Macrophages and Natural Killer Cells Characteristics in Variously Colored Endometriotic Lesions: A Cross-Sectional Analytic Study

Document Type : Original Article

Authors

1 Reproductive Endocrinology and Infertility Unit, Department of Obstetrics and Gynaecology, Faculty of Medicine Ramathibodi Hospital, Mahidol University, Bangkok, Thailand

2 Department of Anatomy, Faculty of Science, Mahidol University, Bangkok, Thailand

3 Chulabhorn International College of Medicine, Thammasat University, Pathumthani, Thailand

4 Office of Research Academic and Innovation, Faculty of Medicine Ramathibodi Hospital, Mahidol University, Bangkok, Thailand

5 Department of Pathology, Faculty of Medicine Ramathibodi Hospital, Mahidol University, Bangkok, Thailand

Abstract

Background: Dysregulation of the immune response contribute to a significant role in endometriosis. This research 
examined macrophages and natural killer (NK) cells numbers in endometriotic lesions and their association with the different lesion colors: red, black, and white. To investigate the amount of the CD68 and CD56 in eutopic endometrium and different type of the endometriotic lesions.
Materials and Methods: A cross-sectional analytic study was conducted. Women suspected endometriosis requiring 
laparoscopic surgery between July 2016 and January 2017 were recruited. Their lesions were classified as red, black, or white and these lesions were excised by standard laparoscopic surgery. Twenty-four endometriotic lesions from each color group were obtained from 45 women who met the inclusion criteria. One type of lesion was collected from 25 women. Two different lesion types and three-color lesion types were collected from the same women in 13 and 7 subjects, respectively. Immunohistochemistry staining with anti-human mouse cluster of differentiation (CD) 68 monoclonal antibody for macrophages and mouse anti-human CD56 monoclonal antibody for NK cells were performed.
Results: The number of CD68 macrophages in red lesions was higher than in black and white lesions [median (25th-75th 
percentile); 10 (5-19.4), 0 (0-6.9), 0 (0-2.5) cells per mm2, respectively, adjusted P=0.001 for red vs. black lesions and red 
vs. white lesions, and adjusted P=1.000 for black and white lesions]. The number of CD56 NK cells was not significantly 
different among red, black, and white lesions [median (25th-75th percentile; 5 (2-16.5), 3.8 (0-14.4), 1.3 (0-6.9) respectively, adjusted P=1.000 for red vs. black lesions and black vs. white lesions, and adjusted P=0.617 for red vs. white lesions].
Conclusion: The dynamic changes in the immune cells in ectopic endometrium were specific to the macrophages but 
not to the NK cells, as demonstrated by the highest number of CD68 macrophages in the red lesion, the earliest established ectopic endometrium. NK cells in endometriosis may have a role in the uterus.

