Identification of a novel monocytic phenotype in Classic Hodgkin Lymphoma tumor microenvironment

Autoři: Ginell R. Post aff001;  Youzhong Yuan aff001;  Emily R. Holthoff aff001;  Charles M. Quick aff001;  Steven R. Post aff001
Působiště autorů: Department of Pathology, University of Arkansas for Medical Sciences, Little Rock, Arkansas, United States of America aff001
Vyšlo v časopise: PLoS ONE 14(11)
Kategorie: Research Article
doi: 10.1371/journal.pone.0224621


Classic Hodgkin lymphoma (CHL) characteristically shows few malignant cells in a microenvironment comprised of mixed inflammatory cells. Although CHL is associated with a high cure rate, recent studies have associated poor prognosis with absolute monocyte count in peripheral blood and increased monocyte/macrophages in involved lymph nodes. Thus, the role of monocytic infiltration and macrophage differentiation in the tumor microenvironment of CHL may be more relevant than absolute macrophage numbers to defining prognosis in CHL patients and potentially have therapeutic implications. Most studies identify tumor-associated macrophages (TAMs) using markers (e.g., CD68) expressed by macrophages and other mononuclear phagocytes, such as monocytes. In contrast, Class A Scavenger Receptor (SR-A/CD204) is expressed by tissue macrophages but not monocytic precursors. In this study, we examined SR-A expression in CHL (n = 43), and compared its expression with that of other macrophage markers. We confirmed a high prevalence of mononuclear cells that stained with CD68, CD163, and CD14 in CHL lymph nodes. However, SR-A protein expression determined by immunohistochemistry was limited to macrophages localized in sclerotic bands characteristic of nodular sclerosis CHL. In contrast, SR-A protein was readily detectable in lymph nodes with metastatic tumor, extra-nodal CHL, T cell/histiocyte-rich large B cell lymphoma, and resident macrophages in non-malignant tissues, including spleen, lymph node, liver and lung. The results of SR-A protein expression paralleled the expression of SR-A mRNA determined by quantitative RT-PCR. These data provide evidence that tumor-infiltrating monocyte/macrophages in CHL have a unique phenotype that likely depends on the microenvironment of nodal CHL.

Klíčová slova:

Alveolar macrophages – Cell staining – Immunohistochemistry techniques – Immunostaining – Lymph nodes – Macrophages – Monocytes – Spleen


1. Kuppers R. The biology of Hodgkin's lymphoma. Nat Rev Cancer. 2009;9(1):15–27. doi: 10.1038/nrc2542 19078975.

2. Aldinucci D, Gloghini A, Pinto A, De Filippi R, Carbone A. The classical Hodgkin's lymphoma microenvironment and its role in promoting tumour growth and immune escape. The Journal of pathology. 2010;221(3):248–63. doi: 10.1002/path.2711 20527019.

3. Guo B, Cen H, Tan X, Ke Q. Meta-analysis of the prognostic and clinical value of tumor-associated macrophages in adult classical Hodgkin lymphoma. BMC Med. 2016;14(1):159. doi: 10.1186/s12916-016-0711-6 27745550.

4. Steidl C, Lee T, Shah SP, Farinha P, Han G, Nayar T, et al. Tumor-associated macrophages and survival in classic Hodgkin's lymphoma. N Engl J Med. 2010;362(10):875–85. doi: 10.1056/NEJMoa0905680 20220182; PubMed Central PMCID: PMC2897174.

5. Koh YW, Park C, Yoon DH, Suh C, Huh J. CSF-1R expression in tumor-associated macrophages is associated with worse prognosis in classical Hodgkin lymphoma. Am J Clin Pathol. 2014;141(4):573–83. Epub 2014/03/13. doi: 10.1309/AJCPR92TDDFARISU 24619759.

6. Zaki MA, Wada N, Ikeda J, Shibayama H, Hashimoto K, Yamagami T, et al. Prognostic implication of types of tumor-associated macrophages in Hodgkin lymphoma. Virchows Arch. 2011;459(4):361–6. Epub 2011/08/30. doi: 10.1007/s00428-011-1140-8 21874508.

