Publications by authors named "Mohammad Reza Mirlashari"

15 Publications

  • Page 1 of 1

HLA class I depletion by citric acid, and irradiation of apheresis platelets for transfusion of refractory patients.

Transfusion 2021 04 13;61(4):1222-1234. Epub 2021 Feb 13.

Department of Immunology and Transfusion Medicine, Oslo University Hospital, Oslo, Norway.

Background: Patients can form antibodies to foreign human leukocyte antigen (HLA) Class I antigens after exposure to allogeneic cells. These anti-HLA class I antibodies can bind transfused platelets (PLTs) and mediate their destruction, thus leading to PLT refractoriness. Patients with PLT refractoriness need HLA-matched PLTs, which require expensive HLA typing of donors, antibody analyses of patient sera and/or crossmatching. An alternative approach is to reduce PLT HLA Class I expression using a brief incubation in citric acid on ice at low pH.

Methods And Materials: Apheresis PLT concentrates were depleted of HLA Class I complexes by 5 minutes incubation in ice-cold citric acid, at pH 3.0. Surface expression of HLA Class I complexes, CD62P, CD63, phosphatidylserine, and complement factor C3c was analyzed by flow cytometry. PLT functionality was tested by thromboelastography (TEG).

Results: Acid treatment reduced the expression of HLA Class I complexes by 71% and potential for C3c binding by 11.5-fold compared to untreated PLTs. Acid-treated PLTs were significantly more activated than untreated PLTs, but irrespective of this increase in steady-state activation, CD62P and CD63 were strongly upregulated on both acid-treated and untreated PLTs after stimulation with thrombin receptor agonist peptide. Acid treatment did not induce apoptosis over time. X-ray irradiation did not significantly influence the expression of HLA Class I complexes, CD62P, CD63, and TEG variables on acid treated PLTs.

Conclusion: The relatively simple acid stripping method can be used with irradiated apheresis PLTs and may prevent transfusion-associated HLA sensitization and overcome PLT refractoriness.
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http://dx.doi.org/10.1111/trf.16282DOI Listing
April 2021

Antitumor, Anti-Inflammatory and Antiallergic Effects of Mushroom Extract and the Related Medicinal Basidiomycetes Mushrooms, and : A Review of Preclinical and Clinical Studies.

Nutrients 2020 May 8;12(5). Epub 2020 May 8.

Institute of Clinical Medicine, University of Oslo, 0318 Oslo, Norway.

Since the 1980s, medicinal effects have been documented in scientific studies with the related mushrooms Murill (AbM), (HE) and (GF) from Brazilian and Eastern traditional medicine. Special focus has been on their antitumor effects, but the mushrooms' anti-inflammatory and antiallergic properties have also been investigated. The antitumor mechanisms were either direct tumor attack, e.g., apoptosis and metastatic suppression, or indirect defense, e.g., inhibited tumor neovascularization and T helper cell (Th) 1 immune response. The anti-inflammatory mechanisms were a reduction in proinflammatory cytokines, oxidative stress and changed gut microbiota, and the antiallergic mechanism was amelioration of a skewed Th1/Th2 balance. Since a predominant Th2 milieu is also found in cancer, which quite often is caused by a local chronic inflammation, the three conditions-tumor, inflammation and allergy-seem to be linked. Further mechanisms for HE were increased nerve and beneficial gut microbiota growth, and oxidative stress regulation. The medicinal mushrooms AbM, HE and GF appear to be safe, and can, in fact, increase longevity in animal models, possibly due to reduced tumorigenesis and oxidation. This article reviews preclinical and clinical findings with these mushrooms and the mechanisms behind them.
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http://dx.doi.org/10.3390/nu12051339DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7285126PMC
May 2020

NETs analysed by novel calprotectin-based assays in blood donors and patients with multiple myeloma or rheumatoid arthritis: A pilot study.

