Tumor infiltrating lymphocytes: an intriguing player in the survival of colorectal cancer patients

Vanessa Deschoolmeester, Marc Baay, Eric Van Marck, Joost Weyler, Peter Vermeulen, Filip Lardon, Jan B Vermorken, Vanessa Deschoolmeester, Marc Baay, Eric Van Marck, Joost Weyler, Peter Vermeulen, Filip Lardon, Jan B Vermorken

Abstract

Background: There is growing evidence that both local and systemic inflammatory responses play an important role in the progression of a variety of solid tumors. Colorectal cancer results from the cumulative effect of sequential genetic alterations, leading to the expression of tumor associated antigens possibly inducing a cellular anti-tumor immune response. It is well recognized that cytotoxic lymphocytes constitute one of the most important effector mechanisms of anti-tumor-immunity. However, their potential prognostic influence in colorectal cancer remains controversial. Aim of the study was to examine infiltration of CD3+ and CD8+ lymphocytes in colorectal cancer and their prognostic potential.Two-hundred-fifteen colorectal cancer cases, previously analyzed for microsatellite instability (MSI), were selected for immunohistochemical detection of CD3+, CD8+ infiltration and the expression of granzyme B. Prognostic relevance was assessed by survival analysis.

Results: Strong correlations were found between the infiltration of lymphocytes and several clinicopathological variables. Survival analysis revealed that intra-epithelial infiltration of CD3+ and CD8+ T lymphocytes and stromal infiltration of CD3+ lymphocytes had a major impact on the patients' overall survival in the univariate analysis, however independent of their association with MSI-status. In addition, it was also demonstrated that there was an important disease specific survival advantage for patients with microsatellite stable (MSS) tumors containing intraepithelial CD8+ tumor infiltrating lymphocytes. When samples were analyzed for colon cancer and rectal cancer separately, the results of the overall population were confirmed in colon cancer only. When entered into a multiple Cox regression analysis adjusting for other possible important confounding factors, the strong impact of lymphocyte infiltration on overall survival was not maintained. Only early stage and young age (borderline significant for overall population only) were associated with a better overall survival (early disease with disease-free survival also).

Conclusions: In conclusion our results suggest a role for infiltrating CD3+ and CD8+ T lymphocytes in colorectal cancer whereby tumor infiltration could reflect a general principle of antitumor immunity, irrespective of the MSI-status.

Figures

Figure 1
Figure 1
Immunohistochemical staining of lymphocyte infiltration in colorectal cancer. Examples of lymphocyte infiltration in colorectal tumors. A and C: CD8+ T cells infiltrating the invasive margin (A), the stroma (A and C) and in the cancer cell nests (C). B and D: CD3+ cells infiltrating the invasive margin (B), the stroma (B and D) and in the cancer cell nests (D).
Figure 2
Figure 2
Semiquantitative scoring of inflammatory infiltration. Semiquantitative scoring of inflammatory infiltration of the three different tumor regions, based on two levels of infiltration (null-mild versus moderate-severe). (IM: invasive margin, ST: stroma, CC: cancer cell nests, GRB: granzyme B, CD8*GRB: CD8 T cells expressing GRB).
Figure 3
Figure 3
Kaplan-Meier survival curves for infiltrating lymphocytes in colorectal cancer. Kaplan-Meier curves for overall survival among colorectal cancer patients according to (A) the infiltration of CD3+ lymphocytes in the cancer cell nests and (B) the stroma, (C) according to the infiltration of CD8+ lymphocytes in the cancer cell nests and (D) according to the infiltration of CD8+ lymphocytes in the cancer cell nests of MSS tumors (IM: invasive margin, ST: stroma, CC: cancer cell nests, MSS: microsatellite stable).
Figure 4
Figure 4
Kaplan-Meier survival curves for infiltrating lymphocytes in colon cancer only. Kaplan-Meier curves for overall survival among colon cancer patients according to (A) the infiltration of CD3+ lymphocytes in the cancer cell nests and (B) the stroma, (C) according to the infiltration of CD8+ lymphocytes in the cancer cell nests and (D) according to the infiltration of CD8+ lymphocytes in the cancer cell nests of MSS tumors (IM: invasive margin, ST: stroma, CC: cancer cell nests, MSS: microsatellite stable).

