ABSTRACT
As main recipient cells for porcine reproductive and respiratory syndrome virus (PRRSV), porcine alveolar macrophage (PAM) are involved in the progress of several highly pathogenic virus infections. However, due to the fact that the PAM cells can only be obtained from primary tissues, research on PAM-based virus-host interactions remains challenging. The improvement of induced pluripotent stem cells (iPSCs) technology provides a new strategy to develop IPSCs-derived PAM cells. Since the CD163 is a macrophage-specific marker and a validated receptor essential for PRRSV infection, generation of stable porcine induced pluripotent stem cells lines containing CD163 reporter system play important roles in the investigation of IPSCs-PAM transition and PAM-based virus-host interaction. Based on the CRISPR/Cas9- mediated gene editing system, we designed a sgRNA targeting CD163 locus and constructed the corresponding donor vectors. To test whether this reporter system has the expected function, the reporter system was introduced into primary PAM cells to detect the expression of RFP. To validate the low effect on stem cell pluripotency, we generated porcine iPSC lines containing CD163 reporter and assessed the pluripotency through multiple assays such as alkaline phosphatase staining, immunofluorescent staining, and EdU staining. The red-fluorescent protein (RFP) expression was detected in CD163-edited PAM cells, suggesting that our reporter system indeed has the ability to reflect the expression of gene CD163. Compared with wild-type (WT) iPSCs, the CD163 reporter-iPSCs display similar pluripotency-associated transcription factors expression. Besides, cells with the reporter system showed consistent cell morphology and proliferation ability as compared to WT iPSCs, indicating that the edited-cells have no effect on stem cell pluripotency. In conclusion, we generated porcine iPSCs that contain a CD163 reporter system. Our results demonstrated that this reporter system was functional and safe. This study provides a platform to investigate the iPS-PAM development and virus-host interaction in PAM cells.
Subject(s)
Animals , Swine , Induced Pluripotent Stem Cells/metabolism , Receptors, Cell Surface/genetics , Antigens, CD/metabolism , Porcine respiratory and reproductive syndrome virus/geneticsABSTRACT
Histiocytic sarcoma is very rare malignant neoplasm showing morphological and immunophenotypic characteristics of histiocyte. Morphologic diagnosis is challenging due to overlapping histologic features with diverse mimics. Histologic diagnosis is confirmed by expression of one or more histiocytic markers, including CD163, CD68, and lysozyme. We present a bona fida case of extranodal histiocytic sarcoma involving the small intestine in a 67-year-old male. The diagnosis is confirmed by diffuse immunoreactivity of CD163, which is believed to be more specific histiocytic marker than CD68.
ABSTRACT
Objective To investigate the expression of macrophages in the bone marrow of chronic lympho-cytic leukemia (CLL) patients and its clinical significance in the pathogenesis of CLL .Methods Fifty-eight patients with newly diagnosed CLL were selected ,including 24 cases of Binet A ,19 cases of Binet B ,15 cases of Binet C ,and 20 patients with iron deficiency anemia were selected as control group .Immunohistochemical staining was used to detect the expression of CD68+ (M1+M2 type) and CD163 (M2 type) in CLL patients . The expression differences in bone marrow tissues of CLL patients at different stages were compared . Results The numbers of CD68+ and CD163+ cells in CLL group were significantly higher than those in con-trol group (P<0 .05) ,while those in Binet C stage patients were higher than in Binet B stage patients (P<0 .05) and those in Binet B stage patients were higher than in Binet A stage patients (P<0 .05) .The progres-sion of CLL was positively correlated with the infiltration density of CD 163+ cells (P<0 .05) .Conclusion Macrophages have a high density of infiltration in the bone marrow of CLL patients ,and the infiltration densi-ty varies in different periods of CLL .With the progresses of the disease ,macrophages infiltrated into bone marrow gradually polarize to M 2 type ,w hich is relevant with the course of CLL progress .
