Publications by authors named "Marie-Caroline Dieu-Nosjean"

63 Publications

Tertiary Lymphoid Structure-B Cells Narrow Regulatory T Cells Impact in Lung Cancer Patients.

Front Immunol 2021 8;12:626776. Epub 2021 Mar 8.

Sorbonne Université, UMRS 1135, Faculté de Médecine Sorbonne Université, Paris, France.

The presence of tertiary lymphoid structures (TLS) in the tumor microenvironment is associated with better clinical outcome in many cancers. In non-small cell lung cancer (NSCLC), we have previously showed that a high density of B cells within TLS (TLS-B cells) is positively correlated with tumor antigen-specific antibody responses and increased intratumor CD4 T cell clonality. Here, we investigated the relationship between the presence of TLS-B cells and CD4 T cell profile in NSCLC patients. The expression of immune-related genes and proteins on B cells and CD4 T cells was analyzed according to their relationship to TLS-B density in a prospective cohort of 56 NSCLC patients. We observed that tumor-infiltrating T cells showed marked differences according to TLS-B cell presence, with higher percentages of naïve, central-memory, and activated CD4 T cells and lower percentages of both immune checkpoint (ICP)-expressing CD4 T cells and regulatory T cells (Tregs) in the TLS-B tumors. A retrospective study of 538 untreated NSCLC patients showed that high TLS-B cell density was even able to counterbalance the deleterious impact of high Treg density on patient survival, and that TLS-B Treg patients had the best clinical outcomes. Overall, the correlation between the density of TLS-B tumors with early differentiated, activated and non-regulatory CD4 T cell cells suggest that B cells may play a central role in determining protective T cell responses in NSCLC patients.
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http://dx.doi.org/10.3389/fimmu.2021.626776DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7983944PMC
March 2021

[Rethinking the place of B lymphocyte and tertiary lymphoid structures in oncoimmunotherapy].

Med Sci (Paris) 2021 02 16;37(2):130-133. Epub 2021 Feb 16.

Laboratoire Microenvironnement immunitaire et immunothérapie, UMRS 1135 Sorbonne Université, Inserm U1135, Centre d'immunologie et des maladies infectieuses (CIMI-Paris), Faculté de médecine Sorbonne Université, 91 boulevard de l'Hôpital, 75013 Paris, France.

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http://dx.doi.org/10.1051/medsci/2020276DOI Listing
February 2021

Natural killer cells in the human lung tumor microenvironment display immune inhibitory functions.

J Immunother Cancer 2020 10;8(2)

Centre de Recherche des Cordeliers, Sorbonne Universite, Inserm, Universite de Paris, Team Inflammation, complement and cancer, F-75006, Paris, France

Background: Natural killer (NK) cells play a crucial role in tumor immunosurveillance through their cytotoxic effector functions and their capacity to interact with other immune cells to build a coordinated antitumor immune response. Emerging data reveal NK cell dysfunction within the tumor microenvironment (TME) through checkpoint inhibitory molecules associated with a regulatory phenotype.

Objective: We aimed at analyzing the gene expression profile of intratumoral NK cells compared with non-tumorous NK cells, and to characterize their inhibitory function in the TME.

Methods: NK cells were sorted from human lung tumor tissue and compared with non- tumoral distant lungs.

Results: In the current study, we identify a unique gene signature of NK cell dysfunction in human non-small cell lung carcinoma (NSCLC). First, transcriptomic analysis reveals significant changes related to migratory pattern with a downregulation of sphingosine-1-phosphate receptor 1 (S1PR1) and CX3C chemokine receptor 1 (CX3CR1) and overexpression of C-X-C chemokine receptor type 5 (CXCR5) and C-X-C chemokine receptor type 6 (CXCR6). Second, cytotoxic T-lymphocyte-associated protein 4 (CTLA-4) and killer cell lectin like receptor (KLRC1) inhibitory molecules were increased in intratumoral NK cells, and CTLA-4 blockade could partially restore MHC class II level on dendritic cell (DC) that was impaired during the DCs/NK cell cross talk. Finally, NK cell density impacts the positive prognostic value of CD8 T cells in NSCLC.

Conclusions: These findings demonstrate novel molecular cues associated with NK cell inhibitory functions in NSCLC.
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http://dx.doi.org/10.1136/jitc-2020-001054DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7570244PMC
October 2020

[The biology of PD1 and CTLA-4 as immunotherapeutic targets and the issue of biomarkers].

Med Sci (Paris) 2019 Dec 6;35(12):957-965. Epub 2020 Jan 6.

UMR Inserm 1052, Centre de Recherche en Cancérologie de Lyon (CRCL), Lyon, France.

The identification in the 1990's of the role of CTLA-4 and PD-1, two inhibitory receptors of T lymphocytes, in the control of the anti-tumor immune responses, led to the awarding of the Nobel Prize in Physiology or Medicine in 2018 to Dr. James Allison and Dr. Tasuku Honjo. These inhibitory receptors called immune checkpoints are essential to prevent any deleterious impact of on-going immune responses against pathogens or cancer cells on healthy tissues and, hence, guarantee the integrity of the host. These major discoveries have led James Allison and Tasuku Honjo to develop anti-CTLA-4 and anti-PD1/L-1 antibodies, respectively, in order to switch off these immune "brakes", making it possible to efficiently attack tumor cells. CTLA-4 regulates the amplitude of the early T-cell activation and inhibits the activity of CD28, a major activating co-receptor expressed by T cells. PD-1 is expressed by memory and effector T lymphocytes and is involved in the regulation of chronically activated cells, as observed during inflammatory processes. Immunotherapeutic treatments resulting from these discoveries have now a major place in the arsenal of anti-cancer therapies. This review presents firstly a synthesis of knowledge on CTLA-4, PD-1 and their ligands, their mechanisms of action and regulation and, secondly, an overview of biomarkers that have been associated with clinical response to anti-PD-1/PD-L1 and anti-CTLA-4 antibody therapies.
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http://dx.doi.org/10.1051/medsci/2019192DOI Listing
December 2019

Automated image analysis of NSCLC biopsies to predict response to anti-PD-L1 therapy.

