Abstract Objective To assess whether reshaping of the immune balance by infusion of autologous natural regulatory T cells (nTregs) in patients after kidney transplantation is safe, feasible, and enables the tapering of lifelong high dose immunosuppression, with its limited efficacy, adverse effects, and high direct and indirect costs, along with addressing several key challenges of nTreg treatment, such as easy and robust manufacturing, danger of over immunosuppression, interaction with standard care drugs, and functional stability in an inflammatory environment in a useful proof-of-concept disease model. Design Investigator initiated, monocentre, nTreg dose escalation, phase I/IIa clinical trial (ONEnTreg13). Setting Charité-University Hospital, Berlin, Germany, within the ONE study consortium (funded by the European Union). Participants Recipients of living donor kidney transplant (ONEnTreg13, n=11) and corresponding reference group trial (ONErgt11-CHA, n=9). Interventions CD4+ CD25+ FoxP3+ nTreg products were given seven days after kidney transplantation as one intravenous dose of 0.5, 1.0, or 2.5-3.0×10 6 cells/kg body weight, with subsequent stepwise tapering of triple immunosuppression to low dose tacrolimus monotherapy until week 48. Main outcome measures The primary clinical and safety endpoints were assessed by a composite endpoint at week 60 with further three year follow-up. The assessment included incidence of biopsy confirmed acute rejection, assessment of nTreg infusion related adverse effects, and signs of over immunosuppression. Secondary endpoints addressed allograft functions. Accompanying research included a comprehensive exploratory biomarker portfolio. Results For all patients, nTreg products with sufficient yield, purity, and functionality could be generated from 40-50 mL of peripheral blood taken two weeks before kidney transplantation. None of the three nTreg dose escalation groups had dose limiting toxicity. The nTreg and reference groups had 100% three year allograft survival and similar clinical and safety profiles. Stable monotherapy immunosuppression was achieved in eight of 11 (73%) patients receiving nTregs, while the reference group remained on standard dual or triple drug immunosuppression (P=0.002). Mechanistically, the activation of conventional T cells was reduced and nTregs shifted in vivo from a polyclonal to an oligoclonal T cell receptor repertoire. Conclusions The application of autologous nTregs was safe and feasible even in patients who had a kidney transplant and were immunosuppressed. These results warrant further evaluation of Treg efficacy and serve as the basis for the development of next generation nTreg approaches in transplantation and any immunopathologies. Trial registration NCT02371434 (ONEnTreg13) and EudraCT:2011-004301-24 (ONErgt11).
Blood pressure has been traditionally measured at peripheral arteries. In the past decade, evidence has grown that central aortic blood pressure may be a more powerful predictor for cardiovascular events, but data on its regulation are rare. The present work examines the impact of microgravity on central blood pressure for the first time.We performed 7 parabolic flights with 22 seconds of weightlessness in each parabola. Hemodynamic parameters including central systolic blood pressure were measured noninvasively in a free-floating position in 20 healthy subjects (19-43 years of age).Arterial elasticity at rest was normal in all participants (augmentation index 14% (interquartile range (IQR) 10-22), pulse wave velocity 5.2 m/s (IQR 5.0-5.4)). Transition of 1g to 0g led to a significant increase of central systolic blood pressure from 124 (IQR 118-133) to 127 (IQR 119-133) mm Hg (P = 0.017). Cardiac index augmented significantly from 2.5 (IQR 2.2-2.8) to 2.7 (IQR 2.3-3.0) l/min/m2 (P < 0.001), while peripheral vascular resistance showed a decrease from 1.30 (IQR 1.14-1.48) to 1.25 (IQR 1.15-1.40) s × mm Hg/ml (P = 0.037). Peripheral systolic blood pressure did not change significantly (P > 0.05).Whereas there is a multitude of studies on the effects of microgravity on peripheral blood pressure, this study provides first data on central aortic blood pressure. An acute loss of gravity leads to a central blood volume shift with an augmentation of cardiac output. In healthy subjects with normal arterial stiffness, the compensatory decrease of peripheral resistance does not outweigh this effect resulting in an increase of central blood pressure.
Abstract The role of autoimmunity in post-acute sequelae of COVID-19 (PASC) is not well explored, although clinicians observe a growing population of convalescent COVID-19 patients with manifestation of post-acute sequelae of COVID-19. We analyzed the immune response in 40 post-acute sequelae of COVID-19 patients with non-specific PASC manifestation and 15 COVID-19 convalescent healthy donors. The phenotyping of lymphocytes showed a significantly higher number of CD8+ T cells expressing the Epstein-Barr virus induced G protein coupled receptor 2, chemokine receptor CXCR3 and C-C chemokine receptor type 5 playing an important role in inflammation and migration in PASC patients compared to controls. Additionally, a stronger, SARS-CoV-2 reactive CD8+ T cell response, characterized by IFNγ production and predominant T EMRA phenotype but low SARS-CoV-2 avidity was detected in PASC patients compared to controls. Furthermore, higher titers of several autoantibodies were detected among PASC patients. Our data suggest that a persistent inflammatory response triggered by SARS-CoV-2 might be responsible for the observed sequelae in PASC patients. These results may have implications on future therapeutic strategies.
