Skip to main content
Key Specialties
Cardiology Diabetology Endocrinology Gastroenterology Geriatrics and Gerontology Gynecology and Obstetrics Hematology Infectious Disease Internal Medicine Nephrology Neurology Oncology Pediatrics Respiratory Medicine Rheumatology Surgery
Congress Coverage
2024 ACC Congress 2023 ESMO Congress 2023 EASD Congress 2023 ERS Congress 2023 ESC Congress
Clinical Collections
Advances in Alzheimer’s

At a glance: The ONWARDS insulin icodec trials

A round-up of the ONWARDS phase 3 clinical trials evaluating the novel weekly insulin icodec in people with diabetes.
ADVANCED SEARCH
Log in
Top
Cancer Immunology, Immunotherapy
Published in: Cancer Immunology, Immunotherapy 8/2019

01-08-2019 | Melanoma | Original Article

Reduced CTL motility and activity in avascular tumor areas

Authors: Yoav Manaster, Zohar Shipony, Anat Hutzler, Masha Kolesnikov, Camila Avivi, Bruria Shalmon, Iris Barshack, Michal J. Besser, Tali Feferman, Guy Shakhar

Published in: Cancer Immunology, Immunotherapy | Issue 8/2019

Login to get access

Abstract

Patchy infiltration of tumors by cytotoxic T cells (CTLs) predicts poorer prognosis for cancer patients. The factors limiting intratumoral CTL dissemination, though, are poorly understood. To study CTL dissemination in tumors, we histologically examined human melanoma samples and used mice to image B16-OVA tumors infiltrated by OT-I CTLs using intravital two-photon microscopy. In patients, most CTLs concentrated around peripheral blood vessels, especially in poorly infiltrated tumors. In mice, OT-I CTLs had to cluster around tumor cells to efficiently kill them in a contact-and perforin-dependent manner and cytotoxicity was strictly antigen-specific. OT-I CTLs as well as non-specific CTLs concentrated around peripheral vessels, and cleared the tumor cells around them. This was also the case when CTLs were injected directly into the tumors. CTLs crawled rapidly only in areas within 50 µm of flowing blood vessels and transient occlusion of vessels immediately, though reversibly, stopped their migration. In vitro, oxygen depletion and blockade of oxidative phosphorylation also reduced CTL motility. Taken together, these results suggest that hypoxia limits CTL migration away from blood vessels, providing immune-privileged niches for tumor cells to survive. Normalizing intratumoral vasculature may thus synergize with tumor immunotherapy.
Appendix
Available only for authorised users
Literature
1.
Vesely MD, Kershaw MH, Schreiber RD, Smyth MJ (2011) Natural innate and adaptive immunity to cancer. Annu Rev Immunol 29:235–271CrossRefPubMed
2.
Coulie PG, Van den Eynde BJ, van der Bruggen P, Boon T (2014) Tumour antigens recognized by T lymphocytes: at the core of cancer immunotherapy. Nat Rev Cancer 14:135–146CrossRefPubMed
3.
Angell H, Galon J (2013) From the immune contexture to the immunoscore: the role of prognostic and predictive immune markers in cancer. Curr Opin Immunol 25:261–267CrossRefPubMed
4.
Aerts JG, Hegmans JP (2013) Tumor-specific cytotoxic T cells are crucial for efficacy of immunomodulatory antibodies in patients with lung cancer. Cancer Res 73:2381–2388CrossRefPubMed
5.
Herbst RS, Soria J-C, Kowanetz M, Fine GD, Hamid O, Gordon MS et al (2014) Predictive correlates of response to the anti-PD-L1 antibody MPDL3280A in cancer patients. Nature 515:563–567CrossRefPubMedPubMedCentral
6.
7.
8.
Voskoboinik I, Whisstock JC, Trapani JA (2015) Perforin and granzymes: function, dysfunction and human pathology. Nat Rev Immunol 15:388–400CrossRefPubMed
9.
Zhang B, Karrison T, Rowley DA, Schreiber H (2008) IFN-gamma- and TNF-dependent bystander eradication of antigen-loss variants in established mouse cancers. J Clin Investig 118:1398–1404CrossRefPubMedPubMedCentral
10.
Schietinger A, Arina A, Liu RB, Wells S, Huang J, Engels B et al (2013) Longitudinal confocal microscopy imaging of solid tumor destruction following adoptive T cell transfer. Oncoimmunology. 2:e26677CrossRefPubMedPubMedCentral
11.
Galon J, Costes A, Sanchez-Cabo F, Kirilovsky A, Mlecnik B, Lagorce-Pagès C et al (2006) Type, density, and location of immune cells within human colorectal tumors predict clinical outcome. Science 313:1960–1964CrossRefPubMed
12.
Erdag G, Schaefer JT, Smolkin ME, Deacon DH, Shea SM, Dengel LT et al (2012) Immunotype and immunohistologic characteristics of tumor-infiltrating immune cells are associated with clinical outcome in metastatic melanoma. Cancer Res 72:1070–1080CrossRefPubMedPubMedCentral
13.
Kiss J, Timar J, Somlai B, Gilde K, Fejos Z, Gaudi I et al (2007) Association of microvessel density with infiltrating cells in human cutaneous malignant melanoma. Pathol Oncol Res 13:21–31CrossRefPubMed
14.
Brizel DM, Scully SP, Harrelson JM, Brizel M, Harrelson M, Layfleld J et al (1996) Tumor oxygenation predicts for the likelihood of distant metastases in human soft tissue sarcoma. Cancer Res 56:941–943PubMed
