| 1. |
- Aljadi, Zenib, et al.
(author)
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Activation of Basophils Is a New and Sensitive Marker of Biocompatibility in Hemodialysis
- 2014
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In: Artificial Organs. - : Wiley. - 0160-564X .- 1525-1594. ; 38:11, s. 945-953
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Journal article (peer-reviewed)abstract
- The hemodialysis procedure involves contact between peripheral blood and the surface of dialyzer membranes, which may lead to alterations in the pathways of innate and adaptive immunity. We aimed to study the effect of blood-membrane interaction on human peripheral basophils and neutrophils in hemodialysis with high- and low-permeability polysulfone dialyzers. The surface expression of CD203c (basophil selection marker) and CD63 (activation marker) after activation by the bacterial peptide formyl-methionyl-leucyl-phenylalanine (fMLP) or anti-Fc epsilon receptor I (Fc epsilon RI) antibody and the absolute number of basophils was investigated before and after hemodialysis with each of the dialyzers. Moreover, the expression on neutrophils of CD11b, the CD11b active epitope, and CD88 was analyzed in the same groups of individuals. The expression of CD63 in basophils following activation by fMLP was significantly higher in the patient group compared with that in healthy controls, but no differences were observed after activation by anti-Fc epsilon RI. During the hemodialysis procedure, the low-flux membrane induced up-regulation of CD63 expression on basophils, while passage through the high-flux membrane did not significantly alter the responsiveness. In addition, the absolute number of basophils was unchanged after hemodialysis with either of the dialyzers and compared with healthy controls. We found no significant differences in the expression of the neutrophil activation markers (CD11b, the active epitope of CD11b, and CD88) comparing the two different dialyzers before and after dialysis and healthy controls. Together, these findings suggest that alterations in basophil activity may be a useful marker of membrane bioincompatibility in hemodialysis.
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| 2. |
- Lindberg, Jenny, et al.
(author)
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Monocyte and Neutrophil Chemotactic Activity at the Site of Interstitial Inflammation in Patients on High-Flux Hemodialysis or Hemodiafiltration
- 2009
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In: Blood Purification. - : S. Karger AG. - 0253-5068 .- 1421-9735. ; 28:1, s. 47-52
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Journal article (peer-reviewed)abstract
- Background/Aims: We have observed a difference between patients on low-flux hemodialysis (HD) or peritoneal dialysis and patients on hemodiafiltration (HDF) or high-flux HD in the capacity of transmigrated leukocytes to mobilize CD11b in response to inflammatory stimuli compared with healthy subjects. This could be due to different interstitial chemokine concentrations. Methods: We measured concentrations of circulating and interstitial macrophage inflammatory protein-1 alpha (MIP-1 alpha), matrix metalloproteinase-9 (MMP-9)/neutrophil gelatinase-associated lipocalin (NGAL), monocyte chemoattractant protein-1 (MCP-1) and interleukin-8 (IL-8) in 10 patients on HDF or high-flux HD and 11 healthy subjects by using immunoassay. Results: The interstitial concentrations of MIP-1 alpha, MMP-9/NGAL and IL-8 were similar in patients and healthy subjects, while the corresponding concentration of MCP-1 was significantly higher in patients on HDF or high-flux HD as compared with healthy subjects (p < 0.01). Conclusion: We suggest that an equal or higher concentration of chemokines in the interstitium in patients with HDF or high-flux HD might be one mechanism responsible for the preserved function of transmigrated leukocytes. Copyright (C) 2009 S. Karger AG, Basel
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| 3. |
- Olsson, Jenny, et al.
