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OBJECTIVES@#Immunophenotyping technique is a powerful tool for the diagnosis and differential diagnosis of chronic lymphocytic leukemia (CLL) and other B-cell chronic lymphoproliferative diseases (B-CLPD). CD200 is strongly expressed in CLL. This study aims to analyze the clinical value of modified Matutes score (MMS) containing CD200 in the diagnosis of CLL.@*METHODS@#We retrospectively analyzed 103 B-CLPD patients diagnosed from January 2020 to July 2021, including 64 CLL patients, 11 follicular lymphoma (FL) patients, 14 mantle cell lymphoma (MCL) patients, 6 marginal zone lymphoma (MZL) patients, 1 hairy cell leukemia (HCL) patient, and 7 lymphoplasmic lymphoma/Waldenstrom macroglobulinemia (LPL/WM) patients. The expression of CD markers between the CLL group and the non-CLL group was compared, and the sensitivity, specificity, and clinical consistency of MMS and Royal Marsden Hospital (RMH) immunophenotyping score system were analyzed.@*RESULTS@#There were significant differences in the expressions of CD5, CD23, FMC7, CD22, CD79b, CD200, and sIg between the CLL group and the non-CLL group (χ2 values were 37.42, 54.98, 30.71, 11.67, 55.26, 68.48, and 17.88, respectively, all P<0.01). When the RMH immunophenotyping score≥4, the sensitivity was 79.7%, and the specificity was 100%. When the MMS≥3, the sensitivity was 95.3%, and the specificity was 100%. The Kappa coefficient of RMH immunophenotyping system was 0.677, and the Kappa coefficient of MMS system was 0.860.@*CONCLUSIONS@#The MMS system containing CD200 has better sensitivity and same specificity compared with RMH immunophenotyping system, and MMS system may be more useful in the diagnosis of CLL.
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Humans , Adult , Leukemia, Lymphocytic, Chronic, B-Cell/pathology , Retrospective Studies , B-Lymphocytes/pathology , Lymphoma, Mantle-Cell/pathology , Diagnosis, Differential , Lymphoma, B-Cell, Marginal Zone , Flow Cytometry/methodsABSTRACT
Objective:To investigate the expressions of CD200 and inducible costimulator (ICOS) protein in angioimmunoblastic T-cell lymphoma (AITL) and the relationship with prognosis as well as their significances in the differential diagnosis of AITL.Methods:A total of 39 AITL patients in the First People's Hospital of Chenzhou, the Fourth People's Hospital of Chenzhou, Xiangnan College Affiliated Hospital and Chenzhou 3rd People's Hospital from June 2012 to December 2019, and 10 patients with classic Hodgkin lymphoma (CHL) and 10 patients with peripheral T cell lymphoma, not otherwise specified (PTCL-NOS) from August 2016 to July 2019 in the First People's Hospital of Chenzhou were selected. Immunohistochemistry was used to detect the expressions of CD200, ICOS, CD10, programmed death 1 (PD-1), bcl-6 and CXC chemokine receptor-13 (CXCL13) proteins, and the correlation of CD200 and ICOS with clinicopathological features and prognosis of AITL patients was analyzed, and the diagnostic significance of both in differentiating AITL from PTCL-NOS and CHL was also analyzed.Results:The positive rates of CD200 and ICOS in 39 AITL patients were 71.79% (28/39) and 61.54% (24/39), respectively. There were 7 cases of CD200 weak and moderate positive in 10 CHL patients, and ICOS proteins were all negative. Among 10 PTCL-NOS patients, 4 patients had CD200 positive and 1 patient had ICOS positive. The differences in positive rates of ICOS protein between AITL patients and CHL, PTCL-NOS patients were statistically significant (all P < 0.05); the differences in positive rates of CD200 protein between AITL patients and CHL, PTCL-NOS patients were not statistically significant ( χ2=0.013, P=0.911; χ2=3.551, P=0.060). The positive rate of CD200 in AITL patients with elevated lactate dehydrogenase (LDH) and international prognostic index (IPI) score of 3-4 was higher than that in AITL patients with normal LDH and IPI score of 0-2 (both P < 0.05); The positive rate of ICOS in AITL patients with elevated LDH and PD-1 positive was higher than that in AITL patients with normal LDH and PD-1 negative (both P < 0.05). CD200 negative AITL patients had better 3-year overall survival (OS) rate (4.2% vs. 66.7%) and 3-year progression-free survival (PFS) rate (5.3% vs. 77.1%) compared with those in CD200 positive AITL patients, and the differences between both groups were statistically significant (both P < 0.01); there was a statistically significant difference in 3-year OS rate between ICOS positive AITL patients and ICOS negative AITL patients (15.3% vs. 38.6%, P=0.011), while there was no statistically significant difference in 3-year PFS rate of both groups (18.6% vs. 41.5%, P=0.059). Multivariate analysis showed CD200 ( HR=0.076, 95% CI 1.555-79.497, P=0.001), extranodal involvement or not ( HR=11.117, 95% CI 1.555-79.497, P=0.016) and LDH ( HR=2.147, 95% CI 0.844-5.459, P=0.109) were independent influencing factors of OS in AITL patients; CD200 ( HR=0.075, 95% CI 0.016-0.357, P=0.001) and LDH ( HR=2.335, 95% CI 0.929-5.870, P=0.071) were independent influencing factors of PFS in AITL patients. Conclusions:CD200 and ICOS can be used as immunohistochemical indicators to assist the diagnosis of AITL patients. ICOS protein helps to differentiate AITL from CHL and PTCL-NOS; CD200 can be used as indicators to judge the prognosis and deterioration of AITL patients.
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Objective To observe the changes of NF-κB and inflammatory cytokine expression after CD200 pretreatment in severe heatstroke rats for exploring whether the molecular mechanism of CD200 inhibition of severe heat stroke inflammatory response is related to NF-κB signaling pathway. Methods Forty male Wistar rats were randomly divided into control group (n=8), severe heatstroke model group (HS group, n=16), and CD200 pretreatment group (CD200 group, n=16). The HS group and the CD200 group were injected with physiological saline and CD200 recombinant fusion protein before heat exposure to prepare a classical rat heat stroke model, and the control group was placed at room temperature of 22.0±1.0 ℃. The expression of NF-κB/ p65 mRNA in lung tissue were detected at 60 min after model establishment, and serum high mobility group protein B1 (HMGB1), tumor necrosis factor alpha (TNF-α) and interleukin 6 (IL-6) were detected. The pathological changes of the lungs were observed, and the survival time of severe heatstroke rats were recorded. Results CD200 pretreatment could inhibit the expression of NF-κB/p65mRNA. Compared with the control group, the expressions of NF-κB/p65 mRNA in the HS group and CD200 group were significantly increased (P<0.05), and the expression of NF-κB/p65 mRNA in CD200 group was lower than that in HS group (P<0.05). Serum HMGB1, TNF-α and IL-6 in HS group and CD200 group were significantly higher than those in the control group (P<0.05). The HMGB1, TNF-α and IL-6 in the HS group were higher than those in the CD200 group (P<0.05). The median survival time of severe heat stroke were prolonged in the CD200 group compared with the HS group (P<0.05), and the pathomorphological changes showed that inflammation and alveolar exudation were significantly reduced in the CD200 group compared with the HS group. Conclusions CD200 pretreatment can alleviate the inflammatory response in severe heatstroke rats. The possible molecular mechanism of CD200 to relieve severe heatstroke inflammatory response may be involved with NF-κB signaling pathway, which can reduce severe disease by inhibiting the activation of NF-κB and the expression of inflammatory factors.