Keywords

Main Subjects

References

1. Symons LK, Miller JE, Kay VR, Marks RM, Liblik K, Koti M, et al. The immunopathophysiology of endometriosis. Trends Mol Med. 2018; 24(9): 748-762.
2. Hogg C, Horne AW, Greaves E. Endometriosis-associated macrophages: origin, phenotype, and function. Front Endocrinol (Lausanne). 2020; 11: 7.
3. Berbic M, Schulke L, Markham R, Tokushige N, Russell P, Fraser IS. Macrophage expression in endometrium of women with and without endometriosis. Hum Reprod. 2009; 24(2): 325-332.
4. Maeda N, Izumiya C, Taniguchi K, Matsushima S, Fukaya T. Role of NK cells and HLA-G in endometriosis. Front Biosci (Schol Ed). 2012; 4: 1568-1581.
5. Jeung I, Cheon K, Kim MR. Decreased cytotoxicity of peripheral and peritoneal natural killer cell in endometriosis. Biomed Res Int. 2016; 2016: 2916070.
6. Laschke MW, Menger MD. Basic mechanisms of vascularization in endometriosis and their clinical implications. Hum Reprod Update. 2018; 24(2): 207-224.
7. Lawson C, Al-Akoum M, Maheux R, Akoum A. Increased expression of interleukin-1 receptor type 1 in active endometriotic lesions. Reproduction. 2007; 133(1): 265-274.
8. Khan KN, Masuzaki H, Fujishita A, Hamasaki T, Kitajima M, Hasuo A, et al. Association of interleukin-6 and estradiol with hepatocyte growth factor in peritoneal fluid of women with endometriosis. Acta Obstet Gynecol Scand. 2002; 81(8): 764-771.
9. Dell RB, Holleran S, Ramakrishnan R. Sample size determination. ILAR J. 2003; 44(3): 207-213.
10. Revised American Society for Reproductive Medicine classification of endometriosis: 1996. Fertil Steril. 1997;67(5): 817-821.
11. Gottfried E, Kunz-Schughart LA, Weber A, Rehli M, Peuker A, Muller A, et al. Expression of CD68 in non-myeloid cell types. Scand J Immunol. 2008; 67(5): 453-463.
12. Thiruchelvam U, Wingfield M, O’Farrelly C. Natural killer cells: key players in endometriosis. Am J Reprod Immunol. 2015; 74(4): 291-301.
13. Aziz A, Ahmad M, Khoo A. A modified automated protocol for EBER in situ hybridization staining of xenograft tissue. J Morphol Sci. 2017; 34(3): 203-206.
14. Matsuzaki S, Canis M, Murakami T, Dechelotte P, Bruhat MA, Okamura K. Immunohistochemical analysis of the role of angiogenic status in the vasculature of peritoneal endometriosis. Fertil Steril. 2001; 76(4): 712-716.
15. Tan XJ, Lang JH, Liu DY, Shen K, Leng JH, Zhu L. Expression of vascular endothelial growth factor and thrombospondin-1 mRNA in patients with endometriosis. Fertil Steril. 2002; 78(1): 148-153.
16. Donnez J, Squifflet J, Casanas-Roux F, Pirard C, Jadoul P, Van Langendonckt A. Typical and subtle atypical presentations of endometriosis. Obstet Gynecol Clin North Am. 2003; 30(1): 83-93.
17. Kokorine I, Nisolle M, Donnez J, Eeckhout Y, Courtoy PJ, Marbaix E. Expression of interstitial collagenase (matrix metalloproteinase-1) is related to the activity of human endometriotic lesions. Fertil Steril. 1997; 
68(2): 246-251.
18. Kim HO, Yang KM, Kang IS, Koong MK, Kim HS, Zhang X, et al. Expression of CD44s, vascular endothelial growth factor, matrix metalloproteinase-2 and Ki-67 in peritoneal, rectovaginal and ovarian endometriosis. J Reprod Med. 2007; 52(3): 207-213.
19. Harirchian P, Gashaw I, Lipskind ST, Braundmeier AG, Hastings JM, Olson MR, et al. Lesion kinetics in a non-human primate model of endometriosis. Hum Reprod. 2012; 27(8): 2341-2351.
20. González-Ramos R, Defrère S, Devoto L. Nuclear factor-kappaB: a main regulator of inflammation and cell survival in endometriosis pathophysiology. Fertil Steril. 2012; 98(3): 520-528.
21. Kats R, Metz CN, Akoum A. Macrophage migration inhibitory factor is markedly expressed in active and early-stage endometriotic lesions. J Clin Endocrinol Metab. 2002; 87(2): 883-889.
22. Omwandho CO, Konrad L, Halis G, Oehmke F, Tinneberg HR. Role of TGF-betas in normal human endometrium and endometriosis. Hum Reprod. 2010; 25(1): 101-109.