J Immunother Cancer 2019 05 6;7(1):121. Epub 2019 May 6.

AstraZeneca, Gaithersburg, MD, 20878, USA.

Background: Immune checkpoint therapies (ICTs) targeting the programmed cell death-1 (PD1)/programmed cell death ligand-1 (PD-L1) pathway have improved outcomes for patients with non-small cell lung cancer (NSCLC), particularly those with high PD-L1 expression. However, the predictive value of manual PD-L1 scoring is imperfect and alternative measures are needed. We report an automated image analysis solution to determine the predictive and prognostic values of the product of PD-L1+ cell and CD8+ tumor infiltrating lymphocyte (TIL) densities (CD8xPD-L1 signature) in baseline tumor biopsies.

Methods: Archival or fresh tumor biopsies were analyzed for PD-L1 and CD8 expression by immunohistochemistry. Samples were collected from 163 patients in Study 1108/NCT01693562, a Phase 1/2 trial to evaluate durvalumab across multiple tumor types, including NSCLC, and a separate cohort of 199 non-ICT- patients. Digital images were automatically scored for PD-L1+ and CD8+ cell densities using customized algorithms applied with Developer XD™ 2.7 software.

Results: For patients who received durvalumab, median overall survival (OS) was 21.0 months for CD8xPD-L1 signature-positive patients and 7.8 months for signature-negative patients (p = 0.00002). The CD8xPD-L1 signature provided greater stratification of OS than high densities of CD8+ cells, high densities of PD-L1+ cells, or manually assessed tumor cell PD-L1 expression ≥25%. The CD8xPD-L1 signature did not stratify OS in non-ICT patients, although a high density of CD8+ cells was associated with higher median OS (high: 67 months; low: 39.5 months, p = 0.0009) in this group.

Conclusions: An automated CD8xPD-L1 signature may help to identify NSCLC patients with improved response to durvalumab therapy. Our data also support the prognostic value of CD8+ TILS in NSCLC patients who do not receive ICT.

Trial Registration: ClinicalTrials.gov identifier: NCT01693562 . Study code: CD-ON-MEDI4736-1108. Interventional study (ongoing but not currently recruiting). Actual study start date: August 29, 2012. Primary completion date: June 23, 2017 (final data collection date for primary outcome measure).
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http://dx.doi.org/10.1186/s40425-019-0589-xDOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6501300PMC
May 2019

Designed Methods for the Sorting of Tertiary Lymphoid Structure-Immune Cell Populations.

Methods Mol Biol 2018 ;1845:189-204

Cordeliers Research Center, Laboratory "Cancer, Immune Control and Escape", Institut National de la Santé et de la Recherche Médicale (INSERM), UMRS 1138, Paris, France.

The tumor microenvironment is a complex network of interacting cells composed of immune and nonimmune cells. It has been reported that the composition of the immune contexture has a significant impact on tumor growth and patient survival in different solid tumors. For instance, we and other groups have previously demonstrated that a strong infiltration of T-helper type 1 (Th1) or memory CD8 T cells is associated with long-term survival of cancer patients. Nevertheless, the prognostic value of the other immune populations, namely regulatory T cells (Treg), B cells, and gamma delta (γδ) T cells, remains a matter of debate. Herein, we describe novel flow cytometry-based strategies to sort out these different immune populations in order to evaluate their role in non-small cell lung cancer (NSCLC).
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http://dx.doi.org/10.1007/978-1-4939-8709-2_11DOI Listing
May 2019

Development of Methods for Selective Gene Expression Profiling in Tertiary Lymphoid Structure Using Laser Capture Microdissection.

Methods Mol Biol 2018 ;1845:119-137

Cordeliers Research Center, Laboratory "Cancer, Immune Control and Escape", Institut National de la Santé et de la Recherche Médicale (INSERM), UMRS 1138, Paris, France.

Tertiary lymphoid structures (TLS) are de novo lymphoid formations that are induced within tissues during inflammatory episodes. TLS have been reported at various anatomic sites and in many different contexts like cancer, infections, autoimmunity, graft rejection, and idiopathic diseases. These inducible, ectopic, and transient lymphoid structures exhibit the prototypical architecture found within secondary lymphoid organs (SLO) and have been recently appreciated as a major driver of the local adaptive immune reaction. As TLS emerge within tissues, the isolation in situ and the molecular characterization of these structures are challenging to operate. Laser capture microdissection (LCM) is a powerful tool to isolate selective structural components and cells from frozen or paraffin-embedded tissues. We and other groups previously applied LCM to decipher the molecular network within TLS and uncover their intrinsic connection with the local microenvironment. In this chapter, we describe a detailed LCM method for selecting and isolating TLS in situ to perform comprehensive downstream molecular analyses.
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http://dx.doi.org/10.1007/978-1-4939-8709-2_8DOI Listing
May 2019

Development of Tools for the Selective Visualization and Quantification of TLS-Immune Cells on Tissue Sections.

Methods Mol Biol 2018 ;1845:47-69

Sorbonne University, UMRS 1138, Cordeliers Research Center, Paris, France.

Tertiary lymphoid structures (TLS) are considered as genuine markers of inflammation. Their presence within inflamed tissues or within the tumor microenvironment has been associated with the local development of an active immune response. While high densities of TLS are correlated with disease severity in autoimmune diseases or during graft rejection, it has been associated with longer patient survival in many cancer types. Their efficient visualization and quantification within human tissues may represent new tools for helping clinicians in adjusting their therapeutic strategy. Some immunohistochemistry (IHC) protocols are already used in the clinic to appreciate the level of immune infiltration in formalin-fixed, paraffin-embedded (FFPE) tissues. However, the use of two or more markers may sometimes be useful to better characterize this immune infiltrate, especially in the case of TLS. Besides the growing development of multiplex labeling approaches, imaging can also be used to overcome some technical difficulties encountered during the immunolabeling of tissues with several markers.This chapter describes IHC methods to visualize in a human tissue (tumoral or not) the presence of TLS. These methods are based on the immunostaining of four TLS-associated immune cell populations, namely follicular B cells, follicular dendritic cells (FDCs), mature dendritic cells (mDCs), and follicular helper T cells (T), together with non-T T cells. Methodologies for subsequent quantification of TLS density are also proposed, as well as a virtual multiplexing method based on image registration using the open-source software ImageJ (IJ), aiming at co-localizing several immune cell populations from different IHC stainings performed on serial tissue sections.
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http://dx.doi.org/10.1007/978-1-4939-8709-2_4DOI Listing
May 2019

, and Mutations Predict Tumor Immune Profile and the Response to Anti-PD-1 in Lung Adenocarcinoma.

