ABSTRACT
BACKGROUND: Human adipose stromal cells-derived extracellular vesicles (haMSC-EVs) have been shown to alleviate inflammation in acute lung injury (ALI) animal models. However, there are few systemic studies on clinical-grade haMSC-EVs. Our study aimed to investigate the manufacturing, quality control (QC) and preclinical safety of clinical-grade haMSC-EVs. METHODS: haMSC-EVs were isolated from the conditioned medium of human adipose MSCs incubated in 2D containers. Purification was performed by PEG precipitation and differential centrifugation. Characterizations were conducted by nanoparticle tracking analysis, transmission electron microscopy (TEM), Western blotting, nanoflow cytometry analysis, and the TNF-α inhibition ratio of macrophage [after stimulated by lipopolysaccharide (LPS)]. RNA-seq and proteomic analysis with liquid chromatography tandem mass spectrometry (LC-MS/MS) were used to inspect the lot-to-lot consistency of the EV products. Repeated toxicity was evaluated in rats after administration using trace liquid endotracheal nebulizers for 28 days, and respiratory toxicity was evaluated 24 h after the first administration. In vivo therapeutic effects were assessed in an LPS-induced ALI/ acute respiratory distress syndrome (ARDS) rat model. RESULTS: The quality criteria have been standardized. In a stability study, haMSC-EVs were found to remain stable after 6 months of storage at - 80°C, 3 months at - 20 °C, and 6 h at room temperature. The microRNA profile and proteome of haMSC-EVs demonstrated suitable lot-to-lot consistency, further suggesting the stability of the production processes. Intratracheally administered 1.5 × 108 particles/rat/day for four weeks elicited no significant toxicity in rats. In LPS-induced ALI/ARDS model rats, intratracheally administered haMSC-EVs alleviated lung injury, possibly by reducing the serum level of inflammatory factors. CONCLUSION: haMSC-EVs, as an off-shelf drug, have suitable stability and lot-to-lot consistency. Intratracheally administered haMSC-EVs demonstrated excellent safety at the tested dosages in systematic preclinical toxicity studies. Intratracheally administered haMSC-EVs improved the lung function and exerted anti-inflammatory effects on LPS-induced ALI/ARDS model rats.
Subject(s)
Acute Lung Injury , Extracellular Vesicles , Mesenchymal Stem Cells , Respiratory Distress Syndrome , Humans , Rats , Animals , Chromatography, Liquid , Proteomics , Lipopolysaccharides/pharmacology , Tandem Mass Spectrometry , Acute Lung Injury/therapy , Respiratory Distress Syndrome/therapy , Obesity , Quality Control , Extracellular Vesicles/physiology , Mesenchymal Stem Cells/physiologyABSTRACT
PURPOSE: To assess diagnostic performance and reproducibility of reduced bowel wall enhancement evaluated by quantitative methods using CT to identify bowel necrosis among closed-loop small bowel obstruction (CL-SBO) patients. METHODS: This retrospective single-center study included patients who diagnosed with CL-SBO caused by adhesion or internal hernia during January 2016 and May 2022. Patients were divided into necrotic group (n = 41) and non-necrotic group (n = 67) according to surgical exploration and postoperative pathology. Two doctors independently measured the attenuation of bowel wall and consensus was reached through panel discussion with a third gastrointestinal radiologist. Reduced bowel wall enhancement was assessed by four quantitative methods. Univariate analyses were used to evaluate the association between each method and bowel necrosis, and kappa/intraclass correlation coefficient values were used to assess interobserver agreement. Diagnostic performance parameters were calculated for each method. RESULTS: Reduced bowel wall enhancement in arterial phase (OR 8.98, P < 0.0001), reduced bowel wall enhancement in portal phase (OR 16.84, P < 0.001), adjusted reduced bowel wall enhancement in arterial phase (OR 29.48, P < 0.001), adjusted reduced bowel wall enhancement in portal phase (OR 145.69, P < 0.001) were significantly associated with bowel necrosis. Adjusted reduced bowel wall enhancement in portal phase had the best diagnostic performance (AUC: 0.92; Youden index: 0.84; specificity: 94.03 %) and interobserver agreement (kappa value of 0.59-0.73) to predict bowel necrosis. CONCLUSION: When assessing reduced bowel enhancement to predict bowel necrosis among CL-SBO patients, using unenhanced CT images and proximal dilated loop as standard references in portal phase is the most accurate quantitative method among those tested.
