Recruitment of Regulatory T Cells with rCCL17 Promotes M2 Microglia/Macrophage Polarization Through TGFβ/TGFβR/Smad2/3 Pathway in a Mouse Model of Intracerebral Hemorrhage

Background Intracerebral hemorrhage (ICH) is a devastating neurological disease with high mortality and morbidity. The microglia activation and peripheral inflammatory cells infiltration play an important role in the ICH prognosis. Previous studies have demonstrated that regulatory T cells (Tregs) ameliorated neuroinflammation following experimental ICH. However, the specific molecular mechanism underlying such effects of Tregs remains unclear. In the present study, our aims were to examine the effect of Tregs recruitment induced by recombinant CC chemokine ligand 17(rCCL17) in an intrastriatal autologous blood mouse model of ICH and to determine whether the TGF-β/TGF-βR/Smad2/3 pathway was involved in Tregs promoted M2 microglia/macrophage polarization. Methods A total of 404 adult CD1 mice (male, eight-week-old) were subject to sham surgery or autologous blood injection of ICH. A CD25-specific mouse antibody or isotype control mAb was injected intraperitoneally 48h prior to ICH induction to deplete Tregs. Recombinant CCL17 (rCCL17), a C-C chemokine receptor 4 (CCR4), was delivered intranasally at 1 h post-ICH. SB431542, a specific inhibitor of TGF-β was administered intraperitoneally 1 h before ICH induction. Post-ICH assessments included neuro-behavior evaluation, brain edema, hematoma volume, hemoglobin content, western blotting, double immunofluorescence staining and immunohistochemistry. Results Endogenous brain expressions of CCL17 and Tregs marker Foxp3 as well as the number of Tregs in the perihematomal region were increased following ICH. The Tregs deletion by a CD25 antibody aggravated short-term neurological deficits and brain edema, increased the level of inflammatory cytokines and peripheral inflammatory cells infiltration, exacerbated hematoma expansion and increased M1phenotypes of microglia/macrophage in ICH mice. The rCCL17 treatment increased the number of Tregs in the brain, reduced hematoma expansion and brain edema, promoted microglia/macrophage polarization toward M2 phenotypes. Moreover, the expressions of brain TGF-β/phosphorylated-Smad2/3 were increased. The neuroprotective effects of rCCL17 were abolished by co-administration of the selective TGF-β inhibitor SB431542. Conclusions Our study demonstrated rCCL17 recruited of Tregs to brain in the autologous blood injection model of ICH. Tregs promoted microglia/macrophages polarization toward M2 phenotype and alleviation early brain injury, at least in part, through the TGFβ/TGFβR/Smad2/3 signaling pathway in ICH mice. Thus, rCCL17-mediated Tregs recruitment may provide a promising therapeutic strategy to reduce early brain injury after ICH.

Early Results of a Novel Treatment Technique with Autologous Blood for Chronic Lateral Epicondylitis

Background: Chronic lateral epicondylitis can be a severe disabling condition. There is still lack of consensus on best treatment, as no single intervention has been proven to be superior regarding pain relief and improvement of function. Due to the self-limiting nature of this elbow condition, we are looking for a fast and safe treatment method to break through this pattern of pain and loss of elbow function. Autologous blood injection therapy by means of an automatic injection system, can be a promising new treatment option for this group of patients suffering from chronic lateral epicondylitis. In this study, we evaluated the short-term results of autologous blood injection therapy in a standardized way by using an automatic injection system (=ITEC device) for the treatment of chronic lateral epicondylitis. Methods: A total of 141 patients with chronic lateral epicondylitis (88 female, 53 male) were enrolled in this clinical treatment evaluation being treated with the ITEC device. The mean age of the patients was 50.0 years (19 years-73 years). Numeric rating scale (NRS) and a patient reported outcome measurement tool (Oxford Elbow Score (OES)) were measured at baseline, six weeks and three months follow-up. Results: Pain (NRS, OES) and elbow function / quality of life (OES) were significantly improved within 6 weeks after ITEC treatment. This improvement in NRS and OES sustained during the 3 months follow-up period. Conclusion: Autologous blood injection therapy by means of a new automatic injection system (ITEC device) is a safe and effective treatment method for patients with chronic lateral epicondylitis. More research is necessary to see if this effectiveness sustains in the long-term follow-up.

AUTOLOGOUS BLOOD INJECTION INTRACORONARY ARTERY FOR TREATING SLOW-FLOW AND NO-REFLOW IN ACUTE CORONARY SYNDROME RELATED TO PRIMARY PCI

Slow flow and no-reflow phenomenon are taken to sudden loss of coronary artery flow, typically after stenting or angioplasty in primary PCI. Otherwise conventional therapy, we report a technique, which autologous blood into intracoronary to supply oxygen and break process thrombosis results in successfully management no-reflow in primary PCI

Divergent Functions of Tissue-Resident and Blood-Derived Macrophages in the Hemorrhagic Brain

