Abstract
Background
Ectopic lymphoid formations are called tertiary lymphoid structures (TLSs). TLSs in cancer have been reported to be associated with good prognosis and immunotherapy response. However, the relationship between TLSs and lymph node (LN) metastasis is unclear.
Methods
We analyzed 218 patients with radically resected lung adenocarcinoma. TLSs were defined as the overlap of T cell zone and B cell zone. Granzyme B + cells were defined as cytotoxic lymphocytes. We evaluated phenotypes of lymphocytes in TLSs, tumor-infiltrating lymphocytes (TILs) and LNs by immunohistochemistry. We divided the patients into mature TLS (DC-Lamp high) and immature TLS (DC-Lamp low) groups. The relationship between TLS maturation and clinicopathological factors was analyzed.
Results
The mature TLS group was associated with significantly lower frequency of LN metastasis (P < 0.0001) and early cancer stage (P = 0.0049). The mature TLS group had significantly more CD8 + (P = 0.0203) and Foxp3 + (P = 0.0141) cells in TILs than the immature TLS group had. Mature TLSs were independently associated with a favorable overall survival (hazard ratio [HR] = 0.17, P = 0.0220) and disease-free survival (HR = 0.54, P = 0.0436). Multivariate analysis showed that mature TLS was an independent low-risk factor for LN metastasis (odds ratio = 0.06, P = 0.0003). The number of cytotoxic lymphocytes in LNs was higher in the mature TLS group than in the immature group (20.0 vs. 15.1, P = 0.017).
Conclusion
Mature TLSs were associated with an increased number of cytotoxic lymphocytes in draining LNs, a lower frequency of LN metastasis, and favorable outcomes. Mature TLSs may support antitumor immunity by lymphocyte activation.
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Data availability
The datasets analyzed during the current study are available from the corresponding author on reasonable request.
Abbreviations
- DC:
-
Dendritic cell
- DFS:
-
Disease-free survival
- HEV:
-
High endothelial venule
- HR:
-
Hazard ratio
- LN:
-
Lymph node
- LT:
-
Lymphotoxin.
- OR:
-
Odds ratio
- OS:
-
Overall survival
- PD-L1:
-
Programmed cell death-ligand 1
- pN:
-
Pathological N status
- pT:
-
Pathological T status
- TIL:
-
Tumor-infiltrating lymphocyte
- TLS:
-
Tertiary lymphoid structure
- TNF:
-
Tumor necrosis factor
References
Reck M, Rodriguez-Abreu D, Robinson AG, Hui R, Csoszi T, Fulop A, Gottfried M, Peled N, Tafreshi A, Cuffe S, O’Brien M, Rao S, Hotta K, Leiby MA, Lubiniecki GM, Shentu Y, Rangwala R, Brahmer JR, Investigators K (2016) Pembrolizumab versus chemotherapy for PD-L1-positive non-small-cell lung cancer. N Engl J Med 375(19):1823–1833. https://doi.org/10.1056/NEJMoa1606774
Gandhi L, Rodriguez-Abreu D, Gadgeel S, Esteban E, Felip E, De Angelis F, Domine M, Clingan P, Hochmair MJ, Powell SF, Cheng SY, Bischoff HG, Peled N, Grossi F, Jennens RR, Reck M, Hui R, Garon EB, Boyer M, Rubio-Viqueira B, Novello S, Kurata T, Gray JE, Vida J, Wei Z, Yang J, Raftopoulos H, Pietanza MC, Garassino MC, Investigators K (2018) Pembrolizumab plus chemotherapy in metastatic non-small-cell lung cancer. N Engl J Med 378(22):2078–2092. https://doi.org/10.1056/NEJMoa1801005
