High Counts of CD68+ and CD163+ Macrophages in Mantle Cell Lymphoma Are Associated With Inferior Prognosis

Philippa Li; Ji Yuan; Fahad Shabbir Ahmed; Austin McHenry; Kai Fu; Guohua Yu; Hongxia Cheng; Mina L Xu; David L Rimm; Zenggang Pan

doi:10.3389/fonc.2021.701492

High Counts of CD68+ and CD163+ Macrophages in Mantle Cell Lymphoma Are Associated With Inferior Prognosis

Front Oncol. 2021 Aug 30:11:701492. doi: 10.3389/fonc.2021.701492. eCollection 2021.

Authors

Philippa Li¹, Ji Yuan^{2

3}, Fahad Shabbir Ahmed^{1

4}, Austin McHenry¹, Kai Fu^{3

5}, Guohua Yu^{3

6}, Hongxia Cheng^{3

7}, Mina L Xu¹, David L Rimm¹, Zenggang Pan¹

Affiliations

¹ Department of Pathology, Yale University School of Medicine, New Haven, CT, United States.
² Department of Laboratory Medicine and Pathology, Mayo Clinic, Rochester, MN, United States.
³ Department of Pathology and Microbiology, University of Nebraska Medical Center, Omaha, NE, United States.
⁴ Department of Pathology, Wayne State University, Detroit, MI, United States.
⁵ Department of Pathology and Laboratory Medicine, Roswell Park Cancer Center, Buffalo, NY, United States.
⁶ Department of Pathology, Yantai Yuhuangding Hospital, Yantai, China.
⁷ Department of Pathology, Shandong Provincial Hospital Affiliated to Shandong First Medical University, Jinan, China.

Abstract

Background: Lymphoma-associated macrophages (LAMs) are key components in the lymphoma microenvironment, which may impact disease progression and response to therapy. There are two major subtypes of LAMs, CD68+ M1 and CD163+ M2. M2 LAMs can be transformed from M1 LAMs, particularly in certain diffuse large B-cell lymphomas (DLBCL). While mantle cell lymphoma (MCL) is well-known to contain frequent epithelioid macrophages, LAM characterization within MCL has not been fully described. Herein we evaluate the immunophenotypic subclassification, the expression of immune checkpoint molecule PD-L1, and the prognostic impact of LAMs in MCL.

Materials and methods: A total of 82 MCL cases were collected and a tissue microarray block was constructed. Immunohistochemical staining was performed using CD68 and CD163, and the positive cells were recorded manually in four representative 400× fields for each case. Multiplexed quantitative immunofluorescence assays were carried out to determine PD-L1 expression on CD68+ M1 LAMs and CD163+ M2 LAMs. In addition, we assessed Ki67 proliferation rate of MCL by an automated method using the QuPath digital imaging analysis. The cut-off points of optimal separation of overall survival (OS) were analyzed using the X-Tile software, the SPSS version 26 was used to construct survival curves, and the log-rank test was performed to calculate the p-values.

Results: MCL had a much higher count of M1 LAMs than M2 LAMs with a CD68:CD163 ratio of 3:1. Both M1 and M2 LAMs were increased in MCL cases with high Ki67 proliferation rates (>30%), in contrast to those with low Ki67 (<30%). Increased number of M1 or M2 LAMs in MCL was associated with an inferior OS. Moreover, high expression of PD-L1 on M1 LAMs had a slightly better OS than the cases with low PD-L1 expression, whereas low expression of PD-L1 on M2 LAMs had a slightly improved OS, although both were not statistically significant.

Conclusions: In contrast to DLBCL, MCL had a significantly lower rate of M1 to M2 polarization, and the high levels of M1 and M2 LAMs were associated with poor OS. Furthermore, differential PD-L1 expressions on LAMs may partially explain the different functions of tumor-suppressing or tumor-promoting of M1 and M2 LAMs, respectively.

Keywords: PD-L1; lymphoma associated macrophage; lymphoma microenvironment; mantle cell lymphoma; quantitative immunofluorescence analysis.