Immunohistochemical markers for tumor associated macrophages and survival in advanced classical Hodgkin's lymphoma

Abstract

A subset of patients with advanced classical Hodgkin's lymphoma is refractory to standard therapies. Therefore, it is relevant to identify new biologically-based prognostic markers. Recently, tumor associated macrophages have been proposed as a factor that predicts survival, although contradictory results have also been reported. Here we analyzed four macrophage markers (CD68, CD163, LYZ, and STAT1) using immunohistochemistry and automated quantification, in two independent series of advanced classical Hodgkin's lymphoma (n=266 and 103 patients, respectively). Our results did not confirm that specific macrophage immunohistochemical markers could be used as surrogates for gene expression profiling studies. Survival analyses did not show correlation between CD163, LYZ or STAT1 and either failure-free or disease-specific survival. There was an association between CD68 and disease-specific survival, but it was not consistent in both series. In conclusion, individual tumor associated macrophage markers cannot be used to predict outcome before technical standardization and prospective validation in independent series of patients with comparable stages and treatments.Key words: Hodgkin's lymphoma, outcome, tumor associated macrophages. Hodgkin's lymphoma. Haematologica 2012;97(7): 1080-1084. doi:10.3324/haematol.2011 This is an open-access paper. Citation: Sánchez-Espiridión B, Martin-Moreno AM, Montalbán C, Medeiros LJ, Vega F, Younes A, Piris MA, and Garcia JF. Immunohistochemical markers for tumor associated macrophages and survival in advanced classical ABSTRACT © F e r r a t a S t o r t i F o u n d a t i o nstage disease who had a survival rate of 100%. These results suggest that semi-quantitative assessment of CD68 + TAMs can be used to identify patients at high risk of disease relapse or progression, as well as patients with early-stage disease with low risk for relapse who are currently over-treated.11 However, this approach has a major drawback as the number of low-risk patients in the Steidl et al. study was low because very few cases of cHL had less than 5% of TAM. Therefore, about 72% of cases in the original report fall into the high-risk group 10 and might be selected for alternative treatments.Similar results about the predictive value of TAM in cHL have recently been reported by other groups. 12,13 However, the consistency of IHC markers in different series has not been clearly confirmed.14,15 Differences in the composition of the case series, technical variability, and different cut offs for expression of TAM associated markers could, in part, explain these diverse results. Also, roles for Epstein-Barr virus (EBV) and age as confounding factors have been claimed, since both variables influence the host immune response and TAM composition. 12,16We recently used GEP to identify specific genes associated with treatment failure in cHL patients 8,17 including gene signatures associated with reactive cells in the microenvironment. This gene signature was related to outcome in a selected series of advanced s...

Overexpression of Semaphorin4D Indicates Poor Prognosis and Prompts Monocyte Differentiation toward M2 Macrophages in Epithelial Ovarian Cancer

“…CD163, more specific than CD68 marker, is expressed in the M2 macrophages. Several studies had already proved that high M2 macrophage count indicated tumor more aggressive and poor prognosis, including breast cancer (Shabo et al, 2008), lung adenocarcinoma (Zhang et al, 2011), and classical Hodgkin's lymphoma (Sanchez-Espiridion et al, 2012). Moreover, Lan et al recently showed that the infiltration of CD163-positive M2 macrophages as well as activation of macrophages towards the M2 phenotype may contribute to poor survival in advanced ovarian cancer (Lan et al, 2013), which was consistent with our results that demonstrated that both high M2 macrophage count in primary tumor and percent in ascites were closely related to EOC patients with advanced stage and platinum resistant and refractory and suggested a shorter OS and PFS.…”

Serum CD163 and TARC as Disease Response Biomarkers in Classical Hodgkin Lymphoma

“…CD163 positivity was scored by 2 independent hematopathologists (C. Keane and L.A. Seymour) using a standardized protocol and the mean value recorded. Samples were delineated as high or low based on a previously published cutoff of 25% (16)(17)(18). In addition, dual staining of CD163 and CD68 (mouse antihuman CD68 clone: PG-M, 40 mg/mL, DAKO) was done on 12 samples to confirm CD163 specificity.…”

“…CD163 is an M2 macrophage marker that is also expressed in a subset of monocytes, and is highly expressed within tumor-infiltrating cells of the malignant node. Tissue levels of CD163 within the cHL node may be associated with survival, but results are conflicting (12)(13)(14)(15)(16)(17)(18). A study of serum CD163 (sCD163) in patients with stage I/ II melanoma found that pretherapy sCD163 was an independent predictor of survival (19).…”

High tumour-associated macrophages infiltration is correlated with poor survival outcome in classical Hodgkin’s lymphoma

“…Different cut-off points were used and accordingly the results are different. While, most studies reported association between high CD68+ TAM infiltration and poor survival outcomes (DFS, [5,19,[35][36][37][38]41] DSS, [13,19,36] or OS), [5,[35][36][37][41][42][43][44][45] fewer studies found that there no relation between high levels of CD68+ TAM infiltration and outcome. [16,26,39,40] The results from our study support the trend that higher CD68+ TAM infiltration in TAM can be used as a biological marker for stratification of patients with risk of adverse outcome.…”

“…The presence of numerous tumour associated macrophages (TAM) in HL correlated with B symptoms and a poor response to therapy. [10][11][12][13][14] Also, TAM had been associated with disease status in non-haematological malignancies. [15][16][17] A gene signature of TAM associated with primary treatment failure in cHL and immunohistochemical detection of TAM was correlated with a shorter disease free survival (DFS).…”