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Long-Term Survival for Patients With Non–Small-Cell Lung Cancer With Intratumoral Lymphoid Structures

Marie-Caroline Dieu-Nosjean, Martine Antoine, Claire Danel, Didier Heudes, Marie Wislez, Virginie Poulot, Nathalie Rabbe, Ludivine Laurans, Eric Tartour, Luc de Chaisemartin, Serge Lebecque, Wolf-Herman Fridman, Jacques Cadranel

From the Laboratoire Microenvironnement Immunitaire et Tumeurs, Institut National de la Santé et de la Recherche Médicale (INSERM) U872, Centre de Recherche des Cordeliers; Université Pierre et Marie Curie, UMR S872; Université Paris Descartes, UMR S872; Service d'Anatomie-Pathologie, Hôpital Tenon, AP-HP; Laboratoire de Biologie Cellulaire et d'Immunopathologie Pulmonaire, UPRES EA3493, Université Paris VI, Hôpital Tenon; Service d'Anatomie-Pathologie, Hôpital Européen Georges Pompidou, AP-HP; Service de Pneumologie et Réanimation Respiratoire, Hôpital Tenon, AP-HP; Laboratoire d'Immunologie, Hôpital Européen Georges Pompidou, AP-HP, Paris; INSERM U503, CERVI; and Centre Hospitalier Lyon Sud, Lyon, France

Corresponding author: Marie-Caroline Dieu-Nosjean, PhD, Laboratoire Microenvironnement Immunitaire et Tumeurs, INSERM UMR S872, Centre de Recherche des Cordeliers, 15 rue de l'école de Médecine, F-75270 Paris cedex 06, France; e-mail: mc.dieu-nosjean{at}crc.jussieu.fr

	ABSTRACT

TOP ABSTRACT INTRODUCTION PATIENTS AND METHODS RESULTS DISCUSSION AUTHORS' DISCLOSURES OF... AUTHOR CONTRIBUTIONS Appendix REFERENCES

 
Purpose It has been established that the immune system plays an important role in tumor rejection. There is also compelling evidence that immune responses can develop independently of secondary lymphoid organs in tertiary lymphoid structures. We studied the presence and the correlation of tertiary lymphoid structures with clinical outcome in non–small-cell lung cancer (NSCLC), as the prognostic value of these structures in patients with cancer had not yet been established.

Patients and Methods This retrospective study was performed by immunohistochemistry on paraffin-embedded tissue specimens from 74 patients with early-stage NSCLC.

Results Tertiary lymphoid structures were detected in some tumors but not in nontumoral lungs. Thus we called these structures tumor-induced bronchus-associated lymphoid tissue (Ti-BALT). As in lymph nodes, Ti-BALTs were composed of mature dendritic cell (DC)/T-cell clusters adjacent to B-cell follicles and had features of an ongoing immune response. Because the quantitative counting of Ti-BALT was difficult to achieve, we used mature DCs that homed exclusively in Ti-BALT as a specific marker of these structures. Univariate analysis showed that the density of mature DCs was highly associated with a favorable clinical outcome (overall, disease-specific, and disease-free survival), suggesting that Ti-BALT may participate in antitumoral immunity. The density of tumor-infiltrating lymphocytes, in particular, CD4⁺ and T-bet⁺ Th1 T cells, was profoundly decreased in tumors weakly infiltrated by mature DCs.

Conclusion The density of mature DCs was found to be a better predictor of clinical outcome than the other parameters tested. The number of tumor-infiltrating mature DCs may identify patients with early-stage NSCLC who have a high risk of relapse.

	INTRODUCTION

TOP ABSTRACT INTRODUCTION PATIENTS AND METHODS RESULTS DISCUSSION AUTHORS' DISCLOSURES OF... AUTHOR CONTRIBUTIONS Appendix REFERENCES

 
Lung cancer is the most common cause of cancer-related death in the world. Approximately 80% to 90% of cases involve non–small-cell lung cancer (NSCLC), which includes adenocarcinoma and squamous cell carcinoma. Only patients whose tumors can be completely resected have a significant chance of increased survival. However, as many as 30% of patients with stage I disease experience recurrence after surgery. The correlation between tumor-infiltrating immune cells and the prognosis of patients with lung cancer is controversial.