References

    1. Jemal A, Siegel R, Ward E, Hao Y, Xu J, Thun MJ. Cancer Statistics, 2009. CA Cancer J Clin. 2009;59:225–249. doi: 10.3322/caac.20006.
    1. Weitz J, Koch M, Debus J, Hohler T, Galle PR, Buchler MW. Colorectal cancer. Lancet. 2005;365:153–165. doi: 10.1016/S0140-6736(05)17706-X.
    1. Lurje G, Zhang W, Lenz HJ. Molecular prognostic markers in locally advanced colon cancer. Clin Colorectal Cancer. 2007;6:683–690. doi: 10.3816/CCC.2007.n.037.
    1. Koch M, Beckhove P, Op den Winkel J, Autenrieth D, Wagner P, Nummer D. Tumor infiltrating T lymphocytes in colorectal cancer: Tumor-selective activation and cytotoxic activity in situ. Ann Surg. 2006;244:986–992. doi: 10.1097/01.sla.0000247058.43243.7b.
    1. NIH consensus conference. Adjuvant therapy for patients with colon and rectal cancer. JAMA. 1990;264:1444–1450. doi: 10.1001/jama.264.11.1444.
    1. Shulman K, Schilsky R. Adjuvant therapy of colon cancer. Semin Oncol. 1995;22:600–10.
    1. Dalerba P, Maccalli C, Casati C, Castelli C, Parmiani G. Immunology and immunotherapy of colorectal cancer. Crit Rev Oncol Hematol. 2003;46:33–57. doi: 10.1016/S1040-8428(02)00159-2.
    1. Canna K, McArdle PA, McMillan DC, McNicol AM, Smith GW, McKee RF. The relationship between tumour T-lymphocyte infiltration, the systemic inflammatory response and survival in patients undergoing curative resection for colorectal cancer. Br J Cancer. 2005;92:651–654. doi: 10.1038/sj.bjc.6602419.
    1. Roxburgh CS, Salmond JM, Horgan PG, Oien KA, McMillan DC. Comparison of the prognostic value of inflammation-based pathologic and biochemical criteria in patients undergoing potentially curative resection for colorectal cancer. Ann Surg. 2009;249:788–793. doi: 10.1097/SLA.0b013e3181a3e738.
    1. Roxburgh CS, Salmond JM, Horgan PG, Oien KA, McMillan DC. Tumour inflammatory infiltrate predicts survival following curative resection for node-negative colorectal cancer. Eur J Cancer. 2009;45:2138–2145. doi: 10.1016/j.ejca.2009.04.011.
    1. Linnebacher M, Gebert J, Rudy W, Woerner S, Yuan Y, Bork P. Frameshift peptide-derived T-cell epitopes: a source of novel tumor-specific antigens. Int J Cancer. 2001;93:6–11. doi: 10.1002/ijc.1298.
    1. Diederichsen AC, Hjelmborg JB, Christensen PB, Zeuthen J, Fenger C. Prognostic value of the CD4+/CD8+ ratio of tumour infiltrating lymphocytes in colorectal cancer and HLA-DR expression on tumour cells. Cancer Immunol Immunother. 2003;52:423–428. doi: 10.1007/s00262-003-0388-5.
    1. Guidoboni M, Gafa R, Viel A, Doglioni C, Russo A, Santini A. Microsatellite instability and high content of activated cytotoxic lymphocytes identify colon cancer patients with a favorable prognosis. Am J Pathol. 2001;159:297–304.
    1. Ishikawa T, Fujita T, Suzuki Y, Okabe S, Yuasa Y, Iwai T. Tumor-specific immunological recognition of frameshift-mutated peptides in colon cancer with microsatellite instability. Cancer Res. 2003;63:5564–5572.
    1. Drescher KM, Sharma P, Watson P, Gatalica Z, Thibodeau SN, Lynch HT. Lymphocyte recruitment into the tumor site is altered in patients with MSI-H colon cancer. Fam Cancer. 2009;8:231–239. doi: 10.1007/s10689-009-9233-0.
    1. Schwitalle Y, Kloor M, Eiermann S, Linnebacher M, Kienle P, Knaebel HP. Immune response against frameshift-induced neopeptides in HNPCC patients and healthy HNPCC mutation carriers. Gastroenterology. 2008;134:988–997. doi: 10.1053/j.gastro.2008.01.015.
    1. Titu LV, Monson JR, Greenman J. The role of CD8(+) T cells in immune responses to colorectal cancer. Cancer Immunol Immunother. 2002;51:235–247. doi: 10.1007/s00262-002-0276-4.