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BACKGROUND: Immune rejection of skin allograft is a clinical problem to be solved. Our previous study has shown that human placenta-derived CD200+ mesenchymal stem cells may have a strong capability of immunoregulation. OBJECTIVE: To investigate the effects of human placenta-derived CD200+ mesenchymal stem cells on rejection of skin allograft. METHODS: Skin allograft models of c57/BL6 mice were established and divided into three groups as control group, human placenta-derived mesenchymal stem cells group (PMSCs group), and CD200+ mesenchymal stem cells group (CD200+-PMSCs group). PBS (control group), normal PMSCs and CD200+-PMSCs were injected into the mice through the tail vein, respectively. The necrotic time, survival time and situation of grafted skin were observed. The number of peripheral white blood cells was counted after 7 days of treatment. The expression levels of interleukin-10, interferon-γ and tumor necrosis factor-α were detected by Q-PCR and ELISA. RESULTS AND CONCLUSION: (1) Compared with the control group, in the PMSCs and CD200+-PMSCs groups, the condition of skin allograft was better, and the survival time was significantly prolonged (P < 0.001). The condition and survival time of skin allograft in the CD200+-PMSCs group were significantly superior to those in the PMSCs group (P < 0.01). (2) After 7 days of treatment, the number of peripheral white blood cells in the PMSCs and CD200+-PMSCs groups was significantly less than that in the control group (P < 0.01). (3) Compared with the control group, the mRNA expression level of interleukin-10 in the spleen was significantly increased in the CD200+-PMSCs group (P < 0.05), and the mRNA expression levels of interferon-γ and tumor necrosis factor-α in the spleen were significantly down-regulated in the PMSCs and CD200+-PMSCs groups (P < 0.05, P < 0.01). The mRNA expression levels of interferon-γ and tumor necrosis factor-α in the spleen in the CD200+-PMSCs group were significantly lower than those in the PMSCs group (P < 0.01, P < 0.05). (4) Compared with the control group, the expression level of interleukin-10 in the blood was significantly increased (P < 0.05, P < 0.001), and the expression levels of interferon-γ and tumor necrosis factor-α in the blood were significantly down-regulated in the CD200+-PMSCs and PMSCs groups (P < 0.05, P < 0.001; P < 0.01, P < 0.001). The expression levels of interferon-γ and tumor necrosis factor-α in the blood in the CD200+-PMSCs group were significantly lower than those in the PMSCs group (P < 0.05). These results indicate that human placenta-derived mesenchymal stem cells have immunosuppressive effect on the rejection of skin allograft, and CD200+ cells may have better immunoregulatory effects by regulating the expressions of interleukin-10, interferon-γ and tumor necrosis factor-α
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Objective To evaluate the role of hippocampal CD200 receptor 1(CD200R1)in peri-operative neurocognitive disorders(PND)in mice.Methods Sixty clean-grade male C57BL/6 mice,aged 9-10 months,weighing 32-38 g,were used in the study.The experiment was performed in two parts.Ex-periment Ⅰ Thirty-six mice were divided into 2 groups(n＝18 each)using a random number table meth-od: control group(group C)and PND group.Group C only received isoflurane anesthesia.Partial left lo-bectomy of the liver was performed under isoflurane anesthesia in group PND.Contextual fear conditioning test was performed at 1,3 and 7 days after surgery,and the freezing time was recorded.The mice were then sacrificed,and the hippocampus was isolated for determination of interleukin-1β(IL-1β)and tumor necrosis factor-α(TNF-α)contents(by enzyme-linked immunosorbent assay)and CD200 and CD200R1 expression(by Western blot).ExperimentⅡ Twenty-four mice were divided into 2 groups(n＝12 each)using a random number table method: CD200-Fc group and IgG1-Fc group.Recombinant proteins CD200-Fc and human IgG1-Fc were injected into the lateral cerebral ventricle in CD200-Fc group and IgG1-Fc group,respectively.Partial left lobectomy of the liver was performed after the end of injection.Contextual fear conditioning test was performed at 1 and 3 days after surgery,and the freezing time was recorded.Re-sults Experiment Ⅰ Compared with group C,the freezing time in the contextual fear conditioning test was significantly shortened,and the contents of IL-1β were increased at 1 and 3 days after surgery,the contents of TNF-α were increased at 3 and 7 days after surgery,and the expression of CD200 and CD200R1 was up-regulated at 1 day after surgery in group PND(P＜0.05).ExperimentⅡ Compared with IgG1-Fc group,the freezing time in the contextual fear conditioning test was significantly prolonged at 1 day after surgery in CD200-Fc group(P＜0.05).Conclusion Up-regulated expression of hippocampal CD200R1 is the endogenous protective mechanism of PND in mice.
