Low-dose interleukin-2 reverses chronic migraine-related sensitizations through peripheral interleukin-10 and transforming growth factor beta-1 signaling

Low-dose interleukin-2 (LD-IL-2) treatment has been shown to effectively reverse chronic migraine-related behaviors and the sensitization of trigeminal ganglion (TG) neurons through expansion and activation of peripheral regulatory T cells (Tregs) in mice. In this study, we investigated the molecular mechanisms underlying the effects of LD-IL-2 and Treg cells. LD-IL-2 treatment increases the production of cytokines interleukin-10 (IL-10) and transforming growth factor beta-1 (TGFβ1) in T cells, especially Treg cells, suggesting that they may mediate the therapeutic effect of LD-IL-2. Indeed, neutralizing antibodies against either IL-10 or TGFβ completely blocked the effects of LD-IL-2 on the facial mechanical hypersensitivity as well as the sensitization of TG neurons resulting from repeated nitroglycerin (NTG, a reliable trigger of migraine in patients) administration in mice, indicating that LD-IL-2 and Treg cells engage both peripheral IL-10 and TGFβ signaling pathways to reverse chronic-migraine related sensitizations. In a

Interleukin-7 restores lymphocytes in septic shock: The IRIS-7 randomized clinical trial

BACKGROUND: A defining pathophysiologic feature of sepsis is profound apoptosis-induced death and depletion of CD4+ and CD8+ T cells. Interleukin-7 (IL-7) is an antiapoptotic common γ-chain cytokine that is essential for lymphocyte proliferation and survival. Clinical trials of IL-7 in over 390 oncologic and lymphopenic patients showed that IL-7 was safe, invariably increased CD4+ and CD8+ lymphocyte counts, and improved immunity. METHODS: We conducted a prospective, randomized, double-blind, placebo-controlled trial of recombinant human IL-7 (CYT107) in patients with septic shock and severe lymphopenia. Twenty-seven patients at academic sites in France and the United States received CYT107 or placebo for 4 weeks. Primary aims were to determine the safety of CYT107 in sepsis and its ability to reverse lymphopenia. RESULTS: CYT107 was well tolerated without evidence of inducing cytokine storm or worsening inflammation or organ dysfunction. CYT107 caused a 3- to 4-fold increase in absolute lymphocyte counts and in circulating CD4+ and CD8+ T cells that persisted for weeks after drug administration. CYT107 also increased T cell proliferation and activation. CONCLUSIONS: This is the first trial of an immunoadjuvant therapy targeting defects in adaptive immunity in patients with sepsis. CYT107 reversed the marked loss of CD4+ and CD8+ immune effector cells, a hallmark of sepsis and a likely key mechanism in its morbidity and mortality. CYT107 represents a potential new way forward in the treatment of patients with sepsis by restoring adaptive immunity. Such immune-based therapy should be broadly protective against diverse pathogens including multidrug resistant bacteria that preferentially target patients with impaired immunity. TRIAL REGISTRATION: Trials registered at clinicaltrials.gov: NCT02640807 and NCT02797431. FUNDING: Revimmune, NIH National Institute of General Medical Sciences GM44118

Blockade ofthe negative co-stimulatory molecules PD-1 and CTLA-4 improves survival in primary and secondary fungal sepsis