Bruno TC (2020) New predictors for immunotherapy responses sharpen our view of the tumour microenvironment. Nature 577(7791):474–476. https://doi.org/10.1038/d41586-019-03943-0
Dieu-Nosjean MC, Goc J, Giraldo NA, Sautes-Fridman C, Fridman WH (2014) Tertiary lymphoid structures in cancer and beyond. Trends Immunol 35(11):571–580. https://doi.org/10.1016/j.it.2014.09.006
Dieu-Nosjean MC, Antoine M, Danel C, Heudes D, Wislez M, Poulot V, Rabbe N, Laurans L, Tartour E, de Chaisemartin L, Lebecque S, Fridman WH, Cadranel J (2008) Long-term survival for patients with non-small-cell lung cancer with intratumoral lymphoid structures. J Clin Oncol 26(27):4410–4417. https://doi.org/10.1200/JCO.2007.15.0284
Colbeck EJ, Ager A, Gallimore A, Jones GW (2017) Tertiary lymphoid structures in cancer: drivers of antitumor immunity, immunosuppression, or bystander sentinels in Disease? Front Immunol 8:1830. https://doi.org/10.3389/fimmu.2017.01830
Gu-Trantien C, Migliori E, Buisseret L, de Wind A, Brohee S, Garaud S, Noel G, Dang Chi VL, Lodewyckx JN, Naveaux C, Duvillier H, Goriely S, Larsimont D, Willard-Gallo K (2017) CXCL13-producing TFH cells link immune suppression and adaptive memory in human breast cancer. JCI Insight. https://doi.org/10.1172/jci.insight.91487
Nayar S, Campos J, Smith CG, Iannizzotto V, Gardner DH, Mourcin F, Roulois D, Turner J, Sylvestre M, Asam S, Glaysher B, Bowman SJ, Fearon DT, Filer A, Tarte K, Luther SA, Fisher BA, Buckley CD, Coles MC, Barone F (2019) Immunofibroblasts are pivotal drivers of tertiary lymphoid structure formation and local pathology. Proc Natl Acad Sci U S A 116(27):13490–13497. https://doi.org/10.1073/pnas.1905301116
Lin L, Hu X, Zhang H, Hu H (2019) Tertiary lymphoid organs in cancer immunology: mechanisms and the new strategy for immunotherapy. Front Immunol 10:1398. https://doi.org/10.3389/fimmu.2019.01398
Nerviani A, Pitzalis C (2018) Role of chemokines in ectopic lymphoid structures formation in autoimmunity and cancer. J Leukoc Biol 104(2):333–341. https://doi.org/10.1002/JLB.3MR0218-062R
Germain C, Gnjatic S, Tamzalit F, Knockaert S, Remark R, Goc J, Lepelley A, Becht E, Katsahian S, Bizouard G, Validire P, Damotte D, Alifano M, Magdeleinat P, Cremer I, Teillaud JL, Fridman WH, Sautes-Fridman C, Dieu-Nosjean MC (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(7):832–844. https://doi.org/10.1164/rccm.201309-1611OC
Hiraoka N, Ino Y, Yamazaki-Itoh R, Kanai Y, Kosuge T, Shimada K (2015) Intratumoral tertiary lymphoid organ is a favourable prognosticator in patients with pancreatic cancer. Br J Cancer 112(11):1782–1790. https://doi.org/10.1038/bjc.2015.145
Koti M, Xu AS, Ren KYM, Visram K, Ren R, Berman DM, Siemens DR (2017) Tertiary lymphoid structures associate with tumour stage in urothelial bladder cancer. Bladder Cancer 3(4):259–267. https://doi.org/10.3233/BLC-170120
Posch F, Silina K, Leibl S, Mundlein A, Moch H, Siebenhuner A, Samaras P, Riedl J, Stotz M, Szkandera J, Stoger H, Pichler M, Stupp R, van den Broek M, Schraml P, Gerger A, Petrausch U, Winder T (2018) Maturation of tertiary lymphoid structures and recurrence of stage II and III colorectal cancer. Oncoimmunology 7(2):e1378844. https://doi.org/10.1080/2162402X.2017.1378844