Accumulating evidence indicates that adaptive immunity can be initiated independent of secondary lymphoid organs. For instance, splenectomized alymphoplastic mice can reject xenografts,¹ clear viral infection,² or mount an allergic airway inflammation after inhalation of allergens.³ The de novo formation of ectopic lymphoid structures (also called tertiary lymphoid structures) at the site of inflammation can arise in potentially all inflamed noncanonical lymphoid organs.⁴ Bronchus-associated lymphoid tissues (BALTs) have been described in the human fetus and infant lung.⁵ They disappear in the normal adult lung,⁶ in contrast with other organs such as the gut, where they are constitutively present (gut-associated lymphoid tissue). BALTs have been observed in several inflammatory lung diseases in humans (fibrosis, pneumonia, hypersensitivity pneumonitis, diffuse pan-bronchiolitis, and tobacco-induced inflammation), although it is still unclear whether these structures are simple lymphoid aggregates or functional immune structures.

A tumor is composed of malignant, stromal, endothelial, and immune cells that form a heterogeneous network and exhibit complex interactions. Although tumor eradication by the immune system is often inefficient, there is evidence that many developing cancers are not ignored by the immune system.⁷ Spontaneous tumor regressions occurring concomitantly with autoimmune manifestations and the higher incidence of tumors in immunosuppressed patients are indications of the involvement of the immune system in tumor rejection. Mice deficient in immune functions spontaneously develop tumors. The density of tumor-infiltrating lymphocytes (TILs) with cytotoxic and memory phenotypes is highly predictive of good clinical outcome.^8-14 However, although prognosis is related to the homing of effector immune cells, it is still unclear where the activation of the specific immune response takes place: in the tumor, the draining lymph node, or both.

The presence of tertiary lymphoid structures has never been reported in lung cancer, and thus far, no studies have been performed investigating putative correlation with clinical outcome. Here we provide evidence that in certain tumors, immune cells are organized in tertiary lymphoid structures and that the density of mature dendritic cells (DCs), a cell population that is exclusively detected in BALT, is correlated with prolonged survival.

	PATIENTS AND METHODS

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Patients
Paraffin-embedded tumor biopsies with representative areas of tumor and adjacent lung parenchyma were retrieved retrospectively from 74 successive patients diagnosed between 1998 and 2002 with early-stage NSCLC.¹⁵ Lung biopsies from five patients taken either at a distance from the primary tumor or from a different lesion-free lobe were used as tissue references. Preoperative evaluation of patients included lung, brain, and adrenal computed tomography scan and liver ultrasound echography. All patients underwent complete surgical resection of their tumors, including multilevel lymph node sampling or lymphadenectomy, but none received preoperative chemotherapy or radiotherapy. Patients with an Eastern Cooperative Oncology Group performance status¹⁶ 1 were eligible. The main clinical and pathologic features of the patients are presented in Table 1. Patients with mixed histologic features, a T3 tumor, or pleural invasion were ineligible. At the completion of the study, the minimal clinical follow-up was 48 months for the last patient included in the cohort. The consent of all patients was obtained, even though French law does not require specific approval of an institutional review board or the consent of patients for retrospective observational, noninterventional analysis of medical records.

Method for Cell Quantification
Stained cells were counted semi-quantitatively (score 0, 1, 2, 3, and 4 for none, very low, weak, intermediate, and high density of positive cells, respectively) in each intermediate-power field (IPF) in the tumoral areas of the entire tissue section (from 19 to 76 fields, original magnification x100), and expressed as mean score per IPF, with SEMs calculated. The numbers of DC-Lamp⁺ mature DCs and Granzyme-B⁺ CD8⁺ T cells were lower than the number of cells described above, allowing us to realize a quantitative counting. Those stained cells were expressed as mean cells per IPF, with SEMs calculated. CD4, CD8, CD45ra, CD45ro, TIA-1 stainings were counted as a percentage of positive cells among CD3⁺ T cells. The necrosis and fibrosis were counted as the percentage of the positive areas among the whole tumor mass section. Both immunostaining and scoring were evaluated according to the criteria described above by four independent observers (M.-C.D.-N., M.A., V.P., and L.D.C.) who were blinded to clinical outcome. The investigators reviewed the few cases with conflicting results to reach a consensus.

