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Glial tumors and the complement cascade
Abstracts / Molecular Immunology 56 (2013) 240–316
Objectives: To assess the relationship between the concentration of the LCP recognition molecules, LCP activation and SAH pathology as well as outcome. Patients and methods: SAH severity was assessed in 39 SAH patients using World Federation of Neurological Surgeons grading scale. VSP was defined as neuro-worsening with angiographic confirmation of vessel narrowing. Cerebral ischemia was defined as a hypodense lesion on computer tomography scan performed before discharge. Outcome at 6 months was assessed by Glasgow Outcome Scale. Twenty healthy subjects served as controls. Plasma concentrations of mannose-binding lectin (MBL); ficolin-1, -2 and -3, C3 activation products and soluble sC5b-9 terminal complement complex (TCC) were measured. In addition ficolin-3 mediated functional LCP activity was assessed. Results: Significantly increased concentrations of C3 activation products and TCC were observed in the SAH patients compared with controls. Overall, ficolin-1 concentration was increased and ficolin-2 was decreased in patients compared to controls, while no changes were detected for MBL and ficolin-3. However, ficolin3 mediated functional LCP activity was reduced. Only low levels of plasma ficolin-3 and ficolin-3 mediated functional LCP activity were related to SAH severity, VSP and cerebral ischemia. In addition ficolin-3 mediated functional LCP activity was decreased in patients with an unfavourable outcome. Conclusion: Our data suggest that ficolin-3 is the main activator of the LCP in SAH. Ficolin-3 is related to severity of brain injury evaluated by clinical and structural parameters. These results indicate that ficolin-3 mediated functional LCP activity may be targeted to control injury progression in SAH. http://dx.doi.org/10.1016/j.molimm.2013.05.108 P CD 14 Glial tumors and the complement cascade T. Bouwens 1,2,∗ , M. Lamfers 1 , R. Veerhuis 3 , C. Dirven 1 , L. Trouw 2 , H. Al-Khawaja 1 1 Erasmus Medical Center Rotterdam, Neurosurgery, Rotterdam, Netherlands 2 Leiden University Medical Center, Rheumatology, Leiden, Netherlands 3 Vrije University Medical Center, Clinical Chemistry, Amsterdam, Netherlands
Introduction: Compared to other types of cancer, Glioblastoma Multiforme (GBM) shows one of the worst survival rates, median survival being less than 15 months and the true 5-year survival only 2%. Components of the adaptive immune system have been identified to be involved in GBM pathology. Whereas, very little is known about the contribution of the innate immune system. The complement cascade forms a vital part of this system. Several molecular pathways that are characteristic for the aggressive nature of GBM are known to be induced or modulated by the activated complement system. Goal: Identify the role of the complement cascade in GBM pathology with the ultimate aim to investigate the complement cascade as a target for treatment of GBM disease. Patients and methods: Tissue and serum samples were obtained during elective surgery of susceptible primary glial tumor patients at the Erasmus Medical Center. A gene expression dataset, consisting of 276 glial tumor samples and 8 normal samples was used. Multiple gene expression analyses were performed on a single-gene and on a pathway level. ELISA and IHC were combined to assess levels and localization of critical components of the initiation pathways and of the effector phase of the complement cascade.