23. Khan KN, Masuzaki H, Fujishita A, Kitajima M, Sekine I, Ishimaru T. Differential macrophage infiltration in early and advanced endometriosis and adjacent peritoneum. Fertil Steril. 2004; 81(3): 652-661.
24. Takebayashi A, Kimura F, Kishi Y, Ishida M, Takahashi A, Yamanaka A, et al. Subpopulations of macrophages within eutopic endometrium of endometriosis patients. Am J Reprod Immunol. 2015; 73(3): 221-231.
25. Collette T, Maheux R, Mailloux J, Akoum A. Increased expression of matrix metalloproteinase-9 in the eutopic endometrial tissue of women with endometriosis. Hum Reprod. 2006; 21(12): 3059-3067.
26. Shao J, Zhang B, Yu JJ, Wei CY, Zhou WJ, Chang KK, et al. Macrophages promote the growth and invasion of endometrial stromal cells by downregulating IL-24 in endometriosis. Reproduction. 2016; 152(6): 673-682.
27. Greaves E, Temp J, Esnal-Zufiurre A, Mechsner S, Horne AW, Saunders PT. Estradiol is a critical mediator of macrophage-nerve cross talk in peritoneal endometriosis. Am J Pathol. 2015; 185(8): 2286-2297.
28. Sekiguchi K, Ito Y, Hattori K, Inoue T, Hosono K, Honda M, et al. VEGF receptor 1-expressing macrophages recruited from bone marrow enhances angiogenesis in endometrial tissues. Sci Rep. 2019; 9(1): 7037.
29. Johan MZ, Ingman WV, Robertson SA, Hull ML. Macrophages infiltrating endometriosis-like lesions exhibit progressive phenotype changes in a heterologous mouse model. J Reprod Immunol. 2019; 132: 1-8.
30. Fukui A, Mai C, Saeki S, Yamamoto M, Takeyama R, Kato T, et al. Pelvic endometriosis and natural killer cell immunity. Am J Reprod Immunol. 2021; 85(4): e13342.
31. Oosterlynck DJ, Meuleman C, Waer M, Vandeputte M, Koninckx PR. The natural killer activity of peritoneal fluid lymphocytes is decreased in women with endometriosis. Fertil Steril. 1992; 58(2): 290-295.
32. Tanaka E, Sendo F, Kawagoe S, Hiroi M. Decreased natural killer cell activity in women with endometriosis. Gynecol Obstet Invest. 1992; 34(1): 27-30.
33. Dias JA, Jr., Podgaec S, de Oliveira RM, Carnevale Marin ML, Baracat EC, Abrao MS. Patients with endometriosis of the rectosigmoid have a higher percentage of natural killer cells in peripheral blood. J Minim Invasive Gynecol. 2012; 19(3): 317-324.
34. Maeda N, Izumiya C, Yamamoto Y, Oguri H, Kusume T, Fukaya T. Increased killer inhibitory receptor KIR2DL1 expression among natural killer cells in women with pelvic endometriosis. Fertil Steril. 2002; 77(2): 297-302.
35. Matsuoka S, Maeda N, Izumiya C, Yamashita C, Nishimori Y, Fukaya T. Expression of inhibitory-motif killer immunoglobulin-like receptor, KIR2DL1, is increased in natural killer cells from women with pelvic endometriosis. Am J Reprod Immunol. 2005; 53(5): 249-254.
36. Zhang C, Maeda N, Izumiya C, Yamamoto Y, Kusume T, Oguri H, et al. Killer immunoglobulin-like receptor and human leukocyte antigen expression as immunodiagnostic parameters for pelvic endometriosis. Am J Reprod Immunol. 2006; 55(2): 106-114.
37. Drury JA, Parkin KL, Coyne L, Giuliani E, Fazleabas AT, Hapangama DK. The dynamic changes in the number of uterine natural killer cells are specific to the eutopic but not to the ectopic endometrium in women 
and in a baboon model of endometriosis. Reprod Biol Endocrinol. 2018; 16(1): 67.
38. Sentman CL, Meadows SK, Wira CR, Eriksson M. Recruitment of uterine NK cells: induction of CXC chemokine ligands 10 and 11 in human endometrium by estradiol and progesterone. J Immunol. 2004; 173(11): 6760-6766.
39. Seshadri S, Sunkara SK. Natural killer cells in female infertility and recurrent miscarriage: a systematic review and meta-analysis. Hum Reprod Update. 2014; 20(3): 429-438.
40. Freitag N, Pour SJ, Fehm TN, Toth B, Markert UR, Weber M, et al. Are uterine natural killer and plasma cells in infertility patients associated with endometriosis, repeated implantation failure, or recurrent pregnancy loss? Arch Gynecol Obstet. 2020; 302(6): 1487-1494

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