15.
Höckel M, Schlenger K, Aral B, Mitze M, Schaffer U, Vaupel P (1996) Association between tumor hypoxia and malignant progression in advanced cancer of the uterine cervix. Cancer Res 56:4509–4515PubMed
16.
Huang JH, Cardenas-Navia LI, Caldwell CC, Plumb TJ, Radu CG, Rocha PN et al (2007) Requirements for T lymphocyte migration in explanted lymph nodes. J Immunol 178:7747–7755CrossRefPubMed
17.
Chang CH, Qiu J, O’Sullivan D, Buck MD, Noguchi T, Curtis JD et al (2015) Metabolic competition in the tumor microenvironment is a driver of cancer progression. Cell 162:1229–1241CrossRefPubMedPubMedCentral
18.
Hatfield SM, Kjaergaard J, Lukashev D, Schreiber TH, Belikoff B, Abbott R et al (2015) Immunological mechanisms of the antitumor effects of supplemental oxygenation. Sci Transl Med 7:1–12CrossRef
19.
Mempel TR, Bauer CA (2009) Intravital imaging of CD8+ T cell function in cancer. Clin Exp Metastasis 26:311–327CrossRefPubMed
20.
Broz ML, Binnewies M, Boldajipour B, Nelson AE, Pollack JL, Erle DJ et al (2014) Dissecting the tumor myeloid compartment reveals rare activating antigen-presenting cells critical for t cell immunity. Cancer Cell 26:638–652CrossRefPubMedPubMedCentral
21.
Breart B, Lemaitre F, Celli S, Bousso P (2008) Two-photon imaging of intratumoral CD8+ T cell cytotoxic activity during adoptive T cell therapy in mice. J Clin Investig 118:1390–1397CrossRefPubMedPubMedCentral
22.
Boissonnas A, Fetler L, Zeelenberg IS, Hugues S, Amigorena S (2007) In vivo imaging of cytotoxic T cell infiltration and elimination of a solid tumor. J Exp Med 204:345–356CrossRefPubMedPubMedCentral
23.
Ruifrok AC, Johnston DA (2001) Quantification of histochemical staining by color deconvolution. Anal Quant Cytol Histol 23:291–299PubMed
24.
Zangi L, Klionsky YZ, Yarimi L, Bachar-Lustig E, Eidelstein Y, Shezen E et al (2012) Deletion of cognate CD8 T cells by immature dendritic cells: a novel role for perforin, granzyme A, TREM-1, and TLR7. Blood 120:1647–1657CrossRefPubMed
25.
Crissman JD, Hatfield J, Schaldenbrand M, Sloane BF, Honn KV (1985) Arrest and extravasation of B16 amelanotic melanoma in murine lungs. A light and electron microscopic study. Lab Investig 53:470–478PubMed
26.
Steff AM, Fortin M, Arguin C, Hugo P (2001) Detection of a decrease in green fluorescent protein fluorescence for the monitoring of cell death: an assay amenable to high-throughput screening technologies. Cytometry 45:237–243CrossRefPubMed
27.
Heitjan DF, Manni A, Santen RJ (1993) Statistical analysis of in vivo tumor growth experiments. Cancer Res 53:6042–6050PubMed
28.
Perez OD, Mitchell D, Jager GC, Nolan GP (2004) LFA-1 signaling through p44/42 is coupled to perforin degranulation in CD56+ CD8+ natural killer cells. Blood 104:1083–1093CrossRefPubMed
29.
Dustin ML, Bromley SK, Kan ZY, Peterson DA, Unanue ER (1997) Antigen receptor engagement delivers a stop signal to migrating T lymphocytes. Proc Natl Acad Sci USA 94:3909–3913CrossRefPubMedPubMedCentral
30.
Caramalho I, Faroudi M, Padovan E, Muller S, Valitutti S, Müller S (2009) Visualizing CTL/melanoma cell interactions: multiple hits must be delivered for tumour cell annihilation. J Cell Mol Med 13:3834–3846CrossRefPubMed
31.
Böhm W, Thoma S, Leithäuser F, Möller P, Schirmbeck R, Reimann J (1998) T cell-mediated, IFN-gamma-facilitated rejection of murine B16 melanomas. J Immunol 161:897–908PubMed
32.
Khazen R, Puissegur M, Muller S, Valitutti S (2015) Dissecting early mechanisms of melanoma cell resistance to cytotoxic T lymphocyte attack (TUM10P.1025). J Immunol 194:211.6
33.
Ivanov VN, Bhoumik A, Ronai Z (2003) Death receptors and melanoma resistance to apoptosis. Oncogene 22:3152–3161CrossRefPubMed
34.
Engelhardt JJ, Boldajipour B, Beemiller P, Pandurangi P, Sorensen C, Werb Z et al (2012) Marginating dendritic cells of the tumor microenvironment cross-present tumor antigens and stably engage tumor-specific T cells. Cancer Cell 21:402–417CrossRefPubMedPubMedCentral
35.
Pentcheva-Hoang T, Simpson TR, Montalvo-Ortiz W, Allison JP (2014) Cytotoxic T lymphocyte antigen-4 (CTLA-4) blockade enhances anti-tumor immunity by stimulating melanoma-specific T cell motility. Cancer Immunol Res 4:970–980CrossRef
36.
Halle S, Keyser KA, Stahl FR, Busche A, Marquardt A, Zheng X et al (2016) In vivo killing capacity of cytotoxic T cells is limited and involves dynamic interactions and T cell cooperativity. Immunity 44:233–245CrossRefPubMedPubMedCentral
37.
Clever D, Roychoudhuri R, Constantinides MG, Askenase MH, Sukumar M, Klebanoff CA et al (2016) Oxygen sensing by T cells establishes an immunologically tolerant metastatic niche. Cell 166(1117–1131):e14
38.
Vogel S, Wottawa M, Farhat K, Zieseniss A, Schnelle M, Le-Huu S et al (2010) Prolyl hydroxylase domain (PHD) 2 affects cell migration and F-actin formation via RhoA/Rho-associated kinase-dependent cofilin phosphorylation. J Biol Chem 285:33756–33763CrossRefPubMedPubMedCentral
