scholarly journals Abnormal directed migration of blood polymorphonuclear leukocytes in rheumatoid arthritis. Potential role in increased susceptibility to bacterial infections

1998 ◽  
Vol 7 (1) ◽  
pp. 19-23 ◽  
Author(s):  
F. Aglas ◽  
J. Hermann ◽  
G. Egger
Keyword(s):  
Rheumatoid Arthritis ◽  
Polymorphonuclear Leukocytes ◽  
Bacterial Infections ◽  
Membrane Filter ◽  
Absolute Number ◽  
Healthy Controls ◽  
Distribution Characteristic ◽  
Migratory Activity ◽  
Directed Migration

Rheumatoid arthritis (RA) patients are at higher risks of bacterial infection than healthy subjects. Polymorphonuclear leukocytes (PMN) are the first line of nonspecific cellular defence against these infections. We tested the hypothesis that abnormal directed migration of PMN may be one reason for the increased infection rate of RA patients. PMN migration was investigated in 68 peripheral blood samples of 15 RA patients compared with 64 samples of healthy controls in a novel whole bloodin vitromembrane filter assay. The migration of PMNs from RA patients and controls was stimulated using the bacterial chemoattractant N-formyl-methionyl-leucylphenylalanine (fMLP). Unstimulated PMN migration of RA patients was increased compared with healthy controls as measured by the following parameters: (a) absolute number of migrant PMNs (1954 ± 87 vs. 1238 ± 58 PMN/mm2), (b) percentage of PMNs migrated into the filter (total migration index, TMI) (28.6 ± 0.9 vs. 24.0 ± 0.8 %), (c) the distance half the migrating PMNs had covered (distribution characteristic, DC) (22.6 ± 1.1 vs. 16.1 ± 0.6 mm) and (d) the product of TMI and DC (neutrophil migratory activity, NMA) (669.0 ± 45.0 vs. 389.0 ± 18.9). fMLP stimulated PMNs of RA patients showed defective migration compared to unstimulated samples as shown by (a) a reduced number of migrant PMNs (1799 ± 93 PMN/mm2), (b) lower TMI (26.1 ± 0.9 %), (c) unremarkable altered distribution characteristic (22.9 ± 0.8 mm) and (d) significant reduced migratory activity (600.0 ± 30.0). Our data suggest that the high incidence of infections in RA patients may partly be caused by defective migratory activity of PMNs to bacterial chemoattractants as demonstrated by fMLP.

scholarly journals Mitochondrial N-formylmethionyl proteins as chemoattractants for neutrophils.

1982 ◽  
Vol 155 (1) ◽  
pp. 264-275 ◽  
Author(s):  
H Carp
Keyword(s):  
Polymorphonuclear Leukocytes ◽  
Inflammatory Cells ◽  
Chemotactic Activity ◽  
Mitochondrial Proteins ◽  
Mitochondrial Enzymes ◽  
Directed Migration ◽  
Formyl Peptide ◽  
Cultured Human Cells ◽  
Chemotactic Factors

Mitochondria synthesize several hydrophobic proteins. Like bacteria, mitochondria initiate protein synthesis with an N-formylmethionine residue. Because N-formylmethionyl peptides have been found to be chemotactic for polymorphonuclear leukocytes (PMN), mitochondria isolated from cultured human cells and purified bovine mitochondrial proteins were tested for PMN chemotactic activity in vitro. Nondisrupted mitochondria were not chemotactic. However, intact mitochondria that had been incubated with a lysosomal lysate did stimulate PMN migration. Antibodies directed against two mitochondrial enzymes, cytochrome oxidase and ATPase, (both of which contain mitochondrially synthesized subunits) but not anti-C3 or anti-C5 decreased mitochondrially derived chemotactic activity. In addition, purified bovine mitochondrial N-formylmethionyl proteins stimulated PMN migration in vitro, whereas nonformylated mitochondrial proteins did not. Furthermore, the chemotactic activity of purified mitochondrial proteins and disrupted mitochondria was decreased by the formyl peptide antagonist butyloxycarbonyl-phenylalanine-leucine-phenylalanine-leucine-phenylalanine. Finally, disrupted mitochondria and purified mitochondrial proteins stimulated PMN-directed migration (chemotaxis), according to accepted criteria. In addition to other chemotactic factors, release of N-formylmethionyl proteins from mitochondria at sites of tissue damage, may play a role in the accumulation of inflammatory cells at these sites.

scholarly journals Increased Concentrations of Antibody-Bound Circulatory Cell-Free DNA in Rheumatoid Arthritis