ABSTRACT
Objective To investigate the relationship between M2 macrophages and prognosis of patients with human colorectal cancer by using CD163 protein as a marker. Methods The clinical data and paraffin samples of 648 patients with newly diagnosed colorectal cancer and the paraffin specimens of the tumor tissues after operation were collected from Jan. 2010 to Dec. 2011 in the Department of Colorectal Surgery, Changhai Hospital, Navy Medical University (Second Military Medical University). All patients were diagnosed by postoperative pathology. Paracancerous tissues of the above 38 patients, normal colorectal mucosal tissues of 37 patients undergoing hemorrhoidectomy (PPH), and colorectal adenoma tissues of 33 patients receiving endoscopic treatment served as controls. All specimens were made into tissue microarrays, and the expression of CD163 protein was detected by immunohistochemistry. The relationships between the expression of CD163 protein and clinicopathological parameters and prognosis of patients with colorectal cancer were analyzed. Results The expression level of CD163 was related to the degree of tumor differentiation, serum carbohydrate antigen 19-9 (CA19-9) and tumor recurrence and metastasis (all P0.05). Survival analysis showed that the high CD163 expression group had lower disease-free survival and overall survival rates and poor prognosis compared with the low CD163 expression group (P0.001). Cox multivariate analysis showed that serum expression levels of carcinoembryonic antigen (CEA) and CD163 were independent prognosis factors of patients with colorectal cancer (P0.05). Conclusion M2 macrophage marker CD163 has a good reference value for the prognosis of patients with colorectal cancer.
ABSTRACT
Objective To investigate the correlation between the number of CD68+ and CD163+ macrophages infiltration and the clinical characteristics of oral squamous cell carcinoma (OSCC),and to investigate the role and significance of CD163+ and CD68+ macrophages in OSCC.Methods Forty-two cases of OSCC resected by operation and diagnosed by postoperative pathology with intact clinical data in our hospitals during 2013-2016 were selected.Other 12 cases of oral mamaxillofacial normal tissue served as controls.The number of CD68+ and CD163+ macrophages infiltration in OSCC was detected by immunohistochemistry.Its correlation with clinical characteristics was analyzed.Results The number of CD68+ and CD163+ macrophages infiltration in OSCC was significantly higher than that in the control group (P<0.05);the infiltration quantity of CD68+ and CD163+ macrophages in poor differentiation and moderate differentiation was significantly higher than that in high differentiation(P<0.05).The number of CD68+ macrophages in lymph node metastasis was significantly lower than that without lymph node metastasis (P<0.05).The number of CD163+ macrophages in the lymph node metastasis was significantly higher than that of CD68+ (P<0.05).The number of CD68+ and CD163+ macrophages had positive correlation in OSCC (r=0.48,P=0.00).Conclusion Tumor-associated macrophages (TAMs) and M2 macrophages may play an important role in OSCC,TAMs may inhibits lymph node metastasis in OSCC,while M2 marcophages may promote the lymphatic matastasis of OSCC.
