Mechanistic roles and clinical applications of tertiary lymphoid structures in colorectal cancer drug resistance

Abstract

Therapeutic resistance represents a formidable bottleneck in colorectal cancer (CRC) management, severely limiting the clinical efficacy of both conventional regimens and novel immunotherapies. Emerging evidence underscores tertiary lymphoid structure (TLS) - ectopic lymphoid aggregates newly sculpted within the tumor microenvironment (TME) - as pivotal orchestrators of localized antitumor immunity and prime therapeutic targets to circumvent resistance. Distinct from secondary lymphoid organs, TLS facilitate in situ immune cell priming, clonal expansion, and functional differentiation, thereby sustaining a robust adaptive immune response. This review systematically dissects the cellular architecture, maturation dynamics, and spatial heterogeneity of TLS in primary and metastatic CRC, clarifying how these attributes dictate clinical outcomes and treatment responsiveness. Mechanistically, we delineate the molecular networks driving TLS neogenesis, encompassing chemokine cascades, tumor necrosis factor (TNF) superfamily signaling, and interactive crosstalk with the gut microbiota. Furthermore, we summarize cutting-edge preclinical strategies - including stimulator of interferon genes agonists, bio-nanovaccines, and hypofractionated radiotherapy combinations - engineered to trigger TLS neogenesis, promote germinal center (GC) maturation, and enrich stem-like effector tumor-infiltrating lymphocytes (TILs) to convert immunologically “cold” tumors into “hot” niches. By integrating mechanistic paradigms with clinical applications, this review provides a definitive framework for leveraging TLS targeting as a transformative modality to modulate drug resistance and optimize therapeutic trajectories in CRC; the graphical abstract is shown below.

Keywords

Tertiary lymphoid structures / colorectal cancer / tumor microenvironment / tumor-infiltrating lymphocytes / drug resistance / immunotherapy

References

[1] Morgan E,Gini A.Global burden of colorectal cancer in 2020 and 2040: incidence and mortality estimates from GLOBOCAN.Gut2023;72:338-44

[2] Dekker E,Vleugels JLA,Wallace MB.Colorectal cancer.Lancet2019;394:1467-80

[3] Biller LH.Diagnosis and treatment of metastatic colorectal cancer: a review.JAMA2021;325:669-85

[4] Le DT,Kim TW.Pembrolizumab for previously treated, microsatellite instability-high/mismatch repair-deficient advanced colorectal cancer: final analysis of KEYNOTE-164.Eur J Cancer2023;186:185-95

[5] Chalabi M,Dijkstra KK.Neoadjuvant immunotherapy leads to pathological responses in MMR-proficient and MMR-deficient early-stage colon cancers.Nat Med2020;26:566-76

[6] Ciardiello F,Martini G,Tabernero J.Clinical management of metastatic colorectal cancer in the era of precision medicine.CA Cancer J Clin2022;72:372-401

[7] Li J,Hu H.Remodeling of the immune and stromal cell compartment by PD-1 blockade in mismatch repair-deficient colorectal cancer.Cancer Cell2023;41:1152-69.e7

[8] Kornepati AVR,Curiel TJ.Programmed death ligand 1 signals in cancer cells.Nat Rev Cancer2022;22:174-89 PMCID:PMC9989967

[9] Sautès-Fridman C,Calderaro J.Tertiary lymphoid structures in the era of cancer immunotherapy.Nat Rev Cancer2019;19:307-25

[10] Dieu-Nosjean MC,Giraldo NA,Fridman WH.Tertiary lymphoid structures in cancer and beyond.Trends Immunol2014;35:571-80

[11] Schumacher TN.Tertiary lymphoid structures in cancer.Science2022;375:eabf9419

[12] Hayase S,Fujino S.Fibroblast activation protein and tertiary lymphoid structure in colorectal cancer recurrence.Anticancer Res2022;42:5897-907

[13] Cui X,Li D.Tertiary lymphoid structures as a biomarker in immunotherapy and beyond: advancing towards clinical application.Cancer Lett2025;613:217491

[14] Petitprez F,Keung EZ.B cells are associated with survival and immunotherapy response in sarcoma.Nature2020;577:556-60

[15] Vanhersecke L,Guégan JP.Mature tertiary lymphoid structures predict immune checkpoint inhibitor efficacy in solid tumors independently of PD-L1 expression.Nat Cancer2021;2:794-802 PMCID:PMC8809887

[16] Gao J,Alhalabi O.Neoadjuvant PD-L1 plus CTLA-4 blockade in patients with cisplatin-ineligible operable high-risk urothelial carcinoma.Nat Med2020;26:1845-51 PMCID:PMC9768836

[17] Germain C,Tamzalit F.Presence of B cells in tertiary lymphoid structures is associated with a protective immunity in patients with lung cancer.Am J Respir Crit Care Med2014;189:832-44

[18] Noël G,Garaud S.Functional Th1-oriented T follicular helper cells that infiltrate human breast cancer promote effective adaptive immunity.J Clin Invest2021;131:e139905 PMCID:PMC8483751

[19] Helmink BA,Gao J.B cells and tertiary lymphoid structures promote immunotherapy response.Nature2020;577:549-55 PMCID:PMC8762581

[20] Fridman WH,Petitprez F,Italiano A.B cells and tertiary lymphoid structures as determinants of tumour immune contexture and clinical outcome.Nat Rev Clin Oncol2022;19:441-57

[21] Ware MB,Goon JB.The role of interleukin-7 in the formation of tertiary lymphoid structures and their prognostic value in gastrointestinal cancers.J Immunother Precis Oncol2022;5:105-17 PMCID:PMC9714415

