Proteomic, biomechanical and functional analyses define neutrophil heterogeneity in systemic lupus erythematosus

Funder: NHLI FoundationFunder: NIHR Imperial Biomedical Research Centre; FundRef: http://dx.doi.org/10.13039/501100013342Funder: National Heart Lung and Blood InstituteFunder: Medical Research Council; FundRef: http://dx.doi.org/10.13039/501100000265Funder: National Institute of Biomedical Imaging and Bioengineering; FundRef: http://dx.doi.org/10.13039/100000070Funder: Gates Cambridge ScholarshipFunder: NIH/OXCAM FellowshipObjectives: Low-density granulocytes (LDGs) are a distinct subset of proinflammatory and vasculopathic neutrophils expanded in systemic lupus erythematosus (SLE). Neutrophil trafficking and immune function are intimately linked to cellular biophysical properties. This study used proteomic, biomechanical and functional analyses to further define neutrophil heterogeneity in the context of SLE. Methods: Proteomic/phosphoproteomic analyses were performed in healthy control (HC) normal density neutrophils (NDNs), SLE NDNs and autologous SLE LDGs. The biophysical properties of these neutrophil subsets were analysed by real-time deformability cytometry and lattice light-sheet microscopy. A two-dimensional endothelial flow system and a three-dimensional microfluidic microvasculature mimetic (MMM) were used to decouple the contributions of cell surface mediators and biophysical properties to neutrophil trafficking, respectively. Results: Proteomic and phosphoproteomic differences were detected between HC and SLE neutrophils and between SLE NDNs and LDGs. Increased abundance of type 1 interferon-regulated proteins and differential phosphorylation of proteins associated with cytoskeletal organisation were identified in SLE LDGs relative to SLE NDNs. The cell surface of SLE LDGs was rougher than in SLE and HC NDNs, suggesting membrane perturbances. While SLE LDGs did not display increased binding to endothelial cells in the two-dimensional assay, they were increasingly retained/trapped in the narrow channels of the lung MMM. Conclusions: Modulation of the neutrophil proteome and distinct changes in biophysical properties are observed alongside differences in neutrophil trafficking. SLE LDGs may be increasingly retained in microvasculature networks, which has important pathogenic implications in the context of lupus organ damage and small vessel vasculopathy

183 A phase 1B/2A trial of tofacitinib, an oral janus kinase inhibitor, in systemic lupus erythematosus

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A unified approach to the economic aspects of statistical quality control and improvement