Statistical Analysis
The variables taken into account for statistical analysis included clinical (age, sex, smoking, tumor relapse, and vital status), histopathologic (histology, pathologic TNM [pTNM], tumor differentiation, localization of the primary tumor, necrosis, fibrosis, and proliferation of the tumor), and immunologic parameters (markers described above). To perform univariate analysis, groups of patients were defined according to the bimodal distribution of the density of positive cells, appointed using the following cut-offs: CD3, 1.5 mean score/tumor IPF; CD20, 1.5 mean score/tumor IPF; DC-Lamp, 1.65 mean cells/tumor IPF.

² test with Yates correction and analysis of variance test (posthoc tests with Fisher's and Bonferroni methods) were used for univariate analysis. Overall survival (OS), disease-specific survival (DSS), and disease-free survival (DFS) curves were estimated by Kaplan-Meier method, and significant differences between the groups of patients were evaluated using the log-rank test. An event affecting OS was defined as death from any cause, DSS as death from NSCLC, and DFS as relapse of the primary tumor. Statistical analysis was performed using StatView and JMP softwares (SAS Institute, Cary, NC). A P value less than .05 was considered statistically significant.

	RESULTS

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Presence of Tumor-Induced BALT in Some Early-Stage Lung Cancers
We first searched for the presence of tertiary lymphoid structures on paraffin-embedded sections of 74 early-stage NSCLCs (46 adenocarcinomas and 28 squamous cell carcinomas) and five nonpathologic lung biopsies. These lymphoid organizations have been found in many tumors (Fig 1A), whereas they had not yet been observed in nontumoral lung. Therefore, we called these lymphoid structures tumor-induced BALT (Ti-BALT). To evaluate the prognostic value of Ti-BALT, we tried to quantify these structures; unfortunately, their size was extremely variable and several Ti-BALT can colocalize, which makes quantification by counting uncertain (Fig 1B). Thus we decided to characterize the cellular composition of these lymphoid structures to identify a specific marker of Ti-BALT. We thus used immunohistochemistry to examine the localization of CD3⁺ T cells, CD20⁺ B cells, and DC-Lamp⁺ mature DCs. The density of adaptive immune cells was heterogeneous between tumors, as well as within a given tumor. In some biopsies, CD3⁺ T lymphocytes were detected in clusters where DC-Lamp⁺ mature DCs exclusively homed (Fig 1C). The presence of mature DCs was confirmed by the expression of CD83, another marker of mature DCs (data not shown). These mature DC/T-cell clusters were often surrounded by CD20⁺ B-cell follicles (Fig 1D) characterized by the presence of both a CD21⁺ follicular dendritic cell network (Fig 1E) and Ki67⁺ proliferating germinal center (GC) B cells (Fig 1F).

View larger version (143K): [in this window] [in a new window] [PowerPoint Slide for Teaching]   Fig 1. Characterization of tumor-induced bronchus-associated lymphoid tissue (Ti-BALT) in non–small-cell lung cancer via staining on paraffin-embedded lung tumor biopsies. (A, B) Presence of Ti-BALT (arrow in A or as limited by dashed line in B) in lung tumor section counterstained with hematoxylin and eosin. (C) DC-Lamp+ mature dendritic cells (red) home exclusively into CD3+ T-cell clusters (blue). (D) Presence of adjacent CD20+ B (red) and CD3+ T (blue) cell rich areas of Ti-BALT. (E) CD20+ B-cell follicles (red) are characterized by the presence of a CD21+ follicular dendritic cell network (blue). (F) Some CD20+ B-cell follicles (blue) contained Ki67+ proliferating germinal center B cells (brown). Original magnification: A, x50; B, x100; C, D, F, x400; E, x200. T, tumor nest.