[22] Ding GY,Yun JP.Distribution and density of tertiary lymphoid structures predict clinical outcome in intrahepatic cholangiocarcinoma.J Hepatol2022;76:608-18

[23] Xia J,Niu G.Single-cell landscape and clinical outcomes of infiltrating B cells in colorectal cancer.Immunology2023;168:135-51

[24] Fridman WH,Pupier G,Hernandez I.Tertiary lymphoid structures and B cells: an intratumoral immunity cycle.Immunity2023;56:2254-69

[25] Cipponi A,Seremet T.Neogenesis of lymphoid structures and antibody responses occur in human melanoma metastases.Cancer Res2012;72:3997-4007

[26] Meylan M,Becht E.Tertiary lymphoid structures generate and propagate anti-tumor antibody-producing plasma cells in renal cell cancer.Immunity2022;55:527-41.e5

[27] Goc J,Vo-Bourgais TKD.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 Res2014;74:705-15

[28] Kroeger DR,Nelson BH.Tumor-infiltrating plasma cells are associated with tertiary lymphoid structures, cytolytic T-cell responses, and superior prognosis in ovarian cancer.Clin Cancer Res2016;22:3005-15

[29] Shalapour S.The neglected brothers come of age: B cells and cancer.Semin Immunol2021;52:101479 PMCID:PMC8545794

[30] Zhang C,Zuo JL.Localization and density of tertiary lymphoid structures associate with molecular subtype and clinical outcome in colorectal cancer liver metastases.J Immunother Cancer2023;11:e006425 PMCID:PMC9923349

[31] Zou X,Cheng H.Characterization of intratumoral tertiary lymphoid structures in pancreatic ductal adenocarcinoma: cellular properties and prognostic significance.J Immunother Cancer2023;11:e006698 PMCID:PMC10410893

[32] Wu Z, Zhou J, Xiao Y, et al. CD20⁺CD22⁺ADAM28⁺ B cells in tertiary lymphoid structures promote immunotherapy response.Front Immunol2022;13:865596 PMCID:PMC9130862

[33] Zhu W,Hu S.Chemokine- and chemokine receptor-based signature predicts immunotherapy response in female colorectal adenocarcinoma patients. Sci Rep 2023;13:21358. PMCID:PMC10695967

[34] Hu C,Kong D.Tertiary lymphoid structure-associated B cells enhance CXCL13⁺CD103⁺CD8⁺ tissue-resident memory T-cell response to programmed cell death protein 1 blockade in cancer immunotherapy.Gastroenterology2024;166:1069-84

[35] Gallimore A.Tumor-associated high endothelial venules: inroads enabling immune control of cancer progression.Cancer Immunol Res2022;10:371

[36] Martinet L,Filleron T.Human solid tumors contain high endothelial venules: association with T- and B-lymphocyte infiltration and favorable prognosis in breast cancer.Cancer Res2011;71:5678-87

[37] Duan Q,Zheng J.Turning cold into hot: firing up the tumor microenvironment.Trends Cancer2020;6:605-18

[38] Li Z,Li B.Development and validation of a machine learning model for detection and classification of tertiary lymphoid structures in gastrointestinal cancers.JAMA Netw Open2023;6:e2252553 PMCID:PMC10408275

[39] Li Q,Wang D.Prognostic value of tertiary lymphoid structure and tumour infiltrating lymphocytes in oral squamous cell carcinoma.Int J Oral Sci2020;12:24 PMCID:PMC7493903

[40] Posch F,Leibl S.Maturation of tertiary lymphoid structures and recurrence of stage II and III colorectal cancer.Oncoimmunology2018;7:e1378844 PMCID:PMC5798199

[41] Baxevanis CN,Tsitsilonis OE.Exploring the pivotal functions of tertiary lymphoid structures in cancer prognosis and immunotherapy outcomes.Cancers2025;17:3754 PMCID:PMC12691252

[42] Cheng N,Cheng H.Prognostic value of tumor-infiltrating lymphocytes and tertiary lymphoid structures in epstein-barr virus-associated and -negative gastric carcinoma.Front Immunol2021;12:692859 PMCID:PMC8281029

[43] Ahmed A.Tertiary lymphoid structures in colorectal cancer liver metastases: association with immunological and clinical parameters and chemotherapy response.Anticancer Res2020;40:6367-73

[44] Sofopoulos M,Vaxevanis CK.The prognostic significance of peritumoral tertiary lymphoid structures in breast cancer.Cancer Immunol Immunother2019;68:1733-45 PMCID:PMC11028375

[45] Nakamura S,Hayashi M.Tertiary lymphoid structures correlate with enhancement of antitumor immunity in esophageal squamous cell carcinoma.Br J Cancer2023;129:1314-26 PMCID:PMC10575855

[46] Sun H,Cheng W.The distribution and maturation of tertiary lymphoid structures can predict clinical outcomes of patients with gastric adenocarcinoma.Front Immunol2024;15:1396808 PMCID:PMC11317265

[47] Xu W,Liu WR.Heterogeneity in tertiary lymphoid structures predicts distinct prognosis and immune microenvironment characterizations of clear cell renal cell carcinoma.J Immunother Cancer2023;11:e006667 PMCID:PMC10693897

[48] Ribeiro V,Dieu-Nosjean MC,Rochefort J.The prognostic significance of tertiary lymphoid structures in oral squamous cell carcinomas: a systematic review.Front Oral Health2024;5:1524313 PMCID:PMC11794802