7. Komohara Y, Niino D, Ohnishi K, Ohshima K, Takeya M. Role of tumor-associated macrophages in hematological malignancies. Pathol Int. 2015;65(4):170–6. doi: 10.1111/pin.12259 25707506.

8. Touati M, Delage-Corre M, Monteil J, Abraham J, Moreau S, Remenieras L, et al. CD68-positive tumor-associated macrophages predict unfavorable treatment outcomes in classical Hodgkin lymphoma in correlation with interim fluorodeoxyglucose-positron emission tomography assessment. Leuk Lymphoma. 2015;56(2):332–41. Epub 2014/04/29. doi: 10.3109/10428194.2014.917636 24766492.

9. Azambuja D, Natkunam Y, Biasoli I, Lossos IS, Anderson MW, Morais JC, et al. Lack of association of tumor-associated macrophages with clinical outcome in patients with classical Hodgkin's lymphoma. Ann Oncol. 2012;23(3):736–42. Epub 2011/05/24. doi: 10.1093/annonc/mdr157 21602260; PubMed Central PMCID: PMC3331732.

10. Sanchez-Espiridion B, Martin-Moreno AM, Montalban C, Medeiros LJ, Vega F, Younes A, et al. Immunohistochemical markers for tumor associated macrophages and survival in advanced classical Hodgkin's lymphoma. Haematologica. 2012;97(7):1080–4. doi: 10.3324/haematol.2011.055459 22315492; PubMed Central PMCID: PMC3396681.

11. Harris JA, Jain S, Ren Q, Zarineh A, Liu C, Ibrahim S. CD163 versus CD68 in tumor associated macrophages of classical Hodgkin lymphoma. Diagnostic pathology. 2012;7:12. Epub 2012/02/01. doi: 10.1186/1746-1596-7-12 22289504; PubMed Central PMCID: PMC3281786.

12. Agur A, Amir G, Paltiel O, Klein M, Dann EJ, Goldschmidt H, et al. CD68 staining correlates with the size of residual mass but not with survival in classical Hodgkin lymphoma. Leuk Lymphoma. 2015;56(5):1315–9. Epub 2014/09/11. doi: 10.3109/10428194.2014.963081 25204373.

13. Scott DW, Steidl C. The classical Hodgkin lymphoma tumor microenvironment: macrophages and gene expression-based modeling. Hematology Am Soc Hematol Educ Program. 2014;2014(1):144–50. Epub 2015/02/20. doi: 10.1182/asheducation-2014.1.144 25696847.

14. Falini B, Flenghi L, Pileri S, Gambacorta M, Bigerna B, Durkop H, et al. PG-M1: a new monoclonal antibody directed against a fixative-resistant epitope on the macrophage-restricted form of the CD68 molecule. Am J Pathol. 1993;142(5):1359–72. Epub 1993/05/01. 7684194; PubMed Central PMCID: PMC1886928.

15. Pulford KA, Rigney EM, Micklem KJ, Jones M, Stross WP, Gatter KC, et al. KP1: a new monoclonal antibody that detects a monocyte/macrophage associated antigen in routinely processed tissue sections. J Clin Pathol. 1989;42(4):414–21. Epub 1989/04/01. doi: 10.1136/jcp.42.4.414 2654191; PubMed Central PMCID: PMC1141915.

16. Kunisch E, Fuhrmann R, Roth A, Winter R, Lungershausen W, Kinne RW. Macrophage specificity of three anti-CD68 monoclonal antibodies (KP1, EBM11, and PGM1) widely used for immunohistochemistry and flow cytometry. Annals of the rheumatic diseases. 2004;63(7):774–84. Epub 2004/06/15. doi: 10.1136/ard.2003.013029 15194571; PubMed Central PMCID: PMC1755048.

17. PrabhuDas MR, Baldwin CL, Bollyky PL, Bowdish DME, Drickamer K, Febbraio M, et al. A Consensus Definitive Classification of Scavenger Receptors and Their Roles in Health and Disease. J Immunol. 2017;198(10):3775–89. Epub 2017/05/10. doi: 10.4049/jimmunol.1700373 28483986; PubMed Central PMCID: PMC5671342.

