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Objective:To investigate the role of CD46 and Nectin-4 on Measles virus (MV) infecting human pulmonary alveolar epithelial cells (HPAEpiC),and the interaction between CD46 and Nectin-4.Methods: Measles virus was divided into pre-infection group and 2 h-infection group,HPAEpiCs treatment with anti-CD46 antibody and/or anti-Nectin-4 antibody as experimental groups,and untreated HPAEpiCs as a control.The variation of viral replication level was detected.A Co-immunoprecipitation assay (Co-IP) was used to explore whether CD46 and Nectin-4 had interactive relationship in MV infection.Results: Compared with the control group,MV titers were reduced in HPAEpiCs of the pre-infection group treated with anti-CD46 and anti-Nectin-4 respectively (48.03% and 49.53%).Furthermore,virus titers showed a more reduction in which treated with anti-CD46 and anti-Nectin-4 antibodies (27.15%,P<0.01).Western blot and Real-time PCR showed that anti-CD46 antibody and anti-Nectin-4 antibodies decreased the rate of MV infection.In the 2 h-infection group,however,the treatment with anti-CD46 and anti-Nectin-4 could significantly reduce the MV titer and NP protein in HPAEpiCs.The Co-IP assay showed that there were interaction between CD46 and Nectin-4.Conclusion: CD46 and Nectin-4 mediated MV infecting HPAEpiCs.Moreover,CD46 and Nectin-4 may play a synergetic role in MV infection,which could enhance the infection effect.
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Objective:To determine whether CD46, CD55, and CD59 are differentially expressed in neoplastic and adjacent nor-mal colon tissues and to investigate their influence on clinicopathologic variables. Methods:Immunohistochemistry (a modified two-step method) was used to detect the expression of CD46, CD55, and CD59 in a tissue microarray of 121 cases of colon cancer and corre-sponding adjacent non-tumor tissues with detailed clinical information, including gender, age, differentiation, TNM classification, tu-mor location, and tumor histotype. The colon carcinoma microarray was constructed from patients' samples obtained from the Depart-ment of Gastrointestinal Surgery of Xijing Hospital of the Fourth Military Medical University between October 2004 and June 2006. The correlation between expression and clinicopathologic features was analyzed. Results:The expression levels of CD46, CD55, and CD59 were significantly higher in colon cancer tissues compared with those in normal adjacent colon tissues (P0.05). The expression levels of CD55 and CD59 were correlated with the grade of colon cancer differentiation. Low levels of CD55 and CD59 were detected in cancer cells of highly differentiated cancer, whereas stronger staining for CD55 and CD59 was mainly observed in cancer cells of moderately and poorly differentiated colon cancer (P<0.05). In addition, the expression levels of CD55 and CD59 were higher in stages III and IV colon cancer than those in stages I and II according to TNM classification (P<0.05). Conclusion:CD46, CD55, and CD59 are up-regulated in colon cancer. Specifically, CD55 and CD59 are of clinical relevance to differentiation and TNM staging of colon cancer, and their expression might be closely related to clinical biological behaviors.
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Objective The purpose of this study is to investigate CD 46 expression in advanced invasive breast cancer and in normal breast tissues , and to analyze the correlation between CD 4 expression and effect of neoadjuvant chemotherapy ,exploring its clinical significance for the prognostic value of neoadjuvant chemotherapy in breast cancer .Methods Immunohistochemistry was employed to detect CD 46 expression of tissue samples from 60 patients with invasive breast cancer and 30 normal breasts .We further analyzed the relationship between effect of neoadjuvant chemotherapy and CD 46 expression ,and the prognostic significance for the survival .Rseults CD46-positive rate in breast cancer was apparently higher than that in normal breast tissues (81.67% vs. 46.67%,P<0.05).After neoadjuvant chemotherapy,the CD46 expression was significantly decreased (P <0 .05).There was a negative correlation between CD 46 expression and efficacy of chemotherapy ( P<0.05). CD46 expression in breast cancer could be affected in cancer invasion ,lymph node metastasis(P<0.05).Con-clusion CD46 expression in breast cancer patients is higher than that in healthy people ,and was negatively cor-related with efficacy of adjuvant chemotherapy .The increasing rate of CD 46 positive expression has predictive val-ue for the poor prognosis of patients .