    1. Evans C, Dalgleish AG, Kumar D. Review article: immune suppression and colorectal cancer. Aliment Pharmacol Ther. 2006;24:1163–1177. doi: 10.1111/j.1365-2036.2006.03075.x.
    1. Loose D, Wiele C Van de. The immune system and cancer. Cancer Biother Radiopharm. 2009;24:369–376. doi: 10.1089/cbr.2008.0593.
    1. Watson NF, Ramage JM, Madjd Z, Spendlove I, Ellis IO, Scholefield JH. Immunosurveillance is active in colorectal cancer as downregulation but not complete loss of MHC class I expression correlates with a poor prognosis. Int J Cancer. 2006;118:6–10. doi: 10.1002/ijc.21303.
    1. Atkinson EA, Bleackley RC. Mechanisms of lysis by cytotoxic T cells. Crit Rev Immunol. 1995;15:359–384.
    1. Parmiani G. Tumor-infiltrating T cells--friend or foe of neoplastic cells? N Engl J Med. 2005;353:2640–1. doi: 10.1056/NEJMp058236.
    1. Klintrup K, Makinen JM, Kauppila S, Vare PO, Melkko J, Tuominen H. Inflammation and prognosis in colorectal cancer. Eur J Cancer. 2005;41:2645–2654. doi: 10.1016/j.ejca.2005.07.017.
    1. Naito Y, Saito K, Shiiba K, Ohuchi A, Saigenji K, Nagura H. CD8+ T cells infiltrated within cancer cell nests as a prognostic factor in human colorectal cancer. Cancer Res. 1998;58:3491–4.
    1. Smyrk TC, Watson P, Kaul K, Lynch HT. Tumor-infiltrating lymphocytes are a marker for microsatellite instability in colorectal carcinoma. Cancer. 2001;91:2417–2422. doi: 10.1002/1097-0142(20010615)91:12<2417::AID-CNCR1276>;2-U.
    1. Takemoto N, Konishi F, Yamashita K, Kojima M, Furukawa T, Miyakura Y. The correlation of microsatellite instability and tumor-infiltrating lymphocytes in hereditary non-polyposis colorectal cancer (HNPCC) and sporadic colorectal cancers: the significance of different types of lymphocyte infiltration. Jpn J Clin Oncol. 2004;34:90–98. doi: 10.1093/jjco/hyh018.
    1. Phillips SM, Banerjea A, Feakins R, Li SR, Bustin SA, Dorudi S. Tumour-infiltrating lymphocytes in colorectal cancer with microsatellite instability are activated and cytotoxic. Br J Surg. 2004;91:469–475. doi: 10.1002/bjs.4472.
    1. Popat S, Hubner R, Houlston R. Systematic review of microsatellite instability and colorectal cancer prognosis. J Clin Oncol. 2005;23:609–18. doi: 10.1200/JCO.2005.01.086.
    1. Chang S, Lin J, Yang S, Wang H, Li A, Chi C. Relationship between genetic alterations and prognosis in sporadic colorectal cancer. Int J Cancer. 2006;118:1721–7. doi: 10.1002/ijc.21563.
    1. Dolcetti R, Viel A, Doglioni C, Russo A, Guidoboni M, Capozzi E. High prevalence of activated intraepithelial cytotoxic T lymphocytes and increased neoplastic cell apoptosis in colorectal carcinomas with microsatellite instability. Am J Pathol. 1999;154:1805–13.
    1. Prall F, Duhrkop T, Weirich V, Ostwald C, Lenz P, Nizze H. Prognostic role of CD8+tumor-infiltrating lymphocytes in stage III colorectal cancer with and without microsatellite instability. Human Path. 2004;35:808–816. doi: 10.1016/j.humpath.2004.01.022.
    1. Vermorken JB, Claessen AM, van Tinteren H, Gall HE, Ezinga R, Meijer S. Active specific immunotherapy for stage II and stage III human colon cancer: a randomised trial. Lancet. 1999;353:345–50. doi: 10.1016/S0140-6736(98)07186-4.
    1. Eertwegh AJ van den. Active specific immunotherapy in colon cancer. Recent Results Cancer Res. 2005;165:260–267. full_text.