Taken together, the presence of Ti-BALT was very heterogeneous between tumors. These structures were composed of mature DCs, T cells, and B cells organized with a distribution reminiscent of the secondary lymphoid organs where mature DCs exclusively home.

Prognostic Value of Ti-BALT
At the completion of the study, 54 patients were alive (73%) and 20 patients had died (27%; Table 1). Seven patients died as a result of postoperative complications at the hospital or within 1 month of leaving the hospital and were therefore considered as having died from cancer progression-unrelated causes. Nine deaths were NSCLC-related and four deaths were NSCLC-unrelated (myeloma, n = 1; sepsis, n = 1; vascular brain damage, n = 1; unknown cause, n = 1).

We analyzed the prognostic value of Ti-BALT. As mentioned previously, as a result of the uncertainty associated with quantification by counting, this question was difficult to answer. As it had already been observed that DC-Lamp⁺ mature DCs are selectively detected in Ti-BALT, DC-Lamp was chosen as a specific marker of Ti-BALT. Except for five tumors, a trend was observed by statistical analysis between these two parameters (Fig 2A; R² = 0.317 for 74 tumors and R² = 0.488 for 69 tumors).

View larger version (12K): [in this window] [in a new window] [PowerPoint Slide for Teaching]   Fig 2. Evaluation of DC-Lamp as a marker of tumor-induced bronchus-associated lymphoid tissue (Ti-BALT) and its prognostic value. (A) Correlation between the density of Ti-BALT and the density of tumor-infiltrating DC-Lamp+ mature dendritic cells (DCs). Kaplan-Meier curves of disease-free survival for 74 patients with non–small-cell lung cancer depending on (B) the density of tumor-infiltrating DC-Lamp+ mature DCs and (C) the pathologic TNM stage.

These results indicate that DC-Lamp, which is a specific marker of Ti-BALT, is the highest predictive marker for survival. Increased density of mature DCs is associated with a favorable clinical outcome for patients with early-stage NSCLC.

Density and the Phenotype of TILs Are Distinct Among DC-Lamp Low Versus DC-Lamp High Tumors
TILs were further compared in DC-Lamp–low versus DC-Lamp–high tumors using immunohistochemistry. A combination of T-cell markers let us identify the following subsets: CD4⁺ T cells, CD8⁺ T cells, CD45ra⁺CD45ro^–-naïve T cells, CD45ro⁺CD45ra^– memory T cells, T-bet [T-box transcription factor 21]⁺ Th1 T cells, and T cytotoxic cells (granzyme-B). As presented in Table 3, the density of T cells was closely correlated with the density of mature DCs (P = .0018), indicating that DC-Lamp–low tumors have fewer TILs than DC-Lamp–high tumors. The main difference between the two groups of DC-Lamp tumors was the CD4⁺ T-cell compartment, which was dramatically collapsed in DC-Lamp–low tumors, whereas the CD8⁺ T-cell subset was less affected (as observed by immunohistochemistry). Thus the ratio of CD4/CD8 in DC-Lamp–low tumors was inverse to the ratio seen in DC-Lamp–high tumors (P = .0056). Most CD4⁺ T cells formed clusters with mature DCs (Fig 3A and inset), in contrast with CD8⁺ T cells, which infiltrated all the areas of the tumor (Fig 3B and inset). CD45ra⁺–naïve T cells were rare in both groups of tumors (< 10% of total CD3⁺ T cells), which is in agreement with the expression of CD45ro by most T cells (Fig 3C). Thus the ratio of CD45ra/CD45ro on T-cell subsets was relatively constant in both tumor groups (Table 3; P = .3693). However, the density of T-bet⁺ cells was highly enhanced in DC-Lamp–high tumors as compared with DC-Lamp–low tumors (P = .0037). Cells were detected in tumor nests (Fig 3D) and the stroma reaction, but never in Ti-BALT. Finally, a modest increase of granzyme-B⁺ cytotoxic T lymphocyte was also seen in DC-Lamp–high tumors, but the difference between the two groups of tumors did not reach significance.

