scholarly journals Profiling Human CD55 Transgene Performance Assist in Selecting Best Suited Specimens and Tissues for Swine Organ Xenotransplantation

Biology ◽  
2021 ◽  
Vol 10 (8) ◽  
pp. 747
Author(s):  
Laura Martínez-Alarcón ◽  
Sergio Liarte ◽  
Juan J. Quereda ◽  
Aida Sáez-Acosta ◽  
Carlos de Torre-Minguela ◽  
...  
Keyword(s):  
Tissue Expression ◽  
Transgenic Pigs ◽  
Complement Regulatory Proteins ◽  
Classical Complement Pathway ◽  
Protein Levels ◽  
Human Sera ◽  
And Performance ◽  
Surrogate Tissues ◽  
The Individual ◽  
Classical Complement

Xenotransplantation of pig organs receives substantial attention for being comparable to human’s. However, compatibility constraints involving hyper-acute rejection (HAR) still block clinical applications. Transgenesis of human complement regulatory proteins has been proposed to overcome xenorejection. Pigs expressing human-CD55 have been widely tested in experimental surgery. Still, no standardized method has been developed to determine tissue expression of human decay-accelerating factor (DAF), hCD55’s product, or to predict the ability to overpass HAR. Here we describe objective procedures addressing this need. Organs and tissues from five hCD55 transgenic pigs were collected and classified according to their xenotransplantation value. The ability to overcome HAR was assessed by classical complement pathway hemolysis assays. Quantitative PCR mRNA expression and Western blot protein level studies were performed. Real-time cytotoxicity assays (RTCA) on fibroblast cultures exposed to baboon and human sera informed on longer-term rejection dynamics. While greater hCD55/DAF expression correlated with better performance, the results obtained varied among specimens. Interestingly, the individual with highest mRNA and protein levels showed positive feedback for hCD55 transcript after challenge with human and baboon sera. Moreover, hCD55 expression correlated to DAF levels in the liver, lung and intestine, but not in the heart. Moreover, we found significant correlations among valuable and non-valuable tissues. In sum, the methodology proposed allows us to characterize the hCD55 transgene functioning and performance. Moreover, the correlations found could allow us to predict hCD55/DAF expression in surrogate tissues, thus eliminating the need for direct biopsies, resulting in preservation of organ integrity before xenotransplantation.

scholarly journals Preparing for Life: Plasma Proteome Changes and Immune System Development During the First Week of Human Life

2020 ◽  
Vol 11 ◽  
Author(s):  
Tue Bjerg Bennike ◽  
Benoit Fatou ◽  
Asimenia Angelidou ◽  
Joann Diray-Arce ◽  
Reza Falsafi ◽  
...  
Keyword(s):  
Immune System ◽  
System Development ◽  
Human Life ◽  
Plasma Concentrations ◽  
Plasma Proteome ◽  
Complement Pathway ◽  
Classical Complement Pathway ◽  
Protein Levels ◽  
Immune System Development ◽  
Classical Complement

Neonates have heightened susceptibility to infections. The biological mechanisms are incompletely understood but thought to be related to age-specific adaptations in immunity due to resource constraints during immune system development and growth. We present here an extended analysis of our proteomics study of peripheral blood-plasma from a study of healthy full-term newborns delivered vaginally, collected at the day of birth and on day of life (DOL) 1, 3, or 7, to cover the first week of life. The plasma proteome was characterized by LC-MS using our established 96-well plate format plasma proteomics platform. We found increasing acute phase proteins and a reduction of respective inhibitors on DOL1. Focusing on the complement system, we found increased plasma concentrations of all major components of the classical complement pathway and the membrane attack complex (MAC) from birth onward, except C7 which seems to have near adult levels at birth. In contrast, components of the lectin and alternative complement pathways mainly decreased. A comparison to whole blood messenger RNA (mRNA) levels enabled characterization of mRNA and protein levels in parallel, and for 23 of the 30 monitored complement proteins, the whole blood transcript information by itself was not reflective of the plasma protein levels or dynamics during the first week of life. Analysis of immunoglobulin (Ig) mRNA and protein levels revealed that IgM levels and synthesis increased, while the plasma concentrations of maternally transferred IgG1-4 decreased in accordance with their in vivo half-lives. The neonatal plasma ratio of IgG1 to IgG2-4 was increased compared to adult values, demonstrating a highly efficient IgG1 transplacental transfer process. Partial compensation for maternal IgG degradation was achieved by endogenous synthesis of the IgG1 subtype which increased with DOL. The findings were validated in a geographically distinct cohort, demonstrating a consistent developmental trajectory of the newborn’s immune system over the first week of human life across continents. Our findings indicate that the classical complement pathway is central for newborn immunity and our approach to characterize the plasma proteome in parallel with the transcriptome will provide crucial insight in immune ontogeny and inform new approaches to prevent and treat diseases.

scholarly journals C1 fixation and classical complement pathway activation by a fragment of the Cmu4 domain of IgM.