    1. Michael-Robinson JM, Biemer-Huttmann AE, Purdie DM, Walsh MD, Simms LA, Biden KG. Tumour infiltrating lymphocytes and apoptosis are independent features in colorectal cancer stratified according to microsatellite instability status. Gut. 2001;48:360–366. doi: 10.1136/gut.48.3.360.
    1. Deschoolmeester V, Baay M, Wuyts W, Van ME, Van DN, Vermeulen P. Detection of microsatellite instability in colorectal cancer using an alternative multiplex assay of quasi-monomorphic mononucleotide markers. J Mol Diagn. 2008;10:154–159. doi: 10.2353/jmoldx.2008.070087.
    1. Lin EY, Pollard JW. Role of infiltrated leucocytes in tumour growth and spread. Br J Cancer. 2004;90:2053–2058. doi: 10.1038/sj.bjc.6601705.
    1. Baker K, Zlobec I, Tornillo L, Terracciano L, Jass JR, Lugli A. Differential significance of tumour infiltrating lymphocytes in sporadic mismatch repair deficient versus proficient colorectal cancers: a potential role for dysregulation of the transforming growth factor-beta pathway. Eur J Cancer. 2007;43:624–631. doi: 10.1016/j.ejca.2006.11.012.
    1. Galon J, Costes A, Sanchez-Cabo F, Kirilovsky A, Mlecnik B, Lagorce-Pages C. Type, density, and location of immune cells within human colorectal tumors predict clinical outcome. Science. 2006;313:1960–1964. doi: 10.1126/science.1129139.
    1. Galon J, Fridman WH, Pages F. The adaptive immunologic microenvironment in colorectal cancer: a novel perspective. Cancer Res. 2007;67:1883–1886. doi: 10.1158/0008-5472.CAN-06-4806.
    1. Camus M, Tosolini M, Mlecnik B, Pages F, Kirilovsky A, Berger A. Coordination of intratumoral immune reaction and human colorectal cancer recurrence. Cancer Res. 2009;69:2685–2693. doi: 10.1158/0008-5472.CAN-08-2654.
    1. Chiba T, Ohtani H, Mizoi T, Naito Y, Sato E, Nagura H. Intraepithelial CD8+ T-cell-count becomes a prognostic factor after a longer follow-up period in human colorectal carcinoma: possible association with suppression of micrometastasis. Br J Cancer. 2004;91:1711–1717.
    1. Banerjea A, Bustin S, Dorudi S. The immunogenicity of colorectal cancers with high-degree microsatellite instability. World J Surg Oncol. 2005;3:26. doi: 10.1186/1477-7819-3-26.
    1. Jass JR, Love SB, Northover JM. A new prognostic classification of rectal cancer. Lancet. 1987;1:1303–1306. doi: 10.1016/S0140-6736(87)90552-6.
    1. Ropponen KM, Eskelinen MJ, Lipponen PK, Alhava E, Kosma VM. Prognostic value of tumour-infiltrating lymphocytes (TILs) in colorectal cancer. J Pathol. 1997;182:318–324. doi: 10.1002/(SICI)1096-9896(199707)182:3<318::AID-PATH862>;2-6.
    1. Nielsen HJ, Hansen U, Christensen IJ, Reimert CM, Brunner N, Moesgaard F. Independent prognostic value of eosinophil and mast cell infiltration in colorectal cancer tissue. J Pathol. 1999;189:487–495. doi: 10.1002/(SICI)1096-9896(199912)189:4<487::AID-PATH484>;2-I.
    1. Funada Y, Noguchi T, Kikuchi R, Takeno S, Uchida Y, Gabbert HE. Prognostic significance of CD8+ T cell and macrophage peritumoral infiltration in colorectal cancer. Oncol Rep. 2003;10:309–313.
    1. Pages F, Berger A, Camus M, Sanchez Cabo F, Costes A, Molidor R. Effector memory T cells, early metastasis, and survival in colorectal cancer. N Engl J Med. 2005;353:2654–66. doi: 10.1056/NEJMoa051424.
    1. Halama N, Michel S, Kloor M, Zoernig I, Pommerencke T, von Knebel DM. The localization and density of immune cells in primary tumors of human metastatic colorectal cancer shows an association with response to chemotherapy. Cancer Immun. 2009;9:1.
    1. Ling KL, Pratap SE, Bates GJ, Singh B, Mortensen NJ, George BD. Increased frequency of regulatory T cells in peripheral blood and tumour infiltrating lymphocytes in colorectal cancer patients. Cancer Immun. 2007;7:7.
