Assessment of baseline CCL19+ dendritic cell infiltration for predicting responses to immunotherapy in lung adenocarcinoma patients

doi:10.12122/j.issn.1673-4254.2024.08.11

Abstract

Abstract:

Objective To explore the correlation of baseline CCL19⁺ dendritic cell (CCL19⁺ DC) infiltration in lung adenocarcinoma microenvironment with immunotherapy efficacy and CD8⁺ T cell infiltration. Methods We retrospectively analyzed the data of patients with lung adenocarcinoma hospitalized at First Affiliated Hospital of Henan University of Science and Technology from January, 2020 to December, 2023, and collected tissue samples from 96 patients undergoing immunotherapy for assessing CCL19⁺ DC and CD8⁺ T cell infiltration using immunofluorescence assay. We evaluated the predictive value of baseline CCL19⁺ DCs for patient responses to immunotherapy using receiver-operating characteristics (ROC) curves and analyzed the correlations of baseline CCL19⁺ DC expression with immunotherapy efficacy and CD8⁺ T cell and cytotoxic T lymphocyte (CTL) infiltrations. In co-culture systems of lung adenocarcinoma PC9 cells, CD8⁺ T cells and DCs (overexpressing CCL19 with or without anti PD-1 antibody treatment), the expressions of granzyme B, perforin, IFN-γ, and Ki-67 in T cells were analyzed using flow cytometry. Results The patients with partial or complete remission following immunotherapy had a significantly higher baseline CCL19⁺ DC infiltration level in lung adenocarcinoma tissues than those with poor responses. CCL19⁺ DC infiltration had an area under ROC curve of 0.785, a sensitivity of 75.6%, and a specificity of 62.8% for predicting immunotherapy efficacy. The expression of CD8⁺ T cell surface molecules Granzyme B (P<0.01), Perforin (P<0.01), IFN-γ (P<0.01) and Ki-67 (P<0.001) in patients with high expression of CCL19⁺ DC were higher than those in patients with low expression of CCL19⁺ DC. The baseline CCL19⁺ DC infiltration level was positively correlated with immunotherapy efficacy (P=0.003), CTL infiltration of (r=0.6657, P<0.001) and CD8⁺ T cell infiltration (P=0.007). In the co-cultured cells, CCL19 overexpression combined with anti-PD1 treatment of the DCs more strongly enhanced cytotoxicity and proliferation of CD8⁺ T lymphocytes than either of the single treatments (P<0.01 or 0.001). Conclusion The baseline CCL19⁺ DC infiltration level in lung adenocarcinoma microenvironment is positively correlated with immunotherapy efficacy and CTL infiltration and can thus predict the response to immunotherapy.

Key words: lung adenocarcinoma, CCL19⁺ dendritic cells, immunotherapy, CD8⁺ T cells

Mingyang ZHU, Bokang WANG, Xiusen ZHANG, Kexu ZHOU, Zeyu MIAO, Jiangtao SUN. Assessment of baseline CCL19⁺ dendritic cell infiltration for predicting responses to immunotherapy in lung adenocarcinoma patients[J]. Journal of Southern Medical University, 2024, 44(8): 1529-1536.

Figures/Tables 6

Tab.1 Correlation between baseline expression of CCL19+ DCs and immunotherapy efficacy in patients with lung adenocarcinoma

Patient type

CCL19⁺DC Low expression

CCL19⁺DC High expression

8.663

Fig. 2 Expression of CD8+ T cell surface molecules granzyme B, perforin, IFN-γ, and Ki-67 in lung adenocarcinoma tissues of patients with high and low levels of CCL19+ DC infiltration. A: Immunofluorescence images of CD8+ T surface molecule expressions in each group (scale bar=10 μm, 5 μm). B: Statistical graph of CD8+ T surface molecule expressions. **P<0.01, ***P<0.001.