Background and Purpose: Brain tissue-resident microglia and monocyte-derived macrophages (MDMs) are innate immune cells that contribute to the inflammatory response, phagocytosis of debris, and tissue repair after injury. We have previously reported that both microglia and MDMs transition from proinflammatory to reparative phenotypes over days after an intracerebral hemorrhage (ICH). However, their individual functional properties in the brain remain largely unknown. Here we characterized the differences between microglia and MDMs and further elucidate their distinct activation states and functional contributions to the pathophysiology and recovery after ICH. Methods: Autologous blood injection was used to model ICH in mice. Longitudinal transcriptomic analyses on isolated microglia and MDMs from mice at days 1, 3, 7 and 10 after ICH and naive controls identified core transcriptional programs that distinguish these cells. Imaging flow cytometry and in vivo phagocytosis assays were used to study phagocytic ability of microglia and MDMs. Antigen presentation was evaluated by ovalbumin-OTII CD4 T-cell proliferation assays with bone marrow–derived macrophages and primary microglia cultures. Results: MDMs had higher phagocytic activity and higher erythrophagocytosis in the ICH brain. Differential gene expression revealed distinct transcriptional signatures in the MDMs and microglia after ICH. MDMs had higher expression of MHCII (major histocompatibility complex class II) genes than microglia at all time points and greater ability to induce antigen-specific T-cell proliferation. Conclusions: The different ontogeny of microglia and MDMs lead to divergent responses and functions in the inflamed brain as these 2 cell populations differ in phagocytic functions and antigen-presenting capabilities in the brain after ICH.

Recombinant CCL17-dependent CCR4 activation alleviates neuroinflammation and neuronal apoptosis through the PI3K/AKT/Foxo1 signaling pathway after ICH in mice

Background Intracerebral hemorrhage (ICH), a devastating subtype of stroke, is associated with high mortality and morbidity. Neuroinflammation is an important factor leading to ICH-induced neurological injuries. C-C Chemokine Receptor 4 (CCR4) plays an important role in enhancing hematoma clearance after ICH. However, it is unclear whether CCR4 activation can ameliorate neuroinflammation and apoptosis of neurons following ICH. The aim of the present study was to examine the effects of recombinant CCL17 (rCCL17)-dependent CCR4 activation on neuroinflammation and neuronal apoptosis in an intrastriatal autologous blood injection ICH model, and to determine whether the PI3K/AKT/Foxo1 signaling pathway was involved. Methods Two hundred twenty-six adult (8-week-old) male CD1 mice were randomly assigned to sham and ICH surgery groups. An intrastriatal autologous blood injection ICH model was used. rCCL17, a CCR4 ligand, was delivered by intranasal administration at 1 h, 3 h, and 6 h post-ICH. CCL17 antibody was administrated by intraventricular injection at 1 h post-ICH. C021, a specific inhibitor of CCR4 and GDC0068, an AKT inhibitor were delivered intraperitoneally 1 h prior to ICH induction. Brain edema, neurobehavioral assessments, western blotting, Fluoro-Jade C staining, terminal deoxynucleotidyl transferase dUTP nick end labeling, and immunofluorescence staining were conducted. Results Endogenous expression of CCL17 and CCR4 were increased following ICH, peaking at 5 days post-induction. CCR4 was found to co-localize with microglia, neurons, and astrocytes. rCCL17 treatment decreased brain water content, attenuated short- and long-term neurological deficits, deceased activation of microglia/macrophages and infiltration of neutrophils, and inhibited neuronal apoptosis in the perihematomal region post-ICH. Moreover, rCCL17 treatment post-ICH significantly increased the expression of CCR4, PI3K, phosphorylated AKT, and Bcl-2, while Foxo1, IL-1β, TNF-α, and Bax expression were decreased. The neuroprotective effects of rCCL17 were reversed with the administration of C021 or GDC0068. Conclusions rCCL17-dependent CCR4 activation ameliorated neurological deficits, reduced brain edema, and ameliorated neuroinflammation and neuronal apoptosis, at least in part, through the PI3K/AKT/Foxo1 signaling pathway after ICH. Thus, activation of CCR4 may provide a promising therapeutic approach for the early management of ICH.

EFFICACY OF ULTRASOUND-GUIDED AUTOLOGOUS BLOOD INJECTION VERSUS EXTRACORPOREAL SHOCKWAVE THERAPY IN CHRONIC ACHILLES TENDINOPATHY: A RANDOMIZED CONTROLLED TRIAL

Objective:To evaluate the efcacy of ultrasound-guided autologous blood injection in chronic Achilles tendinopathy. Design: Prospective randomised controlled trial. Setting:Tertiary care hospital. Participants: 45 patients of chronic Achilles tendinopathy. Intervention: Participants were allocated to 2 groups i.e. autologous blood injection under ultrasound-guidance group (n=23) and extra corporeal shockwave therapy group (n=22) while in both groups eccentric heel drop stretching exercise were added. Outcome measures:VAS, VISA-Aand ultrasonography. Follow up at 4, 12 and 24 weeks. Results: The mean VAS score changes within-subject analysis were signicant in both ABI and ESWT group (p<0.001), however between-group analysis at 4, 12 and 24 weeks were not signicant (p>0.05). The mean VISA-A score within-subject analysis were signicant in both the groups (p<0.001). The between-group analysis show signicant difference at 12 weeks (p=0.006) and 24 weeks (p=0.001). The between-group analysis of mean tendon size were signicant at 24 weeks (p=0.001). Conclusions: This study showed that ultrasound guided autologous blood injection is effective in reducing pain and disability in chronic Achilles tendinopathy at 6 months. ABI might have additive regenerative therapeutic effect.