Helmink BA, Reddy SM, Gao J, Zhang S, Basar R, Thakur R, Yizhak K, Sade-Feldman M, Blando J, Han G, Gopalakrishnan V, Xi Y, Zhao H, Amaria RN, Tawbi HA, Cogdill AP, Liu W, LeBleu VS, Kugeratski FG, Patel S, Davies MA, Hwu P, Lee JE, Gershenwald JE, Lucci A, Arora R, Woodman S, Keung EZ, Gaudreau PO, Reuben A, Spencer CN, Burton EM, Haydu LE, Lazar AJ, Zapassodi R, Hudgens CW, Ledesma DA, Ong S, Bailey M, Warren S, Rao D, Krijgsman O, Rozeman EA, Peeper D, Blank CU, Schumacher TN, Butterfield LH, Zelazowska MA, McBride KM, Kalluri R, Allison J, Petitprez F, Fridman WH, Sautes-Fridman C, Hacohen N, Rezvani K, Sharma P, Tetzlaff MT, Wang L, Wargo JA (2020) B cells and tertiary lymphoid structures promote immunotherapy response. Nature 577(7791):549–555. https://doi.org/10.1038/s41586-019-1922-8
Cabrita R, Lauss M, Sanna A, Donia M, Skaarup Larsen M, Mitra S, Johansson I, Phung B, Harbst K, Vallon-Christersson J, van Schoiack A, Lovgren K, Warren S, Jirstrom K, Olsson H, Pietras K, Ingvar C, Isaksson K, Schadendorf D, Schmidt H, Bastholt L, Carneiro A, Wargo JA, Svane IM, Jonsson G (2020) Tertiary lymphoid structures improve immunotherapy and survival in melanoma. Nature 577(7791):561–565. https://doi.org/10.1038/s41586-019-1914-8
Petitprez F, de Reynies A, Keung EZ, Chen TW, Sun CM, Calderaro J, Jeng YM, Hsiao LP, Lacroix L, Bougouin A, Moreira M, Lacroix G, Natario I, Adam J, Lucchesi C, Laizet YH, Toulmonde M, Burgess MA, Bolejack V, Reinke D, Wani KM, Wang WL, Lazar AJ, Roland CL, Wargo JA, Italiano A, Sautes-Fridman C, Tawbi HA, Fridman WH (2020) B cells are associated with survival and immunotherapy response in sarcoma. Nature 577(7791):556–560. https://doi.org/10.1038/s41586-019-1906-8
Cottrell TR, Thompson ED, Forde PM, Stein JE, Duffield AS, Anagnostou V, Rekhtman N, Anders RA, Cuda JD, Illei PB, Gabrielson E, Askin FB, Niknafs N, Smith KN, Velez MJ, Sauter JL, Isbell JM, Jones DR, Battafarano RJ, Yang SC, Danilova L, Wolchok JD, Topalian SL, Velculescu VE, Pardoll DM, Brahmer JR, Hellmann MD, Chaft JE, Cimino-Mathews A, Taube JM (2018) Pathologic features of response to neoadjuvant anti-PD-1 in resected non-small-cell lung carcinoma: a proposal for quantitative immune-related pathologic response criteria (irPRC). Ann Oncol 29(8):1853–1860. https://doi.org/10.1093/annonc/mdy218
Sautes-Fridman C, Petitprez F, Calderaro J, Fridman WH (2019) Tertiary lymphoid structures in the era of cancer immunotherapy. Nat Rev Cancer 19(6):307–325. https://doi.org/10.1038/s41568-019-0144-6
Chamoto K, Chowdhury PS, Kumar A, Sonomura K, Matsuda F, Fagarasan S, Honjo T (2017) Mitochondrial activation chemicals synergize with surface receptor PD-1 blockade for T cell-dependent antitumor activity. Proc Natl Acad Sci U S A 114(5):E761–E770. https://doi.org/10.1073/pnas.1620433114