Tab.2 Correlation between baseline expression of CCL19+ DCs and CD8+ T cell infiltration

Patient	n	CD8⁺ T cell desert type	CD8⁺ T cell rich type	χ²	P
CCL19⁺DC high expression	53	21	32	7.362	0.007
CCL19⁺DC low expression	43	29	14	7.362	0.007

Fig.3 Infiltration of CTL in different lung adenocarcinoma tissues.

Fig.4 Western blotting of CCL19. A: Expressions of CCL19 protein in different groups detected by Western blotting. B: Statistical analysis of CCL19 expressions. ***P<0.001.

Fig.5 Effect of different treatments on the expression of surface molecules of CD8+ T cells in the co-culture systems. A: Flow cytometry of surface molecule expressions on CD8+ T cells. B: Statistical analysis of surface molecule expression on CD8+ T cells. **P<0.01, ***P<0.001.

References 32

1	Denisenko TV, Budkevich IN, Zhivotovsky B. Cell death-based treatment of lung adenocarcinoma[J]. Cell Death Dis, 2018, 9(2): 117.
2	Garon EB, Hellmann MD, Rizvi NA, et al. Five-year overall survival for patients with advanced non-small-cell lung cancer treated with pembrolizumab: results from the phase I KEYNOTE-001 study[J]. J Clin Oncol, 2019, 37(28): 2518-27.
3	Zhang YY, Zhang ZM. The history and advances in cancer immunotherapy: understanding the characteristics of tumor-infiltrating immune cells and their therapeutic implications[J]. Cell Mol Immunol, 2020, 17(8): 807-21.
4	Yi M, Jiao DC, Xu HX, et al. Biomarkers for predicting efficacy of PD-1/PD-L1 inhibitors[J]. Mol Cancer, 2018, 17(1): 129.
5	Havel JJ, Chowell D, Chan TA. The evolving landscape of biomarkers for checkpoint inhibitor immunotherapy[J]. Nat Rev Cancer, 2019, 19(3): 133-50.
6	Hinshaw DC, Shevde LA. The tumor microenvironment innately modulates cancer progression[J]. Cancer Res, 2019, 79(18): 4557-66.
7	Wculek SK, Cueto FJ, Mujal AM, et al. Dendritic cells in cancer immunology and immunotherapy[J]. Nat Rev Immunol, 2020, 20(1): 7-24.
8	Gardner A, Ruffell B. Dendritic cells and cancer immunity[J]. Trends Immunol, 2016, 37(12): 855-65.
9	Veglia F, Gabrilovich DI. Dendritic cells in cancer: the role revisited[J]. Curr Opin Immunol, 2017, 45: 43-51.
10	Cancel JC, Crozat K, Dalod M, et al. Are conventional type 1 dendritic cells critical for protective antitumor immunity and how?[J]. Front Immunol, 2019, 10: 9.
11	Böttcher JP, Sousa CRE. The role of type 1 conventional dendritic cells in cancer immunity[J]. Trends Cancer, 2018, 4(11): 784-92.
12	Gunn MD, Kyuwa S, Tam C, et al. Mice lacking expression of secondary lymphoid organ chemokine have defects in lymphocyte homing and dendritic cell localization[J]. J Exp Med, 1999, 189(3): 451-60.
13	Viola A, Sarukhan A, Bronte V, et al. The pros and cons of chemokines in tumor immunology[J]. Trends Immunol, 2012, 33(10): 496-504.
14	Nagarsheth N, Wicha MS, Zou WP. Chemokines in the cancer microenvironment and their relevance in cancer immunotherapy[J]. Nat Rev Immunol, 2017, 17(9): 559-72.
15	Gu Q, Zhou SF, Chen C, et al. CCL19: a novel prognostic chemokine modulates the tumor immune microenvironment and outcomes of cancers[J]. Aging, 2023, 15(21): 12369-87.