[49] Meylan M,Lacroix L.Early hepatic lesions display immature tertiary lymphoid structures and show elevated expression of immune inhibitory and immunosuppressive molecules.Clin Cancer Res2020;26:4381-9

[50] Chen Y,Li Y.Spatiotemporal single-cell analysis decodes cellular dynamics underlying different responses to immunotherapy in colorectal cancer.Cancer Cell2024;42:1268-85.e7

[51] Wennhold K,Lehmann J.CD86⁺ antigen-presenting B cells are increased in cancer, localize in tertiary lymphoid structures, and induce specific T-cell responses.Cancer Immunol Res2021;9:1098-108

[52] Wang Q,An R.Peritumoral tertiary lymphoid structure and tumor stroma percentage predict the prognosis of patients with non-metastatic colorectal cancer.Front Immunol2022;13:962056 PMCID:PMC9524924

[53] Mao Y,Xi L.Prediction values of tertiary lymphoid structures in the prognosis of patients with left- and right-sided colon cancer: a multicenter propensity score-matched study.Int J Surg2023;109:2344-58 PMCID:PMC10442147

[54] Calderaro J,Becht E.Intra-tumoral tertiary lymphoid structures are associated with a low risk of early recurrence of hepatocellular carcinoma.J Hepatol2019;70:58-65

[55] Wakasu S,Haratake N.Preventive effect of tertiary lymphoid structures on lymph node metastasis of lung adenocarcinoma.Cancer Immunol Immunother2023;72:1823-34 PMCID:PMC10992259

[56] Di Caro G,Grizzi F.Occurrence of tertiary lymphoid tissue is associated with T-cell infiltration and predicts better prognosis in early-stage colorectal cancers.Clin Cancer Res2014;20:2147-58

[57] Maibach F,Seyed Jafari SM,Schenk M.Tumor-infiltrating lymphocytes and their prognostic value in cutaneous melanoma.Front Immunol2020;11:2105 PMCID:PMC7511547

[58] Wang YQ,Zhou W.Integrated analysis of tertiary lymphoid structures and immune infiltration in ccRCC microenvironment revealed their clinical significances: a multicenter cohort study.J Immunother Cancer2024;12:e008613 PMCID:PMC11331356

[59] Johansson-Percival A.Therapeutic induction of tertiary lymphoid structures in cancer through stromal remodeling.Front Immunol2021;12:674375 PMCID:PMC8191417

[60] Zhang C,Zhu J.Application of spatial and single-cell omics in tumor immunotherapy biomarkers.LabMed Discov2025;2:100076

[61] Zhang Y,Zeng Q.CCL19-producing fibroblasts promote tertiary lymphoid structure formation enhancing anti-tumor IgG response in colorectal cancer liver metastasis.Cancer Cell2024;42:1370-85.e9

[62] Yu A,Ma L.The relationship between the tertiary lymphoid structure and immune-infiltrating cells in gastrointestinal cancers: a systematic review and meta-analysis.Immun Inflamm Dis2024;12:e70003 PMCID:PMC11389262

[63] Mustapha R,Monypenny J.Insights into unveiling a potential role of tertiary lymphoid structures in metastasis.Front Mol Biosci2021;8:661516 PMCID:PMC8455920

[64] Karjula T,Mustonen O.Tertiary lymphoid structures in pulmonary metastases of microsatellite stable colorectal cancer.Virchows Arch2023;483:21-32 PMCID:PMC10326136

[65] Schweiger T,Glogner C.Tumor-infiltrating lymphocyte subsets and tertiary lymphoid structures in pulmonary metastases from colorectal cancer.Clin Exp Metastasis2016;33:727-39 PMCID:PMC5035322

[66] Yang R,Li K.Integrated single-cell and spatial transcriptomics reveal the colorectal cancer microenvironment.Crit Rev Oncol Hematol2026;222:105311

[67] Zhan Z,Yi-Ran Z.High endothelial venules proportion in tertiary lymphoid structure is a prognostic marker and correlated with anti-tumor immune microenvironment in colorectal cancer.Ann Med2023;55:114-26 PMCID:PMC9754014

[68] Ling Y,Weng Z.The prognostic value and molecular properties of tertiary lymphoid structures in oesophageal squamous cell carcinoma.Clin Transl Med2022;12:e1074 PMCID:PMC9574489

[69] Ye L,Kang Z.Tumor-infiltrating immune cells act as a marker for prognosis in colorectal cancer.Front Immunol2019;10:2368 PMCID:PMC6811516

[70] Tokunaga R,Sakamoto Y.12-Chemokine signature, a predictor of tumor recurrence in colorectal cancer.Int J Cancer2020;147:532-41 PMCID:PMC9371443

[71] Li X,Liu X,Yang L.A 12-chemokine gene signature is associated with the enhanced immunogram scores and is relevant for precision immunotherapy.Med Oncol2022;39:43

[72] Coppola D,Khalil F.Unique ectopic lymph node-like structures present in human primary colorectal carcinoma are identified by immune gene array profiling.Am J Pathol2011;179:37-45 PMCID:PMC3123872

[73] Weinstein AM.Biosynthesis and functional significance of peripheral node addressin in cancer-associated TLO.Front Immunol2016;7:301 PMCID:PMC4977569

[74] Chaurio RA,Lee Costich T.TGF-β-mediated silencing of genomic organizer SATB1 promotes Tfh cell differentiation and formation of intra-tumoral tertiary lymphoid structures.Immunity2022;55:115-28.e9 PMCID:PMC8852221

[75] Alexander J,Wu TT,Li S.Histopathological identification of colon cancer with microsatellite instability.Am J Pathol2001;158:527-35 PMCID:PMC1850324