(author)
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Expression of neutrophil SOD2 is reduced after lipopolysaccharide stimulation : A potential cause of neutrophil dysfunction in chronic kidney disease
- 2011
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In: Nephrology Dialysis Transplantation. - : Oxford University Press (OUP). - 0931-0509 .- 1460-2385. ; 26:7, s. 2195-2201
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Journal article (peer-reviewed)abstract
- Background. Neutrophils from patients with chronic kidney disease (CKD) are dysfunctional and thus a contributing factor to the risk of infections. The mechanisms for leucocyte dysfunction in CKD are not fully understood. It is known that lipopolysaccharide (LPS) activates transcription of several genes encoding proinflammatory cytokines. We therefore aimed to study the effect of LPS on neutrophil expression of genes related to the inflammatory response to address the hypothesis that LPS-induced gene transcriptions are altered in CKD patients.Methods. We analysed gene expression of LPS-stimulated neutrophils from 30 patients with CKD and 15 healthy controls. Superoxide dismutase-2 (SOD2), IL1A, IL-1R1, IL-1R2 and IL8RA gene expression from both neutrophils and differentiated HL60 cells were measured by quantitative polymerase chain reaction. Differentiated HL60 cells were stimulated with phorbol-12-myristate-7- acetate (PMA) after inhibition of SOD2 by small interfering RNA followed by respiratory burst assessment using flow cytometry.Results. LPS stimulation induced a significant mobilization of CD11b on neutrophils from CKD and healthy controls. Upregulation of SOD2, IL1A, IL-1R1 and IL-1R2 gene expression in neutrophils from healthy controls after LPS stimulation was contrasted by no change in gene transcription (IL-1R1 and IL-1R2) or even a downregulation in patients with CKD (SOD2 and IL1A). Inhibition of SOD2 reduced the PMA-induced respiratory burst and IL1A, IL-1R1, IL-1R2 and IL8RA gene expression in neutrophil-differentiated HL60 cells.Conclusions. Because of the critical role of SOD2 in the generation of hydrogen peroxide during phagocytosis, downregulation of SOD2 gene expression after LPS stimulation in neutrophils from patients with CKD indicates a potential mechanism for neutrophil dysfunction and cytokine dysregulation in these patients.
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| 4. |
- Paulsson, Josefin, et al.
(author)
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Activation of peripheral and in vivo transmigrated neutrophils in patients with stable coronary artery disease
- 2007
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In: Atherosclerosis. - : Elsevier BV. - 0021-9150 .- 1879-1484. ; 192:2, s. 328-334
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Journal article (peer-reviewed)abstract
- Accumulating evidence support a role of neutrophils in coronary artery disease (CAD). However little is known about the action of neutrophils at a local inflammatory site represented by an atherosclerotic plaque. To gain insight into these issues, we applied a skin blister model that permits analyses of in vivo transmigrated neutrophils. We hypothesised that the chronic inflammation in stable CAD mediates priming of neutrophils that impacts the out-come of neutrophil action at an inflammatory site. Thirteen patients with angiographically verified CAD were eligible for study entry together with 13 age and sex matched controls. Markers of inflammation (IL-6 and CRP), neutrophil activation (IL-8 and MMP-9/NGAL), and functional aspects (CD11b up-regulation and intracellular H(2)O(2) production) of peripheral and in vivo transmigrated neutrophils were studied. Systemic IL-8 and MMP-9/NGAL concentrations were significantly increased in patients indicating a primed state in circulating neutrophils. In vivo transmigrated neutrophils in stable CAD patients had an increased propensity to release MMP-9/NGAL and a reduced capacity to up-regulate CD11b and to produce hydrogen peroxide. These aberrations at the inflammatory site may be a consequence of a primed state of circulating neutrophils and point towards potential mechanisms whereby neutrophils at a local inflammatory site may contribute to the pathogenesis of CAD.