1975 ◽  
Vol 142 (5) ◽  
pp. 1322-1326 ◽  
Author(s):  
M M Hurst ◽  
J E Volanakis ◽  
R M Stroud ◽  
J C Bennett
Keyword(s):  
Complement Fixation ◽  
Marked Decrease ◽  
Classical Complement Pathway ◽  
Amino Terminal ◽  
Pathway Activation ◽  
Normal Human ◽  
Carboxy Terminal Region ◽  
The Individual ◽  
Carboxy Terminal ◽  
Classical Complement

A 56 residue fragment derived from a Waldenströme IgM protein and consisting of 24 residues of the amino-terminal portion of the Cmu4 domain disulfide bonded to 32 residues of the carboxy-terminal region of the loop has been shown to fix active C1 (C1) in a C1-fixation assay. Cleavage of the disulfide bond within the CH4 fragment resulted in a marked decrease of C1-fixing ability, although the isolated A and B fragments did retain a limited ability to fix C1. Upon incubation with normal human serum the intact CH4 fragment and equal molar amounts of the isolated A and B peptides consumed C4 suggesting that the C1-activating determinant of IgM remains intact in these three fragments. Furthermore, on a molar basis the intact or the reduced CH4 fragment consumed C4 as effectively as each of its component chains suggesting that transient binding of C1 by the individual A and B peptide chains is sufficient to activate C1. On the basis of these observations it is proposed that a classical complement fixation function, i.e. C1 binding and activation, can be localized within a region of the IgM molecule corresponding to the Cmu4 domain.

scholarly journals Schizophrenia: Complement Cleaning or Killing

Genes ◽  
2021 ◽  
Vol 12 (2) ◽  
pp. 259
Author(s):  
Jirrine T.T. Hogenaar ◽  
Hans van Bokhoven
Keyword(s):  
Complement System ◽  
Therapeutic Interventions ◽  
Complement Pathway ◽  
Complement Protein ◽  
Major Pathway ◽  
Classical Complement Pathway ◽  
Collective Data ◽  
The Individual ◽  
The Complement System ◽  
Classical Complement

Schizophrenia is a psychiatric disorder with a typical onset occurring during adolescence or young adulthood. The heterogeneity of the disorder complicates our understanding of the pathophysiology. Reduced cortical synaptic densities are commonly observed in schizophrenia and suggest a role for excessive synaptic elimination. A major pathway hypothesised to eliminate synapses during postnatal development is the complement system. This review provides an overview of genetic and functional evidence found for the individual players of the classical complement pathway. In addition, the consequences of the absence of complement proteins, in the form of complement protein deficiencies in humans, are taken into consideration. The collective data provide strong evidence for excessive pruning by the classical complement pathway, contributing to cognitive impairment in schizophrenia. In future studies, it will be important to assess the magnitude of the contribution of complement overactivity to the occurrence and prevalence of phenotypic features in schizophrenia. In addition, more insight is required for the exact mechanisms by which the complement system causes excessive pruning, such as the suggested involvement of microglial engulfment and degradation of synapses. Ultimately, this knowledge is a prerequisite for the development of therapeutic interventions for selective groups of schizophrenia patients.

scholarly journals Heat Pump Bridge Analysis Using the Modified Energy Transfer Diagram

Energies ◽  
2020 ◽  
Vol 14 (1) ◽  
pp. 137
Author(s):  
Florian Schlosser ◽  
Heinrich Wiebe ◽  
Timothy G. Walmsley ◽  
Martin J. Atkins ◽  
Michael R. W. Walmsley ◽  
...  
Keyword(s):  
Energy Transfer ◽  
Heat Pump ◽  
Heat Recovery ◽  
Heat Pumps ◽  
Coefficient Of Performance ◽  
Combined Application ◽  
Recovery Potential ◽  
Composite Curve ◽  
And Performance ◽  
The Individual

Heat pumps are the key technology to decarbonise thermal processes by upgrading industrial surplus heat using renewable electricity. Existing insight-based integration methods refer to the idealised Grand Composite Curve requiring the full exploitation of heat recovery potential but leave the question of how to deal with technical or economic limitations unanswered. In this work, a novel Heat Pump Bridge Analysis (HPBA) is introduced for practically targeting technical and economic heat pump potential by applying Coefficient of Performance curves into the Modified Energy Transfer Diagram (METD). Removing cross-Pinch violations and operating heat exchangers at minimum approach temperatures by combined application of Bridge Analysis increases the heat recovery rate and reduce the temperature lift to be pumped at the same time. The insight-based METD allows the individual matching of heat surpluses and deficits of individual streams with the capabilities and performance of different market-available heat pump concepts. For an illustrative example, the presented modifications based on HPBA increase the economically viable share of the technical heat pump potential from 61% to 79%.

scholarly journals Carbamylation of immunoglobulin abrogates activation of the classical complement pathway

2014 ◽  
Vol 44 (11) ◽  
pp. 3403-3412 ◽  
Author(s):  
Catalin Koro ◽  
Ewa Bielecka ◽  
Anders Dahl‐Knudsen ◽  
Jan J. Enghild ◽  
Carsten Scavenius ◽  
...  
Keyword(s):  
Complement Pathway ◽  
Classical Complement Pathway ◽  
Classical Complement

scholarly journals IL6 sensitizes prostate cancer to the antiproliferative effect of IFNα2 through IRF9