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Objective@#To evaluate the role of hippocampal CD200 receptor 1 (CD200R1) in perioperative neurocognitive disorders (PND) in mice.@*Methods@#Sixty clean-grade male C57BL/6 mice, aged 9-10 months, weighing 32-38 g, were used in the study.The experiment was performed in two parts.Experiment Ⅰ Thirty-six mice were divided into 2 groups (n=18 each) using a random number table method: control group (group C) and PND group.Group C only received isoflurane anesthesia.Partial left lobectomy of the liver was performed under isoflurane anesthesia in group PND.Contextual fear conditioning test was performed at 1, 3 and 7 days after surgery, and the freezing time was recorded.The mice were then sacrificed, and the hippocampus was isolated for determination of interleukin-1β (IL-1β) and tumor necrosis factor-α (TNF-α) contents (by enzyme-linked immunosorbent assay) and CD200 and CD200R1 expression (by Western blot). Experiment Ⅱ Twenty-four mice were divided into 2 groups (n=12 each) using a random number table method: CD200-Fc group and IgG1-Fc group.Recombinant proteins CD200-Fc and human IgG1-Fc were injected into the lateral cerebral ventricle in CD200-Fc group and IgG1-Fc group, respectively.Partial left lobectomy of the liver was performed after the end of injection.Contextual fear conditioning test was performed at 1 and 3 days after surgery, and the freezing time was recorded.@*Results@#Experiment Ⅰ Compared with group C, the freezing time in the contextual fear conditioning test was significantly shortened, and the contents of IL-1β were increased at 1 and 3 days after surgery, the contents of TNF-α were increased at 3 and 7 days after surgery, and the expression of CD200 and CD200R1 was up-regulated at 1 day after surgery in group PND (P<0.05). Experiment Ⅱ Compared with IgG1-Fc group, the freezing time in the contextual fear conditioning test was significantly prolonged at 1 day after surgery in CD200-Fc group (P<0.05).@*Conclusion@#Up-regulated expression of hippocampal CD200R1 is the endogenous protective mechanism of PND in mice.
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Background: Chronic lymphoproliferative disorders (CLPDs) are heterogeneous group of disorders with variable clinical presentations and outcomes. Therefore, accurate classification is crucial for treatment planning. At present, flow cytometry immunophenotyping (FCM-IPT) is a useful tool for diagnosing these diseases. However, overlapping immunophenotypes do exist. Recently, differential expression of CD200 and variation in number of CD20 antibody bound per cell (ABC) in different CLPDs has been reported. Materials and Methods: Seventy-seven CLPD cases were analyzed by FCM-IPT for CD200 expression, and Quantibrite bead was used to calculate CD20 ABC. Results: Variability in CD200 expression can help in the differentiation of chronic lymphocytic leukemia (CLL) and hairy cell leukemia (HCL) from other CLPDs. CD200 was brightly expressed in 100% CLL cases, having homogenous bright (2+) intensity. On the contrary, CD200 was uniformly negative in all Mantle cell lymphoma cases except 1, in which the intensity was dim, and the mean fluorescence intensity was significantly lower than CLL. Furthermore, all HCL cases showed bright expression of CD200, thereby making it useful in differentiation from other CLPD with villous lymphocytes. Evaluation of CD20 ABC showed that it differs among various CLPD and was significantly lowest in CLL and highest in HCL both on peripheral blood and bone marrow samples. Conclusion: Our results support the fact that CD200 can be added to routine CLPD panel as it is useful in subcategorizing them. However, inclusion of CD20 ABC to routine panel does not seem plausible but may be done for difficult diagnostic cases or where anti-CD20 therapy is planned.