Scand J Immunol 2020 May 4;91(5):e12870. Epub 2020 Mar 4.

Department of Immunology and Transfusion Medicine, Oslo University Hospital (OUH), Oslo, Norway.

Two novel enzyme-linked immunosorbent assays (ELISAs), designed to detect complexes containing DNA, leucocyte calprotectin and S100A12 proteins, were generated for improved specificity and rapid measurement of neutrophil extracellular traps (NETs). The assays were applied on plasma and serum samples from blood donors for establishment of reference values, and from patients with multiple myeloma (MM) or rheumatoid arthritis (RA) in order to examine putatively increased values in the two different inflammatory conditions. Although NETs were hardly detectable in healthy individuals, NET levels were as expected highly and statistically significantly increased in RA patients. The detection of statistically significantly increased NET levels in MM is a novel finding.
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http://dx.doi.org/10.1111/sji.12870DOI Listing
May 2020

-Based Mushroom Extract Supplementation to Birch Allergic Blood Donors: A Randomized Clinical Trial.

Nutrients 2019 Oct 2;11(10). Epub 2019 Oct 2.

Department of Immunology and Transfusion Medicine, Oslo University Hospital, 0407 Oslo, Norway.

Since Murill (AbM) extract reduced specific IgE and ameliorated a skewed Th1/Th2 balance in a mouse allergy model, it was tested in blood donors with self-reported, IgE-positive, birch pollen allergy and/or asthma. Sixty recruited donors were randomized in a placebo-controlled, double-blinded study with pre-seasonal, 7-week, oral supplementation with the AbM-based extract Andosan. Before and after the pollen season, questionnaires were answered for allergic rhino-conjunctivitis, asthma, and medication; serum IgE was measured, and Bet v 1-induced basophil activation was determined by CD63 expression. The reported general allergy and asthma symptoms and medication were significantly reduced in the AbM compared to the placebo group during pollen season. During the season, there was significant reduction in specific IgE anti-Bet v 1 and anti-t3 (birch pollen extract) levels in the AbM compared with the placebo group. While the maximal allergen concentrations needed for eliciting basophil activation before the season, changed significantly in the placebo group to lower concentrations (i.e., enhanced sensitization) after the season, these concentrations remained similar in the Andosan AbM extract group. Hence, the prophylactic effect of oral supplementation before the season with the AbM-based Andosan extract on aeroallergen-induced allergy was associated with reduced specific IgE levels during the season and basophils becoming less sensitive to allergen activation.
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http://dx.doi.org/10.3390/nu11102339DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6836217PMC
October 2019

Cytotoxic Effect on Human Myeloma Cells and Leukemic Cells by the Murill Based Mushroom Extract, Andosan™.

Biomed Res Int 2017 7;2017:2059825. Epub 2017 Nov 7.

Institute of Clinical Medicine, University of Oslo, P.O. Box 1171, 0318 Oslo, Norway.

Murill is an edible mushroom of the Basidiomycetes family, which has been found to contain a number of compounds with antitumor properties, such as proteoglycans and ergosterol. In the present investigation, we show that the commercial mushroom product Andosan, which contains 82.4% Murill, together with medicinal mushrooms (14.7%) and (2.9%), has a cytotoxic effect on primary myeloma cells, other myeloma cell lines, and leukemia cell lines Although the exact content and hence the mechanisms of action of the Andosan extract are unknown, we have found in this investigation indications of cell cycle arrest when myeloma cell lines are cultivated with Andosan. This may be one of the possible explanations for the cytotoxic effects of Andosan.
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http://dx.doi.org/10.1155/2017/2059825DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5697368PMC
July 2018

Human Adipose-Derived Mesenchymal Stem Cells Respond to Short-Term Hypoxia by Secreting Factors Beneficial for Human Islets In Vitro and Potentiate Antidiabetic Effect In Vivo.

Cell Med 2017 14;9(3):103-116. Epub 2017 Apr 14.