INTRODUCTION: Fungal sepsis is an increasingly common problem in intensive care unit patients.Mortality from fungal sepsis remains high despite antimicrobial therapy that is highly active against most fungal pathogens, a finding consistent with defective host immunity that is present in many patients with disseminated fungemia.One recently recognized immunologic defect that occurs in patients with sepsis is T cell "exhaustion" due to increased expression of programmed cell death -1 (PD-1).This study tested the ability of anti-PD-1 and anti-programmed cell death ligand -1 (anti-PD-L1) antagonistic antibodies to improve survival and reverse sepsis-induced immunosuppression in two mouse models of fungal sepsis. METHODS: Fungal sepsis was induced in mice using two different models of infection, that is, primary fungal sepsis and secondary fungal sepsis occurring after sub-lethal cecal ligation and puncture (CLP).Anti-PD-1 and anti-PD-L1 were administered 24 to 48 h after fungal infection and effects on survival, interferon gamma production, and MHC II expression were examined. RESULTS: Anti-PD-1 and anti-PD-L1 antibodies were highly effective at improving survival in primary and secondary fungal sepsis.Both antibodies reversed sepsis-induced suppression of interferon gamma and increased expression of MHC II on antigen presenting cells.Blockade of cytotoxic T-lymphocyte antigen-4 (CTLA-4), a second negative co-stimulatory molecule that is up-regulated in sepsis and acts like PD-1 to suppress T cell function, also improved survival in fungal sepsis. CONCLUSIONS: Immuno-adjuvant therapy with anti-PD-1, anti-PD-L1 and anti-CTLA-4 antibodies reverse sepsis-induced immunosuppression and improve survival in fungal sepsis.The present results are consistent with previous studies showing that blockade of PD-1 and CTLA-4 improves survival in bacterial sepsis.Thus, immuno-adjuvant therapy represents a novel approach to sepsis and may have broad applicability in the disorder.Given the relative safety of anti-PD-1 antibody in cancer clinical trials to date, therapy with anti-PD-1 in patients with life-threatening sepsis who have demonstrable immunosuppression should be strongly considered

Entwicklung regulierbarer retro- und adenoviraler Vektoren für die Gentherapie

Die Möglichkeit, die Expression von Genen zu regulieren, ist sowohl in der Forschung als auch in der Gentherapie ein wünschenswertes Ziel. Jedoch unterliegt die Regulation der Limitation, dass alle regulierbaren Expressionssysteme aus zwei Komponenten bestehen, dem Transaktivatorprotein (tTA) und dem tTA-abhängigen Promotor, die in die Zielzellen transferiert und adequat exprimiert werden müssen. In der vorliegenden Arbeit wurden verschiedene autoregulative Expressionskassetten basierend auf dem tet-regulierten System entwickelt. Diese Kassetten wurden so konstruiert, dass sie alle Regulationskomponenten beinhalten und so in einem Schritt transferiert werden können (Single- Step-Systeme). Durch die autoregulative Anordnung des tTA's bleibt seine Expression auf die Phase der Geninduktion beschränkt. Toxische Nebenwirkungen werden so minimiert. Diese Kassetten wurden über Elektrotransformation sowie über retro- und adenovirale Infektion in die Zielzellen eingebracht und das jeweilige Expressions- und Regulationspotential ermittelt. Im Gegensatz zur unidirektionalen Expressionseinheit, boten die autoregulativen bidirektionalen Expressionssysteme nach jedem für die Gentherapie relevanten Transfer hohe und stabile Regulationskapazität des interessierenden Gens. Durch die Konstruktion chimärer retro-/adenoviraler Vektoren wurden Viren generiert, deren Charakteristika eine Kombination der positiven retro- und adenoviralen Eigenschaften darstellen. So war es möglich die effiziente Transduktion der Adenoviren mit der stabilen Intergration der Retroviren zu kombinieren. Dieses chimäre System könnte sich daher zu einem relevanten System in der Gentherapie entwickeln.Regulated transgene expression is increasingly used in research but is also a need for certain therapies. Regulatory systems are usually composed of two expression units, one bearing the gene of interest under the control of a regulatable promotor, the second a constitutively expressed transactivator (tTA) which modulates activity of the regulatable promotor. Since the cotransfer of two independent elements is not efficient in primary cells single transduction step vectors conferring regulatable gene expression cassettes would be of advantage. In the present work different autoregulatable expression cassettes based on the tet-regulatable system were designed. These expression cassettes encode all components necessary for regulated gene expression and can therefore be transferred in a single transduction step (single- step systems). Due to the autoregulative arrangement of the tTA, toxic side effects were limited to the time of the induced gene expression. These cassettes were transferred to the target cells both via electrotransformation and retro- and adenoviral infection. The expression- and regulation potential was determined. In contrast to the unidirectional expression unit, the bidirectional expression system offered high and stable regulation capacity of the gene of interest independent of the transfer system used. The construction of chimeric retro-/adenoviral vectors generated viruses, that possess the positive characteristics of both virus types, the high transduction efficiency of adenoviruses and the stable integration of retroviruses. This chimeric system might evolve to a relevant transfer system in gene therepy applications