Systemic lupus erythematosus is a systemic and chronic autoimmune disease characterized by loss of tolerance towards nuclear antigens with autoreactive CD4+ T cells implicated in disease pathogenesis. However, very little is known about their receptor specificity since the detection of human autoantigen specific CD4+ T cells has been extremely challenging. Here we present an analysis of CD4+ T cells reactive to nuclear antigens using two complementary methods: T cell libraries and antigen-reactive T cell enrichment. The frequencies of nuclear antigen specific CD4+ T cells correlated with disease severity. These autoreactive T cells produce effector cytokines such as interferon-γ, interleukin-17, and interleukin-10. Compared to blood, these cells were enriched in the urine of patients with active lupus nephritis, suggesting an infiltration of the inflamed kidneys. Thus, these previously unrecognized characteristics support a role for nuclear antigen-specific CD4+ T cells in systemic lupus erythematosus.
Adipositas geht mit einer als „Metaflammation“ bezeichneten chronisch-subklinischen Entzündung einher, die als unabhängiger Prädiktor für kardiovaskuläre Ereignisse, Diabetes mellitus Typ 2 und chronische Niereninsuffizienz (CKD) identifiziert wurde. Dabei kommt es auf Basis proinflammatorischer Signale, u. a. durch Adipozytenstress, zur Einwanderung von Immunzellen in das Fettgewebe. Zytotoxische T‑Zellen rekrutieren Makrophagen, die ihrerseits die Entzündung antreiben, sodass ein Circulus vitiosus aus sich verstärkenden Signalen mit einem proinflammatorischen Shift der Immunzellpopulation entsteht. Molekulare Endstrecke dessen ist eine zytokinvermittelte Insulinresistenz, die nicht nur den Grundstein für die Entwicklung eines metabolischen Syndroms legt, sondern die durch ihre sekundäre Hyperinsulinämie auch in der Niere Apoptose, Fibrose und endotheliale Dysfunktion triggert. Dieser Prozess wird durch entzündungsfördernde Mediatoren des Fettgewebes (Adipokine) zusätzlich gefördert. Einige immunsupprimierende Medikamente zeigen im Tierversuch erste positive Effekte auf adipositasassoziierte Folgeerkrankungen, sind bis dato aber für diese Indikation klinisch nicht etabliert.
Resveratrol, a phytochemical present in grapes, has been demonstrated to inhibit tumourigenesis in animal models. However, the specific mechanism by which resveratrol exerts its anticarcinogenic effect has yet to be elucidated. In the present study, the inhibitory effects of resveratrol on cell proliferation and apoptosis were evaluated in the human leukaemia cell line HL-60 and the human hepatoma derived cell line HepG2. We found that after a 2 h incubation period, resveratrol inhibited DNA synthesis in a concentration-dependent manner. The IC50 value was 15 microm in both HL-60 and HepG2 cells. When the time of treatment was extended, an increase in IC50 value was observed; for example, at 24 h the IC50 value was 30 microm for HL-60 cells and 60 microm for HepG2 cells. Flow cytometry revealed that cells accumulated in different phases of the cell cycle depending on the resveratrol concentration. Furthermore, an increase in nuclear size and granularity was observed in the G1 and S phases of HL-60 treated and HepG2-treated cells. Apoptosis was also stimulated by resveratrol in a concentration-dependent manner in HL-60 and HepG2 cells. In conclusion, resveratrol inhibits cell proliferation in a concentration- and time-dependent manner by interfering with different stages of the cell cycle. Furthermore, resveratrol treatment causes stimulation of apoptosis as well as an increase in nuclear size and granularity.
Whole genome sequencing (WGS) is increasingly used to aid in understanding pathogen transmission [1]. Very often the number of single nucleotide polymorphisms (SNPs) separating isolates collected during an epidemiological study are used to identify sets of cases that are potentially linked by direct transmission. However, there is little agreement in the literature as to what an appropriate SNP cut-off threshold should be, or indeed whether a simple SNP threshold is appropriate for identifying sets of isolates to be treated as "transmission clusters". The SNP thresholds that have been adopted for inferring transmission vary widely even for one pathogen. As an alternative to reliance on a strict SNP threshold, we suggest that the key inferential target when studying the spread of an infectious disease is the number of transmission events separating cases. Here we describe a new framework for deciding whether two pathogen genomes should be considered as part of the same transmission cluster, based jointly on the number of SNP differences and the length of time over which those differences have accumulated. Our approach allows us to probabilistically characterize the number of inferred transmission events that separate cases. We show how this framework can be modified to consider variable mutation rates across the genome (e.g. SNPs associated with drug resistance) and we indicate how the methodology can be extended to incorporate epidemiological data such as spatial proximity. We use recent data collected from tuberculosis studies from British Columbia, Canada and the Republic of Moldova to apply and compare our clustering method to the SNP threshold approach. In the British Columbia data, different cases break off from the main clusters as cut-off thresholds are lowered; the transmission-based method obtains slightly different clusters than the SNP cut-offs. For the Moldova data, straightforward application of the methods shows no appreciable difference, but when we take into account the fact that resistance conferring sites likely do not follow the same mutation clock as most sites due to selection, the transmission-based approach differs from the SNP cut-off method. Outbreak simulations confirm that our transmission based method is at least as good at identifying direct transmissions as a SNP cut-off. We conclude that the new method is a promising step towards establishing a more robust identification of outbreaks.