View larger version (126K): [in this window] [in a new window] [PowerPoint Slide for Teaching]   Fig 3. Characterization of T-cell subsets and their activities in DC-Lamp–high tumors. (A, B, C) Double immunostainings and (D) single immunostaining followed by hematoxylin counterstaining on tumors DC-Lamp–high. (A, B) DC-Lamp+ mature dendritic cells (DCs; red) are preferentially in contact with CD4+ T helper cells (blue, A) compared with cytotoxic CD8+ T cells (blue, B). A and B insets represent higher magnification of A and B, respectively. (C) Double staining with anti-CD45ro (red) and anti-CD3 (blue) reveals the presence of a vast majority of memory T cells (gray, arrowhead) versus naïve T cells (blue, arrow) in the stroma reaction and tumor nests. (D) Presence of numerous T-bet+ cells (brown) in both the stroma reaction and tumor nests. Original magnification: A, B, x100; A and B insets, x200; C, D, x400. Abbreviation: T, tumor nest; S, stroma reaction.

	DISCUSSION

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In human lung tumors, we report the presence of tertiary lymphoid structures composed of mature DC/T-cell clusters and B-cell follicles. The B-cell areas include proliferating GC B cells and a follicular DC network, features of an ongoing immune response as observed in canonical lymphoid organs. We did not observe these structures in sites distant from the tumor, suggesting that they are induced in response to the tumor microenvironment. Therefore, we have called these structures Ti-BALT. Nonproliferating GC B cells have also been described in ectopic lymphoid structures of idiopathic lung fibrosis,²⁰ indicating that BALT might not have the same maturation status in different lung diseases and that the local microenvironment influences BALT development and function.

We have shown through univariate analysis that the density of mature DCs, a cell population that homes exclusively to Ti-BALT, is highly predictive of survival (OS, DSS, and DFS) in early-stage NSCLC. In this study, smoking history (never/current smokers and pack-years) was identical in both groups of DC-Lamp patients, which argues again for a major role of the tumor microenvironment itself in BALT neogenesis and immune function. We have shown that CD4⁺ T cells colocalized preferentially with mature DCs and that their density was profoundly affected in DC-Lamp–low tumors. These observations suggest a direct link between low densities of mature DCs and CD4⁺ T cells, rare Ti-BALT, and poor clinical outcome. This conclusion is in agreement with previous data²¹ showing that CD4⁺ TILs are associated with a favorable prognosis in NSCLC and that the ratio of CD4/CD8 T cells is increased in regressing melanoma and basal cell carcinoma.^22-24 However, the prognostic value of DC-Lamp was higher than that of CD4, most likely because CD4⁺ T cells contain several cell subsets (naïve, memory/effector, anergic, and regulatory T cells) with opposite functional activities. In particular, the densities of T and B cells, although correlated with the density of mature DCs, were not associated with clinical outcome. In contrast with mature DCs, lymphocytes do not home exclusively to Ti-BALT. Our data suggest that the more tumors are infiltrated by mature DCs, the more T and B cells are organized and proliferated in Ti-BALT. However in the absence of mature DCs, infiltrating lymphocytes neither cluster nor proliferate because of the absence of lymphoid structures. Moreover, lymphocytes, and especially T cells, contain different cell subsets with opposite functional activities, indicating that the quantification of total T and B cells may not be the only method for evaluating lymphocyte implication in clinical outcome.

We have shown that the T-cell composition of Ti-BALT mimics that of gut-associated lymphoid tissue²⁵ and is completely different from conventional lymphoid organs, where naïve and memory T cells are present in equal proportion, always with more CD4⁺ than CD8⁺ T cells.

