A B cell-driven EAE mouse model reveals the impact of B cell-derived cytokines on CNS autoimmunity.

In multiple sclerosis (MS), pathogenic T cell responses are known to be important drivers of autoimmune inflammation. However, increasing evidence suggests an additional role for B cells, which may contribute to pathogenesis via antigen presentation and production of proinflammatory cytokines. However, these B cell effector functions are not featured well in classical experimental autoimmune encephalomyelitis (EAE) mouse models. Here, we compared properties of myelin oligodendrocyte glycoprotein (MOG)-specific and polyclonal B cells and developed an adjuvant-free cotransfer EAE mouse model, where highly activated, MOG-specific induced germinal center B cells provide the critical stimulus for disease development. We could show that high levels of MOG-specific immunoglobulin G (IgGs) are not required for EAE development, suggesting that antigen presentation and activation of cognate T cells by B cells may be important for pathogenesis. As our model allows for B cell manipulation prior to transfer, we found that overexpression of the proinflammatory cytokine interleukin (IL)-6 by MOG-specific B cells leads to an accelerated EAE onset accompanied by activation/expansion of the myeloid compartment rather than a changed T cell response. Accordingly, knocking out IL-6 or tumor necrosis factor α in MOG-specific B cells via CRISPR-Cas9 did not affect activation of pathogenic T cells. In summary, we generated a tool to dissect pathogenic B cell effector function in EAE development, which should improve our understanding of pathogenic processes in MS.

Conversion of Anergic T Cells Into Foxp3- IL-10+ Regulatory T Cells by a Second Antigen Stimulus In Vivo.

T cell anergy is a common mechanism of T cell tolerance. However, although anergic T cells are retained for longer time periods in their hosts, they remain functionally passive. Here, we describe the induction of anergic CD4+ T cells in vivo by intravenous application of high doses of antigen and their subsequent conversion into suppressive Foxp3- IL-10+ Tr1 cells but not Foxp3+ Tregs. We describe the kinetics of up-regulation of several memory-, anergy- and suppression-related markers such as CD44, CD73, FR4, CD25, CD28, PD-1, Egr-2, Foxp3 and CTLA-4 in this process. The conversion into suppressive Tr1 cells correlates with the transient intracellular CTLA-4 expression and required the restimulation of anergic cells in a short-term time window. Restimulation after longer time periods, when CTLA-4 is down-regulated again retains the anergic state but does not lead to the induction of suppressor function. Our data require further functional investigations but at this stage may suggest a role for anergic T cells as a circulating pool of passive cells that may be re-activated into Tr1 cells upon short-term restimulation with high and systemic doses of antigen. It is tentative to speculate that such a scenario may represent cases of allergen responses in non-allergic individuals.

Multiple Sclerosis-Associated Changes in the Composition and Immune Functions of Spore-Forming Bacteria.

Multiple sclerosis (MS) is an autoimmune disease of the central nervous system characterized by adaptive and innate immune system dysregulation. Recent work has revealed moderate alteration of gut microbial communities in subjects with MS and in experimental, induced models. However, a mechanistic understanding linking the observed changes in the microbiota and the presence of the disease is still missing. Chloroform-resistant, spore-forming bacteria, which primarily belong to the classes Bacilli and Clostridia in the phylum Firmicutes, have been shown to exhibit immunomodulatory properties in vitro and in vivo, but they have not yet been characterized in the context of human disease. This study addresses the community composition and immune function of this bacterial fraction in MS. We identify MS-associated spore-forming taxa (primarily in the class Clostridia) and show that their presence correlates with impaired differentiation of IL-10-secreting, regulatory T lymphocytes in vitro. Colonization of antibiotic-treated mice with spore-forming bacteria allowed us to identify some bacterial taxa favoring IL-10+ lymphocyte differentiation and others inducing differentiation of proinflammatory, IFN-γ+ T lymphocytes. However, when fed into antibiotic-treated mice, both MS and control-derived spore-forming bacteria were able to induce similar IL-10-expressing Treg immunoregulatory responses, thus ameliorating symptoms of experimental allergic encephalomyelitis (EAE). Our analysis also identified Akkermansia muciniphila as a key organism that may interact either directly or indirectly with spore-forming bacteria to exacerbate the inflammatory effects of MS-associated gut microbiota. Thus, changes in the spore-forming fraction may influence T lymphocyte-mediated inflammation in MS. This experimental approach of isolating a subset of microbiota based on its functional characteristics may be useful to investigate other microbial fractions at greater depth. IMPORTANCE To address the impact of microbiome on disease development, it is essential to go beyond a descriptive study and evaluate the physiological importance of microbiome changes. Our study integrates computational analysis with in vitro and in vivo exploration of inflammatory properties of spore-forming microbial communities, revealing novel functional correlations. We specifically show that while small differences exist between the microbiomes of MS patients and healthy subjects, these differences are exacerbated in the chloroform-resistant fraction. We further demonstrate that, when purified from MS patients, this fraction is correlated with impaired immunomodulatory responses in vitro.

Aberrant microRNA expression in patients with painful peripheral neuropathies.