Assignment (MSc)--Stellenbosch University, 2004.ENGLISH ABSTRACT: The design of control charts refers to the selection of the parameters implied, including the sample size n, control limit width parameter k, and the sampling interval h. The design of the X -control chart that is based on economic as well as statistical considerations is presently one of the more popular subjects of research. Two assumptions are considered in the development and use of the economic or economic statistical models. These assumptions are potentially critical. It is assumed that the time between process shifts can be modelled by means of the exponential distribution. It is further assumed that there is only one assignable cause. Based on these assumptions, economic or economic statistical models are derived using a total cost function per unit time as proposed by a unified approach of the Lorenzen and Vance model (1986). In this approach the relationship between the three control chart parameters as well as the three types of costs are expressed in the total cost function. The optimal parameters are usually obtained by the minimization of the expected total cost per unit time. Nevertheless, few practitioners have tried to optimize the design of their X -control charts. One reason for this is that the cost models and their associated optimization techniques are often too complex and difficult for practitioners to understand and apply. However, a user-friendly Excel program has been developed in this paper and the numerical examples illustrated are executed on this program. The optimization procedure is easy-to-use, easy-to-understand, and easy-to-access. Moreover, the proposed procedure also obtains exact optimal design values in contrast to the approximate designs developed by Duncan (1956) and other subsequent researchers. Numerical examples are presented of both the economic and the economic statistical designs of the X -control chart in order to illustrate the working of the proposed Excel optimal procedure. Based on the Excel optimization procedure, the results of the economic statistical design are compared to those of a pure economic model. It is shown that the economic statistical designs lead to wider control limits and smaller sampling intervals than the economic designs. Furthermore, even if they are more costly than the economic design they do guarantee output of better quality, while keeping the number of false alarm searches at a minimum. It also leads to low process variability. These properties are the direct result of the requirement that the economic statistical design must assure a satisfactory statistical performance. Additionally, extensive sensitivity studies are performed on the economic and economic statistical designs to investigate the effect of the input parameters and the effects of varying the bounds on, a, 1-f3 , the average time-to-signal, ATS as well as the expected shift size t5 on the minimum expected cost loss as well as the three control chart decision variables. The analyses show that cost is relatively insensitive to improvement in the type I and type II error rates, but highly sensitive to changes in smaller bounds on ATS as well as extremely sensitive for smaller shift levels, t5 . Note: expressions like economic design, economic statistical design, loss cost and assignable cause may seen linguistically and syntactically strange, but are borrowed from and used according the known literature on the subject.AFRIKAANSE OPSOMMING: Die ontwerp van kontrolekaarte verwys na die seleksie van die parameters geïmpliseer, insluitende die steekproefgrootte n , kontrole limiete interval parameter k , en die steekproefmterval h. Die ontwerp van die X -kontrolekaart, gebaseer op ekonomiese sowel as statistiese oorwegings, is tans een van die meer populêre onderwerpe van navorsing. Twee aannames word in ag geneem in die ontwikkeling en gebruik van die ekonomiese en ekonomies statistiese modelle. Hierdie aannames is potensieel krities. Dit word aanvaar dat die tyd tussen prosesverskuiwings deur die eksponensiaalverdeling gemodelleer kan word. Daar word ook verder aangeneem dat daar slegs een oorsaak kan wees vir 'n verskuiwing, of te wel 'n aanwysbare oorsaak (assignable cause). Gebaseer op hierdie aannames word ekonomies en ekonomies statistiese modelle afgelei deur gebruik te maak van 'n totale kostefunksie per tydseenheid soos voorgestel deur deur 'n verenigende (unified) benadering van die Lorenzen en Vance-model (1986). In hierdie benadering word die verband tussen die drie kontrole parameters sowel as die drie tipes koste in die totale kostefunksie uiteengesit. Die optimale parameters word gewoonlik gevind deur die minirnering van die verwagte totale koste per tydseenheid. Desnieteenstaande het slegs 'n minderheid van praktisyns tot nou toe probeer om die ontwerp van hulle X -kontrolekaarte te optimeer. Een rede hiervoor is dat die kosternodelle en hulle geassosieerde optimeringstegnieke té kompleks en moeilik is vir die praktisyns om te verstaan en toe te pas. 'n Gebruikersvriendelike Excelprogram is egter hier ontwikkel en die numeriese voorbeelde wat vir illustrasie doeleindes getoon word, is op hierdie program uitgevoer. Die optimeringsprosedure is maklik om te gebruik, maklik om te verstaan en die sagteware is geredelik beskikbaar. Wat meer is, is dat die voorgestelde prosedure eksakte optimale ontwerp waardes bereken in teenstelling tot die benaderde ontwerpe van Duncan (1956) en navorsers na hom. Numeriese voorbeelde word verskaf van beide die ekonomiese en ekonomies statistiese ontwerpe vir die X -kontrolekaart om die werking van die voorgestelde Excel optimale prosedure te illustreer. Die resultate van die ekonomies statistiese ontwerp word vergelyk met dié van die suiwer ekomomiese model met behulp van die Excel optimerings-prosedure. Daar word aangetoon dat die ekonomiese statistiese ontwerpe tot wyer kontrole limiete en kleiner steekproefmtervalle lei as die ekonomiese ontwerpe. Al lei die ekonomies statistiese ontwerp tot ietwat hoër koste as die ekonomiese ontwerpe se oplossings, waarborg dit beter kwaliteit terwyl dit die aantal vals seine tot 'n minimum beperk. Hierbenewens lei dit ook tot kleiner prosesvartasie. Hierdie eienskappe is die direkte resultaat van die vereiste dat die ekonomies statistiese ontwerp aan sekere statistiese vereistes moet voldoen. Verder is uitgebreide sensitiwiteitsondersoeke op die ekonomies en ekonomies statistiese ontwerpe gedoen om die effek van die inset parameters sowel as van variërende grense op a, 1- f3 , die gemiddelde tyd-tot-sein, ATS sowel as die verskuiwingsgrootte 8 op die minimum verwagte kosteverlies sowel as die drie kontrolekaart besluitnemingsveranderlikes te bepaal. Die analises toon dat die totale koste relatief onsensitief is tot verbeterings in die tipe I en die tipe II fout koerse, maar dat dit hoogs sensitief is vir wysigings in die onderste grens op ATS sowel as besonder sensitief vir klein verskuiwingsvlakke, 8. Let op: Die uitdrukkings ekonomiese ontwerp (economic design), ekonomies statistiese ontwerp (economic statistical design), verlies kostefunksie (loss cost function) en aanwysbare oorsaak (assignable cause) mag taalkundig en sintakties vreemd voordoen, maar is geleen uit, en word so gebruik in die bekende literatuur oor hierdie onderwerp