Takada K, Okamoto T, Shoji F, Shimokawa M, Akamine T, Takamori S, Katsura M, Suzuki Y, Fujishita T, Toyokawa G, Morodomi Y, Okano S, Oda Y, Maehara Y (2016) Clinical significance of PD-L1 protein expression in surgically resected primary lung adenocarcinoma. J Thorac Oncol 11(11):1879–1890. https://doi.org/10.1016/j.jtho.2016.06.006
Figenschau SL, Fismen S, Fenton KA, Fenton C, Mortensen ES (2015) Tertiary lymphoid structures are associated with higher tumor grade in primary operable breast cancer patients. BMC Cancer 15:101. https://doi.org/10.1186/s12885-015-1116-1
Trajkovski G, Ognjenovic L, Karadzov Z, Jota G, Hadzi-Manchev D, Kostovski O, Volcevski G, Trajkovska V, Nikolova D, Spasevska L, Janevska V, Janevski V (2018) Tertiary lymphoid structures in colorectal cancers and their prognostic value. Open Access Maced J Med Sci 6(10):1824–1828. https://doi.org/10.3889/oamjms.2018.341
Dammeijer F, van Gulijk M, Mulder EE, Lukkes M, Klaase L, van den Bosch T, van Nimwegen M, Lau SP, Latupeirissa K, Schetters S, van Kooyk Y, Boon L, Moyaart A, Mueller YM, Katsikis PD, Eggermont AM, Vroman H, Stadhouders R, Hendriks RW, Thusen JV, Grunhagen DJ, Verhoef C, van Hall T, Aerts JG (2020) The PD-1/PD-L1-checkpoint restrains T cell Immunity in tumor-draining lymph nodes. Cancer Cell. https://doi.org/10.1016/j.ccell.2020.09.001
Engelhard VH, Rodriguez AB, Mauldin IS, Woods AN, Peske JD, Slingluff CL Jr (2018) Immune cell infiltration and Tertiary lymphoid structures as determinants of antitumor immunity. J Immunol 200(2):432–442. https://doi.org/10.4049/jimmunol.1701269
Pesce S, Moretta L, Moretta A, Marcenaro E (2016) Human NK cell subsets redistribution in pathological conditions: a role for CCR7 receptor. Front Immunol 7:414. https://doi.org/10.3389/fimmu.2016.00414
de Chaisemartin L, Goc J, Damotte D, Validire P, Magdeleinat P, Alifano M, Cremer I, Fridman WH, Sautes-Fridman C, Dieu-Nosjean MC (2011) Characterization of chemokines and adhesion molecules associated with T cell presence in tertiary lymphoid structures in human lung cancer. Cancer Res 71(20):6391–6399. https://doi.org/10.1158/0008-5472.CAN-11-0952
Guedj K, Khallou-Laschet J, Clement M, Morvan M, Gaston AT, Fornasa G, Dai J, Gervais-Taurel M, Eberl G, Michel JB, Caligiuri G, Nicoletti A (2014) M1 macrophages act as LTbetaR-independent lymphoid tissue inducer cells during atherosclerosis-related lymphoid neogenesis. Cardiovasc Res 101(3):434–443. https://doi.org/10.1093/cvr/cvt263
Germain C, Gnjatic S, Dieu-Nosjean MC (2015) Tertiary lymphoid structure-associated B cells are key players in anti-tumor immunity. Front Immunol 6:67. https://doi.org/10.3389/fimmu.2015.00067
Goc J, Germain C, Vo-Bourgais TK, Lupo A, Klein C, Knockaert S, de Chaisemartin L, Ouakrim H, Becht E, Alifano M, Validire P, Remark R, Hammond SA, Cremer I, Damotte D, Fridman WH, Sautes-Fridman C, Dieu-Nosjean MC (2014) Dendritic cells in tumor-associated tertiary lymphoid structures signal a Th1 cytotoxic immune contexture and license the positive prognostic value of infiltrating CD8+ T cells. Cancer Res 74(3):705–715. https://doi.org/10.1158/0008-5472.CAN-13-1342