39.
Luo W, Lin B, Wang Y, Zhong J, O’Meally R, Cole RN et al (2014) PHD3-mediated prolyl hydroxylation of nonmuscle actin impairs polymerization and cell motility. Mol Biol Cell 25:2788–2796CrossRefPubMedPubMedCentral
40.
Ho PC, Bihuniak JD, MacIntyre AN, Staron M, Liu X, Amezquita R et al (2015) Phosphoenolpyruvate is a metabolic checkpoint of anti-tumor T cell responses. Cell 162:1217–1228CrossRefPubMedPubMedCentral
41.
Hatfield SM, Kjaergaard J, Lukashev D, Belikoff B, Schreiber TH, Sethumadhavan S et al (2014) Systemic oxygenation weakens the hypoxia and hypoxia inducible factor 1α-dependent and extracellular adenosine-mediated tumor protection. J Mol Med (Berl) 92:1283–1292CrossRef
42.
Fischer K, Hoffmann P, Voelkl S, Meidenbauer N, Ammer J, Edinger M et al (2015) Inhibitory effect of tumor cell-derived lactic acid on human T cells. Blood 109:3812–3820CrossRef
43.
Van Overmeire E, Laoui D, Keirsse J, Van Ginderachter JA, Sarukhan A (2014) Mechanisms driving macrophage diversity and specialization in distinct tumor microenvironments and parallelisms with other tissues. Front Immunol 5:1–16
44.
Casazza A, Laoui D, Wenes M, Rizzolio S, Bassani N, Mambretti M et al (2013) Impeding macrophage entry into hypoxic tumor areas by Sema3A/Nrp1 signaling blockade inhibits angiogenesis and restores antitumor immunity. Cancer Cell 24:695–709CrossRefPubMed
45.
Movahedi K, Laoui D, Gysemans C, Baeten M, Stangé G, Van Den Bossche J et al (2010) Different tumor microenvironments contain functionally distinct subsets of macrophages derived from Ly6C(high) monocytes. Cancer Res 70:5728–5739CrossRefPubMed
46.
Venetz D, Ponzoni M, Schiraldi M, Ferreri AJM, Bertoni F, Doglioni C et al (2010) Perivascular expression of CXCL9 and CXCL12 in primary central nervous system lymphoma: T-cell infiltration and positioning of malignant B cells. Int J Cancer 127:2300–2312CrossRefPubMed
47.
Hagemann IS, Lal P, Feldman MD, Benencia F (2014) Tumor endothelium FasL establishes a selective immune barrier promoting tolerance in tumors. Nat Med 20:607–615CrossRefPubMedPubMedCentral
48.
Jain RK (2005) Normalization of tumor vasculature: an emerging concept in antiangiogenic therapy. Science (80-) 307:58–62CrossRef
49.
Sorensen AG, Emblem KE, Polaskova P, Jennings D, Kim H, Ancukiewicz M et al (2012) Increased survival of glioblastoma patients who respond to antiangiogenic therapy with elevated blood perfusion. Cancer Res 72:402–407CrossRefPubMed
50.
Li X, Kostareli E, Suffner J, Garbi N, Hämmerling GJ (2010) Efficient Treg depletion induces T-cell infiltration and rejection of large tumors. Eur J Immunol 40:3325–3335CrossRefPubMed
51.
Klug F, Prakash H, Huber PE, Seibel T, Bender N, Halama N et al (2013) Low-dose irradiation programs macrophage differentiation to an iNOS+/M1 phenotype that orchestrates effective T cell immunotherapy. Cancer Cell 24:589–602CrossRefPubMed
52.
Manning EA, Ullman JG, Leatherman JM, Asquith JM, Hansen TR, Armstrong TD et al (2007) A vascular endothelial growth factor receptor-2 inhibitor enhances antitumor immunity through an immune-based mechanism. Clin Cancer Res 13:3951–3959CrossRefPubMed
53.
Hamzah J, Jugold M, Kiessling F, Rigby P, Manzur M, Marti HH et al (2008) Vascular normalization in Rgs5-deficient tumours promotes immune destruction. Nature 453:410–414CrossRefPubMed
54.
Huang Y, Yuan J, Righi E, Kamoun WS, Ancukiewicz M, Nezivar J et al (2012) Vascular normalizing doses of antiangiogenic treatment reprogram the immunosuppressive tumor microenvironment and enhance immunotherapy. Proc Natl Acad Sci USA 109:17561–17566CrossRefPubMedPubMedCentral
55.
Caldwell CC, Kojima H, Lukashev D, Armstrong J, Farber M, Apasov SG et al (2001) Differential effects of physiologically relevant hypoxic conditions on T lymphocyte development and effector functions. J Immunol 167:6140–6149CrossRefPubMed
56.
Nakagawa Y, Negishi Y, Shimizu M, Takahashi M, Ichikawa M, Takahashi H (2015) Effects of extracellular pH and hypoxia on the function and development of antigen-specific cytotoxic T lymphocytes. Immunol Lett 167:72–86CrossRefPubMed
57.
Gropper Y, Feferman T, Shalit T, Salame TM, Porat Z, Shakhar G (2017) Culturing CTLs under hypoxic conditions enhances their cytolysis and improves their anti-tumor function. Cell Rep 20:2547–2555CrossRefPubMed
Metadata
Title
Reduced CTL motility and activity in avascular tumor areas
Authors
Yoav Manaster
Zohar Shipony
Anat Hutzler
Masha Kolesnikov
Camila Avivi
Bruria Shalmon
Iris Barshack
Michal J. Besser
Tali Feferman
Guy Shakhar
Publication date
01-08-2019
Publisher
Springer Berlin Heidelberg
Keywords
Melanoma
Melanoma
Published in
Cancer Immunology, Immunotherapy / Issue 8/2019
Print ISSN: 0340-7004
Electronic ISSN: 1432-0851
DOI
https://doi.org/10.1007/s00262-019-02361-5

