A distinct expression of CC chemokines by macrophages in nasopharyngeal carcinoma: Implication for the intense tumor infiltration by T lymphocytes and macrophages

doi:10.1053/hupa.2001.20886

Human Pathology

Volume 32, Issue 1, January 2001, Pages 42-49

https://doi.org/10.1053/hupa.2001.20886 Get rights and content

Abstract

Nasopharyngeal carcinoma (NPC) is characterized by harboring Epstein-Barr virus genes in the tumor cells and an intense infiltration of leukocytes in the tumor tissue. These infiltrating cells are mainly composed of T lymphocytes and macrophages. The mechanism of this intense infiltration has long been a puzzle. We attempted to address this issue by studying the expression of CC chemokines, which are responsible for recruiting both T cells and macrophages, by an immunohistochemical approach. In biopsies obtained from nasopharynx of 17 NPC patients that contained tumor cells, expression of macrophage inflammatory protein 1α (MIP-1α), MIP-1β, macrophage chemoattractant protein-1 (MCP-1), MCP-2, MCP-3, and RANTES was detected in the tumor-infiltrating cells, with MIP-1α and MCP-1 found in nearly all biopsies and the others relatively less frequently. Furthermore, expression of interferon-γ (IFN-γ) was also observed in tumor-infiltrating cells. In contrast, CC chemokines and IFN-γ were rarely expressed in the 13 control biopsies that were either normal or with nonspecific inflammation, and in 4 biopsies from untreated NPC patients that contained no tumor cells. Using an immunofluorescent double-staining method, MIP-1α and MCP-1 were identified to be associated with macrophages, and IFN-γ with T cells. Moreover, expression of CCR2 and CCR5, the receptors for these chemokines, was also detected in the tumor-infiltrating cells. These data indicate that the intense tumor infiltration by T cells and macrophages is a result of active recruitment. It seems possible that the intense infiltration of leukocytes in NPC tumor tissue is initiated by the activated tumor-reactive T cells. T cells migrate into the tumor tissue in an antigen-specific mode, and IFN-γ secreted from these pioneer T cells activates tissue macrophages to express CC chemokines, especially MIP-1α and MCP-1, which consequently recruit more T cells and macrophages into the tumor tissue. HUM PATHOL 32:42-49 Copyright © 2001 by W.B. Saunders Company

Section snippets

Biopsies and serum samples

Biopsies of nasopharynx were surgically removed from 34 untreated patients suspected of having NPC, snap-frozen, and stored in liquid nitrogen until use. The diagnosis of NPC was made on histologic examination of hematoxylin and eosin-stained paraffin sections and immunohistochemical staining with an anticytokeratin antibody on cells of epithelial origin. Biopsies obtained from patients diagnosed as non-NPC (either normal or with nonspecific inflammation) were used as controls. Blood samples

Expression of EBER in NPC tumor cells and the humoral response to EBV

Biopsies were obtained from 34 patients. Among them 21 were diagnosed as having NPC, and 13 were diagnosed as having nonspecific inflammation. NPC tumor cells can be identified by immunohistochemical staining with a mAb directed at cytokeratin. It is used for the visualization of tumor tissue in this study (Fig 1B).

. Identification of NPC tumor cells, the infiltrating T cells and macrophages. Biopsies from patient CB (Table 1) were used for the experiments presented in this figure. (A) Tumor

Discussion

Many reports have described that tumor cells of undifferentiated NPC harbor EBV genomes.1, 2, 3, 4, 5, 7, 23, 24, 25 and this is again confirmed by detecting EBERs in NPC tumor cells in the present study. NPC tumor cells are found to express the EBV genes of both the latent and replication phases. These include EBERs, EBNA-1, LMP-1, LMP-2, BZLF-1, and BRLF-1.2, 3, 4, 5, 6, 7 Among them, LMP-2, BZLF-1, and BRLF-1 can induce strong T-cell response, and relatively high frequencies of specific

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Several possible mechanisms may explain the favorable efficacy of anti-PD-1 therapy in nasopharyngeal cancer. Nasopharynx has abundant lymphoid tissue and high infiltration of T and B cells, making it an ideal tumor environment for anti-PD-1 therapy, the so-called “hot tumor” [16–17]. Moreover, EBV infection induces increased expression of PD-L1 through latent membrane proteins 1 and 2, the EBV oncogene [18–21].
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Nasopharyngeal carcinoma (NPC) arises from a unique anatomical site with heavy infiltration of leucocytes, predominantly T-lymphocytes, together with other stromal cells. Undifferentiated NPC is the prevalent histological subtype (>98%) in endemic areas in Southern China including Hong Kong. Epstein–Barr virus (EBV) infection is closely associated with undifferentiated NPC. EBV infection in NPC is generally latent in nature. Expression of viral gene products may facilitate the malignant transformation of premalignant nasopharyngeal epithelium to NPC cells. Among the latent EBV gene products, the high expression of EBV-encoded BART-microRNAs in NPC is strongly implicated in NPC pathogenesis. A unique tumor microenvironment (TME) infiltrated heavily with T-lymphocytes and other stromal cells is believed to be involved to support growth of NPC in patients and latent EBV infection in NPC. Various cytokines secreted by EBV-infected NPC cells and the infiltrated stromal cells in the TME are involved in the recruitment of the rich cellular components to the tumor stroma. The dynamic and reciprocal interaction of cancer cells with stromal cells in the TME has been postulated to support EBV infection and modulate growth and invasive properties of NPC cells. Delineating the complex cancer cell–stromal cell interaction in NPC may reveal novel therapeutic targets effective for NPC treatment.
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^*: Supported by grants 30030, NMRC/0348/1999, NMRC/0327/1999 (H.Z.H.) from the National Medical Research Council, Singapore.

^**: K.F.T. and S.Y.T. contributed equally to this work.

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Original ContributionsA distinct expression of CC chemokines by macrophages in nasopharyngeal carcinoma: Implication for the intense tumor infiltration by T lymphocytes and macrophages*,**

Abstract

Section snippets

Biopsies and serum samples

Expression of EBER in NPC tumor cells and the humoral response to EBV

Discussion

Virology

Immunol Today

Curr Opin Immunol

Expression of EBER1 in primary and metastatic nasopharyngeal carcinoma tissues using in situ hybridization—A correlation with WHO histologic subtypes

Cancer

Expression of Epstein-Barr virus genes and of lymphocyte activation molecules in undifferentiated nasopharyngeal carcinomas

Am J Pathol

Consistent transcription of the Epstein-Barr virus LMP2 gene in nasopharyngeal carcinoma

J Virol

Epstein-Barr virus latent gene transcription in nasopharyngeal carcinoma cells: Coexpression of EBNA1, LMP1, and LMP2 transcripts

J Virol

Expression of Epstein-Barr virus lytic gene BRLF1 in nasopharyngeal carcinoma: Potential use in diagnosis

J Gen Virol

Expression of immune regulatory molecules in Epstein-Barr virus-associated nasopharyngeal carcinomas with prominent lymphoid stroma: Evidence for a functional interaction between epithelial tumor cells and infiltrating lymphoid cells