We have shown that DC-Lamp–high tumors are highly infiltrated by cells positive for T-bet, the interferon gamma–specific transcription factor. Interferon gamma is known to induce CXCR3 ligands, among which the chemokine CXCL10 has been reported to inhibit NSCLC tumorigenesis and spontaneous metastases.²⁶

Cumulative data report the participation of extranodal lymphoid structures in the development of specific immune responses.^1,27 In mice deficient in secondary lymphoid organs, Moyron-Quiroz et al^2,28 demonstrated that robust primary T- and B-cell responses to influenza virus and the maintenance of immunologic memory are developed in inducible BALT, which argues for a crucial role of BALT in protective immunity. Thus the high density of mature DCs (a population that homes selectively to Ti-BALT) in patients with a better survival outcome suggests that the first step of specific immune responses might be initiated in the tumor itself. Tertiary lymphoid structures have been reported to be harmful in autoimmunity, because they are involved in the exacerbation of the local immune response and thus participate in the aggravation of pathogenesis. In the present study, no patient with NSCLC had clinical manifestations of autoimmune or infectious diseases, indicating that lymphoid structures were instead induced in response to the tumor microenvironment. In lung cancer, we propose that the absence of Ti-BALT leads to poor T cell priming, with improper T cell localization and/or activity, and thereby resulting in inefficient antitumor immunity. Tumor-associated antigens, which are continuously sampled and processed by DCs, would potentially be in direct contact with specific T cells, thereby increasing the efficiency and specificity of the priming. During tumor progression, Ti-BALT may thus allow T cells to react more rapidly to the shifting expression profile of tumor antigens in situ.

The presence of proliferating GC B cells in some Ti-BALTs suggests that they may be an active partner in the local initiation of potential antitumoral immunity. As previously described in chronic inflammatory diseases,^29-31 Ti-BALT GCs may support antigen-driven clonal expansion and extensive diversification. Ti-BALT B cells may also play an important role in T-cell–mediated immunity against the tumor.

Because clinical outcome is obviously established after the resection of Ti-BALT–containing tumor, these structures are likely not directly involved in survival. We speculate that Ti-BALT–derived memory cells migrate into the draining lymph nodes to develop a central memory and robust systemic response against micro-metastasis. Finally, DC-Lamp, a marker of Ti-BALT, may have a direct application in the clinical setting, as it could be used to identify patients with early-stage NSCLC who have a high risk of relapse.

	AUTHORS’ DISCLOSURES OF POTENTIAL CONFLICTS OF INTEREST

TOP ABSTRACT INTRODUCTION PATIENTS AND METHODS RESULTS DISCUSSION AUTHORS' DISCLOSURES OF... AUTHOR CONTRIBUTIONS Appendix REFERENCES

 
The author(s) indicated no potential conflicts of interest.

	AUTHOR CONTRIBUTIONS

TOP ABSTRACT INTRODUCTION PATIENTS AND METHODS RESULTS DISCUSSION AUTHORS' DISCLOSURES OF... AUTHOR CONTRIBUTIONS Appendix REFERENCES

 
Conception and design: Marie-Caroline Dieu-Nosjean, Martine Antoine, Claire Danel, Eric Tartour, Serge Lebecque, Wolf-Herman Fridman, Jacques Cadranel

Financial support: Marie-Caroline Dieu-Nosjean, Martine Antoine, Wolf-Herman Fridman, Jacques Cadranel

Administrative support: Marie-Caroline Dieu-Nosjean, Wolf-Herman Fridman, Jacques Cadranel

Provision of study materials or patients: Martine Antoine, Claire Danel, Marie Wislez, Virginie Poulot, Nathalie Rabbe, Jacques Cadranel

Collection and assembly of data: Marie-Caroline Dieu-Nosjean, Martine Antoine, Claire Danel, Didier Heudes, Virginie Poulot, Nathalie Rabbe, Ludivine Laurans

Data analysis and interpretation: Marie-Caroline Dieu-Nosjean, Martine Antoine, Claire Danel, Didier Heudes, Marie Wislez, Ludivine Laurans, Eric Tartour, Luc De Chaisemartin, Wolf-Herman Fridman, Jacques Cadranel

Manuscript writing: Marie-Caroline Dieu-Nosjean, Serge Lebecque, Wolf-Herman Fridman, Jacques Cadranel

Final approval of manuscript: Marie-Caroline Dieu-Nosjean, Martine Antoine, Claire Danel, Didier Heudes, Marie Wislez, Virginie Poulot, Nathalie Rabbe, Ludivine Laurans, Eric Tartour, Luc De Chaisemartin, Serge Lebecque, Wolf-Herman Fridman, Jacques Cadranel

	Appendix

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	ACKNOWLEDGMENTS

 
We thank M. Riquet, MD, and S. Saeland, MD, PhD, for helpful scientific discussions, P. Bruneval, MD, PhD, for support in this project, M. Douheret and C. Rismondo for excellent technical assistance, and A.M. Laval for technical support.