18. Lau SK, Chu PG, Weiss LM. CD163: a specific marker of macrophages in paraffin-embedded tissue samples. Am J Clin Pathol. 2004;122(5):794–801. Epub 2004/10/20. doi: 10.1309/QHD6-YFN8-1KQX-UUH6 15491976.

19. Hogger P, Dreier J, Droste A, Buck F, Sorg C. Identification of the integral membrane protein RM3/1 on human monocytes as a glucocorticoid-inducible member of the scavenger receptor cysteine-rich family (CD163). J Immunol. 1998;161(4):1883–90. Epub 1998/08/26. 9712057.

20. Stanley ER, Guilbert LJ, Tushinski RJ, Bartelmez SH. CSF-1—a mononuclear phagocyte lineage-specific hemopoietic growth factor. J Cell Biochem. 1983;21(2):151–9. doi: 10.1002/jcb.240210206 6309875

21. Sweet MJ, Hume DA. CSF-1 as a regulator of macrophage activation and immune responses. Arch Immunol Ther Exp (Warsz). 2003;51(3):169–77. Epub 2003/08/05. 12894871.

22. Landmann R, Muller B, Zimmerli W. CD14, new aspects of ligand and signal diversity. Microbes Infect. 2000;2(3):295–304. Epub 2000/04/12. doi: 10.1016/s1286-4579(00)00298-7 10758406.

23. Marmey B, Boix C, Barbaroux JB, Dieu-Nosjean MC, Diebold J, Audouin J, 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. Human pathology. 2006;37(1):68–77. Epub 2005/12/20. doi: 10.1016/j.humpath.2005.09.016 16360418.

24. Wu H, Moulton K, Horvai A, Parik S, Glass CK. Combinatorial interactions between AP-1 and ets domain proteins contribute to the developmental regulation of the macrophage scavenger receptor gene. Mol Cell Biol. 1994;14(3):2129–39. Epub 1994/03/01. doi: 10.1128/mcb.14.3.2129 8114743; PubMed Central PMCID: PMC358573.

25. Geng Y, Kodama T, Hansson GK. Differential expression of scavenger receptor isoforms during monocyte-macrophage differentiation and foam cell formation. Arterioscler Thromb. 1994;14(5):798–806. Epub 1994/05/01. doi: 10.1161/01.atv.14.5.798 8172856.

26. Horvai A, Palinski W, Wu H, Moulton KS, Kalla K, Glass CK. Scavenger receptor A gene regulatory elements target gene expression to macrophages and to foam cells of atherosclerotic lesions. Proc Natl Acad Sci U S A. 1995;92(12):5391–5. Epub 1995/06/06. doi: 10.1073/pnas.92.12.5391 7777517; PubMed Central PMCID: PMC41700.

27. Moulton KS, Wu H, Barnett J, Parthasarathy S, Glass CK. Regulated expression of the human acetylated low density lipoprotein receptor gene and isolation of promoter sequences. Proc Natl Acad Sci U S A. 1992;89(17):8102–6. Epub 1992/09/01. doi: 10.1073/pnas.89.17.8102 1518836; PubMed Central PMCID: PMC49864.

28. Moulton KS, Semple K, Wu H, Glass CK. Cell-specific expression of the macrophage scavenger receptor gene is dependent on PU.1 and a composite AP-1/ets motif. Mol Cell Biol. 1994;14(7):4408–18. Epub 1994/07/01. doi: 10.1128/mcb.14.7.4408 8007948; PubMed Central PMCID: PMC358812.

29. Gough PJ, Greaves DR, Suzuki H, Hakkinen T, Hiltunen MO, Turunen M, et al. Analysis of macrophage scavenger receptor (SR-A) expression in human aortic atherosclerotic lesions. Arterioscler Thromb Vasc Biol. 1999;19(3):461–71. Epub 1999/03/12. doi: 10.1161/01.atv.19.3.461 10073945.

30. Naito M, Kodama T, Matsumoto A, Doi T, Takahashi K. Tissue distribution, intracellular localization, and in vitro expression of bovine macrophage scavenger receptors. Am J Pathol. 1991;139(6):1411–23. Epub 1991/12/01. 1750511; PubMed Central PMCID: PMC1886471.