Department of Transplant Medicine, Oslo University Hospital, Oslo, Norway.

Adipose-derived mesenchymal stem cells (ASCs) release factors beneficial for islets in vitro and protect against hyperglycemia in rodent models of diabetes. Oxygen tension has been shown to induce metabolic changes and alter ASCs' release of soluble factors. The effects of hypoxia on the antidiabetic properties of ASCs have not been explored. To investigate this, we incubated human ASCs for 48 h in 21% (normoxia) or 1% O (hypoxia) and compared viability, cell growth, surface markers, differentiation capability, and soluble factors in the conditioned media (CM). Human islets were exposed to CM from ASCs incubated in either normoxia or hypoxia, and islet function and apoptosis after culture with or without proinflammatory cytokines were measured. To test hypoxic preconditioned ASCs' islet protective effects in vivo, ASCs were incubated for 48 h in normoxia or hypoxia before being injected into Balb/c Rag 1 immunodeficient mice with streptozotocin-induced insulitis. Progression of diabetes and insulin content of pancreas were measured. We found that incubation in hypoxia was well tolerated by ASCs and that levels of VEGF-A, FGF-2, and bNGF were elevated in CM from ASCs incubated in hypoxia compared to normoxia, while levels of HGF, IL-8, and CXCL1 were reduced. CM from ASCs incubated in hypoxia significantly improved human islet function and reduced apoptosis after culture, and reduced cytokine-induced apoptosis. In our mouse model, pancreas insulin content was higher in both groups receiving ASCs compared to control, but the mice receiving preconditioned ASCs had lower random and fasting blood glucose, as well as improved oral glucose tolerance compared to untreated mice. In conclusion, our in vitro results indicate that the islet protective potential of ASCs improves in hypoxia, and we give insight into factors involved in this. Finally we show that hypoxic preconditioning potentiates ASCs' antidiabetic effect in vivo.
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http://dx.doi.org/10.3727/215517917X693401DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5509020PMC
April 2017

3',4'-Dimethoxyflavone and valproic acid promotes the proliferation of human hematopoietic stem cells.

Stem Cell Res Ther 2013 May 24;4(3):60. Epub 2013 May 24.

Introduction: Human hematopoietic stem cells (HSCs) have been clinically used for transplantation and gene and cellular therapy for more than 4 decades. However, this use is limited because of the challenges in the ex vivo culturing of HSCs. The major hurdle is to amplify these cells without losing their self-renewing property.

Methods: In our study, we tested 3',4'-dimethoxyflavone (3'4'-DMF) and valproic acid (VPA) on the ex vivo expansion of HSCs under both normoxic (20% O2) and hypoxic (1% O2) conditions. 3'4'-DMF is a widely used anticancer drug that acts as a competitive antagonist of the aryl hydrocarbon receptor. VPA is a potent inhibitor of histone deacetylase and is used in the treatment of neurologic disorders.

Results: Culturing HSCs (from mobilized peripheral blood) under normoxia, with 3'4'-DMF and VPA, highly preserved the CD34 positivity (3'4'-DMF, 22.1%, VPA, 20.3%) after 1 week and strongly enhanced the CD34(+) cells (3'4'-DMF, 27.8 fold; VPA, 34.1 fold) compared with the control cultures (11.6% and 14.4 fold). Addition of 3'4'-DMF and VPA also resulted in more primary colonies and replating efficiency compared with control cultures. Although no significant effect was observed on the enhancement of CD34(+) cells under hypoxia, the number of primary colonies was significantly higher than the control cultures.

Conclusions: Based on these findings, this study presents, for the first time, in vitro evidence for a new and relevant effect of 3'4'-DMF on human HSCs. In addition, the results suggest a potential clinical use of 3'4'-DMF and VPA in HSC therapy.
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http://dx.doi.org/10.1186/scrt208DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3706763PMC
May 2013

Biological response modifiers in photochemically pathogen-reduced versus untreated apheresis platelet concentrates.