Changes in the neuro-immune balance play a major role in the induction and maintenance of neuropathic pain. We recently reported pathophysiologically relevant alterations in skin and sural nerve cytokine expression in peripheral neuropathies of different etiologies. Immune processes and cytokine expression are under tight control of microRNAs (miRNAs). To identify potential master switches in the neuro-immune balance, we aimed at characterizing inflammation-regulating miRNA profiles in patients with peripheral neuropathies. In an unselected patient cohort with polyneuropathies of different etiologies seen at our neuromuscular center between 2014 and 2015, we determined the systemic and local relative expression of miR-21-5p, miR-146a, and miR-155. In white blood cells we found higher miR-21 (p<0.001) and miR-146a (p<0.001) expression and lower miR-155 (p<0.001) expression when compared to healthy controls. In sural nerve, miR-21 (p<0.02) was increased in painful compared to painless neuropathies. In painful neuropathies, skin biopsies from the lower leg had reduced miR-146a (p<0.001) and miR-155 (p<0.001) expression compared to the thigh. Thus, peripheral neuropathies are associated with aberrant miRNA expression in white blood cells, sural nerve, and skin. These miRNA patterns may help to identify factors that determine the painfulness of peripheral neuropathies and lead to druggable targets.

[Treatment Options for Neurogenic Drop Foot: A Systematic Literature Research]. / Behandlungsoptionen beim neurogenen Lähmungsfuß ­ eine systematische Literaturrecherche.

Background Neurogenic drop foot may be caused by central or peripheral lesions of the nervous system. Depending on whether the first or second motor neuron is damaged, a flaccid or spastic drop foot develops. Spastic drop foot persists as a residual long-term complication after stroke in about 14 % of patients. Various conservative and surgical treatment options are available. Methods This article is based on a systematic literature review for medical evidence of functional electrical stimulation (FES) with the keywords "functional electrical stimulation AND drop foot" and "functional electrical stimulation AND gait AND stroke" in PubMed and Cochrane databases. Randomised controlled trials and cohort studies of the past 10 years were selected according to specific criteria. Additionally, four guidelines were included. Results Current guidelines provide little guidance for practical treatment and fail to give due consideration to new surgical procedures such as neural implants. In 18 randomised controlled trials on FES in stroke patients with drop foot, the FES-treated population showed either significant superiority or non-inferiority versus control. Two cohort studies confirmed significant improvements by FES. Conclusion The analysis demonstrates the importance of considering causes and severity of drop foot as well as patients' pre- and post-operative conditions for choosing treatment options. For active, high-demand patients, neural implants are valuable treatment options.

Does a foot-drop implant improve kinetic and kinematic parameters in the foot and ankle?

INTRODUCTION: Unlike the drop foot therapy with ortheses, the therapeutic effect of an implantable peroneus nerve stimulator (iPNS) is not well described. IPNS is a dynamic therapy option which is placed directly to the motoric part of the peroneal nerve and evokes a dorsiflexion of the paralysed foot. This retrospective study evaluates the kinematics and kinetics in drop foot patients who were treated with an iPNS. MATERIALS AND METHODS: 18 subjects (mean age 51.3 years) with a chronic stroke-related drop foot were treated with an implantable peroneal nerve stimulator. After a mean follow-up from 12.5 months, kinematics and kinetics as well as spatiotemporal parameters were evaluated and compared in activated and deactivated iPNS. Therefore, a gait analysis with motion capture system (Vicon Motion System Ltd®, Oxford, UK) and Plug-in-Gait model was performed. RESULTS: The study showed significantly improved results in ankle dorsiflexion from 6.8° to 1.8° at the initial contact and from -7.3° to 0.9° during swing phase (p ≤ 0.004 and p ≤ 0.005, respectively). Likewise, we could measure improved kinetics, i.a. with a statistically significant improvement in vertical ground reaction force at loading response from 99.76 to 106.71 N/kg (p = 0.043). Enhanced spatiotemporal results in cadence, douple support, stride length, and walking speed could also be achieved, but without statistical significance (p > 0.05). CONCLUSIONS: The results show statistically significant improvement in ankle dorsiflexion and vertical ground reaction forces. These facts indicate a more gait stability and gait efficacy. Therefore, the use of an iPNS appears an encouraging therapeutic option for patients with a stroke-related drop foot.

Hip and knee effects after implantation of a drop foot stimulator.

BACKGROUND: An active ankle dorsiflexion is essential for a proper gait pattern. If there is a failure of the foot lifting, considerable impairments occur. The therapeutic effect of an implantable peroneus nerve stimulator (iPNS) for the ankle dorsiflexion is already approved by recent studies. However, possible affection for knee and hip motion after implantation of an iPNS is not well described. OBJECTIVE: The objective of this retrospective study was to examine with a patient cohort whether the use of iPNS induces a lower-extremity flexion withdrawal response in the form of an increased knee and hip flexion during swing phase. METHODS: Eighteen subjects (12 m/6 w) treated with an iPNS (ActiGait®, Otto Bock, Duderstadt, Germany) were examined in knee and hip motion by gait analysis with motion capture system (Vicon Motion System Ltd®, Oxford, UK) and Plug-in-Gait model after a mean follow up from 12.5 months. The data were evaluated and compared in activated and deactivated iPNS. RESULTS: Only little changes could be documented, as a slight average improvement in peak knee flexion during stand phase from 1.0° to 2.5° and peak hip flexion in stance from 3.1° to 2.1° In contrast, peak knee flexion during swing appeared similar (25.3° to 25.7°) same as peak hip flexion during swing. In comparison with the healthy extremity, a more symmetric course of the knee flexion during stand phase could be shown. CONCLUSIONS: No statistical significant improvements or changes in hip and knee joint could be shown in this study. Only a more symmetric knee flexion during stand phase and a less hip flexion during stand phase might be hints for a positive affection of iPNS for knee and hip joint. It seems that the positive effect of iPNS is only based on the improvement in ankle dorsiflexion according to the recent literature.