	NOTES

 
Supported by Grant No. ARC n°C01-010 from Association pour la Recherche sur le Cancer through the Alliance pour la Recherche sur le Cancer network.

M.A. and C.D. contributed equally to this study.

Authors’ disclosures of potential conflicts of interest and author contributions are found at the end of this article.

	REFERENCES

TOP ABSTRACT INTRODUCTION PATIENTS AND METHODS RESULTS DISCUSSION AUTHORS' DISCLOSURES OF... AUTHOR CONTRIBUTIONS Appendix REFERENCES

 
1. Tesar BM, Chalasani G, Smith-Diggs L, et al: Direct antigen presentation by a xenograft induces immunity independently of secondary lymphoid organs. J Immunol 173:4377-4386, 2004[Abstract/Free Full Text]

2. Moyron-Quiroz JE, Rangel-Moreno J, Kusser K, et al: Role of inducible bronchus associated lymphoid tissue (iBALT) in respiratory immunity. Nat Med 10:927-934, 2004[CrossRef][Medline]

3. Gajewska BU, Alvarez D, Vidric M, et al: Generation of experimental allergic airways inflammation in the absence of draining lymph nodes. J Clin Invest 108:577-583, 2001[CrossRef][Medline]

4. Drayton DL, Liao S, Mounzer RH, et al: Lymphoid organ development: From ontogeny to neogenesis. Nat Immunol 7:344-353, 2006[CrossRef][Medline]

5. Gould SJ, Isaacson PG: Bronchus-associated lymphoid tissue (BALT) in human fetal and infant lung. J Pathol 169:229-234, 1993[CrossRef][Medline]

6. Tschernig T, Pabst R: Bronchus-associated lymphoid tissue (BALT) is not present in the normal adult lung but in different diseases. Pathobiology 68:1-8, 2000[CrossRef][Medline]

7. Dunn GP, Old LJ, Schreiber RD: The three Es of cancer immunoediting. Annu Rev Immunol 22:329-360, 2004[CrossRef][Medline]

8. Pagès F, Berger A, Camus M, et al: Effector memory T cells, early metastasis, and survival in colorectal cancer. N Engl J Med 353:2654-2666, 2005[Abstract/Free Full Text]

9. Clemente CG, Mihm MC Jr, Bufalino R, et al: Prognostic value of tumor infiltrating lymphocytes in the vertical growth phase of primary cutaneous melanoma. Cancer 77:1303-1310, 1996[CrossRef][Medline]

10. Zhang L, Conejo-Garcia JR, Katsaros D, et al: Intratumoral T cells, recurrence, and survival in epithelial ovarian cancer. N Engl J Med 348:203-213, 2003[Abstract/Free Full Text]

11. Tartour E, Gey A, Sastre-Garau X, et al: Prognostic value of intratumoral interferon gamma messenger RNA expression in invasive cervical carcinomas. J Natl Cancer Inst 90:287-294, 1998[CrossRef][Medline]

12. Dunn GP, Koebel CM, Schreiber RD: Interferons, immunity and cancer immunoediting. Nat Rev Immunol 6:836-848, 2006[CrossRef][Medline]

13. Dalerba P, Maccalli C, Casati C, et al: Immunology and immunotherapy of colorectal cancer. Crit Rev Oncol Hematol 46:33-57, 2003[Medline]

14. Galon J, Costes A, Sanchez-Cabo F, et al: Type, density, and location of immune cells within human colorectal tumors predict clinical outcome. Science 313:1960-1964, 2006[Abstract/Free Full Text]

