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ORIGINAL ARTICLE
Year : 2021 | Volume
: 46
| Issue : 3 | Page : 155-159
Hemoglobin scavenger receptor (cluster of differentiation 163) role in acute leukemia
Hossam El Din Salah Mohamed1, Mohamed Eissa MD 2
1 Department of Clinical Pathology, Faculty of Medicine, Zagazig University, Zagazig, Egypt
2 Department of Clinical Pathology, Faculty of Medicine, Zagazig University, Zagazig; Department of Pathology, College of Medicine, King Khalid University, Abha, KSA, Egypt
Correspondence Address:
Mohamed Eissa
Kingdom Of Saudi Arabia, College of Medicine, King Khaled University, Abha, 61412, Pathology Department, P.O. Box 641, KSA
Egypt
Source of Support: None, Conflict of Interest: None
CheckDOI: 10.4103/ejh.ejh_41_20
Background Cluster of differentiation 163 (CD163) is a biomarker correlated with several normal and pathological states.
Aim This work was carried out to evaluate the expression of CD163 in patients with acute leukemia.
Patients and methods The study was carried out on 50 participants divided into three groups: 10 apparently normal healthy individuals, 30 patients with acute myeloid leukemia (AML), and 10 patients with acute lymphoid leukemia. All participants were subjected to a thorough history and clinical examination. Becton–Dickinson Calibur FACScan color multiparameter flow cytometry was used for the detection of CD163 expression in patients with acute leukemia.
Results There was a significant difference in CD163 expression between AML and acute lymphoid leukemia (F=7.83) (P=0.001). CD163 expression was not observed in patients with acute lymphoblastic leukemia. However, it was expressed in 14 patients with acute myeloblastic leukemia. Five patients were diagnosed as M4, and all of them (100%) showed positive expression of CD163. Eight patients were diagnosed as M5, and all of them (100%) showed positive expression of CD163. However, CD163 was expressed in only one case among 17 patients with AML subtypes other than M4/M5. There was a significant difference between monocytic and nonmonocytic leukemic patients regarding CD163 expression (P<0.001). There was a strong correlation between CD163 and other markers predominantly found in monocytic leukemia such as CD14 (r=0.8) (P<0.001), CD15 (r=6.43) (P<0.001), and CD64 (r=0.82) (P<0.001).
Conclusion CD163 was exclusively expressed on the monocytic and myelomonocytic leukemia, so it can be used for the diagnosis of the monocytic type of AML. Although it cannot be used as a prognostic marker, it could be a novel immunotherapeutic intervention for acute monocytic and myelomonocytic leukemia.
Keywords: acute lymphocytic leukemia, acute myeloid leukemia, cluster of differentiation 163, diagnosis, monocytic, prognostic
Acute myeloid leukemia (AML) or acute nonlymphocytic leukemia is a clonal proliferation of immature myeloid precursors with an arrest in the maturation [1].
AML is the predominant form of leukemia during the neonatal period but represents less than 15% of cases of leukemia in children under 10 years and 25–35% between ages 10 and 15 years. However, in adults, it represents 80–90% of cases of acute leukemia [2].
The scavenger receptor cysteine-rich superfamily is a family of structurally related transmembrane glycoproteins of soluble or membrane-bound protein receptors. No integrated function has been described for the scavenger receptor cysteine-rich domains, but it could result in diagnostic and/or therapeutic service for a number of physiologic and pathologic conditions [3].
Cluster of differentiation 163 (CD163) is a scavenger receptor of the complex of haptoglobin (Hp) and hemoglobin (Hb) (Hp–Hb) formed after the lysis of red blood cells. It functions as an acute-phase-regulated receptor involved in the clearance and endocytosis of Hb/Hp complexes by macrophages and may thereby protect tissues from free Hb-mediated oxidative damage [4].
This protein may also function as an innate immune sensor for bacteria (Staphylococcus mutans, Staphylococcus aureus, and Escherichia More Details coli) and virus strains and an inducer of local inflammation and apoptosis [5].
CD163 is a potential inflammation biomarker, as the mature tissue macrophages express high levels of CD163, including Kupffer cells in the liver, red pulp macrophages in the spleen, cortical macrophages of the thymus, resident mature bone marrow (BM) macrophages, and scattered macrophages in different tissues [6].
Hb scavenger receptor was discovered in 1987. This protein is encoded by the CD163 gene on chromosome 12 P13 [7]. The most potent stimulators of CD163 expression known are glucocorticoid, interleukin (IL)-6, IL-10, and heme/Hb, whereas IL-4, lipopolysaccharide, tumor necrosis factor-α, interferon γ, and granulocyte–macrophage colony-stimulating factor downregulate CD163 expression [8].
Senescent or malformed red blood cells are cleared by tissue macrophages in the spleen, liver, and BM. Clearance of free Hb by the scavenger receptor CD163 is one of the physiologic functions of peripheral blood monocytes [9].