Transfusion 2013 Jan 7;53(1):147-55. Epub 2012 May 7.

Department of Immunology and Transfusion Medicine, Oslo University Hospital, Ullevaal, Oslo, Norway.

Background: Lipids and other biologically active substances accumulate in platelet concentrates (PCs) during storage. Some of these substances have been suggested to modulate immune responses and to play a pathogenic role in the development of transfusion-related acute lung injury. This study compared the content and impact of some biological response modifiers in PCs treated with pathogen reduction (PR) technology and nontreated PCs.

Study Design And Methods: Apheresis PCs (n = 12) were split in two: one split was subjected to PR treatment (INTERCEPT, Cerus Corp.) and the other split was left untreated. Basic characterization and content of vascular endothelial growth factor (VEGF) and sCD154 were measured. Lipopolysaccharide (LPS)-induced secretion of interleukin-10 (IL-10) and tumor necrosis factor-α (TNF-α) was measured after incubation of heparinized whole blood with platelet (PLT) supernatants. The supernatants' neutrophil (PMN)-priming capacity, and thereby activation of the NADPH oxidase, was measured as the rate of superoxide anion production after formyl-Met-Leu-Phe activation. Lipids were extracted from the supernatants on Day 6 and tested for PMN-priming activity.

Results: Supernatants from PR-treated PCs demonstrated significantly higher mean PLT volume (MPV) and O(2) , lower pH, CO(2) , and HCO(3-) , and significantly less LPS-induced TNF-α secretion compared to untreated PCs. No differences in swirling, PLT count, potassium levels, glucose consumption, lactate production, IL-10, VEGF, sCD154, or PMN-priming activity were found between the groups over time.

Conclusion: INTERCEPT PR treatment caused no substantial differences in PCs, except for minor changes in MPV and metabolic variables. Further studies are needed to explain the differences in the LPS-induced TNF-α secretion.
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http://dx.doi.org/10.1111/j.1537-2995.2012.03681.xDOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3690765PMC
January 2013

Glycogen synthase kinase-3 (GSK-3) inhibition induces apoptosis in leukemic cells through mitochondria-dependent pathway.

Leuk Res 2012 Apr 15;36(4):499-508. Epub 2011 Dec 15.

Department of Immunology and Transfusion Medicine, Oslo University Hospital, Oslo, Norway.

The roles of glycogen synthase kinase-3 (GSK-3) in cell survival and apoptosis are controversial. We examined the effect of a specific GSK-3 inhibitor (SB-415286) on the regulation of leukemic cells proliferation and apoptosis. SB-415286 (40 μM) induced cell growth inhibition, β-catenin stabilization, cell cycle arrest in G(2)/M phase, cyclin B1 downregulation, and apoptosis in leukemic cell lines KG1a, K562, and CMK. Blocking the death receptor pathway by using a specific inhibitor of caspase-8, did not inhibit SB-415286-induced apoptosis. This indicates that activation of caspase-8 is part of the intrinsic apoptotic pathway and occurs downstream of mitochondria membrane potential depolarization mediated by other caspases. Furthermore, we found that depolarization of mitochondria membrane caused by GSK-3 inhibition is regulated by dephosphorylation of anti-apoptotic protein Bcl-2 and downregulation of Bcl-xL. Thus, inhibition of GSK-3-induced apoptosis of leukemic cells could be an attractive target for treatment of leukemia.
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http://dx.doi.org/10.1016/j.leukres.2011.11.013DOI Listing
April 2012

Evaluation of nonleukoreduced red blood cell transfusion units collected at delivery from the placenta.

Transfusion 2007 Aug;47(8):1481-7

Departments of Pediatrics and Immunology & Transfusion Medicine, Ullevål University Hospital, University of Oslo, Kirkeveien 166, N-0407 Oslo, Norway.