31. Komohara Y, Ohnishi K, Kuratsu J, Takeya M. Possible involvement of the M2 anti-inflammatory macrophage phenotype in growth of human gliomas. The Journal of pathology. 2008;216(1):15–24. Epub 2008/06/17. doi: 10.1002/path.2370 18553315.

32. Yoshikawa K, Mitsunaga S, Kinoshita T, Konishi M, Takahashi S, Gotohda N, et al. Impact of tumor-associated macrophages on invasive ductal carcinoma of the pancreas head. Cancer science. 2012;103(11):2012–20. Epub 2012/08/31. doi: 10.1111/j.1349-7006.2012.02411.x 22931216.

33. Komohara Y, Hasita H, Ohnishi K, Fujiwara Y, Suzu S, Eto M, et al. Macrophage infiltration and its prognostic relevance in clear cell renal cell carcinoma. Cancer science. 2011;102(7):1424–31. Epub 2011/04/02. doi: 10.1111/j.1349-7006.2011.01945.x 21453387.

34. Shigeoka M, Urakawa N, Nakamura T, Nishio M, Watajima T, Kuroda D, et al. Tumor associated macrophage expressing CD204 is associated with tumor aggressiveness of esophageal squamous cell carcinoma. Cancer science. 2013;104(8):1112–9. Epub 2013/05/08. doi: 10.1111/cas.12188 23648122.

35. Hirayama S, Ishii G, Nagai K, Ono S, Kojima M, Yamauchi C, et al. Prognostic impact of CD204-positive macrophages in lung squamous cell carcinoma: possible contribution of Cd204-positive macrophages to the tumor-promoting microenvironment. Journal of thoracic oncology: official publication of the International Association for the Study of Lung Cancer. 2012;7(12):1790–7. Epub 2012/11/17. doi: 10.1097/JTO.0b013e3182745968 23154550.

36. Saito Y, Komohara Y, Niino D, Horlad H, Ohnishi K, Takeya H, et al. Role of CD204-positive tumor-associated macrophages in adult T-cell leukemia/lymphoma. J Clin Exp Hematop. 2014;54(1):59–65. Epub 2014/06/20. 24942947.

37. Takeya M, Komohara Y. Role of tumor-associated macrophages in human malignancies: friend or foe? Pathol Int. 2016;66(9):491–505. Epub 2016/07/23. doi: 10.1111/pin.12440 27444136.

38. Jaffe ES, Stein H, Swerdlow SH. Classic Hodgkin lymphoma. In: Swerdlow SH, Campo E, Harris NL, Jaffe ES, Pileri SA, Stein H, et al., editors. WHO Classification of Tumors of Haematopoietic and Lymphoid Tissues. 4th ed. Lyon: IARC; 2017. p. 435–42.

39. Ott G, Delabie J, Gascoyne RD, Campo E, Stein H, Jaffe ES. T-cell/histiocytes-rich large B-cell lymphoma. In: Swerdlow SH, Campo E, Harris NL, Jaffe ES, Pileri SA, Stein H, et al., editors. WHO Classification of Tumors of Haematopoietic and Lymphoid Tissues. 4th ed. Lyon: IARC; 2017. p. 289–99.

40. Cattaruzza L, Gloghini A, Olivo K, Di Francia R, Lorenzon D, De Filippi R, et al. Functional coexpression of Interleukin (IL)-7 and its receptor (IL-7R) on Hodgkin and Reed-Sternberg cells: Involvement of IL-7 in tumor cell growth and microenvironmental interactions of Hodgkin's lymphoma. Int J Cancer. 2009;125(5):1092–101. Epub 2009/04/25. doi: 10.1002/ijc.24389 19391137.

41. Liao HS, Matsumoto A, Itakura H, Doi T, Honda M, Kodama T, et al. Transcriptional inhibition by interleukin-6 of the class A macrophage scavenger receptor in macrophages derived from human peripheral monocytes and the THP-1 monocytic cell line. Arterioscler Thromb Vasc Biol. 1999;19(8):1872–80. Epub 1999/08/14. doi: 10.1161/01.atv.19.8.1872 10446065.