2007 ◽  
Vol 53 (9) ◽  
pp. 1609-1614 ◽  
Author(s):  
Xiao-Yan Zhong ◽  
Ines von Mühlenen ◽  
Ying Li ◽  
Anjeung Kang ◽  
Anurag Kumar Gupta ◽  
...  
Keyword(s):  
Rheumatoid Arthritis ◽  
Animal Experiments ◽  
Basic Research ◽  
Healthy Controls ◽  
Serum Samples ◽  
Cell Free Dna ◽  
Free Dna ◽  
New Evidence

Abstract Background: Increased concentrations of cell-free DNA have been found in several disorders and have been interpreted as evidence of increased rates of cell death or turnover. Evidence from in vitro and animal experiments suggests that DNA may play a role in the pathogenesis of rheumatoid arthritis (RA). Methods: We measured cell-free DNA in plasma and serum from patients with RA and healthy controls by use of quantitative PCR for glyceraldehyde-3-phosphate dehydrogenase (GAPDH) DNA. We used protein G Sepharose™ bead adsorption of plasma and elution to isolate antibody-bound DNA. Results: In paired plasma and serum samples of 16 healthy controls the median GAPDH copies were 4500 genome equivalents (GE)/mL plasma (range 319–21 000) and in 26 RA patients 17 000 GE/mL plasma (2100–2 375 000, P = 0.0001). In the serum from normal controls the median GAPDH copies were 35 000 GE/mL (1700–239 000) and from RA patients 222 000 GE/mL (21 000–2 375 000, P = 0.004). A median of 81% of the cell-free DNA in RA was associated with antibody compared with 9% in healthy controls (P = 0.001). The concentrations of DNA did not vary with the type of therapy patients received. Conclusions: These results provide new evidence for a role of cell-free DNA-antibody complexes in the etiology of RA, suggest new avenues for basic research, and may prove to be relevant to diagnosis and assessment of therapy.

scholarly journals Evaluation of Th9 lymphocytes in peripheral blood of rheumatoid arthritis patients and correlation with anti-tumor necrosis factor therapy: results from an in vitro pivotal study

Reumatismo ◽  
2016 ◽  
Vol 68 (2) ◽  
pp. 83 ◽  
Author(s):  
R. Talotta ◽  
A. Berzi ◽  
F. Atzeni ◽  
D. Dell'Acqua ◽  
P. Sarzi Puttini ◽  
...  
Keyword(s):  
Rheumatoid Arthritis ◽  
Tumor Necrosis Factor ◽  
Peripheral Blood ◽  
Tumor Necrosis ◽  
Mononuclear Cells ◽  
Healthy Controls ◽  
T Helper ◽  
Th9 Cells ◽  
Necrosis Factor

The aim of this study was to determine the prevalence of T helper 9 (Th9) lymphocytes in rheumatoid arthritis (RA) patients and to identify a possible association between the percentage of Th9 and the discontinuation of a biological treatment with an anti-tumor necrosis factor (TNF) (infliximab). We collected peripheral blood mononuclear cells (PBMCs) from 55 consecutive RA outpatients and 10 healthy controls. Among RA patients, 15 were not receiving any immunosuppressive drug, 20 were successfully treated with infliximab and 20 discontinued infliximab because of adverse events or inefficacy and were treated with other biological agents. PBMCs were cultured with/without infliximab 50 mg/L for 18 h, and the percentage of Th9 cells was assessed by means of flow cytometry. Th9 lymphocytes were identified as interferon gamma, interleukin (IL)4-, IL17-, IL9-secreting cluster of differentiation 4 (CD4)+ T cells. Cytometric analysis revealed no significant decrease in the percentage of Th9 cells after infliximab exposure in any of the groups, although it was lower in healthy controls than RA patients either before and after the infliximab stimulation assay. Th9 cells are IL-9-secreting T helper lymphocytes whose role in RA is still poorly known. IL-9 levels are increased in RA patients, in whom this cytokine plays a crucial role. Th9 cells are the major producers of IL-9, and their prevalence is higher in RA patients than in healthy subjects; however our experiment <em>in vitro</em> does not demonstrate an association between Th9 lymphocytes and the response to infliximab. Further studies are required to evaluate the real involvement of Th9 population in the immunogenicity of anti-TNF agents.

scholarly journals THU0104 THE GUT MICROBIOTA AND ITS RELEVANCE TO PERIPHERAL T REGULATORY CELLS AND T HELPER 17 IN PATIENTS WITH RHEUMATOID ARTHRITIS

2020 ◽  
Vol 79 (Suppl 1) ◽  
pp. 266.1-266
Author(s):  
Y. LI ◽  
S. X. Zhang ◽  
X. F. Yin ◽  
Z. Mingxing ◽  
J. Luo ◽  
...  
Keyword(s):  
Rheumatoid Arthritis ◽  
T Cells ◽  
Gut Microbiota ◽  
Lymphocyte Subsets ◽  
Joint Destruction ◽  
Absolute Number ◽  
P Value ◽  
Rrna Gene ◽  
Healthy Controls ◽  
The Absolute