[76] Zhang L,Zheng L.Lineage tracking reveals dynamic relationships of T cells in colorectal cancer.Nature2018;564:268-72

[77] Bugatti M,Missale F.A population of TIM4+FOLR2+ macrophages localized in tertiary lymphoid structures correlates to an active immune infiltrate across several cancer types.Cancer Immunol Res2022;10:1340-53

[78] Mei Y,Hu H.Single-cell analyses reveal suppressive tumor microenvironment of human colorectal cancer.Clin Transl Med2021;11:e422 PMCID:PMC8181206

[79] Becht E,Giraldo NA.Immune and stromal classification of colorectal cancer is associated with molecular subtypes and relevant for precision immunotherapy.Clin Cancer Res2016;22:4057-66

[80] Kim JH,Lee JA.Genomic and transcriptomic characterization of heterogeneous immune subgroups of microsatellite instability-high colorectal cancers.J Immunother Cancer2021;9:e003414 PMCID:PMC8672019

[81] Wang H,Zhang Y.Subtyping of microsatellite stability colorectal cancer reveals guanylate binding protein 2 (GBP2) as a potential immunotherapeutic target.J Immunother Cancer2022;10:e004302 PMCID:PMC8984016

[82] Masuda K,Miller J.Multiplexed single-cell analysis reveals prognostic and nonprognostic T cell types in human colorectal cancer.JCI Insight2022;7:e154646 PMCID:PMC9057629

[83] Mlecnik B,Angell HK.Integrative analyses of colorectal cancer show immunoscore is a stronger predictor of patient survival than microsatellite instability.Immunity2016;44:698-711

[84] Xu J,Jiang K,Yuan Y.Combining single-cell sequencing to identify key immune genes and construct the prognostic evaluation model for colon cancer patients.Clin Transl Med2021;11:e465 PMCID:PMC8288000

[85] Yang Y,Zhou H,Wu S.Integrative single-cell and spatial transcriptomics reveals the crosstalk of CTHRC1+ CAF and MMP7+ epithelial axis as a potential therapeutic target and predicts poor clinical outcomes in colorectal cancer.Mediators Inflamm2026;2026:e9314553 PMCID:PMC13061587

[86] Jia H,Wang G.Spatial and single-cell transcriptomic analysis reveals fibroblasts dependent immune environment in colorectal cancer.Biofactors2025;51:e70012

[87] Bhatt D,Sawant D.STARTRAC analyses of scRNAseq data from tumor models reveal T cell dynamics and therapeutic targets.J Exp Med2021;218:e20201329 PMCID:PMC8077174

[88] Wang M,Xu K.T cell exhaustion and dendritic cell-mediated tertiary lymphoid structures (TLSs) modulation affect response to neoadjuvant chemoradiotherapy in microsatellite stable rectal cancer.Adv Sci2026;13:e14332 PMCID:PMC12822459

[89] Kang J,Cha H.Systematic dissection of tumor-normal single-cell ecosystems across a thousand tumors of 30 cancer types.Nat Commun2024;15:4067 PMCID:PMC11094150

[90] Xu Z,Zhang Y.Exploiting tertiary lymphoid structures gene signature to evaluate tumor microenvironment infiltration and immunotherapy response in colorectal cancer.Front Oncol2024;14:1383096 PMCID:PMC11153738

[91] Liu X,Zhang Y.Multi-algorithm-integrated tertiary lymphoid structure gene signature for immune landscape characterization and prognosis in colorectal cancer patients.Biomedicines2024;12:2644 PMCID:PMC11592260

[92] Lu H,Wengert G.Tumor and local lymphoid tissue interaction determines prognosis in high-grade serous ovarian cancer.Cell Rep Med2023;4:101092 PMCID:PMC10394173

[93] Leko V.Identifying and targeting human tumor antigens for T cell-based immunotherapy of solid tumors.Cancer Cell2020;38:454-72 PMCID:PMC7737225

[94] Kalaora S,Nejman D.Identification of bacteria-derived HLA-bound peptides in melanoma.Nature2021;592:138-43 PMCID:PMC9717498

[95] Pelka K,Chen JH.Spatially organized multicellular immune hubs in human colorectal cancer.Cell2021;184:4734-52.e20 PMCID:PMC8772395

[96] Kinker GS,Diniz AB.Mature tertiary lymphoid structures are key niches of tumour-specific immune responses in pancreatic ductal adenocarcinomas.Gut2023;72:1927-41

[97] Yoshitomi H.Shared and distinct roles of T peripheral helper and T follicular helper cells in human diseases.Cell Mol Immunol2021;18:523-7 PMCID:PMC8027819

[98] Cui C,Joshi NS.T follicular helper cells in cancer, tertiary lymphoid structures, and beyond.Semin Immunol2023;69:101797

[99] Crotty S.T follicular helper cell biology: a decade of discovery and diseases.Immunity2019;50:1132-48 PMCID:PMC6532429

[100] Bindea G,Tosolini M.Spatiotemporal dynamics of intratumoral immune cells reveal the immune landscape in human cancer.Immunity2013;39:782-95

[101] Rodriguez AB,Woods AN.Immune mechanisms orchestrate tertiary lymphoid structures in tumors via cancer-associated fibroblasts.Cell Rep2021;36:109422 PMCID:PMC8362934

[102] O’Connor RA,Koppensteiner L,Akram AR.Cancer-associated fibroblasts drive CXCL13 production in activated T cells via TGF-beta.Front Immunol2023;14:1221532 PMCID:PMC10373066