Background: The objective of this study was to evaluate the suitability of cord blood (CB) as a source of red blood cells (RBCs) for autologous transfusion.

Study Design And Methods: CB was collected in 150-mL storage containers with citrate phosphate dextrose (CPD) as anticoagulant and stored in either saline, adenine, glucose, and mannitol (SAG-M; n = 18) or phosphate, adenine, glucose, guanosine, saline, and mannitol (PAGGS-M; n = 18) for 35 days at 4 degrees C. Hematologic status and hemolysis were studied. The lipopolysaccharide (LPS)-induced production of tumor necrosis factor (TNF)-alpha and transforming growth factor (TGF)-beta1 from CB monocytes was analyzed after incubation with addition of weekly sampled supernatants from the CB RBC units. Five additional units (PAGGS-M) were leukoreduced and thereafter analyzed as indicated above.

Results: Hemolysis increased significantly over time, in SAG-M more than in PAGGS-M. During storage in both media, the number of white blood cells (WBCs) decreased, and the LPS-induced production of TNF-alpha and TGF-beta1 decreased and increased, respectively. There were no significant changes in the LPS-induced production of TNF-alpha and TGF-beta1 in the leukoreduced CB RBC units.

Conclusion: Hemolysis in CB RBC units increased significantly over time, and PAGGS-M appears to be superior to SAG-M as a preservation solution for CB RBC. The changes in LPS-induced TNF-alpha and TGF-beta1 production over time were probably caused by substances released from apoptotic and/or necrotic WBCs. Further studies are needed to identify both which substances are responsible for the changes in LPS-induced cytokine release and the clinical significance hereof.
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http://dx.doi.org/10.1111/j.1537-2995.2007.01287.xDOI Listing
August 2007

Evaluation of platelet activation and cytokine release during storage of platelet concentrates processed from buffy coats either manually or by the automated OrbiSac system.

Transfusion 2007 Jan;47(1):126-32

Faculty Division Ullevål University Hospital, University of Oslo, Oslo, Norway.

Background: This study aimed to compare platelet (PLT) quality during storage of buffy coat (BC) PLT concentrates (PCs), prepared either manually or by the automated OrbiSac system (Gambro BCT).

Study Design And Methods: Following overnight storage at 20 to 22 degrees C, five BCs were pooled with 300 mL of PLT additive solution. Twenty-one PCs were produced manually (M-PCs) and 21 by the automated OrbiSac system (A-PCs). Swirling, PLT count, mean PLT volume, blood gas analyses, potassium, glucose, and lactate were assessed. Expression of the activation markers CD42a, CD62P, and PAC-1 was analyzed by flow cytometry on resting PLTs and PLTs stimulated with thrombin receptor agonist peptide (TRAP). Levels of CCL5 and transforming growth factor-beta1 (TGF-beta1) were measured by enzyme-linked immunosorbent assay.

Results: The A-PCs had significantly larger volume and higher PLT yield, PLT recovery, and white blood cell concentration than the M-PCs, whereas the red blood cell content was significantly highest in the M-PCs. pH levels were between 6.9 and 7.2 in all PCs. Neither glucose consumption nor lactate production differed significantly over time. A-PCs had, compared to M-PCs, significantly higher expression of CD62P on resting PLTs, lower capacity for up regulating CD62P on TRAP-stimulated PLTs, and higher levels of CCL5 during storage. TRAP-stimulated A-PCs had a significantly higher potential for down regulation of CD42a than M-PCs. No difference was found in TGF-beta1 levels or TRAP-induced up regulation of PAC-1.

Conclusion: The levels of CCL5 and the expression of CD62P in resting and stimulated PLTs indicate that PLTs in A-PCs are slightly more activated than in M-PCs, but the clinical importance of this finding is yet unknown.
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http://dx.doi.org/10.1111/j.1537-2995.2007.01075.xDOI Listing
January 2007

Platelet activation and residual activation potential during storage of hyperconcentrated platelet products in two different platelet additive solutions.

