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01-01-2016 | Original Article

Metastatic spread in patients with non-small cell lung cancer is associated with a reduced density of tumor-infiltrating T cells

Authors: Philipp Müller, Sacha I. Rothschild, Walter Arnold, Petra Hirschmann, Lukas Horvath, Lukas Bubendorf, Spasenija Savic, Alfred Zippelius

Published in: Cancer Immunology, Immunotherapy | Issue 1/2016

Abstract

Tumor-infiltrating lymphocytes play an important role in cell-mediated immune destruction of cancer cells and tumor growth control. We investigated the heterogeneity of immune cell infiltrates between primary non-small cell lung carcinomas (NSCLC) and corresponding metastases. Formalin-fixed, paraffin-embedded primary tumors and corresponding metastases from 34 NSCLC patients were analyzed by immunohistochemistry for CD4, CD8, CD11c, CD68, CD163 and PD-L1. The percentage of positively stained cells within the stroma and tumor cell clusters was recorded and compared between primary tumors and metastases. We found significantly fewer CD4⁺ and CD8⁺ T cells within tumor cell clusters as compared with the stromal compartment, both in primary tumors and corresponding metastases. CD8⁺ T cell counts were significantly lower in metastatic lesions than in the corresponding primary tumors, both in the stroma and the tumor cell islets. Of note, the CD8/CD4 ratio was significantly reduced in metastatic lesions compared with the corresponding primary tumors in tumor cell islets, but not in the stroma. We noted significantly fewer CD11c⁺ cells and CD68⁺ as well as CD163⁺ macrophages in tumor cell islets compared with the tumor stroma, but no difference between primary and metastatic lesions. Furthermore, the CD8/CD68 ratio was higher in primary tumors than in the corresponding metastases. We demonstrate a differential pattern of immune cell infiltration in matched primary and metastatic NSCLC lesions, with a significantly lower density of CD8⁺ T cells in metastatic lesions compared with the primary tumors. The lower CD8/CD4 and CD8/CD68 ratios observed in metastases indicate a rather tolerogenic and tumor-promoting microenvironment at the metastatic site.

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Khong HT, Restifo NP (2002) Natural selection of tumor variants in the generation of “tumor escape” phenotypes. Nat Immunol 3:999–1005. doi:10.1038/ni1102-999 PubMedPubMedCentralCrossRef

Lin A, Schildknecht A, Nguyen LT, Ohashi PS (2010) Dendritic cells integrate signals from the tumor microenvironment to modulate immunity and tumor growth. Immunol Lett 127:77–84. doi:10.1016/j.imlet.2009.09.003 PubMedCrossRef

Chen JJ, Lin YC, Yao PL, Yuan A, Chen HY, Shun CT, Tsai MF, Chen CH, Yang PC (2005) Tumor-associated macrophages: the double-edged sword in cancer progression. J Clin Oncol 23:953–964. doi:10.1200/JCO.2005.12.172 PubMedCrossRef

Loose D, Van de Wiele C (2009) The immune system and cancer. Cancer Biother Radiopharm 24:369–376. doi:10.1089/cbr.2008.0593 PubMedCrossRef

Audia S, Nicolas A, Cathelin D et al (2007) Increase of CD4⁺ CD25⁺ regulatory T cells in the peripheral blood of patients with metastatic carcinoma: a Phase I clinical trial using cyclophosphamide and immunotherapy to eliminate CD4⁺ CD25⁺ T lymphocytes. Clin Exp Immunol 150:523–530. doi:10.1111/j.1365-2249.2007.03521.x PubMedPubMedCentralCrossRef

Theoharides TC, Conti P (2004) Mast cells: the Jekyll and Hyde of tumor growth. Trends Immunol 25:235–241. doi:10.1016/j.it.2004.02.013 PubMedCrossRef

Gavert N, Ben-Ze’ev A (2008) Epithelial-mesenchymal transition and the invasive potential of tumors. Trends Mol Med 14:199–209. doi:10.1016/j.molmed.2008.03.004 PubMedCrossRef

Solinas G, Germano G, Mantovani A, Allavena P (2009) Tumor-associated macrophages (TAM) as major players of the cancer-related inflammation. J Leukoc Biol 86:1065–1073. doi:10.1189/jlb.0609385 PubMedCrossRef

Galon J, Costes A, Sanchez-Cabo F et al (2006) Type, density, and location of immune cells within human colorectal tumors predict clinical outcome. Science 313:1960–1964. doi:10.1126/science.1129139 PubMedCrossRef

10.

Pagès F, Berger A, Camus M et al (2005) Effector memory T cells, early metastasis, and survival in colorectal cancer. N Engl J Med 353:2654–2666. doi:10.1056/NEJMoa051424 PubMedCrossRef

11.