Clin Cancer Res 2018 11 15;24(22):5710-5723. Epub 2018 May 15.

Institut National de la Santé et de la Recherche Médicale (INSERM), UMRS 1138, Cordeliers Research Center, Team Cancer, Immune Control and Escape, Paris, France.

By unlocking antitumor immunity, antibodies targeting programmed cell death 1 (PD-1) exhibit impressive clinical results in non-small cell lung cancer, underlining the strong interactions between tumor and immune cells. However, factors that can robustly predict long-lasting responses are still needed. We performed in-depth immune profiling of lung adenocarcinoma using an integrative analysis based on immunohistochemistry, flow-cytometry, and transcriptomic data. Tumor mutational status was investigated using next-generation sequencing. The response to PD-1 blockers was analyzed from a prospective cohort according to tumor mutational profiles and PD-L1 expression, and a public clinical database was used to validate the results obtained. We showed that distinct combinations of , and mutations were major determinants of the tumor immune profile (TIP) and of the expression of PD-L1 by malignant cells. Indeed, the presence of mutations without co-occurring or alterations (-mut/--WT), independently of mutations, identified the group of tumors with the highest CD8 T-cell density and PD-L1 expression. In this tumor subtype, pathways related to T-cell chemotaxis, immune cell cytotoxicity, and antigen processing were upregulated. Finally, a prolonged progression-free survival (PFS: HR = 0.32; 95% CI, 0.16-0.63, < 0.001) was observed in anti-PD-1-treated patients harboring -mut/--WT tumors. This clinical benefit was even more remarkable in patients with associated strong PD-L1 expression. Our study reveals that different combinations of , and mutations, together with PD-L1 expression by tumor cells, represent robust parameters to identify best responders to PD-1 blockade. .
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http://dx.doi.org/10.1158/1078-0432.CCR-18-0163DOI Listing
November 2018

Expression of LLT1 and its receptor CD161 in lung cancer is associated with better clinical outcome.

Oncoimmunology 2018;7(5):e1423184. Epub 2018 Jan 29.

Laboratory "Immune Microenvironment and Tumors", Department "Cancer, Immunology, Immunotherapy", INSERM UMRS 1138, Cordeliers Research Center, Paris, France.

Co-stimulatory and inhibitory receptors expressed by immune cells in the tumor microenvironment modulate the immune response and cancer progression. Their expression and regulation are still not fully characterized and a better understanding of these mechanisms is needed to improve current immunotherapies. Our previous work has identified a novel ligand/receptor pair, LLT1/CD161, that modulates immune responses. Here, we extensively characterize its expression in non-small cell lung cancer (NSCLC). We show that LLT1 expression is restricted to germinal center (GC) B cells within tertiary lymphoid structures (TLS), representing a new hallmark of the presence of active TLS in the tumor microenvironment. CD161-expressing immune cells are found at the vicinity of these structures, with a global enrichment of NSCLC tumors in CD161 CD4 and CD8 T cells as compared to normal distant lung and peripheral blood. CD161 CD4 T cells are more activated and produce Th1-cytokines at a higher frequency than their matched CD161-negative counterparts. Interestingly, CD161 CD4 T cells highly express OX40 co-stimulatory receptor, less frequently 4-1BB, and display an activated but not completely exhausted PD-1-positive Tim-3-negative phenotype. Finally, a meta-analysis revealed a positive association of (coding for LLT1) and (coding for CD161) gene expression with favorable outcome in NSCLC, independently of the size of T and B cell infiltrates. These data are consistent with a positive impact of LLT1/CD161 on NSCLC patient survival, and make CD161-expressing CD4 T cells ideal candidates for efficient anti-tumor recall responses.
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http://dx.doi.org/10.1080/2162402X.2017.1423184DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5927544PMC
January 2018

Impaired Tumor-Infiltrating T Cells in Patients with Chronic Obstructive Pulmonary Disease Impact Lung Cancer Response to PD-1 Blockade.

Am J Respir Crit Care Med 2018 10;198(7):928-940

1 Institut National de la Santé et de la Recherche Médicale (INSERM), UMRS 1138, Cordeliers Research Center, Team Cancer, Immune Control and Escape, Paris, France.

Rationale: Patients with chronic obstructive pulmonary disease (COPD) have a higher prevalence of lung cancer. The chronic inflammation associated with COPD probably promotes the earliest stages of carcinogenesis. However, once tumors have progressed to malignancy, the impact of COPD on the tumor immune microenvironment remains poorly defined, and its effects on immune-checkpoint blockers' efficacy are still unknown.

Objectives: To study the impact of COPD on the immune contexture of non-small cell lung cancer.

Methods: We performed in-depth immune profiling of lung tumors by immunohistochemistry and we determined its impact on patient survival (n = 435). Tumor-infiltrating T lymphocyte (TIL) exhaustion by flow cytometry (n = 50) was also investigated. The effectiveness of an anti-PD-1 (programmed cell death-1) treatment (nivolumab) was evaluated in 39 patients with advanced-stage non-small cell lung cancer. All data were analyzed according to patient COPD status.