Other articles of this Issue 8/2019

Cancer Immunology, Immunotherapy 8/2019 Go to the issue

Meeting Report

4th Symposium on Advances in Cancer Immunology and Immunotherapy, November 29–December 1, 2018, Athens, Greece

Original Article

Histopathology-based immunoscore predicts recurrence for intrahepatic cholangiocarcinoma after hepatectomy

Original Article

High levels of tissue inhibitor of metalloproteinase-1 (TIMP-1) in the serum are associated with poor prognosis in HPV-negative squamous cell oropharyngeal cancer

Original Article

Preclinical assessment of transiently TCR redirected T cells for solid tumour immunotherapy

Original Article

Serum PCSK9 levels at the second nivolumab cycle predict overall survival in elderly patients with NSCLC: a pilot study

Original Article

CTLA-4 antibody ipilimumab negatively affects CD4+ T-cell responses in vitro
Webinar | 19-02-2024 | 17:30 (CET)

Keynote webinar | Spotlight on antibody–drug conjugates in cancer

Watch now

Antibody–drug conjugates (ADCs) are novel agents that have shown promise across multiple tumor types. Explore the current landscape of ADCs in breast and lung cancer with our experts, and gain insights into the mechanism of action, key clinical trials data, existing challenges, and future directions.

Dr. Véronique Diéras
Prof. Fabrice Barlesi
Developed by: Springer Medicine
Image Credits