Bindea G, Mlecnik B, Tosolini M et al (2013) Spatiotemporal dynamics of intratumoral immune cells reveal the immune landscape in human cancer. Immunity 39:782–795. doi:10.1016/j.immuni.2013.10.003 PubMedCrossRef

12.

Al-Shibli KI, Donnem T, Al-Saad S, Persson M, Bremnes RM, Busund LT (2008) Prognostic effect of epithelial and stromal lymphocyte infiltration in non-small cell lung cancer. Clin Cancer Res 14:5220–5227. doi:10.1158/1078-0432.CCR-08-0133 PubMedCrossRef

13.

Dieu-Nosjean MC, Antoine M, Danel C et al (2008) Long-term survival for patients with non-small-cell lung cancer with intratumoral lymphoid structures. J Clin Oncol 26:4410–4417. doi:10.1200/JCO.2007.15.0284 PubMedCrossRef

14.

Hiraoka K, Miyamoto M, Cho Y et al (2006) Concurrent infiltration by CD8⁺ T cells and CD4⁺ T cells is a favourable prognostic factor in non-small-cell lung carcinoma. Br J Cancer 94:275–280. doi:10.1038/sj.bjc.6602934 PubMedPubMedCentralCrossRef

15.

Zhuang X, Xia X, Wang C, Gao F, Shan N, Zhang L, Zhang L (2010) A high number of CD8⁺ T cells infiltrated in NSCLC tissues is associated with a favorable prognosis. Appl Immunohistochem Mol Morphol 18:24–28. doi:10.1097/PAI.0b013e3181b6a741 PubMedCrossRef

16.

Takanami I, Takeuchi K, Giga M (2001) The prognostic value of natural killer cell infiltration in resected pulmonary adenocarcinoma. J Thorac Cardiovasc Surg 121:1058–1063. doi:10.1067/mtc.2001.113026 PubMedCrossRef

17.

Villegas FR, Coca S, Villarrubia VG, Jiménez R, Chillón MJ, Jareño J, Zuil M, Callol L (2002) Prognostic significance of tumor infiltrating natural killer cells subset CD57 in patients with squamous cell lung cancer. Lung Cancer 35:23–28. doi:10.1016/S0169-5002(01)00292-6 PubMedCrossRef

18.

Hasegawa T, Suzuki H, Yamaura T et al (2014) Prognostic value of peripheral and local forkhead box P3 regulatory T cells in patients with non-small-cell lung cancer. Mol Clin Oncol 2:685–694. doi:10.3892/mco.2014.299 PubMedPubMedCentral

19.

Hodi FS, O’Day SJ, McDermott DF et al (2010) Improved survival with ipilimumab in patients with metastatic melanoma. N Engl J Med 363:711–723. doi:10.1056/NEJMoa1003466 PubMedPubMedCentralCrossRef

20.

Robert C, Thomas L, Bondarenko I et al (2011) Ipilimumab plus dacarbazine for previously untreated metastatic melanoma. N Engl J Med 364:2517–2526. doi:10.1056/NEJMoa1104621 PubMedCrossRef

21.

Robert C, Long GV, Brady B et al (2014) Nivolumab in previously untreated melanoma without BRAF mutation. N Engl J Med 372:320–330. doi:10.1056/NEJMoa1412082 PubMedCrossRef

22.

Brahmer J, Reckamp KL, Baas P et al (2015) Nivolumab versus docetaxel in advanced squamous-cell non-small-cell lung cancer. N Engl J Med 373:123–135. doi:10.1056/NEJMoa1504627 PubMedCrossRef

23.

Topalian SL, Hodi FS, Brahmer JR et al (2012) Safety, activity, and immune correlates of anti-PD-1 antibody in cancer. N Engl J Med 366:2443–2454. doi:10.1056/NEJMoa1200690 PubMedPubMedCentralCrossRef

24.

Brahmer JR, Tykodi SS, Chow LQ et al (2012) Safety and activity of anti-PD-L1 antibody in patients with advanced cancer. N Engl J Med 366:2455–2465. doi:10.1056/NEJMoa1200694 PubMedPubMedCentralCrossRef

25.

Siegel R, Naishadham D, Jemal A (2013) Cancer statistics, 2013. CA Cancer J Clin 63:11–30. doi:10.3322/caac.21166 PubMedCrossRef

26.

Fidler IJ, Hart IR (1982) Biological diversity in metastatic neoplasms: origins and implications. Science 217:998–1003. doi:10.1126/science.7112116 PubMedCrossRef

27.

Talmadge JE, Wolman SR, Fidler IJ (1982) Evidence for the clonal origin of spontaneous metastases. Science 217:361–363. doi:10.1126/science.6953592 PubMedCrossRef

28.