[103] Horeweg N, Workel HH, Loiero D, et al.; TransPORTEC consortium. Tertiary lymphoid structures critical for prognosis in endometrial cancer patients.Nat Commun2022;13:1373 PMCID:PMC8927106

[104] Thommen DS,Herzig P.A transcriptionally and functionally distinct PD-1⁺ CD8⁺ T cell pool with predictive potential in non-small-cell lung cancer treated with PD-1 blockade.Nat Med2018;24:994-1004 PMCID:PMC6110381

[105] Workel HH,Arnold R.A transcriptionally distinct CXCL13⁺CD103⁺CD8⁺ T-cell population is associated with B-cell recruitment and neoantigen load in human cancer.Cancer Immunol Res2019;7:784-96

[106] Remark R,Cremer I.Characteristics and clinical impacts of the immune environments in colorectal and renal cell carcinoma lung metastases: influence of tumor origin.Clin Cancer Res2013;19:4079-91

[107] van de Pavert SA,Goverse G.Chemokine CXCL13 is essential for lymph node initiation and is induced by retinoic acid and neuronal stimulation.Nat Immunol2009;10:1193-9 PMCID:PMC2771164

[108] Borelli A.Lymphotoxin: from the physiology to the regeneration of the thymic function.Cell Death Differ2021;28:2305-14 PMCID:PMC8329281

[109] Ansel KM,Hyman PL.A chemokine-driven positive feedback loop organizes lymphoid follicles.Nature2000;406:309-14

[110] Barone F,Nayar S,Buckley CD.Stromal fibroblasts in tertiary lymphoid structures: a novel target in chronic inflammation.Front Immunol2016;7:477 PMCID:PMC5100680

[111] Nayar S,Smith CG.Immunofibroblasts are pivotal drivers of tertiary lymphoid structure formation and local pathology.Proc Natl Acad Sci U S A2019;116:13490-7 PMCID:PMC6613169

[112] de Chaisemartin L,Damotte D.Characterization of chemokines and adhesion molecules associated with T cell presence in tertiary lymphoid structures in human lung cancer.Cancer Res2011;71:6391-9

[113] Nayar S,Chung MM.Bimodal expansion of the lymphatic vessels is regulated by the sequential expression of IL-7 and lymphotoxin α1β2 in newly formed tertiary lymphoid structures.J Immunol2016;197:1957-67 PMCID:PMC4991245

[114] Drayton DL,Lee J,Ruddle NH.Ectopic LT alpha beta directs lymphoid organ neogenesis with concomitant expression of peripheral node addressin and a HEV-restricted sulfotransferase.J Exp Med2003;197:1153-63 PMCID:PMC2193975

[115] Mempel TR,Altenburger LM.How chemokines organize the tumour microenvironment.Nat Rev Cancer2024;24:28-50 PMCID:PMC11480775

[116] Asrir A,Coudert J.Tumor-associated high endothelial venules mediate lymphocyte entry into tumors and predict response to PD-1 plus CTLA-4 combination immunotherapy.Cancer Cell2022;40:318-34.e9

[117] Yang M,Li K.Detection and quantitative analysis of tumor-associated tertiary lymphoid structures.J Zhejiang Univ Sci B2023;24:779-95 PMCID:PMC10500099

[118] Vinuesa CG.How T cells earn the follicular rite of passage.Immunity2011;35:671-80

[119] Yeh CH,Okada T,Kelsoe G.Primary germinal center-resident T follicular helper cells are a physiologically distinct subset of CXCR5^hiPD-1^hi T follicular helper cells.Immunity2022;55:272-89.e7 PMCID:PMC8842852

[120] Yuan H,Yan Y.Single-cell sequencing reveals the heterogeneity of B cells and tertiary lymphoid structures in muscle-invasive bladder cancer.J Transl Med2024;22:48 PMCID:PMC10787393

[121] Zhang Q.Tertiary lymphoid structures are critical for cancer prognosis and therapeutic response.Front Immunol2022;13:1063711 PMCID:PMC9875059

[122] Cha HR,Kim ED.Mucosa-associated epithelial chemokine/CCL28 expression in the uterus attracts CCR10+ IgA plasma cells following mucosal vaccination via estrogen control.J Immunol2011;187:3044-52

[123] Xiong N,Hu S,Yang J.CCR10 and its ligands in regulation of epithelial immunity and diseases.Protein Cell2012;3:571-80 PMCID:PMC4430102

[124] Mergia Terefe E,Rakhshani A.Roles of CCR10/CCL27-CCL28 axis in tumour development: mechanisms, diagnostic and therapeutic approaches, and perspectives.Expert Rev Mol Med2022;24:e37

[125] deLuca L, Ng D, Gao Y, Wortzman ME, Watts TH, Gommerman JL. LTβR signaling in dendritic cells induces a type I IFN response that is required for optimal clonal expansion of CD8+ T cells.Proc Natl Acad Sci U S A2011;108:2046-51 PMCID:PMC3033245

[126] Summers-DeLuca LE,Cosovic B.Expression of lymphotoxin-alphabeta on antigen-specific T cells is required for DC function.J Exp Med2007;204:1071-81 PMCID:PMC2118582

[127] Gommerman JL,Ware CF.The Lymphotoxin Network: orchestrating a type I interferon response to optimize adaptive immunity.Cytokine Growth Factor Rev2014;25:139-45 PMCID:PMC4081445

[128] León B,Browning JL,Randall TD.Regulation of T(H)2 development by CXCR5+ dendritic cells and lymphotoxin-expressing B cells.Nat Immunol2012;13:681-90 PMCID:PMC3548431