ABSTRACT
Objective To investigate the diagnostic and prognostic value of serum soluble CD163 (sCD163 )and the positive rate of membrane -bound CD163 (mCD163 )in peripheral blood mononuclear cells (PBMC)in children with infection -associated hemophagocytic syndrome (IAHS).Methods Between July 2012 and June 2016,26 pediatric patients with IAHS (IAHS group)and 28 pediatric patients with sepsis(sepsis group)admitted to Children′s Hospital Affiliated to Shanghai Jiaotong University were selected,and 20 healthy children were taken as healthy control group. Sandwich enzyme linked immunosorbent assay was used to detect serum sCD163 .The population of circulating mCD163 positive monocytes was determined by using flow cytometry.Receiver operating characteristic (ROC)curves were used to evaluate the diagnostic and prognostic values of sCD163 and mCD163 in children with IAHS compared with the diagnos-tic and prognostic values of plasma ferritin,and so on.Results The serum levels of sCD163 in patients of IAHS group, sepsis group and healthy control group were (1264 ±538)mg/L,(862 ±332)mg/L,(610 ±316)mg/L,respective-ly.And the population of mCD163 -positive PBMC in patients of IAHS group,sepsis group and healthy control group was (88.3 ±9.7)%,(68.5 ±18.3)%,(28.9 ±5.2)%,respectively.Both serum sCD163 and the population of mCD163 -positive PBMC were significantly higher in IAHS group compared with those of sepsis group (t =2.031 ,P =0.048;t =3.191 ,P =0.002,respectively).The serum sCD163 and population of mCD163 -positive PBMC in sepsis group were higher than controls (t =3.848,P =0.002;t =4.049,P =0.000,respectively).Moreover,the areas under the ROC curve (AUC)for the mCD163 ,sCD163 ,were 0.853(P =0.013),0.762(P =0.004),0.755(P =0.049),respec-tively.mCD163 at a cutoff of 83.7% had a high diagnosis sensitivity (81 .8%)and specificity (72.4%).The optimal cutoff values of sCD163 and ferritin for predicting IAHS was 888 mg/L (sensitivity 66.7% and specificity 63.3%)and 2880 μg/L (sensitivity 80.0% and specificity 54.5%).In addition,the serum level of sCD163 and the population of mCD163 -positive PBMCs were significantly increased in acute phase and decreased in recovery phase[(1553 ±542) mg/L vs.(866 ±92)mg/L,(91 .0 ±6.4)% vs.(79.0 ±4.6)%,t =2.450,χ2 =3.419,P =0.036,0.007]in IAHS group.Furthermore,subgroup analysis indicated that the serum level of sCD163 and the population of mCD163 -positive PBMCs were significantly higher in dead patients than those in survived patients [(1748.91 ±518.17)mg/L vs. (909.69 ±171 .35)mg/L,t =3.070,P =0.011 ;(93.50 ±8.42)% vs.(77.30 ±3.28)%,χ2 =3.005,P =0.024, respectively].Conclusion Serum sCD163 and the population of mCD163 -positive PMSCs are specific and validity bio-markers for early diagnosis of IAHS,which also are associated with treatment response assessment and prognostic analy-sis in IAHS.
ABSTRACT
Objective To study the expression of CD163 in macrophages and sCD163 level in the serum of neonatal rat model of Escherichia coli (E. coli) sepsis. Methods A total of 72 specific-pathogen-free (SPF) neonatal rats (P7) were randomly and equally assigned into experiment group and control group. E. coli was injected peritoneally and the sepsis model was established in the experiential group while normal saline (NS) was injected in the control group. Samples were collected at 2, 4, 6, 12, 24 h and 48 h after the treatment. CD163 expression in macrophages of lung and liver tissues were tested using immunohistochemical(IHC) method, and the dynamic changes of sCD163 concentration in the serum were monitored usingenzyme-linked immunosorbent assay (ELISA) method. Results In the experiment group, CD163 expression in macrophages of lung and liver were gradually decreased at eachtime point (P 0. 05). At 4 h and later timepoints, the differences were statistically significant (P 0. 05). At 4 h and later timepoints, the differences were statistically significant (P <0. 001). Conclusions CD163 plays an important role in sepsis.
ABSTRACT
Objective:To investigate the expression of CD 163 in the plasma and tumor tissue of non-small cell lung cancer (NSCLC)patients,and the effects of cancer cell migration and proliferation .Methods:35 cases of NSCLC patients in our hospital as the study group,and 35 healthy volunteers as control group ,we used Real-time RCR to detect the expression of CD163 in the plasma and tumor tissue of NSCLC patients.Using small interfering RNA (siRNA) to inhibit the expression of CD163 in MH-2,and co-culture with A549.The level of cancer cell migration was observed by transwell and the level of cell proliferation was observed by CCK -8.Results:The expression of CD163 in the plasma and tumor tissue of NSCLC patients was significantly higher than the control group , the difference was statistically significant (P<0.05);after the expression of CD163 was suppressed in MH-2,the levels of cancer cell migration and proliferation were significantly decreased , the difference was statistically significant ( P<0.05 ) .Conclusion: The expression of CD163 in the plasma and tumor tissue of NSCLC patients is increased ,and may elevate the levels of cancer cell migration and proliferation.