Dunn GP, Old LJ, Schreiber RD (2004) The immunobiology of cancer immunosurveillance and immunoediting. Immunity 21:137–148. doi:10.1016/j.immuni.2004.07.017 PubMedCrossRef

29.

Schreiber RD, Old LJ, Smyth MJ (2011) Cancer immunoediting: integrating immunity’s roles in cancer suppression and promotion. Science 331:1565–1570. doi:10.1126/science.1203486 PubMedCrossRef

30.

Smyth MJ, Dunn GP, Schreiber RD (2006) Cancer immunosurveillance and immunoediting: the roles of immunity in suppressing tumor development and shaping tumor immunogenicity. Adv Immunol 90:1–50. doi:10.1016/S0065-2776(06)90001-7 PubMedCrossRef

31.

Obeid M, Tesniere A, Ghiringhelli F et al (2007) Calreticulin exposure dictates the immunogenicity of cancer cell death. Nat Med 13:54–61. doi:10.1038/nm1523 PubMedCrossRef

32.

Apetoh L, Ghiringhelli F, Tesniere A et al (2007) Toll-like receptor 4-dependent contribution of the immune system to anticancer chemotherapy and radiotherapy. Nat Med 13:1050–1059. doi:10.1038/nm1622 PubMedCrossRef

33.

Ghiringhelli F, Apetoh L, Tesniere A et al (2009) Activation of the NLRP3 inflammasome in dendritic cells induces IL-1beta-dependent adaptive immunity against tumors. Nat Med 15:1170–1178. doi:10.1038/nm.2028 PubMedCrossRef

34.

Müller P, Martin K, Theurich S et al (2014) Microtubule-depolymerizing agents used in antibody-drug conjugates induce antitumor immunity by stimulation of dendritic cells. Cancer Immunol Res 2:741–755. doi:10.1158/2326-6066.CIR-13-0198 PubMedCrossRef

35.

Martin K, Müller P, Schreiner J, Prince SS, Lardinois D, Heinzelmann-Schwarz VA, Thommen DS, Zippelius A (2014) The microtubule-depolymerizing agent ansamitocin P3 programs dendritic cells toward enhanced anti-tumor immunity. Cancer Immunol Immunother 63:925–938. doi:10.1007/s00262-014-1565-4 PubMedCrossRef

36.

Zhang W, Guo N, Yu C, Wang H, Zhang Y, Xia H, Yu J, Lu J (2012) Differential expression of ERCC-1 in the primary tumors and metastatic lymph nodes of patients with non-small cell lung cancer adenocarcinoma. Tumour Biol 33:2209–2216. doi:10.1007/s13277-012-0482-4 PubMedPubMedCentralCrossRef

37.

Shimizu K, Yukawa T, Hirami Y, Okita R, Saisho S, Maeda A, Yasuda K, Nakata M (2013) Heterogeneity of the EGFR mutation status between the primary tumor and metastatic lymph node and the sensitivity to EGFR tyrosine kinase inhibitor in non-small cell lung cancer. Target Oncol 8:237–242. doi:10.1007/s11523-012-0241-x PubMedCrossRef

38.

Tanoue LT, Detterbeck FC (2009) New TNM classification for non-small-cell lung cancer. Expert Rev Anticancer Ther 9:413–423. doi:10.1586/era.09.11 PubMedCrossRef

39.

Ries CH, Cannarile MA, Hoves S et al (2014) Targeting tumor-associated macrophages with anti-CSF-1R antibody reveals a strategy for cancer therapy. Cancer Cell 25:846–859. doi:10.1016/j.ccr.2014.05.016 PubMedCrossRef

40.

Ngiow SF, von Scheidt B, Akiba H, Yagita H, Teng MW, Smyth MJ (2011) Anti-TIM3 antibody promotes T cell IFN-γ-mediated antitumor immunity and suppresses established tumors. Cancer Res 71:3540–3551. doi:10.1158/0008-5472.CAN-11-0096 PubMedCrossRef

41.

Quatromoni JG, Eruslanov E (2012) Tumor-associated macrophages: function, phenotype, and link to prognosis in human lung cancer. Am J Transl Res 4:376–389PubMedPubMedCentral

42.

Franklin RA, Liao W, Sarkar A, Kim MV, Bivona MR, Liu K, Pamer EG, Li MO (2014) The cellular and molecular origin of tumor-associated macrophages. Science 344:921–925. doi:10.1126/science.1252510 PubMedPubMedCentralCrossRef

43.

Zitvogel L, Kepp O, Kroemer G (2011) Immune parameters affecting the efficacy of chemotherapeutic regimens. Nat Rev Clin Oncol 8:151–160. doi:10.1038/nrclinonc.2010.223 PubMedCrossRef

44.