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Objective To establish and observe the effects of CD46-siRNA on proliferation,in-vasion and metastasis of MCF-7 human breast cancer cell.Methods The pEGFP-C1-CD46 siRNA was established and transfected into the MCF-7 cell.Changes in CD46 mRNA and protein expression were test-ed and variation of invasion and migration capacity in MCF-7 /CD46-siRNA cell was also observed.Results Compared with the control group and scrambled group,the expression of CD46 mRNA was significantly decreased(P <0.05)and CD46 protein expression was also suppressed in the MCF-7 cells of the silence group.Proliferation activity of MCF-7 cell in the control group and scrambled group was gradually in-creased with the time,but this phenomenon was not obvious in the silence group.At the 3th and 4th day af-ter transcription,differences in proliferation activity were significant among the groups,and activity in the silent group was significantly lower than the other two groups(P <0.05).Counts in invasion and migration of MCF-7 cell in the control group and scrambled group were significantly higher than the silent group (P <0.05 ).Conclusion The expression vector of pEGFP-C1-CD46 siRNA can suppress the CD46 mRNA and protein expression in MCF-7 human breast cancer cell and reduce the capacity in proliferation activity,invasion and migration.
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CD46 is not only identified as a complement regulatory protein which protects host cells from complement attack,but also a new co-stimulatory molecule for human T cells.CD3/CD46 co-stimulation can induce a T-regulatory 1 cell(Tr1)-specific cytokine phenotype in human CD4+T cells.However,the role of CD46 as a co-stimulatory molecule in the modulation of the acquired immunity,such as transplant immunology,remains unclear.In this study,CD4+T cells were isolated from human CD46-transgenic C57BL/6 mice by magnetic-activated cell sorting,and further induced by anti-CD3,anti-CD28 and anti-CD46 antibodies respectively,and anti-CD3/anti-CD28 antibodies,anti-CD3/anti-CD46 antibodies,or the monoclonal antibody panel against CD3/CD28/CD46.The levels of interleukin-2(IL-2),γ-interferon(γ-IFN),interleukin-10(IL-10)and transforming growth factor-β(TGF-β)were detected in the supematants of different groups.Suppression of allogeneic T cell proliferation were assessed by using mixed lymphocyte reaction(MLR)assay,in which monoclonal antibodies against CD46 were added to the culture.The results showed that CD3/CD28,CD3/CD46 and CD3/CD28/CD46 co-stimulation could significantly induce stronger proliferation of T cells than CD3 stimulation(P＜0.05),and CD3/CD28/CD46 co-stimulation significantly increased the proliferation of T cells when compared with CD3/CD28 or CD3/CD46 co-stimulation(P＜0.05 for each).IL-2 and γ-IFN levels were much higher in CD3/CD28 co-stimulation group than in CD3,CD28,CD46 and CD3/CD46 groups(P＜0.05 for each).IL-10 and TGF-β levels were dramatically increased in CD3/CD46 co-stimulation group as compared with those in the CD3,CD28,CD46 and CD3/CD28 groups(P＜0.05 for each).CD3/CD46 co-stimulation significantly inhibited the T cell proliferation and allogenic immune responses through the secretion of IL-10 and TGF-β in MLR (P＜0.05).These results suggested that CD3/CD46 can induce Tr1 cells to modulate allogenic immune responses,and it may become a novel target for the development of new therapeutic approach for T-cell-mediated diseases.CD46 plays an important role in regulating the T cell-mediated immune responses by bridging innate and acquired immunity.