ABSTRACT
BACKGROUND: Tumor associated macrophages (TAMs) and CXC chemokine receptor 4 (CXCR4) have emerged as potential biomarkers in various human cancers. The aims of this study were to investigate the clinical characteristics of anaplastic thyroid cancer (ATC) patients according to the TAM numbers in the tumor tissue, and to evaluate the associations between CXCR4 expressions and macrophage densities in ATC tumor microenvironment. METHODS: Total 14 ATC samples from thyroid tissue microarray were used. Immunohistochemical staining was performed using anti-CD163 and anti-CXCR4 antibodies. According to the immunoreactivity of CD163, all subjects were divided into two groups: low-CD163 (n=8) and high-CD163 (n=6) groups. RESULTS: The mean diagnostic age was 65±7 years and the median tumor size was 4.3 cm, ranging 2.5 to 15 cm. Clinicopathological characteristics were not significantly different between low-CD163 and high-CD163 groups, while age of diagnosis was younger in high-CD163 group than that of low-CD163 group with marginal significance (56.9±5.5 years vs. 67.5±6.8 years, P=0.09). However, overall survival was significantly reduced in high-CD163 group (5.5 months [range, 1 to 10]) compared with low-CD163 groups (8.8 months [range, 6 to 121); log-rank test, P=0.0443). Moreover, high-CD163 group showed strong CXCR4 expressions in both cancer and stromal compartments, while low-CD163 group showed relatively weak, stromal-dominant CXCR4 expressions. Additionally, CD163 and CXCR4 expressions showed a strong positive correlation (γ²=0.432, P=0.013). CONCLUSION: Increased number of TAMs showed poor overall survival in ATC, suggesting TAMs are potentially a prognostic biomarker for ATC. CXCR4 expression was significantly correlated with CD163-positive TAM densities, which suggest the possible role of CXCR4 in TAM recruitments.
Subject(s)
Humans , Antibodies , Biomarkers , Diagnosis , Macrophages , Receptors, CXCR , Receptors, CXCR4 , Thyroid Carcinoma, Anaplastic , Thyroid Gland , Tumor MicroenvironmentABSTRACT
Objective To analyze the correlation between Doppler ultrasound parameters, blood biochemical indexes and the severity of esophageal varices (EV), and to explore the values of them in the diagnosis of severe EV.Methods A total of 102 hospitalized patients with liver cirrhosis was consecutively collected.All patients underwent endoscopic examination to determine the presence and severity of EV, then they were divided into moderate/severe EV group (n=78) and non/mild EV group (n=24).Congestion index of the portal vein (PV-CI), damping index of hepatic vein(HV-DI), hepatic vein arrival time(HVAT) and intrahepatic circulatory time (IHCT) were measured by Doppler ultrasound and contrast-enhanced ultrasound.And at same time, the level of yon Willebrand factor antigen (vWF-Ag) and soluble CD163 (sCD163) in peripheral blood were detected by enzyme linked immunosorbent assay (ELISA) method.The differences of these parameters between two groups were compared by independent samples t-test or Mann-Whitney rank sum test.The sensitivity, specificity, positive predictive value (PPV), negative predictive value (NPV) and diagnostic accuracy (DA) of related parameters in the prediction of EV were calculated according to receiver operating characteristic(ROC) curve.Results Ultrasound parameters PV-CI, HV-DI and peripheral blood vWF-Ag, sCD163 levels of moderate/severe EV group were all significantly higher than those of none/mild EV group 0.11 (0.05) cm · s vs 0.07(0.03) cm· s;0.72±0.11 vs0.52±0.12;1 824.00(558.00) U/Lvs 1 533.80(311.50) U/L;(72.57±10.94) μg/L vs (57.91±10.40) μg/L;Z=-4.949, t=-7.759, Z=-5.420, t=-5.804, all P＜0.05).However HVAT and IHCT were significantly shorter than those of non/mild EV group (17.00(4.25) s vs 27.78(6.75) s;(6.62±1.85) s vs (9.33±2.26) s, Z=-3.822, t=5.820, both P＜0.05).ROC curve analysis showed that the areas under the ROC curve of HVDI, vWF-Ag and sCD163 predicting moderate/severe EV were all more than 0.8, the appropriate cutoff value of HV-DI, vWF-Ag and sCD163 was 0.64, 1 693.8 U/L and 63.98 μg/L, respectively.The sensitivity were all more than 75％,PPV were all greater than 90％ and DA were all more than 80％.Conclusion HV-DI, peripheral blood vWF-Ag and sCD163 levels can be taken as noninvasive parameters, which can effectively predict the presence of severe EV, facilitate early detection of these high-risk bleeding patients and prevent bleeding.