Transfusion 2005 Aug;45(8):1349-55

Department of Immunology and Transfusion Medicine, Ullevål University Hospital, Oslo, Norway.

Background: To improve platelet (PLT) quality, hyperconcentrated PLT concentrates (hcPCs) were compared to standard PLT concentrates (stdPCs) in two different PLT additive solutions, T-Sol and PAS-27a. PAS-27a differs from T-Sol by containing glucose, phosphate, potassium, magnesium, and bicarbonate.

Study Design And Methods: PLTs were harvested by apheresis twice from 14 donors; each unit was divided into two. Four units from each donor were produced: hcPCs, 2000 x 10(9) per L in T-Sol or PAS-27a; and stdPCs, 1400 x 10(9) per L in 65 percent T-Sol or PAS-27a and 35 percent acid citrate dextrose-plasma. On Days 1 through 4, swirling was scored and PLT count, mean PLT volume, pH, blood gas, glucose, and lactate were measured. Expression of CD42a, CD62P, CD63, and PAC-1 was analyzed by flow cytometry on resting PLTs and PLTs stimulated with thrombin receptor agonist peptide (TRAP).

Results: Glucose consumption and lactate production were significantly higher in hcPCs stored in PAS-27a than in T-Sol. Both stdPC and hcPC PLTs in T-Sol expressed CD62P and PAC-1 significantly higher than in PAS-27a. Over time the T-Sol hcPCs revealed highest expression of CD62P and CD63. A significantly higher capacity for up regulation of CD62P, CD63, and PAC-1 upon TRAP stimulation was found for stdPCs and hcPCs in PAS-27a compared to PLTs in T-Sol. TRAP-stimulated PLTs in stdPCs and hcPCs suspended in PAS-27a showed significantly higher potential for down regulation of CD42a than the T-Sol concentrates.

Conclusions: PLTs appear better preserved in vitro in PAS-27a than in T-Sol, and this suggests that storage of hcPCs in PAS-27a could be extended beyond 24 hours.
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http://dx.doi.org/10.1111/j.1537-2995.2005.00218.xDOI Listing
August 2005

Expression and involvement of Toll-like receptors (TLR)2, TLR4, and CD14 in monocyte TNF-alpha production induced by lipopolysaccharides from Neisseria meningitidis.

Med Sci Monit 2003 Aug;9(8):BR316-24

Research Forum, Ullevaal University Hospital, Oslo, Norway.

Background: The present study was undertaken to examine the ability of lipopolysaccharide-containing outer membrane vesicles (OMV-LPS) and purified LPS (P-LPS) from the same meningococcal strain to induce the expression of Toll-like receptors (TLR2 and TLR4) and TNF-alpha production in leukocytes, and further to study the involvement of TLRs, and CD14 in monocyte TNF- alpha production in an ex vivo human whole blood system.

Material/methods: OMV-LPS or P-LPS were added to human whole blood and expression of TLR2/4 and production of TNF- alpha in leukocytes were measured by flow cytometry. To study involvement of TLRs and CD14 in monocyte cytokine production we used monoclonal antibodies against TLR2/4 and CD14.

Results: OMV-LPS and P-LPS induced surface expression (maximal at 120 min) of TLR2 and TLR4 on granulocytes and monocytes. LPS incorporated in OMV was less potent (weight basis) than P-LPS in inducing monocyte TNF- alpha production. When inducing monocyte TNF-alpha by OMV-LPS, antibodies directed against TLR2 and TLR4 caused 45 and 78% inhibition, respectively. When inducing TNF- alpha by P-LPS, antibodies against TLR2 had no effect, whereas anti-TLR4 antibodies caused 63% inhibition. Antibodies against CD14 inhibited nearly completely the monocyte TNF- alpha response induced by meningococcal LPS irrespective of whether LPS was presented in purified form or incorporated in membrane vesicles.