Measurements And Main Results: Remarkably, COPD severity is positively correlated with the coexpression of PD-1/TIM-3 (T-cell immunoglobulin and mucin domain-containing molecule-3) by CD8 T cells. In agreement, we observed a loss of CD8 T cell-associated favorable clinical outcome in COPD patients. Interestingly, a negative prognostic value of PD-L1 (programmed cell death ligand 1) expression by tumor cells was observed only in highly CD8 T cell-infiltrated tumors of COPD patients. Finally, data obtained on 39 patients with advanced-stage non-small cell lung cancer treated by an anti-PD-1 antibody showed longer progression-free survival in COPD patients, and also that the association between the severity of smoking and the response to nivolumab was preferentially observed in COPD patients.

Conclusions: COPD is associated with an increased sensitivity of CD8 tumor-infiltrating T lymphocytes to immune escape mechanisms developed by tumors, thus suggesting a higher sensitivity to PD-1 blockade in patients with COPD.
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http://dx.doi.org/10.1164/rccm.201706-1110OCDOI Listing
October 2018

[Which future for B lymphocytes infiltrating solid tumors: prognostic biomarker and/or therapeutic target?]

Med Sci (Paris) 2018 Jan 31;34(1):72-78. Epub 2018 Jan 31.

Inserm, UMRS 1138, Centre de recherche des cordeliers, laboratoire « Cancer et Immunité anti-tumorale », F-75006, Paris, France - Université Paris Descartes, Sorbonne Paris Cité, UMRS 1138, Centre de recherche des cordeliers, 15, rue de l'École de Médecine, F-75006, Paris, France - Université Pierre et Marie Curie (UPMC), Paris 06, Sorbonne Université, UMRS 1138, Centre de recherche des cordeliers, 15, rue de l'École de Médecine, F-75006, Paris, France.

The role of B lymphocytes in tumor immuno-surveillance has been neglected for a long time because it has been often considered to be ineffective if not pro-tumoral. Extensive studies of the tumor immune microenvironment, particularly in humans, has now made it possible to specify the nature of the interactions between B cells and their cellular partners. This review will present a number of parameters that dictate the fate of B cells toward a pro-tumor versus an anti-tumor function. Thus, the ability to elicit a B cell-and/or an antibody-dependent anti-tumor immunity involves a wide variety of molecular and cellular mechanisms, some of which may represent novel therapeutic targets in oncology.
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http://dx.doi.org/10.1051/medsci/20183401016DOI Listing
January 2018

Tertiary Lymphoid Structures: An Anti-tumor School for Adaptive Immune Cells and an Antibody Factory to Fight Cancer?

Front Immunol 2017 21;8:830. Epub 2017 Jul 21.

INSERM, UMRS 1138, Cordeliers Research Center, Team "Cancer, Immune Control and Escape", Paris, France.

Tertiary lymphoid structures (TLS) present in human solid tumors are essential for the shaping of a favorable immune micro-environment to control tumor development in most cases. They represent a formidable school for T-cell priming, B cell activation, and differentiation into plasma cells and an exquisitely located factory for antibody production. The manipulation of TLS neogenesis and maintenance represents, therefore, an exciting task to set up efficient anti-cancer vaccine strategies leading to long-lasting anti-tumor adaptive responses. To achieve this goal, a number of important issues are still pending. How TLS-T and -B cells and antibodies locally produced are related to the improved survival of cancer patients with high density of TLS is still unclear. In addition, the mechanisms by which tumors escape the immune surveillance exerted by TLS are still poorly understood and the role of immune suppressive cytokines, regulatory T cells, and/or antibodies in this process remains to be explored. The identification of the key parameters that distinguish TLS with anti- or possible pro-tumor activity is also essential to make the therapeutic targeting of TLS a success. Finally, how TLS-based therapeutic approaches can be associated with targeted therapies or immunointerventions, such as the use of ICP blockers to improve anti-tumor responses, is an open question. We will discuss these different issues in the present review.
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http://dx.doi.org/10.3389/fimmu.2017.00830DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5519532PMC
July 2017

Key Features of Gamma-Delta T-Cell Subsets in Human Diseases and Their Immunotherapeutic Implications.

Front Immunol 2017 30;8:761. Epub 2017 Jun 30.

Cordeliers Research Center, UMRS 1138, Team "Cancer, Immune Control and Escape", INSERM, Paris, France.

The unique features of gamma-delta (γδ) T cells, related to their antigen recognition capacity, their tissue tropism, and their cytotoxic function, make these cells ideal candidates that could be targeted to induce durable immunity in the context of different pathologies. In this review, we focus on the main characteristics of human γδ T-cell subsets in diseases and the key mechanisms that could be explored to target these cells.
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http://dx.doi.org/10.3389/fimmu.2017.00761DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5491929PMC
June 2017

Immune contexture and histological response after neoadjuvant chemotherapy predict clinical outcome of lung cancer patients.

Oncoimmunology 2016;5(12):e1255394. Epub 2016 Dec 8.

INSERM U1138, Team "Cancer, Immune Control, and Escape" Cordeliers Research Center, Paris, France; Pierre et Marie Curie-Paris 6 University, Paris, France; Paris Descartes-Paris 5 University, Paris, France; Pathology Department, Cochin hospital, AP-HP, Paris, France.

There is now growing evidence that the immune contexture influences cancer progression and clinical outcome of patients with non-small cell lung cancer (NSCLC). If chemotherapy is widely used to treat patients with advanced-stage NSCLC, it remains unclear how it could modify the immune contexture and impact its prognostic value. Here, we analyzed two retrospective cohorts, respectively composed of 122 stage III-N2 NSCLC patients treated with chemotherapy before surgery and 39 stage-matched patients treated by surgery only. In patients treated with neoadjuvant chemotherapy, the histological characteristics, the expression of PD-L1 protein, and the tumor immune microenvironment (CD8 T cells, DC-LAMP mature dendritic cells, and CD68 macrophages) were evaluated and their prognostic value assessed together with standard clinical parameters. By analyzing pre- and post-treatment specimens, we did not find any changes in the PD-L1 expression. We also found that the tumor immune contexture in patients treated with neoadjuvant chemotherapy exhibited a similar pattern that the one found in chemotherapy-naive patients, with comparable densities of tumor-infiltrating CD8 and DC-LAMP cells and a similar spatial organization. The percentage of residual viable tumor cells and the immune pattern (CD8 and DC-LAMP cell densities) were significantly associated with the clinical outcome and allowed the identification of short- and long-term survivors, respectively. In multivariate analysis, the immune pattern was found to be the strongest independent prognostic factor. In conclusion, this study decrypts the complex interplay between cancer and immune cells in patients undergoing chemotherapy and supports potential beneficial synergistic effect of immunotherapy and chemotherapy.
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http://dx.doi.org/10.1080/2162402X.2016.1255394DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5213838PMC
December 2016

Tertiary Lymphoid Structures in Cancers: Prognostic Value, Regulation, and Manipulation for Therapeutic Intervention.