Pagès F, Galon J, Dieu-Nosjean MC, Tartour E, Sautès-Fridman C, Fridman WH (2010) Immune infiltration in human tumors: a prognostic factor that should not be ignored. Oncogene 29:1093–1102. doi:10.1038/onc.2009.416 PubMedCrossRef

45.

Zitvogel L, Kepp O, Aymeric L, Ma Y, Locher C, Delahaye NF, André F, Kroemer G (2010) Integration of host-related signatures with cancer cell-derived predictors for the optimal management of anticancer chemotherapy. Cancer Res 70:9538–9543. doi:10.1158/0008-5472.CAN-10-1003 PubMedCrossRef

46.

Germain C, Gnjatic S, Tamzalit F et al (2014) Presence of B cells in tertiary lymphoid structures is associated with a protective immunity in patients with lung cancer. Am J Respir Crit Care Med 189:832–844. doi:10.1164/rccm.201309-1611OC PubMedCrossRef

47.

Halama N, Michel S, Kloor M et al (2011) Localization and density of immune cells in the invasive margin of human colorectal cancer liver metastases are prognostic for response to chemotherapy. Cancer Res 71:5670–5677. doi:10.1158/0008-5472.CAN-11-0268 PubMedCrossRef

48.

Ladoire S, Mignot G, Dabakuyo S et al (2011) In situ immune response after neoadjuvant chemotherapy for breast cancer predicts survival. J Pathol 224:389–400. doi:10.1002/path.2866 PubMedCrossRef

49.

Galon J, Fridman WH, Pagès F (2007) The adaptive immunologic microenvironment in colorectal cancer: a novel perspective. Cancer Res 67:1883–1886. doi:10.1158/0008-5472.CAN-06-4806 PubMedCrossRef

50.

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 25:261–267. doi:10.1016/j.coi.2013.03.004 PubMedCrossRef

51.

Restifo NP (2013) A “big data” view of the tumor “immunome”. Immunity 39:631–632. doi:10.1016/j.immuni.2013.10.002 PubMedPubMedCentralCrossRef

52.

Beyer I, van Rensburg R, Lieber A (2013) Overcoming physical barriers in cancer therapy. Tissue Barriers 1:e23647. doi:10.4161/tisb.23647 PubMedPubMedCentralCrossRef

53.

Wang A, Wang HY, Liu Y, Zhao MC, Zhang HJ, Lu ZY, Fang YC, Chen XF, Liu GT (2015) The prognostic value of PD-L1 expression for non-small cell lung cancer patients: a meta-analysis. Eur J Surg Oncol 41:450–456. doi:10.1016/j.ejso.2015.01.020 PubMedCrossRef

54.

Tumeh PC, Harview CL, Yearley JH et al (2014) PD-1 blockade induces responses by inhibiting adaptive immune resistance. Nature 515:568–571. doi:10.1038/nature13954 PubMedPubMedCentralCrossRef

55.

Harlin H, Meng Y, Peterson AC, Zha Y, Tretiakova M, Slingluff C, McKee M, Gajewski TF (2009) Chemokine expression in melanoma metastases associated with CD8⁺ T-cell recruitment. Cancer Res 69:3077–3085. doi:10.1158/0008-5472.CAN-08-2281 PubMedCrossRef

56.

Bald T, Landsberg J, Lopez-Ramos D et al (2014) Immune cell-poor melanomas benefit from PD-1 blockade after targeted type I IFN activation. Cancer Discov 4:674–687. doi:10.1158/2159-8290.CD-13-0458 PubMedCrossRef

57.

Woo EY, Yeh H, Chu CS, Schlienger K, Carroll RG, Riley JL, Kaiser LR, June CH (2002) Cutting edge: regulatory T cells from lung cancer patients directly inhibit autologous T cell proliferation. J Immunol 168:4272–4276. doi:10.4049/jimmunol.168.9.4272 PubMedCrossRef

58.

Mantovani A, Germano G, Marchesi F, Locatelli M, Biswas SK (2011) Cancer-promoting tumor-associated macrophages: new vistas and open questions. Eur J Immunol 41:2522–2525. doi:10.1002/eji.201141894 PubMedCrossRef

Title: Metastatic spread in patients with non-small cell lung cancer is associated with a reduced density of tumor-infiltrating T cells
Authors: Philipp Müller
Sacha I. Rothschild
Walter Arnold
Petra Hirschmann
Lukas Horvath
Lukas Bubendorf
Spasenija Savic
Alfred Zippelius
Publication date: 01-01-2016
Publisher: Springer Berlin Heidelberg
Published in: Cancer Immunology, Immunotherapy / Issue 1/2016
Print ISSN: 0340-7004
Electronic ISSN: 1432-0851
DOI: https://doi.org/10.1007/s00262-015-1768-3

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