2013 ◽  
Vol 20 (5) ◽  
pp. 677-689 ◽  
Author(s):  
Holger H H Erb ◽  
Regina V Langlechner ◽  
Patrizia L Moser ◽  
Florian Handle ◽  
Tineke Casneuf ◽  
...  
Keyword(s):  
Prostate Cancer ◽  
Cell Lines ◽  
Cellular Proliferation ◽  
Tissue Expression ◽  
Type I ◽  
Regulatory Factor ◽  
Quantitative Real Time Pcr ◽  
Antiproliferative Effects ◽  
Protein Levels

Development and progression of prostate cancer (PCa) are associated with chronic inflammation. The cytokine interleukin 6 (IL6) can influence progression, differentiation, survival, and angiogenesis of PCa. To identify novel pathways that are triggered by IL6, we performed a gene expression profiling of two PCa cell lines, LNCaP and MDA PCa 2b, treated with 5 ng/ml IL6. Interferon (IFN) regulatory factor 9 (IRF9) was identified as one of the most prevalent IL6-regulated genes in both cell lines. IRF9 is a mediator of type I IFN signaling and acts together with STAT1 and 2 to activate transcription of IFN-responsive genes. The IL6 regulation of IRF9 was confirmed at mRNA and protein levels by quantitative real-time PCR and western blot respectively in both cell lines and could be blocked by the anti-IL6 antibody Siltuximab. Three PCa cell lines, PC3, Du-145, and LNCaP-IL6+, with an autocrine IL6 loop displayed high expression of IRF9. A tissue microarray with 36 PCa tissues showed that IRF9 protein expression is moderately elevated in malignant areas and positively correlates with the tissue expression of IL6. Downregulation and overexpression of IRF9 provided evidence for an IFN-independent role of IRF9 in cellular proliferation of different PCa cell lines. Furthermore, expression of IRF9 was essential to mediate the antiproliferative effects of IFNα2. We concluded that IL6 is an inducer of IRF9 expression in PCa and a sensitizer for the antiproliferative effects of IFNα2.

scholarly journals Rotor Interaction Noise in Counter-Rotating Propfan Propulsion Systems

2011 ◽  
Vol 134 (1) ◽  
Author(s):  
Andreas Peters ◽  
Zoltán S. Spakovszky
Keyword(s):  
Polar Angle ◽  
Secondary Flows ◽  
Tip Vortex ◽  
System Level ◽  
Analysis Tool ◽  
Acoustic Analogy ◽  
Wake Interaction ◽  
Open Rotor ◽  
And Performance ◽  
The Individual

Due to their inherent noise challenge and potential for significant reductions in fuel burn, counter-rotating propfans (CRPs) are currently being investigated as potential alternatives to high-bypass turbofan engines. This paper introduces an integrated noise and performance assessment methodology for advanced propfan powered aircraft configurations. The approach is based on first principles and combines a coupled aircraft and propulsion system mission and performance analysis tool with 3D unsteady, full-wheel CRP computational fluid dynamics computations and aeroacoustic simulations. Special emphasis is put on computing CRP noise due to interaction tones. The method is capable of dealing with parametric studies and exploring noise reduction technologies. An aircraft performance, weight and balance, and mission analysis was first conducted on a candidate CRP powered aircraft configuration. Guided by data available in the literature, a detailed aerodynamic design of a pusher CRP was carried out. Full-wheel unsteady 3D Reynolds-averaged Navier-Stokes (RANS) simulations were then used to determine the time varying blade surface pressures and unsteady flow features necessary to define the acoustic source terms. A frequency domain approach based on Goldstein’s formulation of the acoustic analogy for moving media and Hanson’s single rotor noise method was extended to counter-rotating configurations. The far field noise predictions were compared to measured data of a similar CRP configuration and demonstrated good agreement between the computed and measured interaction tones. The underlying noise mechanisms have previously been described in literature but, to the authors’ knowledge, this is the first time that the individual contributions of front-rotor wake interaction, aft-rotor upstream influence, hub-endwall secondary flows, and front-rotor tip-vortices to interaction tone noise are dissected and quantified. Based on this investigation, the CRP was redesigned for reduced noise incorporating a clipped rear-rotor and increased rotor-rotor spacing to reduce upstream influence, tip-vortex, and wake interaction effects. Maintaining the thrust and propulsive efficiency at takeoff conditions, the noise was calculated for both designs. At the interaction tone frequencies, the redesigned CRP demonstrated an average reduction of 7.25 dB in mean sound pressure level computed over the forward and aft polar angle arcs. On the engine/aircraft system level, the redesigned CRP demonstrated a reduction of 9.2 dB in effective perceived noise (EPNdB) and 8.6 EPNdB at the Federal Aviation Regulations (FAR) 36 flyover and sideline observer locations, respectively. The results suggest that advanced open rotor designs can possibly meet Stage 4 noise requirements.

Sign in / Sign up

Export Citation Format

Share Document