Front Immunol 2016 3;7:407. Epub 2016 Oct 3.

INSERM, UMR_S 1138, Team "Cancer, Immune Control and Escape", Cordeliers Research Center, Paris, France; UMR_S 1138, Centre de Recherche des Cordeliers, University Paris Descartes, Paris, France; UMR_S 1138, Centre de Recherche des Cordeliers, Sorbonne University, UPMC University Paris 06, Paris, France.

Tertiary lymphoid structures (TLS) are ectopic lymphoid aggregates that reflect lymphoid neogenesis occurring in tissues at sites of inflammation. They are detected in tumors where they orchestrate local and systemic anti-tumor responses. A correlation has been found between high densities of TLS and prolonged patient's survival in more than 10 different types of cancer. TLS can be regulated by the same set of chemokines and cytokines that orchestrate lymphoid organogenesis and by regulatory T cells. Thus, TLS offer a series of putative new targets that could be used to develop therapies aiming to increase the anti-tumor immune response.
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http://dx.doi.org/10.3389/fimmu.2016.00407DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5046074PMC
October 2016

Intratumoral Immune Cell Densities Are Associated with Lung Adenocarcinoma Gene Alterations.

Am J Respir Crit Care Med 2016 12;194(11):1403-1412

1 INSERM UMRS 1138, Centre de Recherche des Cordeliers, Paris, France.

Rationale: Tumor-infiltrating immune cells affect lung cancer outcome. However, the factors that influence the composition and function of the tumor immune environment remain poorly defined and need investigation, particularly in the era of immunotherapy.

Objectives: To determine whether the tumoral immune environment is related to lung adenocarcinoma mutations.

Methods: This retrospective cohort included 316 consecutive patients with lung adenocarcinoma (225 men; 258 smokers) studied from 2001 to 2005 in a single center. We investigated the association of densities of intratumoral mature dendritic cells (mDCs), CD8 T cells, neutrophils, and macrophages with clinical and pathological variables and tumor cell mutation profiles obtained by next-generation sequencing.

Measurements And Main Results: In 282 tumors, we found 460 mutations, mainly in TP53 (59%), KRAS (40%), STK11 (24%), and EGFR (14%). Intratumoral CD8 T-cell density was high in smokers (P = 0.02) and TP53-mutated tumors (P = 0.02) and low in BRAF-mutated tumors (P = 0.005). Intratumoral mDC density was high with low pathological tumor stage (P = 0.01) and low with STK11 mutation (P = 0.004). Intratumoral neutrophil density was high and low with BRAF mutation (P = 0.04) and EGFR mutation (P = 0.02), respectively. Intratumoral macrophage density was low with EGFR mutation (P = 0.01). Intratumoral CD8 T-cell and mDC densities remained strong independent markers of overall survival (P = 0.001 and P = 0.02, respectively).

Conclusions: Intratumoral immune cell densities (mDCs, CD8 T cells, neutrophils, macrophages) were significantly associated with molecular alterations in adenocarcinoma underlying the interactions between cancer cells and their microenvironment.
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http://dx.doi.org/10.1164/rccm.201510-2031OCDOI Listing
December 2016

Tertiary lymphoid structures, drivers of the anti-tumor responses in human cancers.

Immunol Rev 2016 May;271(1):260-75

INSERM, UMR_S 1138, Cordeliers Research Center, Team 13 Cancer, Immune Control and Escape, Paris, France.

The characterization of the microenvironment of human tumors led to the description of tertiary lymphoid structures (TLS) characterized by mature dendritic cells in a T-cell zone adjacent to B-cell follicle including a germinal center. TLS represent sites of lymphoid neogenesis that develop in most solid cancers. Analysis of the current literature shows that the TLS presence is associated with a favorable clinical outcome for cancer patients, regardless of the approach used to quantify TLS and the stage of the disease. Using several approaches that combine immunohistochemistry, gene expression assays, and flow cytometry on large series of lung tumors, our work demonstrated that TLS are important sites for the initiation and/or maintenance of the local and systemic T- and B-cell responses against tumors. Surrounded by high endothelial venules, they represent a privileged area for the recruitment of lymphocytes into tumors and generation of central-memory T and B cells that circulate and limit cancer progression. TLS can be considered as a novel biomarker to stratify the overall survival risk of untreated cancer patients and as a marker of efficient immunotherapies. The induction and manipulation of cancer-associated TLS using drug agonists and/or biotherapies should open new avenues to treat cancer patients.
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http://dx.doi.org/10.1111/imr.12405DOI Listing
May 2016

Immune Contexture, Immunoscore, and Malignant Cell Molecular Subgroups for Prognostic and Theranostic Classifications of Cancers.

Adv Immunol 2016 29;130:95-190. Epub 2016 Jan 29.