    1. Salahshor S, Kressner U, Fischer H, Lindmark G, Glimelius B, Pahlman L. Microsatellite instability in sporadic colorectal cancer is not an independent prognostic factor. Br J Cancer. 1999;81:190–3. doi: 10.1038/sj.bjc.6690676.
    1. Fujiwara T, Stolker J, Watanabe T, Rashid A, Longo P, Eshleman J. Accumulated clonal genetic alterations in familial and sporadic colorectal carcinomas with widespread instability in microsatellite sequences. Am J Pathol. 1998;153:1063–78.
    1. Feeley KM, Fullard JF, Heneghan MA, Smith T, Maher M, Murphy RP. Microsatellite instability in sporadic colorectal carcinoma is not an indicator of prognosis. J Pathol. 1999;188:14–17. doi: 10.1002/(SICI)1096-9896(199905)188:1<14::AID-PATH323>;2-Q.
    1. Messerini L, Ciantelli M, Baglioni S, Palomba A, Zampi G, Papi L. Prognostic significance of microsatellite instability in sporadic mucinous colorectal cancers. Hum Pathol. 1999;30:629–34. doi: 10.1016/S0046-8177(99)90086-2.
    1. Curran B, Lenehan K, Mulcahy H, Tighe O, Bennett MA, Kay EW. Replication error phenotype, clinicopathological variables, and patient outcome in Dukes' B stage II (T3, N0, M0) colorectal cancer. Gut. 2000;46:200–4. doi: 10.1136/gut.46.2.200.
    1. Wang C, van Rijnsoever M, Grieu F, Bydder S, Elsaleh H, Joseph D. Prognostic significance of microsatellite instability and Ki-ras mutation type in stage II colorectal cancer. Oncology. 2003;64:259–65. doi: 10.1159/000069311.
    1. Emterling A, Wallin A, Arbman G, Sun XF. Clinicopathological significance of microsatellite instability and mutated RIZ in colorectal cancer. Ann Oncol. 2004;15:242–246. doi: 10.1093/annonc/mdh045.
    1. Storojeva I, Boulay J, Heinimann K, Ballabeni P, Terracciano L, Laffer U. Prognostic and predictive relevance of microsatellite instability in colorectal cancer. Oncol Rep. 2005;14:241–9.
    1. Buckowitz A, Knaebel H, Benner A, Blaker H, Gebert J, Kienle P. Microsatellite instability in colorectal cancer is associated with local lymphocyte infiltration and low frequency of distant metastases. Br J Cancer. 2005;92:1746–53. doi: 10.1038/sj.bjc.6602534.
    1. Menon AG, Janssen-van Rhijn CM, Morreau H, Putter H, Tollenaar RA, Velde CJH van de. Immune system and prognosis in colorectal cancer: a detailed immunohistochemical analysis. Lab Invest. 2004;84:493–501. doi: 10.1038/labinvest.3700055.
    1. Speetjens FM, de Bruin EC, Morreau H, Zeestraten EC, Putter H, van Krieken JH. Clinical impact of HLA class I expression in rectal cancer. Cancer Immunol Immunother. 2008;57:601–609. doi: 10.1007/s00262-007-0396-y.
    1. Duffy MJ, van DA, Haglund C, Hansson L, Holinski-Feder E, Klapdor R. Tumour markers in colorectal cancer: European Group on Tumour Markers (EGTM) guidelines for clinical use. Eur J Cancer. 2007;43:1348–1360. doi: 10.1016/j.ejca.2007.03.021.
    1. Deschoolmeester V, Van Damme N, Baay M, Claes K, Van ME, Baert FJ. Microsatellite instability in sporadic colon carcinomas has no independent prognostic value in a Belgian study population. Eur J Cancer. 2008;44:2288–2295. doi: 10.1016/j.ejca.2008.06.043.
    1. Deschoolmeester V, Baay M, Wuyts W, Van ME, Pelckmans P, Lardon F. Comparison of three commonly used PCR-based techniques to analyze MSI status in sporadic colorectal cancer. J Clin Lab Anal. 2006;20:52–61. doi: 10.1002/jcla.20097.

Source: PubMed

Patrocinadores e Colaboradores

Condições médicas

Intervenções de drogas

3
Se inscrever