Zeng DQ, Yu YF, Ou QY, Li XY, Zhong RZ, Xie CM, Hu QG (2016) Prognostic and predictive value of tumor-infiltrating lymphocytes for clinical therapeutic research in patients with non-small cell lung cancer. Oncotarget 7(12):13765–13781. https://doi.org/10.18632/oncotarget.7282
Silina K, Soltermann A, Attar FM, Casanova R, Uckeley ZM, Thut H, Wandres M, Isajevs S, Cheng P, Curioni-Fontecedro A, Foukas P, Levesque MP, Moch H, Line A, van den Broek M (2018) Germinal centers determine the prognostic relevance of tertiary lymphoid structures and are impaired by corticosteroids in lung squamous cell carcinoma. Cancer Res 78(5):1308–1320. https://doi.org/10.1158/0008-5472.CAN-17-1987
Solinas C, Garaud S, De Silva P, Boisson A, Van den Eynden G, de Wind A, Risso P, Rodrigues Vitoria J, Richard F, Migliori E, Noel G, Duvillier H, Craciun L, Veys I, Awada A, Detours V, Larsimont D, Piccart-Gebhart M, Willard-Gallo K (2017) Immune checkpoint molecules on tumor-infiltrating lymphocytes and their association with tertiary lymphoid structures in human breast cancer. Front Immunol 8:1412. https://doi.org/10.3389/fimmu.2017.01412
Collins M, Ling V, Carreno BM (2005) The B7 family of immune-regulatory ligands. Genome Biol 6(6):223. https://doi.org/10.1186/gb-2005-6-6-223
Hodge G, Barnawi J, Jurisevic C, Moffat D, Holmes M, Reynolds PN, Jersmann H, Hodge S (2014) Lung cancer is associated with decreased expression of perforin, granzyme B and interferon (IFN)-gamma by infiltrating lung tissue T cells, natural killer (NK) T-like and NK cells. Clin Exp Immunol 178(1):79–85. https://doi.org/10.1111/cei.12392
Johansson-Percival A, He B, Li ZJ, Kjellen A, Russell K, Li J, Larma I, Ganss R (2017) De novo induction of intratumoral lymphoid structures and vessel normalization enhances immunotherapy in resistant tumors. Nat Immunol 18(11):1207–1217. https://doi.org/10.1038/ni.3836
Martin C, Thevenot G, Danel S, Chapron J, Tazi A, Macey J, Dusser DJ, Fajac I, Burgel PR (2011) Pseudomonas aeruginosa induces vascular endothelial growth factor synthesis in airway epithelium in vitro and in vivo. Eur Respir J 38(4):939–946. https://doi.org/10.1183/09031936.00134910
Acknowledgements
We thank MN and YK for her invaluable help with tissue processing. We also thank CK, BSc, PhD, from Edanz Group (https://en-author-services.edanzgroup.com/ac) for editing a draft of this manuscript.
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Conception and design: SW, TT, MM. Development of methodology: SW, FK, TT, YO. Acquisition of data: SW, YO, YO, FK. Analysis and interpretation of data: SW, FK, NH, TT, TT, YO, MS, MM. Writing, review, and/or revision of manuscript: SW, FK, NH, TT, YO, MM. Study supervision: TT, MS, YO, MM.
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This study was approved by our Institutional Review Board (Kyushu University, IRB No. 2019–232).
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Wakasu, S., Tagawa, T., Haratake, N. et al. Preventive effect of tertiary lymphoid structures on lymph node metastasis of lung adenocarcinoma. Cancer Immunol Immunother 72, 1823–1834 (2023). https://doi.org/10.1007/s00262-022-03353-8
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DOI: https://doi.org/10.1007/s00262-022-03353-8