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Objective: To observe the effects of recombinant adenovirus TRAIL (AdS-TRAIL & Ad5F35-TRAIL) on apoptosis of non-small cell lung (NSCLC) cells, so as to assess the value of Ad-TRAIL in gene therapy of NSCLC. Meth-ods: CAR and CD46 expression levels in lung cancer cell lines (A549, Z793, QG56 and NCI-H520) and the primary lung cancer cells from samples of 10 NSCLC patients were assayed by flow cytometry analysis. The lung cancer cell lines and primary lung cancer cells were infected with Ad5-TRAIL & Ad5F35-TRAIL adenoviral vectors at MOI 10 or 50, re-spectively; the percentage of apoptosis cells labeled by Annexin V-FITC in different cells were measured by flow cytometry 48 h after transfection. Results: The expression of CD46 were higher than that of CAR in all the lung cancer lines (A549, Z793, QG56 and NCI-H520) and the primary lung cancer cells. Significant apoptosis was observed in Z793 and QG56 cells transfected with Ad5-TRAIL or Ad5F35-TRAIL at MOI 10, with the apoptosis rate being (1.76±2.10)% (Ad5-TRAIL), (15.96±2.89) % (Ad5F35-THAIL) and (6.05±1.58) % (Ad5-TRAIL), (10.11±1.26) % (Ad5F35-TRAIL), respectively, compared to no adenovirus-transfected cells ([2.33±0.37] % and [5.95±1.89]%, respectively, P < 0.05). Less than 10% of apoptosis cells were detected in NCI-H520 cells transfected with Ad5- or Ad5F35-TRAIL at MOI 50 ([12.89±3.2] % for AdS-TRAIL and [9.08±1.35]% for Ad5F35-TRAIL, respectively) compared to no adenovirus-transfected cells ([7.04±2.17] %, P > 0.05). Moreover, apoptosis induced by Ad5- or Ad5F35-TRAIL transfection in A549 cells was not detected both at MOI 10 and 50. About half of the primary lung cancer cells from 10 patients induced apoptosis after transfected with Ad5-TRAIL or Ad5F35-TRAIL vector. A higher percentage of apoptotic cells were found in Ad5F35-TRAIL group than those in Ad5-TRAIL and control groups. Conclusion: Ad5-TRAIL can induce apoptosis of NSCLC cells in vitro, and Ad5F35-TRAIL is more potent than Ad5-TRAIL, so Ad5F35-TRAIL is more suitable for gene therapy of NSCLC.
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Objective To identify the expression of complement regulatory protein CD46, CD55, and CD59 on primary murine pallium astrocytes and the effect of inflammatory factors on it in order to lay the foundation for studying the complement system in AD. Methods The primary murine astrocytes were cultured and purified. The expression of CD46, CD55, and CD59 on the levels of mRNA and protein was assayed by immunofluorescence before and after the stimulation of LPS and IFN-?. Results The expression of CD59 mRNA was confirmed, but the expression of CD46 and CD55 was indefinite. There was no significant difference between stimulation and non-stimulation groups. Immunofluorescence results indicated that CD59 was positive, while CD46 and CD55 were weakly positive. Conclusion Protectin CD59 expresses copiously on primary murine astrocytes, which presumably protects astrocytes from the lysis of complement.
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Objective:To determine the expression of complement regulatory proteins(CRPs) C_1-INH,MCP(CD46) in the nasal mucosa of allergic rhinitis rats and to study the role of CRPs in the pathogenesis of allergic rhinitis.Methods: Nine healthy SD rats were sensitized and intranasally challenged with ovalbumin and Al(OH)_3(as supplement) to establish allergic rhinitis models and another 9 SD rats treated with saline were taken as control.The nasal mucosa in respiratory area of both groups was obtained.Then Western blotting was performed to investigate the expression levels of C_1-INH and CD46.Results: Western blotting showed that both C_1-INH and CD46 had been detected in rat nasal mucosa.Expression level of CD46 in the nasal mucosa of allergic rhinitis was significantly higher than that in control rats(P