Background:Rheumatoid arthritis (RA) is a common autoimmune disorder with joint destruction and synovial inflammation characterized by abnormal immune responses to autoantigens. Our previous studies have demonstrated that impaired peripheral lymphocytes especially insufficiency of regulatory T cells (Tregs) played an important role in pathogenesis of RA1 2. Interestingly, the dysbiosis of gut microbiota triggers several types of autoimmune diseases through the imbalance of T lymphocyte subsets3. However, the detailed gut microbiota of RA patients and its correlation with Tregs and helper T cells 17 (Th17) are unclear up until now.Objectives:To compare the difference of gut microbiota between RA and healthy controls (HCs), and to investigate the relevance of gut microbiota with circulating Tregs and Th17 in patients with RA.Methods:From December 2018 to August 2019, a total of 205 diagnosed patients with RA and 199 age and sex-matched HCs were enrolled in this study. Stool of Every participant was collected for bacterial DNA extraction and 16S ribosomal RNA (rRNA) gene sequencing. The absolute numbers of Tregs and Th17 in PB of these individuals were measured by Flow Cytometer (FCM) combined with standard absolute counting beads. Data were expressed as mean ± standard deviation to the distribution. Independent-samples T test and Spearman rank correlation test. P value <0.05 were considered statistically significant.Results:Patients with RA had a significantly difference of diversity and abundance of intestinal microbiota compared with those of HCs (P< 0.05). Detailedly, the abundance of Proteobacteria was significantly increased in RA patients (P< 0.05), and the abundance of Firmicutes, Fusobacteria and Verrucomicrobia were significantly reduced (P<0.05) at the level of Phylum (Figure 1). At the genus level, in the RA group, the abundance of Escherichia, Ruminococcus2 and Clostridium_sensu_stricto were significantly increased (P< 0.05), but the abundance of Lachnospiracea_incertae_sedis, Prevotella, Clostridium_XlVa, Roseburia, Dialister, Blautia, Megamonas and Gemmiger were significantly lower than the healthy controls (P< 0.05) (Figure 2). Moreover, Blautia, Anaerostipes and Ruminococcus2 have negative correlation with the absolute number of Tregs, and Cloacibacillus and Streptophyta have positive correlation with the absolute number of Th17.Conclusion:Patients with RA had a dysbiosis of the gut microbiota in both diversity and abundance, which is closely related to the impaired peripheral lymphocyte subsets, that may be related to the pathogenesis of RA, which might provide a new idea for RA treatment.References:[1]Wen HY, Wang J, Zhang SX, et al. Low-Dose Sirolimus Immunoregulation Therapy in Patients with Active Rheumatoid Arthritis: A 24-Week Follow-Up of the Randomized, Open-Label, Parallel-Controlled Trial.J Immunol Res2019;2019:7684352. doi: 10.1155/2019/7684352 [published Online First: 2019/11/30][2]Niu HQ, Li ZH, Zhao WP, et al. Sirolimus selectively increases circulating Treg cell numbers and restores the Th17/Treg balance in rheumatoid arthritis patients with low disease activity or in DAS28 remission who previously received conventional disease-modifying anti-rheumatic drugs.Clin Exp Rheumatol2019 [published Online First: 2019/05/11][3]Lee N, Kim WU. Microbiota in T-cell homeostasis and inflammatory diseases.Exp Mol Med2017;49(5):e340. doi: 10.1038/emm.2017.36 [published Online First: 2017/05/27]Acknowledgments:NoneDisclosure of Interests:None declared

Revival of Krebs–Ringer balanced salt solution for the investigation of polymorphonuclear leukocytes and Pseudomonas aeruginosa biofilm interaction

2019 ◽  
Vol 77 (5) ◽  
Author(s):  
Thomas Bjarnsholt ◽  
Peter Østrup Jensen ◽  
Maria Alhede
Keyword(s):  
Pseudomonas Aeruginosa ◽  
Salt Solution ◽  
Polymorphonuclear Leukocytes ◽  
Bacterial Infections ◽  
Treatment Strategies ◽  
Term Survival ◽  
Lactate Dehydrogenase Activity ◽  
Bacterial Aggregation ◽  
Novel Treatment Strategies