ABSTRACT
Genetic polymorphisms within immunity-related candidate genes in pigs have been identified to control variations in immune functions and/or disease resistance. It has become necessary to evaluate the effects of other genetic markers of economically important traits prior to introducing them into marker-assisted selection programs. In this study, polymorphisms of porcine genes coding Interferon-induced Gunylate binding protein 1 (GBP1), GBP2, CD163, and CD169 were investigated for their association with growth and meat quality traits in a Korean native pig breed-Yorkshire inter-crossed F2 pig population (KY-F2). KY-F2 animals (n=346) have been successfully used for linkage mapping to identify quantitative loci that control meat quality, growth, and immunity traits. In our results, polymorphisms in genes GBP1 and GBP2 showed association with pig growth rate as well as meat quality traits such as crude fat, drip loss, and meat color (yellowness) in the KY-F2 population. The polymorphism in gene CD163 only showed association with crude fat, as a meat quality trait. CD169 gene was associated with pork tenderness. In conclusion, four immune-related genetic markers were validated for their association with growth and meat quality traits to gauge their potential use in a swine selection program. The results warrant further studies in other commercial pig populations.
Subject(s)
Animals , Carrier Proteins , Chromosome Mapping , Clinical Coding , Disease Resistance , DNA , Genetic Markers , Meat , Polymorphism, Genetic , SwineABSTRACT
Objective To investigate the expression and clinical significance of tumor-associated macrophages (TAMs) in invasive micropapillary carcinoma (IMPC) of breast.Methods Immunohistochemical streptavidin-biotin method(LSAB) method for CD68 and CD163 were performed on 68 cases of IMPC and 72 cases of invasive ductal carcinoma,not otherwise specified(IDC-NOS).Results CD68 and CD163 were mainly expressed in IMPC interstitial macrophages in the cytoplasm or on the membrane,and occasionally expressed in tumor nest.CD68 positive rate in tumor stroma in IMPC (47/68,69.1％) was higher than in IDC-NOS group (37/72,51.5％),and the difference between the two groups was statistically significant (P =0.022).CD163 positive rate in tumor stroma between the two groups was not statistically significant(P =0.682).CD68 + macrophages in IMPC stroma were positively correlated with pathological stage,histological grade,lymph node metastasis,vascular invasion,and Ki67 expression(P ＜ 0.05),and inversely correlated with the expression of ER(P =0.037).Univariate analysis showed that CD68 + macrophages in the stroma of IMPC were significantly associated with progression-free survival(P =0.027),Conclusion The expression of TAMs was different in different types of breast carcinoma and widely expressed in IMPC,which may play a significant role in high invasion and metastasis behavior of IMPC.