42. Geng YJ, Hansson GK. Interferon-gamma inhibits scavenger receptor expression and foam cell formation in human monocyte-derived macrophages. J Clin Invest. 1992;89(4):1322–30. Epub 1992/04/01. doi: 10.1172/JCI115718 1556191; PubMed Central PMCID: PMC442994.

43. Hsu HY, Nicholson AC, Hajjar DP. Inhibition of macrophage scavenger receptor activity by tumor necrosis factor-alpha is transcriptionally and post-transcriptionally regulated. J Biol Chem. 1996;271(13):7767–73. doi: 10.1074/jbc.271.13.7767 8631819

44. Bottalico LA, Wager RE, Agellon LB, Assoian RK, Tabas I. Transforming growth factor-beta 1 inhibits scavenger receptor activity in THP-1 human macrophages. J Biol Chem. 1991;266(34):22866–71. Epub 1991/12/05. 1744079.

45. Aldinucci D, Lorenzon D, Olivo K, Rapana B, Gattei V. Interactions between tissue fibroblasts in lymph nodes and Hodgkin/Reed-Sternberg cells. Leuk Lymphoma. 2004;45(9):1731–9. Epub 2004/06/30. doi: 10.1080/10428190410001683633 15223630.

46. Doi T, Higashino K, Kurihara Y, Wada Y, Miyazaki T, Nakamura H, et al. Charged collagen structure mediates the recognition of negatively charged macromolecules by macrophage scavenger receptors. J Biol Chem. 1993;268(3):2126–33. 8380589

47. Mazur A, Holthoff E, Vadali S, Kelly T, Post SR. Cleavage of Type I Collagen by Fibroblast Activation Protein-alpha Enhances Class A Scavenger Receptor Mediated Macrophage Adhesion. PloS one. 2016;11(3):e0150287. Epub 2016/03/05. doi: 10.1371/journal.pone.0150287 26934296; PubMed Central PMCID: PMC4774960.

48. Neyen C, Pluddemann A, Roversi P, Thomas B, Cai L, van der Westhuyzen DR, et al. Macrophage scavenger receptor A mediates adhesion to apolipoproteins A-I and E. Biochemistry. 2009;48(50):11858–71. Epub 2009/11/17. doi: 10.1021/bi9013769 19911804; PubMed Central PMCID: PMC2793687.

49. Santiago-Garcia J, Kodama T, Pitas RE. The class A scavenger receptor binds to proteoglycans and mediates adhesion of macrophages to the extracellular matrix. J Biol Chem. 2003;278(9):6942–6. Epub 2002/12/19. doi: 10.1074/jbc.M208358200 12488451.

50. Kirkham PA, Spooner G, Ffoulkes-Jones C, Calvez R. Cigarette smoke triggers macrophage adhesion and activation: role of lipid peroxidation products and scavenger receptor. Free radical biology & medicine. 2003;35(7):697–710. Epub 2003/10/30. doi: 10.1016/S0891-5849(03)00390-3 14583334.

51. Santiago-Garcia J, Mas-Oliva J, Innerarity TL, Pitas RE. Secreted forms of the amyloid-beta precursor protein are ligands for the class A scavenger receptor. J Biol Chem. 2001;276(33):30655–61. Epub 2001/06/05. doi: 10.1074/jbc.M102879200 11389145.

52. Gowen BB, Borg TK, Ghaffar A, Mayer EP. The collagenous domain of class A scavenger receptors is involved in macrophage adhesion to collagens. J Leukoc Biol. 2001;69(4):575–82. 11310843

53. Gowen BB, Borg TK, Ghaffar A, Mayer EP. Selective adhesion of macrophages to denatured forms of type I collagen is mediated by scavenger receptors. Matrix biology: journal of the International Society for Matrix Biology. 2000;19(1):61–71. doi: 10.1016/s0945-053x(99)00052-9 10686426.