ABSTRACTTo study the interaction between aggregating bacteria and polymorphonuclear leukocytes (PMNs) in vitro, the chosen medium must favor both the isolated PMNs and the bacteria. To investigate the best-suited medium for the in vitro survival of isolated unactivated human PMNs, we compared three different mammalian cell media: Krebs–Ringer balanced salt solution (BSS), Hanks’ BSS (HBSS) and Roswell Park Memorial Institute (RPMI) 1640. The death of PMNs was estimated by the release of lactate dehydrogenase activity. Furthermore, two types of serum, human (HS) and fetal bovine (FBS), were compared at different concentrations (0%, 2%, 5%, 10%) and at three different time points (2, 4, 20 h). We show that Krebs–Ringer BSS prolonged the survival of PMNs compared to HBSS and RPMI 1640 and that the addition of 10% FBS significantly enhanced the long-term survival (20 h) compared to HS. Furthermore, we observed aggregation of Pseudomonas aeruginosa when grown in the presence of either a mixture of histones, histone H3, arginine or lysine. In this study, we show that the use of Krebs–Ringer BSS is highly relevant for the study of the interaction of bacteria and PMNs in relation to novel treatment strategies of biofilm infections due to the reproduction of bacterial aggregation as seen in chronic bacterial infections.

scholarly journals Therapeutic applications of mannan-binding lectin

2003 ◽  
Vol 25 (4) ◽  
pp. 33-34 ◽  
Author(s):  
David Kilpatrick
Keyword(s):  
Rheumatoid Arthritis ◽  
Cystic Fibrosis ◽  
Innate Immunity ◽  
Bacterial Infections ◽  
Recurrent Miscarriage ◽  
Association Studies ◽  
Disease Association ◽  
Mannan Binding Lectin ◽  
Binding Lectin

Mannan-binding lectin (MBL) is perhaps the best known of the collectins, a subfamily of C-type lectins possessing an additional collagen-like domain. MBL is thought to be an important component of innate immunity. Since the discovery, around 15 years ago, that MBL was identical to a factor responsible for the opsonization of baker's yeast in vitro, a large number of disease-association studies have been conducted. Most of these investigations support the view that MBL can influence susceptibility to, or affect the course of, many diseases. These include bacterial infections, rheumatoid arthritis, cystic fibrosis and recurrent miscarriage. This meeting was arranged to bring together scientists and clinicians to review the clinical significance of MBL, and to consider the most appropriate target disorders for MBL-replacement therapy.

Inflammatory cytokines shape a changing DNA methylome in monocytes mirroring disease activity in rheumatoid arthritis

2019 ◽  
Vol 78 (11) ◽  
pp. 1505-1516 ◽  
Author(s):  
Javier Rodríguez-Ubreva ◽  
Carlos de la Calle-Fabregat ◽  
Tianlu Li ◽  
Laura Ciudad ◽  
Maria L Ballestar ◽  
...  
Keyword(s):  
Rheumatoid Arthritis ◽  
Dna Methylation ◽  
Disease Activity ◽  
Inflammatory Diseases ◽  
Systemic Autoimmune Disease ◽  
Healthy Controls ◽  
Blood Monocytes ◽  
Necrosis Factor Alpha ◽  
Extracellular Signals

ObjectiveRheumatoid arthritis (RA) is a chronic systemic autoimmune disease that mainly targets joints. Monocytes and macrophages are critical in RA pathogenesis and contribute to inflammatory lesions. These extremely plastic cells respond to extracellular signals which cause epigenomic changes that define their pathogenic phenotype. Here, we interrogated how DNA methylation alterations in RA monocytes are determined by extracellular signals.MethodsHigh-throughput DNA methylation analyses of patients with RA and controls and in vitro cytokine stimulation were used to investigate the underlying mechanisms behind DNA methylation alterations in RA as well as their relationship with clinical parameters, including RA disease activity.ResultsThe DNA methylomes of peripheral blood monocytes displayed significant changes and increased variability in patients with RA with respect to healthy controls. Changes in the monocyte methylome correlate with DAS28, in which high-activity patients are divergent from healthy controls in contrast to remission patients whose methylome is virtually identical to healthy controls. Indeed, the notion of a changing monocyte methylome is supported after comparing the profiles of same individuals at different stages of activity. We show how these changes are mediated by an increase in disease activity-associated cytokines, such as tumour necrosis factor alpha and interferons, as they recapitulate the DNA methylation changes observed in patients in vitro.ConclusionWe demonstrate a direct link between RA disease activity and the monocyte methylome through the action of inflammation-associated cytokines. Finally, we have obtained a DNA methylation-based mathematical formula that predicts inflammation-mediated disease activity for RA and other chronic immune-mediated inflammatory diseases.

Migratory activity of blood polymorphnuclear leukocytes during juvenile rheumatoid arthritis, demonstrated with a new wholeblood membrane filter assay

Inflammation ◽  
1994 ◽  
Vol 18 (4) ◽  
pp. 427-441 ◽  
Author(s):  
Gerd Egger ◽  
Christiane Klemt ◽  
Stefan Spendel ◽  
Wilhelm Kaulfersch ◽  
Harald Kenzian
Keyword(s):  
Rheumatoid Arthritis ◽  
Juvenile Rheumatoid Arthritis ◽  
Membrane Filter ◽  
Migratory Activity ◽  
Filter Assay
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