15. Mountain CF: Revisions in the International System for Staging Lung Cancer. Chest 111:1710-1717, 1997[CrossRef][Medline]

16. Finkelstein DM, Cassileth BR, Bonomi PD, et al: A pilot study of the Functional Living Index-Cancer (FLIC) Scale for the assessment of quality of life for metastatic lung cancer patients: An Eastern Cooperative Oncology Group study. Am J Clin Oncol 11:630-633, 1988[Medline]

17. Piemonte M: International Union Against Cancer: TNM Classification of Malignant Tumors (ed 6). New York, NY, Wiley-Liss, 2002

18. Brambilla E, Travis WD, Colby TV, et al: The new World Health Organization classification of lung tumours. Eur Respir J 18:1059-1068, 2001[Abstract/Free Full Text]

19. Marmey B, Boix C, Barbaroux JB, et al: CD14 and CD169 expression in human lymph nodes and spleen: Specific expansion of CD14+CD169- monocyte-derived cells in diffuse large B-cell lymphomas. Hum Pathol 37:68-77, 2006[CrossRef][Medline]

20. Marchal-Sommé J, Uzunhan Y, Marchand-Adam S, et al: Cutting edge: Nonproliferating mature immune cells form a novel type of organized lymphoid structure in idiopathic pulmonary fibrosis. J Immunol 176:5735-5739, 2006[Abstract/Free Full Text]

21. Wakabayashi O, Yamazaki K, Oizumi S, et al: CD4+ T cells in cancer stroma, not CD8+ T cells in cancer cell nests, are associated with favorable prognosis in human non-small cell lung cancers. Cancer Sci 94:1003-1009, 2003[CrossRef][Medline]

22. Tefany FJ, Barnetson RS, Halliday GM, et al: Immunocytochemical analysis of the cellular infiltrate in primary regressing and non-regressing malignant melanoma. J Invest Dermatol 97:197-202, 1991[CrossRef][Medline]

23. Hunt MJ, Halliday GM, Weedon D, et al: Regression in basal cell carcinoma: An immunohistochemical analysis. Br J Dermatol 130:1-8, 1994[Medline]

24. Patel A, Halliday GM, Barnetson RS: CD4+ T lymphocyte infiltration correlates with regression of a UV-induced squamous cell carcinoma. J Dermatol Sci 9:12-19, 1995[CrossRef][Medline]

25. Suzuki A, Masuda A, Nagata H, et al: Mature dendritic cells make clusters with T cells in the invasive margin of colorectal carcinoma. J Pathol 196:37-43, 2002[CrossRef][Medline]

26. Arenberg D: Chemokines in the biology of lung cancer. J Thorac Oncol 1:287-288, 2006[CrossRef][Medline]

27. Kirk CJ, Hartigan-O'Connor D, Mule JJ: The dynamics of the T-cell antitumor response: Chemokine-secreting dendritic cells can prime tumor-reactive T cells extranodally. Cancer Res 61:8794-8802, 2001[Abstract/Free Full Text]

28. Moyron-Quiroz JE, Rangel-Moreno J, Hartson L, et al: Persistence and responsiveness of immunologic memory in the absence of secondary lymphoid organs. Immunity 25:643-654, 2006[CrossRef][Medline]

29. Gause A, Gundlach K, Zdichavsky M, et al: The B lymphocyte in rheumatoid arthritis: Analysis of rearranged V kappa genes from B cells infiltrating the synovial membrane. Eur J Immunol 25:2775-2782, 1995[CrossRef][Medline]

30. Schröder AE, Greiner A, Seyfert C, et al: Differentiation of B cells in the nonlymphoid tissue of the synovial membrane of patients with rheumatoid arthritis. Proc Natl Acad Sci U S A 93:221-225, 1996[Abstract/Free Full Text]

31. Thaunat O, Field AC, Dai J, et al: Lymphoid neogenesis in chronic rejection: Evidence for a local humoral alloimmune response. Proc Natl Acad Sci U S A 102:14723-14728, 2005[Abstract/Free Full Text]

Submitted November 13, 2007; accepted April 29, 2008.

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