INSERM UMR_S 1138, Cancer, Immune Control and Escape, Cordeliers Research Centre, Paris, France; Université Paris-Descartes, Sorbonne Paris Cité, UMR_S 1338, Cordeliers Research Centre, Paris, France; Sorbonne Universités, UPMC Univ Paris 06, UMR_S 1138, Cordeliers Research Centre, Paris, France. Electronic address:

The outcome of tumors results from genetic and epigenetic modifications of the transformed cells and also from the interactions of the malignant cells with their tumor microenvironment (TME), which includes immune and inflammatory cells. For a given cancer type, the composition of the immunological TME is not homogeneous. Heterogeneity is found between different cancer types and also between tumors from patients with the same type of cancer. Some tumors exhibit a poor infiltration by immune cells, and others are highly infiltrated by lymphocytes. Among the latter, the architecture of the TME, with the localization of immune cells in the invasive front and the center of the tumor, the presence of tumor-adjacent organized lymphoid aggregates, and the type of inflammatory context, determines the prognostic impact of the infiltrating cells. The description and the understanding of the immune and inflammatory landscape in human tumors are of paramount importance at different levels of patient's care. It completes the mutational, transcriptional, and epigenetic patterns of the malignant cells and open paths to understand how tumor cells shape their immune microenvironment and are shaped by the immune reaction. It provides prognostic and theranostic markers, as well as novel targets for immunotherapies.
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http://dx.doi.org/10.1016/bs.ai.2015.12.002DOI Listing
October 2016

Calreticulin Expression in Human Non-Small Cell Lung Cancers Correlates with Increased Accumulation of Antitumor Immune Cells and Favorable Prognosis.

Cancer Res 2016 04 3;76(7):1746-56. Epub 2016 Feb 3.

Institut National de la Santé et de la Recherche Médicale (INSERM), UMRS1138, Centre de Recherche des Cordeliers, Paris, France. Université Pierre et Marie Curie-Paris 6, UMRS1138, Paris, France. Université Paris Descartes-Paris 5, UMRS1138, Paris, France.

A high density of tumor-infiltrating mature dendritic cells (DC) and CD8(+) T cells correlates with a positive prognosis in a majority of human cancers. The recruitment of activated lymphocytes to the tumor microenvironment, primed to recognize tumor-associated antigens, can occur in response to immunogenic cell death (ICD) of tumor cells. ICD is characterized by the preapoptotic translocation of calreticulin (CRT) from the endoplasmic reticulum (ER) to the cell surface as a result of an ER stress response accompanied by the phosphorylation of eukaryotic initiation factor 2α (eIF2α). We conducted a retrospective study on two independent cohorts of patients with non-small cell lung cancer (NSCLC) to investigate the prognostic potential of CRT. We report that the level of CRT expression on tumor cells, which correlated with eIF2α phosphorylation, positively influenced the clinical outcome of NSCLC. High CRT expression on tumor cells was associated with a higher density of infiltrating mature DC and effector memory T-cell subsets, suggesting that CRT triggers the activation of adaptive immune responses in the tumor microenvironment. Accordingly, patients with elevated CRT expression and dense intratumoral infiltration by DC or CD8(+) T lymphocytes had the best prognosis. We conclude that CRT expression constitutes a new powerful prognostic biomarker that reflects enhanced local antitumor immune responses in the lung. Cancer Res; 76(7); 1746-56. ©2016 AACR.
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http://dx.doi.org/10.1158/0008-5472.CAN-15-1142DOI Listing
April 2016

Cancer immune contexture and immunotherapy.

Curr Opin Immunol 2016 Apr 17;39:7-13. Epub 2015 Dec 17.

INSERM UMR_S 1138, Cancer, Immune Control and Escape, Cordeliers Research Centre, Paris, France; Université Paris Descartes, Paris, France; Université Pierre et Marie Curie, Paris, France. Electronic address:

The immune contexture that characterizes the density, the location, the organization and the functional orientation of tumor-infiltrating immune cells in cancers has a clinical impact on patient's outcome. It is, in great part, shaped by the malignant cells, as in a given cancer type, tumors presenting different oncogenic processes have different immune contextures. Moreover, the immune contexture in metastatic sites reflects that of the corresponding primary tumors. Finally, the components forming the immune contexture represent targets and markers of efficient anti-cancer immunotherapies.
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http://dx.doi.org/10.1016/j.coi.2015.11.009DOI Listing
April 2016

A high density of tertiary lymphoid structure B cells in lung tumors is associated with increased CD4 T cell receptor repertoire clonality.

Oncoimmunology 2015 Dec 1;4(12):e1051922. Epub 2015 Jun 1.

Translational Sciences; MedImmune ; Gaithersburg, MD USA.

T and B cell receptor (TCR and BCR, respectively) Vβ or immunoglobulin heavy chain complementarity-determining region 3 sequencing allows monitoring of repertoire changes through recognition, clonal expansion, affinity maturation, and T or B cell activation in response to antigen. TCR and BCR repertoire analysis can advance understanding of antitumor immune responses in the tumor microenvironment. TCR and BCR repertoires of sorted CD4, CD8 or CD19 cells in tumor, non-tumoral distant tissue (NT), and peripheral compartments (blood/draining lymph node [P]) from 47 non-small cell lung cancer (NSCLC) patients (age = 68 y) were sequenced. The clonotype spectra were assessed among different tissues and correlated with clinical and immunological parameters. In all tissues, CD4 and CD8 TCR repertoires had greater clonality relative to CD19 BCR. CD4 T cells exhibited greater clonality in NT compared to tumor ( = 0.002) and P ( < 0.001), concentrated among older patients (age > 68). Younger patients exhibited greater CD4 T cell diversity in P compared to older patients ( = 0.05), and greater CD4 T cell clonality in tumor relative to P ( < 0.001), with fewer shared clonotypes between tumor and P than older patients ( = 0.04). More interestingly, greater CD4 and CD8 T cell clonality in tumor and P, respectively (both = 0.05), correlated with high density of tumor-associated tertiary lymphoid structure (TLS) B cells, a biomarker of higher overall survival in NSCLC. Results indicate distinct adaptive immune responses in NSCLC, where peripheral T cell diversity is modulated by age, and tumor T cell clonal expansion is favored by the presence of TLSs in the tumor microenvironment.
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http://dx.doi.org/10.1080/2162402X.2015.1051922DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4635865PMC
December 2015

Tertiary Lymphoid Structure-Associated B Cells are Key Players in Anti-Tumor Immunity.

Front Immunol 2015 23;6:67. Epub 2015 Feb 23.