The macrophage CD163-mediated endocytosis of Hb–Hp represents also a significant pathway for the uptake of iron, which in turn may be a remarkable function for this protein. The larger part of Hb iron present in elderly red blood cells is recycled for new erythropoiesis after the old cells are phagocytosed by macrophages [10].
CD163 works in two modes: high-affinity scavenger receptor for the Hb–Hp complex and with lower affinity for Hb alone. Competent extracellular hemoglobin clearance is critical to prevent cellular oxidative-mediated and nitrosative-mediated toxicity [11].
The biomarker form of this protein is the soluble plasma CD163, commonly denoted as soluble CD163, which arises from the increased shedding of the membrane-bound receptor, which may represent a form of modulatory CD163 mediated by the tumor necrosis factor-α cleaving enzyme upon activation of cell surface Toll-like receptors [12].
In addition, soluble CD163 is suggested to have an immunomodulatory role by two distinct mechanisms. The first mechanism is achieved by clearance of the Hb, which results in conversion of the heme molecule to carbon monooxide, biliverdin, and Fe. The second is the ligand binding to CD163 to induce tyrosine kinase- and calcium-dependent signaling and increased secretion of IL-6 and IL-10 [13].
Moreover, the soluble CD163 is a biomarker correlated with several normal and pathological states [14]. It acts as a class of macrophage-specific biomarkers relevant to the detection of coronary artery disease, cardiac surgery, transplantation, atherosclerosis, lymphoma, reactive hemophagocytic syndrome, histiocystic neoplasm, myeloproliferative diseases and myelomonocytic leukemia, rheumatoid arthritis, hepatitis, fulminant hepatic failure, liver cirrhosis, malaria, storage diseases, hemophagocytosis, cancer, and even in multiple sclerosis [15].
CD163 could be a novel therapeutic target. Cheng et al. [16] confirmed that CD163 was a novel target gene of signal transducer and activator of transcription 3 in gastric cancer, and CD163 may be a poor prognostic marker and therapeutic target for gastric cancer. Moreover, Chen et al. [17], indicated that CD163 contributed to gliomagenesis via CK2 and provides preclinical evidence that CD163 and the CD163 pathway might serve as a therapeutic target for glioma. Recently, other studied supported the significance of the monocyte/macrophage-related markers for prediction of sepsis in children and highlighted the potential diagnostic use of their soluble forms as new pediatric sepsis biomarkers [18].
This work was done to evaluate the role of CDI63 expression in acute leukemia.
Patients and methodsThis prospective study was conducted according to the principles of the Declaration of Helsinki and was ethically approved by the Medical Ethical Committee, Faculty of Medicine, Zagazig University. The authors declare that they have no competing interests. It was carried out in Medical Oncology and Clinical Pathology Department, Zagazig University hospitals. The control group included 10 apparently normal healthy individuals. The patient group included 30 patients with de novo AML and 10 patients with de novo acute lymphocytic leukemia (ALL). The control group included five females and five males. Their ages ranged from 25 to 65 years, with a mean age of 38±21.6 years. The patient group included 40 patients with de novo acute leukemia, comprising 26 males and 14 females. The age of patients with AML ranged between 25 and 80 years (the mean age was 45.2±14.4 years) whereas that of patients with ALL ranged between 19 and 45 years (the mean of age 21.8±14.1). Patients were followed up throughout the period of the study. All cases were subjected to detailed history, through clinical examination, and routine investigations. Complete blood count was done using Sysmex XS500i. Liver function tests, kidney function tests, erythrocyte sedimentation rate, and serum lactate dehydrogenase were done for all participants. BM aspirates, cytochemical staining for peripheral blood smears and BM aspirate films, and immunophenotyping were done for the patient group.
Sampling
Overall, 2-ml EDTA peripheral blood sample was collected for complete blood count from all participants. BM aspirate specimens were collected from patients for morphologic and immunophenotypic diagnosis.
Cluster of differentiation 163 expression by multiparameter flow cytometry
EDTA-whole fresh peripheral blood and/or BM samples were used. Overall, five μl of selected proper monoclonal antibodies was added to each tube containing 100 μl blood or BM and incubated for 20–30 min in dark room temperature, washed with phosphate-buffered saline three times, then added 300 μl phosphate-buffered saline for analysis. Isotopic control using immunoglobulin G1 and immunoglobulin G2 on each sample was done to exclude autofluorescent. To establish the blast phenotype in every acute leukemia case, a panel of mouse monoclonal antibodies directed against different hemopoietic and differentiation antigens, directly conjugated with fluorescein isothiocyanate, phycoerythrin, and peridinin–chlorophyll proteins, was used. For the detection of CD163 expression, BV510 mouse anti-human CD163 clone GHI/61(RUO) was used. All monoclonal antibodies were purchased from DAKO (Denmark). The samples were analyzed on a Calibur FACScan flow cytometry (Becton–Dickinson, San Jose, California, USA). The leukemic cell population was identified by gating the typical formation in the forward-and sideward-scatter projection, with (residual) lymphocytes and monocytes included. CellQuest software (BD Biosciences, San Jose, California, USA) was used to calculate positive events in the defined gate compared with the isotype control. If the percentage of positive events was 20%, the leukemic sample was considered to be positive for that surface marker.