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Abstract Background Distinction between mature B-cell neoplasms can be difficult due to overlapping of immunologic features and clinical manifestations. This study investigated whether quantifying mean fluorescence intensity of four monoclonal antibodies in a flow cytometry panel is useful for the differential diagnosis and characterization of these disorders. Methods The expressions of CD52, CD200, CD123 and CD43 were analyzed in samples from 124 patients with mature B-cell neoplasms. The quantitative estimation of these antigens was assessed by mean fluorescence intensity. Results The cases included were 78 chronic lymphocytic leukemias, three atypical chronic lymphocytic leukemias, six marginal zone lymphomas, 11 splenic marginal zone lymphomas, nine lymphoplasmacytic lymphomas, six mantle cell lymphomas, two hairy cell leukemias, two hairy cell leukemias variant, five follicular lymphomas, one Burkitt lymphoma and one diffuse large B-cell lymphoma. The mean fluorescence intensity of CD200 was higher in atypical chronic lymphocytic leukemia, chronic lymphocytic leukemia and hairy cell leukemia cases. CD123 showed higher mean fluorescence intensities in hairy cell leukemia cells. Chronic lymphocytic leukemia, atypical chronic lymphocytic leukemia and mantle cell lymphoma had higher expression of CD43 and all follicular lymphoma cases had very low mean fluorescence intensity values. CD52 expression was consistently positive among all cases. Conclusion Quantitative evaluation of these markers can be a useful additional tool to better identify some types of mature B-cell neoplasms.
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Humans , B-Lymphocytes , Leukemia, Lymphocytic, Chronic, B-Cell , Immunophenotyping , Lymphoma, B-Cell , Flow CytometryABSTRACT
Inflammation plays an important role in the pathophysiological mechanism of acute ischemic stroke.CD200 expressed in neurons interacts with CD200 receptor (CD200R) on microglia cells.It can inhibit microglia activation and alleviate the inflammation after cerebral ischemic injury.This article reviews the roles of CD200 and CD200R in the activation of microglia after cerebral ischemia.
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Objective To investigate the expression of CD200 in peripheral lymphocytes and hair follicles from patients with alopecia areata (AA).Methods Flow cytometry was used to detect the expression of CD200 in peripheral lymphocytes from 43 patients with AA and 43 healthy controls.Immunohistochemistry was carried out to quantify the expression of CD200 and cytokeratin 15(CK15,a marker for basal cells of the outer root sheath) in resected scalp specimens from 8 patients with AA and 8 healthy controls.Differences in the expression of CD200 and CK15 between the patients and controls were assessed by independent-samples t test and rank sum test.Data were processed by the software SPSS17.0.Results The percentage of CD200-expressing cells in peripheral blood lymphocytes and T lymphocytes was significantly lower in the patients with AA than in the healthy controls (5.73％ ± 3.46％ vs.12.01％ ± 4.90％,8.85％ ± 4.80％ vs.12.31％ ± 3.12％,t =6.865,3.964,respectively,both P ＜ 0.05).However,no significant difference was observed in the percentage of CD200-expressing cells in peripheral blood B lymphocytes between the patients and controls (74.68％ ± 8.12％ vs.75.75％ ± 9.45％,t =0.570,P ＞ 0.05).Further more,the patients showed a lower expression of CD200 (P ＜ 0.05),but a similar expression of CK15 (P ＞ 0.05) in hair follicles compared with the controls.Conclusion The decrease in CD200 expression in peripheral lymphocytes and hair follicles may be involved in the pathogenesis of AA.
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Objective:To establish a human monocyte cell line U937 stably expressing CD200.Methods:The eukaryotic expression vector pcDNA3-CD200 containing human CD200 cDNA and vector pcDNA3 were transferred into human monocyte U937 cell line by electrotransfer.These two cell lines were selected by G418, and the selected cell lines were subcultured. U937 cell line was enclosed as control group.The expression of CD200 mRNA and protein was detected by RT-PCR and flow cytometry.Results:After G418 selection,U937 cell line in control group was died, and the cell lines transferring pcDNA3 and recombined pcDNA3-CD200 were subcultured over 30 generations.The CD200 mRNA expression in pcDNA3-CD200 group was confirmed with RT-PCR.The CD200 expression in U937-pcDNA3-CD200,U937-pcDNA3 and U937 were 77.20%,3.20% and 2.10%,compared with other two groups (F=133 996.40,P0.05).Conclusion:Our study provides foundation for mechanism of action for CD200 in future.The human monocyte series U937 cell line that expresses CD200 protein stably is established successfully by gene transfection method.