Proteomic, biomechanical and functional analyses define neutrophil heterogeneity in systemic lupus erythematosus

Funder: NHLI FoundationFunder: NIHR Imperial Biomedical Research Centre; FundRef: http://dx.doi.org/10.13039/501100013342Funder: National Heart Lung and Blood InstituteFunder: Medical Research Council; FundRef: http://dx.doi.org/10.13039/501100000265Funder: National Institute of Biomedical Imaging and Bioengineering; FundRef: http://dx.doi.org/10.13039/100000070Funder: Gates Cambridge ScholarshipFunder: NIH/OXCAM FellowshipObjectives: Low-density granulocytes (LDGs) are a distinct subset of proinflammatory and vasculopathic neutrophils expanded in systemic lupus erythematosus (SLE). Neutrophil trafficking and immune function are intimately linked to cellular biophysical properties. This study used proteomic, biomechanical and functional analyses to further define neutrophil heterogeneity in the context of SLE. Methods: Proteomic/phosphoproteomic analyses were performed in healthy control (HC) normal density neutrophils (NDNs), SLE NDNs and autologous SLE LDGs. The biophysical properties of these neutrophil subsets were analysed by real-time deformability cytometry and lattice light-sheet microscopy. A two-dimensional endothelial flow system and a three-dimensional microfluidic microvasculature mimetic (MMM) were used to decouple the contributions of cell surface mediators and biophysical properties to neutrophil trafficking, respectively. Results: Proteomic and phosphoproteomic differences were detected between HC and SLE neutrophils and between SLE NDNs and LDGs. Increased abundance of type 1 interferon-regulated proteins and differential phosphorylation of proteins associated with cytoskeletal organisation were identified in SLE LDGs relative to SLE NDNs. The cell surface of SLE LDGs was rougher than in SLE and HC NDNs, suggesting membrane perturbances. While SLE LDGs did not display increased binding to endothelial cells in the two-dimensional assay, they were increasingly retained/trapped in the narrow channels of the lung MMM. Conclusions: Modulation of the neutrophil proteome and distinct changes in biophysical properties are observed alongside differences in neutrophil trafficking. SLE LDGs may be increasingly retained in microvasculature networks, which has important pathogenic implications in the context of lupus organ damage and small vessel vasculopathy

LOX-1: A potential driver of cardiovascular risk in SLE patients.

Traditional cardiovascular disease (CVD) risk factors, such as hypertension, dyslipidemia and diabetes do not explain the increased CVD burden in systemic lupus erythematosus (SLE). The oxidized-LDL receptor, LOX-1, is an inflammation-induced receptor implicated in atherosclerotic plaque formation in acute coronary syndrome, and here we evaluated its role in SLE-associated CVD. SLE patients have increased sLOX-1 levels which were associated with elevated proinflammatory HDL, oxLDL and hsCRP. Interestingly, increased sLOX-1 levels were associated with patients with early disease onset, low disease activity, increased IL-8, and normal complement and hematological measures. LOX-1 was increased on patient-derived monocytes and low-density granulocytes, and activation with oxLDL and immune-complexes increased membrane LOX-1, TACE activity, sLOX-1 release, proinflammatory cytokine production by monocytes, and triggered the formation of neutrophil extracellular traps which can promote vascular injury. In conclusion, perturbations in the lipid content in SLE patients' blood activate LOX-1 and promote inflammatory responses. Increased sLOX-1 levels may be an indicator of high CVD risk, and blockade of LOX-1 may provide a therapeutic opportunity for ameliorating atherosclerosis in SLE patients

Using the circulating proteome to assess type I interferon activity in systemic lupus erythematosus.

Type I interferon (IFN) drives pathology in systemic lupus erythematosus (SLE) and can be tracked via IFN-inducible transcripts in blood. Here, we examined whether measurement of circulating proteins, which enter the bloodstream from inflamed tissues, also offers insight into global IFN activity. Using a novel protocol we generated 1,132 aptamer-based protein measurements from anti-dsDN

Phase 1 double-blind randomized safety trial of the Janus kinase inhibitor tofacitinib in systemic lupus erythematosus

Increased risk of premature cardiovascular disease in systemic lupus erythematosus (SLE) is not well understood, but in animal models, the Janus kinase inhibitor tofacitinib improves related phenotypes. Here the authors report a Phase 1 double-blind randomized trial that shows tofacitinib is safe and well tolerated in in patients with SLE