Laboratory Cancer, Immune Control and Escape, Cordeliers Research Center, INSERM UMRS1138 , Paris , France ; UMRS1138, University Pierre and Marie Curie , Paris , France ; UMRS1138, University Paris Descartes , Paris , France.

It is now admitted that the immune system plays a major role in tumor control. Besides the existence of tumor-specific T cells and B cells, many studies have demonstrated that high numbers of tumor-infiltrating lymphocytes are associated with good clinical outcome. In addition, not only the density but also the organization of tumor-infiltrating immune cells has been shown to determine patient survival. Indeed, more and more studies describe the development within the tumor microenvironment of tertiary lymphoid structures (TLS), whose presence has a positive impact on tumor prognosis. TLS are transient ectopic lymphoid aggregates displaying the same organization and functionality as canonical secondary lymphoid organs, with T-cell-rich and B-cell-rich areas that are sites for the differentiation of effector and memory T cells and B cells. However, factors favoring the emergence of such structures within tumors still need to be fully characterized. In this review, we survey the state of the art of what is known about the general organization, induction, and functionality of TLS during chronic inflammation, and more especially in cancer, with a particular focus on the B-cell compartment. We detail the role played by TLS B cells in anti-tumor immunity, both as antigen-presenting cells and tumor antigen-specific antibody-secreting cells, and raise the question of the capacity of chemotherapeutic and immunotherapeutic agents to induce the development of TLS within tumors. Finally, we explore how to take advantage of our knowledge on TLS B cells to develop new therapeutic tools.
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http://dx.doi.org/10.3389/fimmu.2015.00067DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4337382PMC
March 2015

Orchestration and Prognostic Significance of Immune Checkpoints in the Microenvironment of Primary and Metastatic Renal Cell Cancer.

Clin Cancer Res 2015 Jul 16;21(13):3031-40. Epub 2015 Feb 16.

INSERM, UMRS 1138, Cordeliers Research Center, Team 13 Cancer, Immune Control and Escape, Paris, France. Université Paris Descartes, Sorbonne Paris Cité, UMRS 1138, Centre de Recherche des Cordeliers, Paris, France. UPMC Univ Paris 06, Sorbonne Universités, UMRS 1138, Centre de Recherche des Cordeliers, Paris, France.

Purpose: Clear cell renal cell carcinoma (ccRCC) has shown durable responses to checkpoint blockade therapies. However, important gaps persist in the understanding of its immune microenvironment. This study aims to investigate the expression and prognostic significance of immune checkpoints in primary and metastatic ccRCC, in relation with mature dendritic cells (DC) and T-cell densities.

Experimental Design: We investigated the infiltration and the localization of CD8(+) T cells and mature DC, and the expression of immune checkpoints (PD-1, LAG-3, PD-L1, and PD-L2) in relation with prognosis, in 135 primary ccRCC tumors and 51 ccRCC lung metastases. RNA expression data for 496 primary ccRCC samples were used as confirmatory cohort.

Results: We identify two groups of tumors with extensive CD8(+) T-cell infiltrates. One group, characterized by high expression of immune checkpoints in the absence of fully functional mature DC, is associated with increased risk of disease progression. The second group, characterized by low expression of immune checkpoints and localization of mature DC in peritumoral immune aggregates (tertiary lymphoid structures), is associated with good prognosis.

Conclusions: The expression of the immune checkpoints and the localization of DC in the tumor microenvironment modulate the clinical impact of CD8(+) T cells in ccRCC.
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http://dx.doi.org/10.1158/1078-0432.CCR-14-2926DOI Listing
July 2015

Tertiary lymphoid structures in cancer and beyond.

Trends Immunol 2014 Nov 22;35(11):571-80. Epub 2014 Oct 22.

Laboratory 'Cancer, Immune Control and Escape', INSERM U1138, Cordeliers Research Centre, Paris, France; University Pierre and Marie Curie, UMRS 1138, Paris, France; University Paris Descartes, UMRS 1138, Paris, France. Electronic address:

Tertiary lymphoid structures (TLS) are ectopic lymphoid formations found in inflamed, infected, or tumoral tissues. They exhibit all the characteristics of structures in the lymph nodes (LN) associated with the generation of an adaptive immune response, including a T cell zone with mature dendritic cells (DC), a germinal center with follicular dendritic cells (FDC) and proliferating B cells, and high endothelial venules (HEV). In this review, we discuss evidence for the roles of TLS in chronic infection, autoimmunity, and cancer, and address the question of whether TLS present beneficial or deleterious effects in these contexts. We examine the relationship between TLS in tumors and patient prognosis, and discuss the potential role of TLS in building and/or maintaining local immune responses and how this understanding may guide therapeutic interventions.
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http://dx.doi.org/10.1016/j.it.2014.09.006DOI Listing
November 2014

The non-small cell lung cancer immune contexture. A major determinant of tumor characteristics and patient outcome.

Am J Respir Crit Care Med 2015 Feb;191(4):377-90

1 Division of Hematology and Medical Oncology and.

Solid tumors, beyond mere accumulation of cancer cells, form a complex ecosystem consisting of normal epithelial cells, fibroblasts, blood and lymphatic vessels, structural components, and infiltrating hematopoietic cells including myeloid and lymphoid elements that impact tumor growth, tumor spreading, and clinical outcome. The composition of the immune microenvironment is diverse, including various populations of T cells, B cells, dendritic cells, natural killer cells, myeloid-derived suppressor cells, neutrophils, or macrophages. The immune contexture describes the density, location, and organization of these immune cells within solid tumors. In lung cancer, which is the deadliest type of cancer, and particularly in non-small cell lung cancer, its most prevalent form, reports have described some of the interactions between the tumor and the host. These data, in addition to articles on various types of tumors, provide a greater understanding of the tumor-host microenvironment interaction and stimulate the development of prognostic and predictive biomarkers, the identification of novel target antigens for therapeutic intervention, and the implementation of tools for long-term management of patients with cancer.
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http://dx.doi.org/10.1164/rccm.201409-1671PPDOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5447326PMC
February 2015

Systemic inflammation, nutritional status and tumor immune microenvironment determine outcome of resected non-small cell lung cancer.