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Results: Our preliminary results of gene-expression analysis of 276 low to high-grade gliomas revealed that complement gene expression is elevated in GBM patients who showed worsened survival. On protein level, the abundant presence of the key complement components C1q, C3, C5aR and C3aR in GBM tumor tissue was observed. Evidence for successful complement cascade termination was found through the presence of the C5b-9 complex. Analysis of the serum concentration of C1q and factor B in glial tumor patients (n = 131) revealed a significant decrease when compared to healthy controls (n = 209). In addition, in parallel analysis of MBL, the frequency of MBL deficiency among GBM patients was found to be significantly (19%) smaller when compared to healthy controls (32%). Conclusion: Using gene expression analyses, immunohistochemistry and serum protein analyses our studies indicate that complement is involved in GBM’s pathophysiology. Future studies in vitro and in vivo are needed to unravel the exact pathophysiological role of complement in glial tumors. http://dx.doi.org/10.1016/j.molimm.2013.05.109 P CD 16 Expression of membrane complement regulators in patients on peritoneal dialysis therapy Y. Sei 1,∗ , M. Mizuno 1,∗ , M. Imai 2 , Y. Suzuki 1 , K. Higashide 1 , N. Okada 2 , C. Harris 3 , S. Matsuo 1 , Y. Ito 1 1
Nagoya University Graduate School of Medicine, Nagoya, Japan Nagoya City University Graduate School of Medicine, Nagoya, Japan 3 Cardiff University, Cardiff, United Kingdom 2
Background: Peritoneal dialysis (PD) therapy is one of the most important renal replacement therapies. Impairment of peritoneal function can limit the long-term efficacy of PD therapy and is caused by several factors that occur during PD therapy, including exposure to peritoneal dialysate, catheter trauma and peritonitis. We previously showed in animal models that inhibition of membrane complement regulators (CRegs) could induce peritoneal inflammation and that dysregulated complement activation induced severe peritoneal injuries. Objective and methods: We investigated the expression of CD46, CD55 and CD59 on peritoneal mesothelial cells and levels of the complement activation marker sC5b-9 in PD fluids (PDF) to clarify influence of complement activation and CRegs expression in PD patients. Primary cell cultures of mesothelial cells were obtained from PDF of 27 PD patients and from omentum of 3 non-chronic kidney disease patients undergoing laparoscopic operations and cells were analysed for expression of CRegs. sC5b-9 levels were measured in the PDF of the PD patients, and background history, including complications of diabetes, usage of icodextrin as PDF, PD history, and dialysate-to-plasma creatinine concentration ratio (D/P Cre), an indicator of peritoneal function, were noted. Results: In PD patients, expression of CD55 but not CD46 and CD59 on mesothelial cells was significantly correlated to peritoneal function (D/P Cre; p < 0.05). Levels of sC5b-9 in the PDF showed weak inverse correlation with expression levels of CD46 and CD59 on mesothelial cells. Loss of expression of CD55 on mesothelial cells correlated with reduced mRNA for CD55 (p < 0.0001); however, there was no correlation between CD46/CD59 protein expression and levels of respective mRNA. Use of icodextrin correlated with decreased levels of sC5b-9 in PDF (p < 0.005). Complication such as diabetes, usage of icodextin, or PD history did not affect CD55 expression. Conclusion: Our results show that PD therapy alters expression of CRegs and complement regulation in the peritoneum. These data
Objectives: To assess the relationship between the concentration of the LCP recognition molecules, LCP activation and SAH pathology as well as outcome. Patients and methods: SAH severity was assessed in 39 SAH patients using World Federation of Neurological Surgeons grading scale. VSP was defined as neuro-worsening with angiographic confirmation of vessel narrowing. Cerebral ischemia was defined as a hypodense lesion on computer tomography scan performed before discharge. Outcome at 6 months was assessed by Glasgow Outcome Scale. Twenty healthy subjects served as controls. Plasma concentrations of mannose-binding lectin (MBL); ficolin-1, -2 and -3, C3 activation products and soluble sC5b-9 terminal complement complex (TCC) were measured. In addition ficolin-3 mediated functional LCP activity was assessed. Results: Significantly increased concentrations of C3 activation products and TCC were observed in the SAH patients compared with controls. Overall, ficolin-1 concentration was increased and ficolin-2 was decreased in patients compared to controls, while no changes were detected for MBL and ficolin-3. However, ficolin3 mediated functional LCP activity was reduced. Only low levels of plasma ficolin-3 and ficolin-3 mediated functional LCP activity were related to SAH severity, VSP and cerebral ischemia. In addition ficolin-3 mediated functional LCP activity was decreased in patients with an unfavourable outcome. Conclusion: Our data suggest that ficolin-3 is the main activator of the LCP in SAH. Ficolin-3 is related to severity of brain injury evaluated by clinical and structural parameters. These results indicate that ficolin-3 mediated functional LCP activity may be targeted to control injury progression in SAH. http://dx.doi.org/10.1016/j.molimm.2013.05.108 P CD 14 Glial tumors and the complement cascade T. Bouwens 1,2,∗ , M. Lamfers 1 , R. Veerhuis 3 , C. Dirven 1 , L. Trouw 2 , H. Al-Khawaja 1 1 Erasmus Medical Center Rotterdam, Neurosurgery, Rotterdam, Netherlands 2 Leiden University Medical Center, Rheumatology, Leiden, Netherlands 3 Vrije University Medical Center, Clinical Chemistry, Amsterdam, Netherlands
Introduction: Compared to other types of cancer, Glioblastoma Multiforme (GBM) shows one of the worst survival rates, median survival being less than 15 months and the true 5-year survival only 2%. Components of the adaptive immune system have been identified to be involved in GBM pathology. Whereas, very little is known about the contribution of the innate immune system. The complement cascade forms a vital part of this system. Several molecular pathways that are characteristic for the aggressive nature of GBM are known to be induced or modulated by the activated complement system. Goal: Identify the role of the complement cascade in GBM pathology with the ultimate aim to investigate the complement cascade as a target for treatment of GBM disease. Patients and methods: Tissue and serum samples were obtained during elective surgery of susceptible primary glial tumor patients at the Erasmus Medical Center. A gene expression dataset, consisting of 276 glial tumor samples and 8 normal samples was used. Multiple gene expression analyses were performed on a single-gene and on a pathway level. ELISA and IHC were combined to assess levels and localization of critical components of the initiation pathways and of the effector phase of the complement cascade.