[129] Hashimoto M,Honma R.A single center study of genes involved in synchronous and metachronous multiple early-stage gastric cancers in Japanese patients with current or former helicobacter pylori infection.Cancers2025;17:464 PMCID:PMC11815868

[130] Ji Y,Yin X.Multi-omics profiling of sodium-overload (NECSO) programs identifies NEK8 as a central driver of colorectal cancer progression through single-cell and spatial transcriptomics.Front Immunol2026;17:1765055 PMCID:PMC12929418

[131] Shaul ME,Tidhar E.Tumor-associated neutrophils drive B-cell recruitment and their differentiation to plasma cells.Cancer Immunol Res2021;9:811-24

[132] Liu W,Xu HC.BAFF attenuates immunosuppressive monocytes in the melanoma tumor microenvironment.Cancer Res2022;82:264-77 PMCID:PMC9397630

[133] Johansson-Percival A,Li ZJ.De novo induction of intratumoral lymphoid structures and vessel normalization enhances immunotherapy in resistant tumors.Nat Immunol2017;18:1207-17

[134] Schmidleithner L.Tfh cells induce intratumoral tertiary lymphoid structures.Trends Immunol2022;43:274-6

[135] Furtado GC,Bongers G.TNFα-dependent development of lymphoid tissue in the absence of RORγt⁺ lymphoid tissue inducer cells.Mucosal Immunol2014;7:602-14 PMCID:PMC4264842

[136] Peske JD,Gemta L,Fu YX.Effector lymphocyte-induced lymph node-like vasculature enables naive T-cell entry into tumours and enhanced anti-tumour immunity.Nat Commun2015;6:7114 PMCID:PMC4435831

[137] Guedj K,Clement M.M1 macrophages act as LTβR-independent lymphoid tissue inducer cells during atherosclerosis-related lymphoid neogenesis.Cardiovasc Res2014;101:434-43

[138] Liu X,Huang D.The predictive role of tertiary lymphoid structures in the prognosis and response to immunotherapy of lung cancer patients: a systematic review and meta-analysis.BMC Cancer2025;25:87 PMCID:PMC11734324

[139] Franciszkiewicz K,Jalil A.Intratumoral induction of CD103 triggers tumor-specific CTL function and CCR5-dependent T-cell retention.Cancer Res2009;69:6249-55

[140] Franciszkiewicz K,Boutet M,Schmitt A.CD103 or LFA-1 engagement at the immune synapse between cytotoxic T cells and tumor cells promotes maturation and regulates T-cell effector functions.Cancer Res2013;73:617-28

[141] Qian J,Wu Q.Nanosecond pulsed electric field stimulates CD103⁺ DC accumulation in tumor microenvironment via NK-CD103⁺ DC crosstalk.Cancer Lett2024;593:216514

[142] Konkel JE,Carpenter AC.Control of the development of CD8αα+ intestinal intraepithelial lymphocytes by TGF-β.Nat Immunol2011;12:312-9 PMCID:PMC3062738

[143] Han J,Jin W.TGF-β controls development of TCRγδ⁺CD8αα⁺ intestinal intraepithelial lymphocytes.Cell Discov2023;9:52 PMCID:PMC10229584

[144] Vella G,Bergers G.High endothelial venules: a vascular perspective on tertiary lymphoid structures in cancer.Front Immunol2021;12:736670 PMCID:PMC8416033

[145] Hiraoka N,Yamazaki-Itoh R,Kosuge T.Intratumoral tertiary lymphoid organ is a favourable prognosticator in patients with pancreatic cancer.Br J Cancer2015;112:1782-90 PMCID:PMC4647237

[146] Arasa J,Halin C.Structure and immune function of afferent lymphatics and their mechanistic contribution to dendritic cell and T cell trafficking.Cells2021;10:1269 PMCID:PMC8161367

[147] Girard JP,Förster R.HEVs, lymphatics and homeostatic immune cell trafficking in lymph nodes.Nat Rev Immunol2012;12:762-73

[148] Gunn MD,Tam C,Rosen SD.A chemokine expressed in lymphoid high endothelial venules promotes the adhesion and chemotaxis of naive T lymphocytes.Proc Natl Acad Sci U S A1998;95:258-63 PMCID:PMC18193

[149] Ager A.High endothelial venules and other blood vessels: critical regulators of lymphoid organ development and function.Front Immunol2017;8:45 PMCID:PMC5289948

[150] Held W,Petrova TV.Stem-cell-like T cells have a specific entry gate to the tumor.Cancer Cell2022;40:243-5

[151] Blanchard L.High endothelial venules (HEVs) in immunity, inflammation and cancer.Angiogenesis2021;24:719-53 PMCID:PMC8487881

[152] Sawada J,Qi R.Molecular signature of tumor-associated high endothelial venules that can predict breast cancer survival.Cancer Immunol Res2022;10:468-81 PMCID:PMC8976767

[153] Chen H,Cui Y,Li B.High endothelial venules in intracranial germinomas: implications for lymphocytes infiltration.Cancer Med2023;12:5450-60 PMCID:PMC10028053

[154] Overacre-Delgoffe AE,Cillo AR.Microbiota-specific T follicular helper cells drive tertiary lymphoid structures and anti-tumor immunity against colorectal cancer.Immunity2021;54:2812-24.e4 PMCID:PMC8865366

[155] Honda K.The microbiota in adaptive immune homeostasis and disease.Nature2016;535:75-84

[156] Li Z,Liang Z.Critical role of the gut microbiota in immune responses and cancer immunotherapy.J Hematol Oncol2024;17:33 PMCID:PMC11094969