54. van Velzen AG, Suzuki H, Kodama T, van Berkel TJ. The role of scavenger receptor class A in the adhesion of cells is dependent on cell type and cellular activation state. Exp Cell Res. 1999;250(1):264–71. doi: 10.1006/excr.1999.4530 10388540

55. El Khoury J, Thomas CA, Loike JD, Hickman SE, Cao L, Silverstein SC. Macrophages adhere to glucose-modified basement membrane collagen IV via their scavenger receptors. J Biol Chem. 1994;269(14):10197–200. 8144597

56. Fraser I, Hughes D, Gordon S. Divalent cation-independent macrophage adhesion inhibited by monoclonal antibody to murine scavenger receptor. Nature. 1993;364(6435):343–6. doi: 10.1038/364343a0 8332192

57. Kuppers R, Engert A, Hansmann ML. Hodgkin lymphoma. J Clin Invest. 2012;122(10):3439–47. Epub 2012/10/02. doi: 10.1172/JCI61245 23023715; PubMed Central PMCID: PMC3534167.

58. de Villiers WJ, Fraser IP, Hughes DA, Doyle AG, Gordon S. Macrophage-colony-stimulating factor selectively enhances macrophage scavenger receptor expression and function. J Exp Med. 1994;180(2):705–9. doi: 10.1084/jem.180.2.705 8046345

59. Nikolic D, Calderon L, Du L, Post SR. SR-A ligand and M-CSF dynamically regulate SR-A expression and function in primary macrophages via p38 MAPK activation. BMC Immunol. 2011;12:37. Epub 2011/07/09. doi: 10.1186/1471-2172-12-37 21736734; PubMed Central PMCID: PMC3141791.

60. Hamilton JA. CSF-1 signal transduction. J Leukoc Biol. 1997;62(2):145–55. doi: 10.1002/jlb.62.2.145 9261328

61. Pixley FJ, Stanley ER. CSF-1 regulation of the wandering macrophage: complexity in action. Trends Cell Biol. 2004;14(11):628–38. Epub 2004/11/03. doi: 10.1016/j.tcb.2004.09.016 15519852.

62. Xu WY, Wang L, Wang HM, Wang YQ, Liang YF, Zhao TT, et al. TLR2 and TLR4 agonists synergistically up-regulate SR-A in RAW264.7 through p38. Molecular immunology. 2007;44(9):2315–23. Epub 2006/12/19. doi: 10.1016/j.molimm.2006.11.013 17173973.

63. Fukuhara-Takaki K, Sakai M, Sakamoto Y, Takeya M, Horiuchi S. Expression of class A scavenger receptor is enhanced by high glucose in vitro and under diabetic conditions in vivo: one mechanism for an increased rate of atherosclerosis in diabetes. J Biol Chem. 2005;280(5):3355–64. Epub 2004/11/24. doi: 10.1074/jbc.M408715200 15556945.

64. van der Kooij MA, Morand OH, Kempen HJ, van Berkel TJ. Decrease in scavenger receptor expression in human monocyte-derived macrophages treated with granulocyte macrophage colony-stimulating factor. Arterioscler Thromb Vasc Biol. 1996;16(1):106–14. Epub 1996/01/01. doi: 10.1161/01.atv.16.1.106 8548409.

65. Tadmor T, Bari A, Marcheselli L, Sacchi S, Aviv A, Baldini L, et al. Absolute Monocyte Count and Lymphocyte-Monocyte Ratio Predict Outcome in Nodular Sclerosis Hodgkin Lymphoma: Evaluation Based on Data From 1450 Patients. Mayo Clin Proc. 2015;90(6):756–64. Epub 2015/06/06. doi: 10.1016/j.mayocp.2015.03.025 26046410.

66. Crane GM, Samols MA, Morsberger LA, Yonescu R, Thiess ML, Batista DA, et al. Tumor-Infiltrating Macrophages in Post-Transplant, Relapsed Classical Hodgkin Lymphoma Are Donor-Derived. PloS one. 2016;11(9):e0163559. doi: 10.1371/journal.pone.0163559 27685855; PubMed Central PMCID: PMC5042490.

67. Vari F, Arpon D, Keane C, Hertzberg MS, Talaulikar D, Jain S, et al. Immune evasion via PD-1/PD-L1 on NK cells and monocyte/macrophages is more prominent in Hodgkin lymphoma than DLBCL. Blood. 2018;131(16):1809–19. Epub 2018/02/17. doi: 10.1182/blood-2017-07-796342 29449276; PubMed Central PMCID: PMC5922274.

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