRI), Providence Cancer Center, Portland, OR, USA
AuthorAffiliation_xml	– name: 1 VU University medical center, Cancer Center Amsterdam, Amsterdam, Netherlands – name: 2 Robert W. Franz Cancer Research center at the Earle A. Chiles Research Institute (EACRI), Providence Cancer Center, Portland, OR, USA – name: 5 Oregon Health and Sciences University (OHSU), Portland, OR, USA – name: 6 Earle A. Chiles Research Institute, Portland, OR, USA – name: 4 Spaarne Gasthuis, Haarlem, Netherlands – name: 3 Antoni van Leeuwenhoek Hospital, Netherlands Cancer Institute, Amsterdam, Netherlands
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Copyright	van de Ven et al. 2015 COPYRIGHT 2015 BioMed Central Ltd. 2015 van de Ven et al. This article is published under license to BioMed Central Ltd. This is an Open Access article distributed under the terms of the Creative Commons Attribution License ( http://creativecommons.org/licenses/by/4.0 ), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. The Creative Commons Public Domain Dedication waiver ( http://creativecommons.org/publicdomain/zero/1.0/ ) applies to the data made available in this article, unless otherwise stated. Notwithstanding the ProQuest Terms and Conditions, you may use this content in accordance with the terms of the License. Copyright © 2015 van de Ven et al. 2015 van de Ven et al.
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Snippet	Breast cancer (BrC)-derived factors inhibit proper development and activation of dendritic cells (DC), which in turn promote the expansion of regulatory T...
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SubjectTerms	Antigens Breast cancer Complications and side effects Health aspects Immunology Immunosuppression Immunotherapy Lymph nodes Lymphatic system Lymphocytes Medicine Medicine & Public Health Metastasis Oncology Poster Presentation Prevention
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Title	Counteracting breast-cancer induced immune suppression by reactivating lymph node-resident conventional dendritic cells (LNR-cDC)
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