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| 5. |
- Paulsson, Josefin
(author)
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The inflammatory response in extravasated leukocytes in patients with coronary artery disease
- 2010
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Doctoral thesis (other academic/artistic)abstract
- Coronary artery disease (CAD) is a manifestation of a chronic inflammation in the coronary arteries. The inflammatory process results in accumulation of monocyte derived cells and formation of atherosclerotic plaques in the intima of the vessel wall. Neutrophils are mainly associated with ruptured plaques and the precise role in CAD is not fully known. Extravasation into local inflammatory sites is coordinated by adhesion molecules and chemokines and transforms the leukocytes into activated tissue dwelling cells. The aim of this thesis was to investigate extravasated monocytes and neutrophils in patients with stable CAD. A skin chamber method was applied in order to induce a local inflammation from which extravasated cells were collected. Paper I and II describes extravasated monocytes. Patients with CAD had a similar number of extravasated monocytes in the chamber exudate compared to healthy subjects. The expression of CD11b following extravasation was lower in patients with CAD compared to healthy controls. This might result in an increased retention of monocytes at a local inflammatory site. Other markers associated with monocyte extravasation, VLA-4 and CX3CR1, were not altered. Extravasated monocytes were further analyzed for functional alterations. Markers associated with antigen presentation, HLA-DR and CD86, and binding of modified cholesterol, CD36 and scavenger receptor A1 (SR-A1) had an increased expression following extravasation compared to in circulation. Furthermore, the binding of acetylated low density lipoprotein (acLDL) increased following extravasation. Monocytes from patients and controls had a similar functional response. However, the chamber fluid from patients with CAD enhanced the expression of CD36 following in vitro stimulation of mononuclear cells. Paper III and IV describes extravasated neutrophils. Extravasated neutrophils from patients with CAD had a significantly lower expression of CD11b and a lower production of reactive oxygen species (ROS) following stimulation compared to healthy controls. This might indicate a refractory stage following extravasation in patients with CAD. The gene expression in extravasated neutrophils was assessed by a gene array. A general induction in the IL-1 axis was seen following extravasation and was associated with an increased expression of chemokines. Expression of IL-1R on human neutrophils was confirmed with flow cytometry and electron microscopy and stimulation with IL-1 resulted in CCL and CXCL chemokine gene and protein expressions. Compared to healthy controls, extravasated neutrophils from patients with CAD had significantly increased expressions of CCL20 and CXCL2. This finding indicates that neutrophils may have an immuno-modulatory role at local inflammatory sites and that patients with CAD have a chemokine profile that could enhance the pathological processes in atherosclerosis. The major findings indicate a potential mechanism for monocyte entrapment at local inflammatory sites. In addition, the local inflammatory milieu in patients with CAD might be pro-atherosclerotic. Neutrophils from patients with CAD had an altered responsiveness and could be refractory. Furthermore, neutrophils may alter the localinflammatory milieu by the production of chemokines.
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| 6. |
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| 7. |
- Wahlberg, Patrik, et al.
(author)
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Functional corpora lutea are formed in matrix metalloproteinase inhibitor-treated plasminogen-deficient mice.
- 2007
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In: Endocrinology. - 0013-7227. ; 148:3, s. 1226-34
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Journal article (peer-reviewed)abstract
- The extended Förster theory (EFT) is for the first time applied to the quantitative determination of the intramolecular distances in proteins. It is shown how the EFT (J. Chem. Phys., 1996, 105, 10896) can be adapted to the analyses of fluorescence depolarisation experiments based on the time-correlated single photon counting technique (TCSPC). The protein system studied was the latent form of plasminogen activator inhibitor type I (PAI-1), which was mutated and labelled by the thiol reactive BODIPY® derivative {N-(4,4-difluoro-5,7-dimethyl-4-bora-3a,4a-diaza-s-indacene-3-yl)methyl iodoacetamide}. The energy migration occurs within pairs of photophysically identical donor groups that undergo reorientational motions on the timescales of energy migration and fluorescence relaxation. Unlike all models currently used for analysing fluorescence TCSPC data, the EFT explicitly accounts for the time-dependent reorientations that influence the rate of electronic energy transfer/migration in a complex manner. The complexity is related to the 2 problem, which has been discussed for years. The EFT brings the analyses of DDEM data to the same level of molecular description as in ESR and NMR spectroscopy, i.e. it yields microscopic information about the reorientation correlation times, the order parameters, as well as inter-chromophoric distances.
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