Conclusions: OMV-LPS and P-LPS from the same meningococcal strain induced expression of TLR2/4 on monocytes and granulocytes. Surface receptors TLR2/4 and CD14 are essential for in vitro cellular activation induced by OMV-LPS and P-LPS, but the functional significance of these receptors during meningococcal infections remains elusive.
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August 2003

Outer membrane vesicles from Neisseria meningitidis.

APMIS 2002 Mar;110(3):193-204

Research Forum, Ullevaal University Hospital, Oslo, Norway.

Flow cytometry was used to study the expression of leukocyte adhesion molecules CD11a, CD11b, CD11c, CD14, and CD62L (L-selectin) and production of reactive oxygen species (ROS) in an ex vivo human whole-blood system stimulated with lipopolysaccharide-containing outer membrane vesicles (LPS-OMV) from N. meningitidis. Results demonstrated a dose-dependent increase in surface expression of CD11a, CD11b, CD11c and CD14 in granulocytes and monocytes (maximal at 30-120 min) upon OMV-LPS challenge, whereas CD62L expression was heavily downregulated (maximal at 30-120 min). The OMV-associated LPS was almost as potent (on a weight basis) as purified LPS from E. coli in inducing adhesion molecule modulation but the response was delayed. Upon stimulation with OMV-LPS or E. coli-LPS, the production of intracellular ROS increased in both granulocytes and monocytes when dihydroethidium (DHE, mainly reflecting superoxide anion) was used as a probe, whereas peroxynitrite production monitored with dihydrorhodamine 123 (DHR) was not significantly changed. The OMV-mediated modulation of leukocyte adhesion molecule expression and increased ROS production may certainly lead to increased entrapment of leukocytes in the microcirculation and contribute to untoward inflammatory reactions as seen in systemic meningococcal disease.
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http://dx.doi.org/10.1034/j.1600-0463.2002.100301.xDOI Listing
March 2002

Platelet-platelet and platelet-leukocyte interactions induced by outer membrane vesicles from N. meningitidis.

Platelets 2002 Mar;13(2):91-9

Research Forum, Ullevaal University Hospital, Oslo, Norway.

Unlabelled: A large part of native meningococcal lipopolysaccharide (LPS), i.e., LPS integrated in the outer cell membrane, is released in the form of 'blebs' from surplus outer membrane material. In the present study we investigated the effects of purified outer membrane vesicles (OMVs) on blood platelet-platelet and platelet-leukocyte interactions. Citrated whole blood was stimulated in vitro with equal amounts (on a weight basis) of OMV-integrated LPS, purified LPS (P-LPS) from the same meningococcal strain and purified E. coli-LPS. The samples were analyzed by flow cytometry. Upon OMV stimulation platelet aggregation increased 2.1-fold, platelet degranulation 1.8-fold, (measured as CD62P expression), platelet binding to monocytes 2.6-fold, whereas platelet binding to granulocytes increased 2.8-fold. Also, the fraction of large heteroconjugates, i.e., large CD45-positive cell aggregates increased 15.7-fold compared to control. P-LPS and E. coli-LPS also significantly increased platelet aggregation and heteroconjugate formation but did not influence platelet degranulation and binding of platelets to leukocytes in whole blood. When using platelet-rich plasma (PRP), OMVs increased platelet aggregation 2.1-fold and CD62P expression 1.9-fold. P-LPS and E. coli-LPS also significantly increased platelet aggregation in PRP but did not influence platelet degranulation. None of the LPS preparations induced platelet microvesiculation, either in whole blood or in PRP.

Conclusion: Meningococcal-derived OMVs as well as purified meningococcal LPS, contribute to increased platelet-platelet and platelet-leukocyte aggregation and may thus be of great importance in the development of microthrombosis and organ dysfunction related to fulminant meningococcal septicemia.
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http://dx.doi.org/10.1080/09537100220122448DOI Listing
March 2002
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