ABSTRACT
There is a higher sepsis rate in the intensive care unit (ICU) patients, which is one of the most important causes for patient death, but the sepsis lacks specific clinical manifestations. Exploring sensitive and specific molecular markers for infection that accurately reflect infection severity and prognosis is very clinically important. In this article, based on our previous study, we introduce some new biomarkers with high sensitivity and specificity for the diagnosis and predicting the prognosis and severity of sepsis. Increase of serum soluble(s) triggering receptor expressed on myeloid cells-1 (sTREM-1) suggests a poor prognosis of septic patients, and changes of locus rs2234237 of sTREM-1 may be the one of important mechanisms. Additionally, urine sTREM-1 can provide an early warning of possible secondary acute kidney injury (AKI) in sepsis patients. Serum sCD163 level was found to be a more important factor than procalcitonin (PCT) and C-reactive protein (CRP) in prognosis of sepsis, especially severe sepsis. Moreover, urine sCD163 also shows excellent performance in the diagnosis of sepsis and sepsis-associated AKI. Circulating microRNAs, such as miR-150, miR-297, miR-574-5p, miR-146a, miR-223, miR -15a and miR-16, also play important roles in the evaluation of status of septic patients. In the foreseeable future, newly-emerging technologies, including proteomics, metabonomics and trans-omics, may exert profound effects on the discovery of valuable biomarkers for sepsis.
ABSTRACT
BACKGROUND: Advances in the understanding of Hodgkin's lymphoma (HL) show various functions of infiltrating immune cells and cytokines in relation to clinical outcomes. The expression of CD163 and c-Met has been suggested to have a role in lymphoid malignancy. Thus, we evaluated the expressions of CD163, c-Met, and serum free light chain (sFLC) in relation to the clinicopathological features of patients with advanced classical HL (cHL). METHODS: We assessed the expression of CD163 and c-Met in 34 patients with cHL through immunohistochemistry on the lymph node biopsy sections and the levels of pretreatment sFLC were estimated using ELISA. RESULTS: High CD163 expression correlated with increased age, B symptoms, International Prognostic Score (IPS) > or =3, mixed cellularity subtype, and low response to treatment. Further, high c-Met expression correlated with increased age at diagnosis, leukocytosis, B symptoms, and lower chance to achieve complete remission. The sFLC levels correlated with increased age at diagnosis, lymphopenia, IPS > or =3, B symptoms, and lower complete remission rates. CONCLUSION: In advanced cHL, increased expression of CD163 and c-Met showed a significant association with adverse prognostic parameters and poor response to treatment. Pretreatment high sFLC level also correlated with poor risk factors, suggesting its use as a candidate prognostic marker. A comprehensive approach for prognostic markers might represent a step towards developing a tailored therapeutic approach for HL.
Subject(s)
Humans , Biopsy , Cytokines , Hodgkin Disease , Immunohistochemistry , Leukocytosis , Light , Lymph Nodes , Lymphopenia , Risk FactorsABSTRACT
CD163 is one of transmembrane glycoproteins expressed only in activated monocyte-macrophage cell membrane, which acts as a marker of activated macrophages. In sepsis, CD163 activates the intracellular signal transduction pathways in macrophages,regulates TNF-α, IL-10 and IL-6 expression, have antiinflammatory and antioxidant effects in the inflammatory process. Soluble CD163 (sCD163) is a soluble form of CD163 ,which could play anti-inflammatory and immunoregulatory effects by inhibiting T cells proliferation and activation. Serum concentration of sCD163 may be highly increased when severe infection. Detection of serum sCD163 is helpful to diagnose sepsis,and to evaluate the prognosis of severe infection and sepsis.
ABSTRACT
AIM:To investigate the influence of free hemoglobin on the initiation and progression of atherosclerosis and the role of CD163 in this process.METHODS:Thirty male SD rats were randomly divided into three groups:control group(group C),atherosclerosis group(group A),atherosclerosis and hemolysis group(group P).The hemolysis and atherosclerosis animal model was established.The free hemoglobin(FHb)and MDA levels in plasma,(RAAPIs)and intima area/midmembrane area(I/M)of each group were measured.The expressions of CD163 and heme oxygenase-1(HO-1)in atherosclerosis plaques in group A and P were detected and measured by means of immunohistochemistry and Western blotting.RESULTS:Compared with group C,the FHb,MDA,CD163 and HO-1 in group A and group P increased significantly(P0.05).The FHb level in plasma and the expressions of CD163 and HO-1 in atherosclerotic plaques correlated with each other(r=0.526,r=0.498,r=0.653;P