277
Results: Our preliminary results of gene-expression analysis of 276 low to high-grade gliomas revealed that complement gene expression is elevated in GBM patients who showed worsened survival. On protein level, the abundant presence of the key complement components C1q, C3, C5aR and C3aR in GBM tumor tissue was observed. Evidence for successful complement cascade termination was found through the presence of the C5b-9 complex. Analysis of the serum concentration of C1q and factor B in glial tumor patients (n = 131) revealed a significant decrease when compared to healthy controls (n = 209). In addition, in parallel analysis of MBL, the frequency of MBL deficiency among GBM patients was found to be significantly (19%) smaller when compared to healthy controls (32%). Conclusion: Using gene expression analyses, immunohistochemistry and serum protein analyses our studies indicate that complement is involved in GBM’s pathophysiology. Future studies in vitro and in vivo are needed to unravel the exact pathophysiological role of complement in glial tumors. http://dx.doi.org/10.1016/j.molimm.2013.05.109 P CD 16 Expression of membrane complement regulators in patients on peritoneal dialysis therapy Y. Sei 1,∗ , M. Mizuno 1,∗ , M. Imai 2 , Y. Suzuki 1 , K. Higashide 1 , N. Okada 2 , C. Harris 3 , S. Matsuo 1 , Y. Ito 1 1
Nagoya University Graduate School of Medicine, Nagoya, Japan Nagoya City University Graduate School of Medicine, Nagoya, Japan 3 Cardiff University, Cardiff, United Kingdom 2
Background: Peritoneal dialysis (PD) therapy is one of the most important renal replacement therapies. Impairment of peritoneal function can limit the long-term efficacy of PD therapy and is caused by several factors that occur during PD therapy, including exposure to peritoneal dialysate, catheter trauma and peritonitis. We previously showed in animal models that inhibition of membrane complement regulators (CRegs) could induce peritoneal inflammation and that dysregulated complement activation induced severe peritoneal injuries. Objective and methods: We investigated the expression of CD46, CD55 and CD59 on peritoneal mesothelial cells and levels of the complement activation marker sC5b-9 in PD fluids (PDF) to clarify influence of complement activation and CRegs expression in PD patients. Primary cell cultures of mesothelial cells were obtained from PDF of 27 PD patients and from omentum of 3 non-chronic kidney disease patients undergoing laparoscopic operations and cells were analysed for expression of CRegs. sC5b-9 levels were measured in the PDF of the PD patients, and background history, including complications of diabetes, usage of icodextrin as PDF, PD history, and dialysate-to-plasma creatinine concentration ratio (D/P Cre), an indicator of peritoneal function, were noted. Results: In PD patients, expression of CD55 but not CD46 and CD59 on mesothelial cells was significantly correlated to peritoneal function (D/P Cre; p < 0.05). Levels of sC5b-9 in the PDF showed weak inverse correlation with expression levels of CD46 and CD59 on mesothelial cells. Loss of expression of CD55 on mesothelial cells correlated with reduced mRNA for CD55 (p < 0.0001); however, there was no correlation between CD46/CD59 protein expression and levels of respective mRNA. Use of icodextrin correlated with decreased levels of sC5b-9 in PDF (p < 0.005). Complication such as diabetes, usage of icodextin, or PD history did not affect CD55 expression. Conclusion: Our results show that PD therapy alters expression of CRegs and complement regulation in the peritoneum. These data