Etude des voies de recrutement des cellules dendritiques dans une tumeur solide

Study of the recruitment pathways of dendritic cells in a solid tumor

• Auteur / Autrice : Delphine Boulet
• Direction : Sylvie Guerder
• Type : Thèse de doctorat
• Discipline(s) : Immunologie
• Date : Soutenance le 22/11/2017
• Etablissement(s) : Toulouse 3
• Ecole(s) doctorale(s) : École doctorale Biologie Santé Biotechnologies (Toulouse)
• Partenaire(s) de recherche : Laboratoire : Centre de Physiopathologie de Toulouse-Purpan (Toulouse ; 2002-2020)

Mots clés

Mots clés contrôlés

Réponse immunitaire

Tumeurs

Cellules dendritiques

Mots clés libres

Cellules dendritiques

Tumeur sous-cutanéetumeur sous-cutanée

Tumeur intra-dermique

Derme

Origine

Chimiokine

Dendritic cells

Subcutaneous tumor

Intra-dermal tumor

Dermis

Origin

Chemokine

Résumé

The concept of immunosurveillance suggests that the innate and adaptative immune system eliminate developing tumors. However, tumor development is associated with important modifications of the stroma which, by multiple mechanisms, restrain the immune response notably by affecting dendritic cells (DC) recruitment and functions. My thesis project aims at deciphering how the tumor environment alters the mechanisms and pathways of DC recruitment and impairs their functions. First, we determine if the site of tumor transplantation affects tumor immunogenicity. We show that tumors transplanted in the dermis (i.d.), an environment containing multiple DC subsets, induce a protective anti-tumoral immune response and tumor rejection. By contrast, the same tumor implanted in the subcutaneous tissue (s.c), mainly containing monocytes, is not rejected. Rejection of i.d. tumor is associated with a rapid (within 2 days) recruitment of DC within the tumor and rapid migration of DC towards tumor draining lymph nodes (dLN) where they present the tumor antigens (TAA) to CD4 and CD8 T lymphocyte. These events also occur for s.c. tumors but with a delayed kinetic. Thus the kinetic of DC mobilisation is decisive for tumor immunogenicity. Analysis of the DC subpopulations (TIDC) infiltrating the i.d. or s.c. tumors at 4 days (D4) and 8 days (D8) post-tumor transplantation, revealed that the different DC subpopulations are present at similar frequencies. Based on these findings, we proposed that i.d. tumor are rapidly infiltrated by dermal DC (DDC), whereas in s.c. tumor, the absence of inflammatory signals would limit DDC recruitment. In this latter case, DC would mainly come from local differentiation of blood-born precursors of DC (pre-cDC). Local differentiation of pre-cDC within the immunosuppressor tumor environment may affect their differentiation program and functions. We found that pre-cDC infiltrate i.d. and s.c. tumors starting at D4 and their frequency increases at D8. To determine the DC origin in tumors, we use CD11c-DTR-GFP mice in which CD11c+ cells express a fusion protein constituted by the diphtheria toxin receptor (DTR) and GFP. To track DDC trafficking within tumors, we injected anti-MHCII antibody before tumor implantation and analysed MHCII+ CD11c+ DDC infiltration in tumor by biphotonic microscopy. At D3 post-tumor transplantation, the DDC infiltration was higher in i.d. than s.c. tumors. To analyse the impact of this early DDC recruitment on anti-tumor immunity, we inhibit early recruitment of DC by injection of pertussis toxin (PTX), a chemokine receptor protein G inhibitor, during the three first days of tumor development. For i.d. tumors, PTX treatment induced 60% reduction of DC recruitment starting at D4. In s.c. tumor, while this effect was observable at D3 (60% reduction) and increased to 80% at D4. These results suggest that early recruitment of DDC to i.d. tumors may be be chemokine independent. PTX treatment, which inhibits DDC migration from tumors to dLN inhibits the early TAA presentation to CD4 and CD8 T cells but did not impaired i.d. tumor rejection. Collectively, these results suggest that a first wave of DDC may infiltrate i.d. tumor. The initial wave of DDC may rapidly activate the adaptive immune system and induce protective anti-tumoral immune response. For s.c. tumor, this first wave in delayed or limited. Tumor neoangioenesis would permit an input of pre-cDC which would differentiate locally into cDC1 and cDC2. To consolidate this model we are developing new protocols to efficiently inhibit early recruitment of DDC in i.d. tumor. Moreover, to determine the DC origin and pathways of DC recruitment in tumors, we exploit several experimental approaches to directly analyse DDC migration toward i.d. and s.c. tumor.

Le concept d’immunosurveillance postule que le système immunitaire inné et adaptatif élimine les tumeurs naissantes. Cependant, le développement de tumeurs est associé à des modifications importantes du stroma qui, par des mécanismes multiples, inhibent la réponse immunitaire en affectant notamment le recrutement et la fonction des cellules dendritiques (DC). Mon projet de thèse vise à préciser comment l’environnement tumoral affecte les mécanismes et les voies de recrutement des DC dans les tumeurs et leurs fonctions. Tout d’abord, nous avons déterminé si la composition en DC du tissu d’implantation affecte l’immunogénicité tumorale. Nous avons montré que les tumeurs implantées dans le derme (i.d.), un environnement riche en DC dermales (DDC), induisent une réponse anti-tumorale protectrice. En revanche, une même tumeur transplantée dans le tissu sous-cutané (s.c.), contenant principalement des monocytes, n’est pas rejetée. Le rejet des tumeurs i.d. est associé à un recrutement précoce et rapide des DC dans la tumeur (dès 2 jours post-injection) et une migration, dans les ganglions drainants (dLN), de DC qui présentent les antigènes tumoraux (TAA) aux lymphocytes T (LT) CD4+ et CD8+. Dans les tumeurs s.c. ces événements sont présents mais retardés. Ceci indique que la cinétique de mobilisation des DC est déterminante pour l’immunogénicité tumorale.