[157] Routy B,Derosa L.Gut microbiome influences efficacy of PD-1-based immunotherapy against epithelial tumors.Science2018;359:91-7

[158] Xu X,Guo F.Gut microbiome influences the efficacy of PD-1 antibody immunotherapy on MSS-type colorectal cancer via metabolic pathway.Front Microbiol2020;11:814 PMCID:PMC7212380

[159] Lynch KT,Meneveau MO.Heterogeneity in tertiary lymphoid structure B-cells correlates with patient survival in metastatic melanoma.J Immunother Cancer2021;9:e002273 PMCID:PMC8190052

[160] Lee SK,Cha PH.A small molecule approach to degrade RAS with EGFR repression is a potential therapy for KRAS mutation-driven colorectal cancer resistance to cetuximab.Exp Mol Med2018;50:1-12 PMCID:PMC6244225

[161] Yang F,Huang X.Interactions between Salmonella and host macrophages - dissecting NF-κB signaling pathway responses.Microb Pathog2021;154:104846

[162] Cao Y,Li T,Li C.Review of metabolomics and microbiomics in esophageal cancer: from pathogenesis to prognosis, LabMed Discov 2025;2:100045.

[163] Liu Z,Tang X,He Y.Intratumoral tertiary lymphoid structures promote patient survival and immunotherapy response in head neck squamous cell carcinoma.Cancer Immunol Immunother2023;72:1505-21 PMCID:PMC10198854

[164] Lei JX,Hu C.Deciphering tertiary lymphoid structure heterogeneity reveals prognostic signature and therapeutic potentials for colorectal cancer: a multicenter retrospective cohort study.Int J Surg2024;110:5627-40 PMCID:PMC11392219

[165] Tian N,Lv Y.Mature tertiary lymphoid structures support B cell-mediated antitumour immunity and are disrupted by neoadjuvant therapy in rectal cancer: a multicentre, retrospective study.EBioMedicine2025;122:106030 PMCID:PMC12670569

[166] Qin C,Ma J.A multi-omic analysis reveals a predictive value of tertiary lymphoid structures in improving the prognosis of colorectal cancer patients with BRAF mutation.Front Immunol2025;16:1662573 PMCID:PMC12434122

[167] Amodio V,Arcella P.EGFR blockade reverts resistance to KRAS^G12C inhibition in colorectal cancer.Cancer Discov2020;10:1129-39 PMCID:PMC7416460

[168] Krauß D,Adachi-Fernandez E.EGFR controls transcriptional and metabolic rewiring in KRAS^G12D colorectal cancer.EMBO Mol Med2025;17:1355-92 PMCID:PMC12162862

[169] Hsu WH,Hsieh MS.The prognostic role of tertiary lymphoid structures and immune microenvironment signatures in early-stage EGFR-mutant lung adenocarcinoma.Cancers2025;17:2379 PMCID:PMC12293609

[170] Xie M,Ma X.The radiological characteristics, tertiary lymphoid structures, and survival status associated with EGFR mutation in patients with subsolid nodules like stage I-II LUAD.BMC Cancer2024;24:372 PMCID:PMC10962174

[171] Chen Z,Cai K.Dynamics of tertiary lymphoid structures and immune cross talk in early versus advanced colorectal cancer: potential implications for immunotherapy.Cancer Immunol Immunother2025;74:185 PMCID:PMC12033131

[172] Lanickova T,Kasikova L.Chemotherapy drives tertiary lymphoid structures that correlate with ICI-responsive TCF1+CD8+ T cells in metastatic ovarian cancer.Clin Cancer Res2025;31:164-80 PMCID:PMC11701433

[173] Ángeles Gil González MDL,Sierra-Rodero B.734 Tertiary lymphoid structures as hallmarks of response to neoadjuvant chemoimmunotherapy in locally advanced non-small cell lung cancer NADIM trial.J ImmunoTher Cancer2025;13:

[174] Zhai K,Ru K.Tertiary lymphoid structures correlate with the therapeutic efficacy and prognosis of resectable esophageal squamous cell carcinoma undergoing neoadjuvant chemoradiotherapy plus immunotherapy.Front Immunol2025;16:1616247 PMCID:PMC12411778

[175] Zhao K,Si Y,Huang Z.TIGIT blockade enhances tumor response to radiotherapy via a CD103 + dendritic cell-dependent mechanism.Cancer Immunol Immunother2023;72:193-209 PMCID:PMC9813151

[176] Hosoi N,Dorjkhorloo G.Mature tertiary lymphoid structures indicate good chemotherapy response and prognosis in advanced colorectal cancer.Ann Gastroenterol Surg2026;10:710-21 PMCID:PMC13178286

[177] Guo Z,Jia W.Tertiary lymphoid structures signature predicts prognosis and clinical benefits from neoadjuvant chemotherapy and PD-1 blockade in colorectal adenocarcinoma.Int J Surg2025;111:5088-104

[178] Zhao Z,Lin ZB.Relationship between tertiary lymphoid structure and the prognosis and clinicopathologic characteristics in solid tumors.Int J Med Sci2021;18:2327-38 PMCID:PMC8100653

[179] Gunderson AJ,Bui C.Germinal center reactions in tertiary lymphoid structures associate with neoantigen burden, humoral immunity and long-term survivorship in pancreatic cancer.Oncoimmunology2021;10:1900635 PMCID:PMC7993148

[180] Onyshchenko K,Rao X.Hypofractionated radiotherapy combined with lenalidomide improves systemic antitumor activity in mouse solid tumor models.Theranostics2024;14:2573-88 PMCID:PMC11024858