16	Maier B, Leader AM, Chen ST, et al. A conserved dendritic-cell regulatory program limits antitumour immunity[J]. Nature, 2020, 580(7802): 257-62.
17	Smalley I, Chen ZH, Phadke M, et al. Single-cell characterization of the immune microenvironment of melanoma brain and lepto-meningeal metastases[J]. Clin Cancer Res, 2021, 27(14): 4109-25.
18	Chen YP, Yin JH, Li WF, et al. Single-cell transcriptomics reveals regulators underlying immune cell diversity and immune subtypes associated with prognosis in nasopharyngeal carcinoma[J]. Cell Res, 2020, 30(11): 1024-42.
19	Guo QQ, Liu LW, Chen ZL, et al. Current treatments for non-small cell lung cancer[J]. Front Oncol, 2022, 12: 945102.
20	Doroshow DB, Sanmamed MF, Hastings K, et al. Immunotherapy in non-small cell lung cancer: facts and hopes[J]. Clin Cancer Res, 2019, 25(15): 4592-602.
21	Collin M, Bigley V. Human dendritic cell subsets: an update[J]. Immunology, 2018, 154(1): 3-20.
22	Garris CS, Arlauckas SP, Kohler RH, et al. Successful anti-PD-1 cancer immunotherapy requires T cell-dendritic cell crosstalk involving the cytokines IFN-γ and IL-12[J]. Immunity, 2022, 55(9): 1749.
23	Salmon H, Idoyaga J, Rahman A, et al. Expansion and activation of CD103⁺ dendritic cell progenitors at the tumor site enhances tumor responses to therapeutic PD-L1 and BRAF inhibition[J]. Immunity, 2016, 44(4): 924-38.
24	Gowhari Shabgah A, Haleem Al-Qaim Z, Markov A, et al. Chemokine CXCL14; a double-edged sword in cancer development[J]. Int Immunopharmacol, 2021, 97: 107681.
25	Gowhari Shabgah A, Qasim MT, Mojtaba Mostafavi S, et al. CXC chemokine ligand 16: a Swiss army knife chemokine in cancer[J]. Expert Rev Mol Med, 2021, 23: e4.
26	Artinger M, Matti C, Gerken OJ, et al. A versatile toolkit for semi-automated production of fluorescent chemokines to study CCR7 expression and functions[J]. Int J Mol Sci, 2021, 22(8): 4158.
27	Hillinger S, Yang SC, Zhu L, et al. EBV-induced molecule 1 ligand chemokine (ELC/CCL19) promotes IFN-gamma-dependent antitumor responses in a lung cancer model[J]. J Immunol, 2003, 171(12): 6457-65.
28	Lu J, Ma JJ, Cai W, et al. CC motif chemokine ligand 19 suppressed colorectal cancer in vivo accompanied by an increase in IL-12 and IFN-γ[J]. Biomedecine Pharmacother, 2015, 69: 374-9.
29	Liu XX, Wang BL, Li YY, et al. Powerful anticolon tumor effect of targeted gene immunotherapy using folate-modified nanoparticle delivery of CCL19 to activate the immune system[J]. ACS Cent Sci, 2019, 5(2): 277-89.
30	Cheng HW, Onder L, Cupovic J, et al. CCL19-producing fibroblastic stromal cells restrain lung carcinoma growth by promoting local antitumor T-cell responses[J]. J Allergy Clin Immunol, 2018, 142(4): 1257-71.e4.
31	Iida Y, Yoshikawa R, Murata A, et al. Local injection of CCL19-expressing mesenchymal stem cells augments the therapeutic efficacy of anti-PD-L1 antibody by promoting infiltration of immune cells[J]. J Immunother Cancer, 2020, 8(2): e000582.
32	Li P, Liu FY, Sun LY, et al. Chemokine receptor 7 promotes cell migration and adhesion in metastatic squamous cell carcinoma of the head and neck by activating integrin αvβ3[J]. Int J Mol Med, 2011, 27(5): 679-87.

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