Statistical methods
All statistical analyses were performed using Statistical Package for the Social Sciences (SPSS), version 21 (SPSSVR; IBM, Armonk, NY, USA). P value less than 0.05 was considered significant. Data were compared between cases and controls using χ2 tests, Mann–Whitney test, Student t test, Kruskal–Wallis Z test, ‘analysis of variance’ test, and Spearman r correlation coefficient, which were used as tests of significance.
Results[Table 1] shows the significant correlation between CD163 expression and myelomonocytic markers.
Table 1 Correlation between cluster of differentiation 163 and acute leukemia markers[Table 2] shows a significant positive expression of CD163 in myelomonocytic leukemias ([Figure 1],[Figure 2],[Figure 3]).
Table 2 Relation between acute myeloid leukemia FAB subtypes and expression of cluster of differentiation 163
Figure 1 Correlation between CD163 and CD14. CD, cluster of differentiation.
Figure 2 Correlation between CD163 and CD15. CD, cluster of differentiation.
Figure 3 Correlation between CD163 and CD64. CD, cluster of differentiation. Discussion In the present work, we studied the expression of the Hb scavenger receptor (CD163/HbSR) in acute leukemia. There was a significant difference between AML and ALL cases (F=7.83) (P=0.001), as CD163 expression was not observed in patients with ALL [17].
AML is a diverse cancer involving an aggregation of immature myeloid cells in the peripheral blood, BM, and other tissues with a fluctuating depression in the production of normal blood cells with a variety of systemic manifestations including anemia, bleeding, and an increased risk of infection as a result of BM failure [18].
Prognostic factors in AML are subdivided into those that are related to the patient and those that are related to the disease. Patient-associated factors are increasing age, sex, coexisting diseases, and poor performance condition. Disease-related factors are white-cell count, pre-leukemic syndromes, cytotoxic therapy for another malignancy, and genetic changes [19].
In this study, the expression of CD163 in 30 patients with AML was assessed. Three (10%) patients with AML/M1 were negative for CD163 expression. Eight (26.7%) AML/M2 cases were included; one of them was positive for CD163, whereas seven cases were negative. Six (20%) cases were AML/M3, and all of them were negative for CD163 expression. These results showed that one (5.9%) patient of 17 nonmonocytic leukemia cases was positive for CD163. Our finding was in agreement with Garcia et al. [20].
Five (16.6%) patients were diagnosed as M4, and all of them (100%) showed positive expression of CD163 (the mean of CD163 expression was 34.8±8.95). Eight (26.7%) patients were diagnosed as M5, and all of them (100%) showed positive expression of CD163 (the mean of CD163 expression was 45.9±16.5). These results showed that all of the patients with monocytic leukemia were positive for CD163 (100%). This finding coincided with Garcia et al. [21] who found that CD163 and CD14 were the most specific markers of monocytic differentiation.
There was a highly significant difference between monocytic and nonmonocytic leukemic patients (F=16.87) (P<0.001). CD163 was expressed in all cases of M4 and M5 (13/13 cases), whereas it was expressed in one of 17 patients with nonmonocytic AML. Our findings were in agreement with Klco et al. [22].
There is a strong correlation between CD163 and other markers predominantly found in monocytic leukemia such as CD14 (r=0.8) (P<0.001), CD15 (r=6.43) (P<0.001), and CD64 (r=0.82) (P<0.001). Our results are in agreement with Bächli et al. [23], who demonstrated that expression of CD163 was positive in 84% (16/19 cases) of M4 patients and 87% (41/47 cases) of M5, whereas other AML subtypes without monocytic markers had no or minimal expression of CD163. The findings were also in agreement with Walter et al. [24], who demonstrated a strong correlation between CD163 expression and markers predominately found in monocytic leukemia.These findings suggest that CD163 can help in the diagnosis of these cases and that anti-CD163 could be a suitable target for immune-therapeutic intervention in monocytic leukemias. Our result was in agreement with Gorczyca et al. [25], and others who demonstrated significant expression of CD163 on leukemic blasts of monocytic and myelomonocytic AML cases, so they implicated it as immunophenotypic differentiation marker for these cells and as a putative target for therapy in patients with AML types M4/M5.
We found that CD163 is one of the most specific markers of monocytic differentiation, but on analyzing the association between CD163 expression and the studied standard prognostic factors of AML, no significant association existed between CD163 expression and any of them in AML group.
ConclusionCD163 expression is a specific marker for monocytes and macrophages. It was exclusively expressed on the monocytic and myelomonocytic leukemia, so it can be used for diagnosis of monocytic type of AML, but it cannot be used as a prognostic marker. CD163 could be a novel therapeutic target for acute monocytic and myelomonocytic leukemia.
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Conflicts of interest
There are no conflicts of interest.
References
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