PLoS One 2014 19;9(9):e106914. Epub 2014 Sep 19.

University Paris Descartes; Paris, France; Deparments of Pathology, Paris Centre University Hospitals, AP-HP, Paris, France; INSERM U1138, Cancer and Immune Escape, Cordeliers Research Center, Paris, France; University Pierre and Marie Curie, UMRS U1138, Paris, France.

Background: Hypothesizing that nutritional status, systemic inflammation and tumoral immune microenvironment play a role as determinants of lung cancer evolution, the purpose of this study was to assess their respective impact on long-term survival in resected non-small cell lung cancers (NSCLC).

Methods And Findings: Clinical, pathological and laboratory data of 303 patients surgically treated for NSCLC were retrospectively analyzed. C-reactive protein (CRP) and prealbumin levels were recorded, and tumoral infiltration by CD8+ lymphocytes and mature dendritic cells was assessed. We observed that factors related to nutritional status, systemic inflammation and tumoral immune microenvironment were correlated; significant correlations were also found between these factors and other relevant clinical-pathological parameters. With respect to outcome, at univariate analysis we found statistically significant associations between survival and the following variables: Karnofsky index, American Society of Anesthesiologists (ASA) class, CRP levels, prealbumin concentrations, extent of resection, pathologic stage, pT and pN parameters, presence of vascular emboli, and tumoral infiltration by either CD8+ lymphocytes or mature dendritic cells and, among adenocarcinoma type, tumor grade (all p<0.05). In multivariate analysis, prealbumin levels (Relative Risk (RR): 0.34 [0.16-0.73], p = 0.0056), CD8+ cell count in tumor tissue (RR = 0.37 [0.16-0.83], p = 0.0162), and disease stage (RR 1.73 [1.03-2.89]; 2.99[1.07-8.37], p = 0.0374- stage I vs II vs III-IV) were independent prognostic markers. When taken together, parameters related to systemic inflammation, nutrition and tumoral immune microenvironment allowed robust prognostic discrimination; indeed patients with undetectable CRP, high (>285 mg/L) prealbumin levels and high (>96/mm2) CD8+ cell count had a 5-year survival rate of 80% [60.9-91.1] as compared to 18% [7.9-35.6] in patients with an opposite pattern of values. When stages I-II were considered alone, the prognostic significance of these factors was even more pronounced.

Conclusions: Our data show that nutrition, systemic inflammation and tumoral immune contexture are prognostic determinants that, taken together, may predict outcome.
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http://journals.plos.org/plosone/article?id=10.1371/journal.pone.0106914PLOS
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4169516PMC
June 2015

Shaping of an effective immune microenvironment to and by cancer cells.

Cancer Immunol Immunother 2014 Oct 12;63(10):991-7. Epub 2014 Aug 12.

Laboratory Cancer, Immune Control and Escape, UMRS 1138 INSERM, Cordeliers Research Center, 15, rue de l'école de Médecine, 75270, Paris Cedex 06, France.

A high density of intratumoral effector memory CD8+/Th1 T cells is associated with favorable prognosis in most cancers and may be induced or increased by immunotherapy. Efficient adaptive immune reactions are shaped in tumor adjacent tertiary lymphoid structures, which exhibit all characteristics of immunity generating lymphoid formations in reactive lymph nodes. Malignant tumor cells impact favorably or deleteriously their immune microenvironment if they bear genetic mutations that result in neo-antigens or by producing chemokines and cytokines that recruit lymphocytes and myeloid cells or increase inflammation and neo-angiogenesis. This intricate network of interactions results in control or escape of tumors, and its understanding will help define goals to monitor efficiency of immunotherapies.
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http://dx.doi.org/10.1007/s00262-014-1590-3DOI Listing
October 2014

Tertiary lymphoid structures in human lung cancers, a new driver of antitumor immune responses.

Oncoimmunology 2014;3:e28976. Epub 2014 May 29.

The Laboratory of Cancer and Immune escape; INSERM U1138; Cordeliers Research Center; Paris, France ; University Pierre and Marie Curie; UMRS1138; Paris, France ; University Paris Descartes; UMRS1138; Paris, France.

T-cell infiltrates are robust prognostic biomarkers in the majority of human cancers. However, the mechanisms that shape a protective T-cell response remain unclear. Our recent study implicates tertiary lymphoid structures in the shaping of an efficient and beneficial immune response in lung cancer patients.
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http://dx.doi.org/10.4161/onci.28976DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4106161PMC
May 2014

TLR7 promotes tumor progression, chemotherapy resistance, and poor clinical outcomes in non-small cell lung cancer.

Cancer Res 2014 Sep 29;74(18):5008-18. Epub 2014 Jul 29.

Institut National de la Santé et de la Recherche Médicale (INSERM), UMRS1138, Centre de Recherche des Cordeliers, Paris, France. Université Pierre et Marie Curie-Paris 6, UMRS1138, Paris, France. Université Paris Descartes, UMRS1138, Paris, France.

Toll-like receptors (TLR) recognize pathogen molecules and danger-associated signals that stimulate inflammatory processes. TLRs have been studied mainly in antigen-presenting cells, where they exert important immune regulatory functions, but they are also expressed by epithelial tumor cells, where they have been implicated in tumor progression. In this study, we demonstrate that the injection of TLR7 agonist in NOD/SCID mice, in C57BL/6 wild-type, and TLR7-deficient mice grafted with lung adenocarcinoma tumor cells leads to increased tumor progression and chemotherapeutic resistance. In patients with non-small cell lung cancer, expression analyses revealed that high TLR7 expression was strongly associated with resistance to neoadjuvant chemotherapy and poor clinical outcomes. Our findings delineate a crucial role for TLR7 in lung cancer physiopathology. Cancer Res; 74(18); 5008-18. ©2014 AACR.
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http://dx.doi.org/10.1158/0008-5472.CAN-13-2698DOI Listing
September 2014