[181] Petroni G,Antonuzzo L.Exploiting tertiary lymphoid structures to stimulate antitumor immunity and improve immunotherapy efficacy.Cancer Res2024;84:1199-209 PMCID:PMC11016894

[182] Oba T,Kajihara R.In situ delivery of iPSC-derived dendritic cells with local radiotherapy generates systemic antitumor immunity and potentiates PD-L1 blockade in preclinical poorly immunogenic tumor models.J Immunother Cancer2021;9:e002432 PMCID:PMC8166607

[183] Guo H,Mao D.Homologous-magnetic dual-targeted metal-organic framework to improve the anti-hepatocellular carcinoma efficacy of PD-1 inhibitor.J Nanobiotechnology2024;22:206 PMCID:PMC11044376

[184] Bai X,Yokota Y.Adaptive antitumor immune response stimulated by bio-nanoparticle based vaccine and checkpoint blockade.J Exp Clin Cancer Res2022;41:132 PMCID:PMC8991500

[185] He A,Dai Z.Nanovaccine-based strategies for lymph node targeted delivery and imaging in tumor immunotherapy.J Nanobiotechnology2023;21:236 PMCID:PMC10364424

[186] Li Y,Jiang Z.Targeting lymph node delivery with nanovaccines for cancer immunotherapy: recent advances and future directions.J Nanobiotechnology2023;21:212 PMCID:PMC10327386

[187] Zhang Y,Fei Z.3D printing scaffold vaccine for antitumor immunity.Adv Mater2021;33:e2106768

[188] Xie D,Chen C.A scaffold vaccine to promote tumor antigen cross-presentation via sustained toll-like receptor-2 (TLR2) activation.Bioact Mater2024;37:315-30 PMCID:PMC11061615

[189] Sinha A,Nguyen MH,Choi SW.A 3D macroporous alginate graphene scaffold with an extremely slow release of a loaded cargo for in situ long-term activation of dendritic cells.Adv Healthc Mater2019;8:e1800571

[190] Chelvanambi M,Taylor JL.STING agonist-based treatment promotes vascular normalization and tertiary lymphoid structure formation in the therapeutic melanoma microenvironment.J Immunother Cancer2021;9:e001906 PMCID:PMC7852948

[191] Lemos H,McCardle C.Overcoming resistance to STING agonist therapy to incite durable protective antitumor immunity.J Immunother Cancer2020;8:e001182 PMCID:PMC7451475

[192] Jin XK,Zhang SM.Engineering metal-based hydrogel-mediated tertiary lymphoid structure formation via activation of the STING pathway for enhanced immunotherapy.Mater Horiz2023;10:4365-79

[193] Zhang S,Wang K.Chitosan-based nano-micelles for potential anti-tumor immunotherapy: synergistic effect of cGAS-STING signaling pathway activation and tumor antigen absorption.Carbohydr Polym2023;321:121346

[194] Hu HG,Zhang BD,Li YM.Pam₃CSK₄-CDG^SF augments antitumor immunotherapy by synergistically activating TLR1/2 and STING.Bioconjug Chem2020;31:2499-503

[195] Lorkowski ME,Bielecki PA.Immunostimulatory nanoparticle incorporating two immune agonists for the treatment of pancreatic tumors.J Control Release2021;330:1095-105 PMCID:PMC7906920

[196] A phase I/II study of Regorafenib plus Avelumab in solid tumors (REGOMUNE). ClinicalTrials.gov ID: NCT03475953. 2023. Available from: https://clinicaltrials.gov/study/NCT03475953?cond=tertiary%20lymphoid%20structure&limit=25&page=1&rank=12. [Last accessed on 21 Jul 2026]

[197] Cousin S,Guegan JP.Regorafenib-avelumab combination in patients with biliary tract cancer (REGOMUNE): a single-arm, open-label, phase II trial.Eur J Cancer2022;162:161-9

[198] Targeting MDMD and PD1 in tumors with tertiary lymphoid structures (EMPIRE). ClinicalTrials.gov ID: NCT06084689. More Information. 2025. Available from: https://clinicaltrials.gov/study/NCT06084689?cond=tertiary%20lymphoid%20structure&limit=25&page=1&rank=2#more-information. [Last accessed on 21 Jul 2026]

[199] Combining epigenetic and immune therapy to beat cancer. (CAIRE). ClinicalTrials.gov ID: NCT04705818. Collaborators and Investigators. 2026. Available from: https://clinicaltrials.gov/study/NCT04705818?cond=tertiary%20lymphoid%20structure&limit=25&page=1&rank=11#collaborators-and-investigators. [Last accessed on 21 Jul 2026]

[200] MEDI5752 in patients with mature tertiary lymphoid structures solid tumors. (TAYLOR). ClinicalTrials.gov ID: NCT05888857. Study Plan. 2025. Available from: https://clinicaltrials.gov/study/NCT05888857?cond=tertiary%20lymphoid%20structure&limit=25&page=1&rank=1#study-plan. [Last accessed on 21 Jul 2026]

[201] Ogino S,Ugai S,Ugai T.Unique role of the prospective cohort incident-tumor biobank method in etiological research.Lancet Reg Health Am2026;53:101290 PMCID:PMC12926294

[202] Ogino S,Hamada T.The early-onset cancer epidemics: evidence synthesis using the prospective cohort incident-tumor biobank method.Eur J Epidemiol2025;40:1405-17

[203] Hamada T,Gurjao C.Smoking habit and long-term colorectal cancer incidence by exome-wide mutational and neoantigen loads: evidence based on the prospective cohort incident-tumour biobank method.BMJ Oncol2025;4:e000787 PMCID:PMC12164326