Subject(s)
Abdominal Injuries , Intestinal Obstruction , Vascular Diseases , Humans , Tomography, X-Ray Computed/methods , Retrospective Studies , Reproducibility of Results , Intestine, Small/diagnostic imaging , Sensitivity and Specificity , Intestinal Obstruction/diagnostic imaging , Intestinal Obstruction/surgery , Vascular Diseases/pathology , Necrosis/diagnostic imaging , Necrosis/pathology , Abdominal Injuries/complicationsABSTRACT
OBJECTIVE: To discuss the management of sigmoid sinus thrombophlebitis secondary to middle ear cholesteatoma. METHODS: We retrospectively analyzed all cases of sigmoid sinus thrombophlebitis caused by middle ear cholesteatoma over a period of 7 years. 7 male and 2 female patients, ranging in age from 9 to 66 years, were diagnosed with sigmoid sinus thrombophlebitis by clinical presentation and radiological examination. By executing a modified mastoidectomy and tympanoplasty (canal wall-down tympanoplasty) to entirely remove the cholesteatoma-like mastoid epithelium, all patients were effectively treated surgically without opening the sigmoid sinus. All patients were treated with broad-spectrum antibiotics, but no anticoagulants were used. RESULTS: 9 patients had otogenic symptoms such as ear pus, tympanic membrane perforation, and hearing loss. In the initial stage of the surgery, modified mastoidectomy and tympanoplasty were performed on 8 of the 9 patients. 1 patient with a brain abscess underwent puncturing (drainage of the abscess) to relieve cranial pressure, and 4 months later, a modified mastoidectomy and tympanoplasty were carried out. Following surgery and medication, the clinical symptoms of every patient improved. After the follow-up of 6 months to 7 years, 3 patients were re-examined for MRV and showed partial sigmoid sinus recovery with recanalization. 4 months following middle ear surgery, the extent of a patient's brain abscess lesions was significantly reduced. 1 patient experienced facial paralysis after surgery and recovered in 3 months. None of the patients had a secondary illness, an infection, or an abscess in a distant organ. CONCLUSION: The key to a better prognosis is an adequate course of perioperative antibiotic medication coupled with surgical treatment. A stable sigmoid sinus thrombus can remain for a long time after middle ear lesions have been removed, and it is less likely to cause infection and abscesses in the distant organs. The restoration of middle ear ventilation is facilitated by tympanoplasty. It is important to work more closely with multidisciplinary teams such as neurology and neurosurgery when deciding whether to perform lateral sinusotomies to remove thrombus or whether to administer anticoagulation.
Subject(s)
Brain Abscess , Cholesteatoma, Middle Ear , Thrombophlebitis , Humans , Male , Female , Child , Adolescent , Young Adult , Adult , Middle Aged , Aged , Cholesteatoma, Middle Ear/complications , Cholesteatoma, Middle Ear/surgery , Retrospective Studies , Treatment Outcome , Ear, Middle/surgery , Ear, Middle/pathology , Tympanoplasty , Mastoid/surgery , Thrombophlebitis/surgery , Thrombophlebitis/complications , Brain Abscess/complications , Brain Abscess/pathology , Brain Abscess/surgeryABSTRACT
The assembled camshaft is a novel manufacturing product which connects the cam and the mandrel by tube hydroforming (THF) technology after they are processed separately. However, in the process of THF, the structure of the cam-bores has a crucial influence on the connection strength of the assembled camshafts. Therefore, three kinds of cam-bores with circular structure, isometric-trilateral profile and logarithmic spiral profile are selected for hydroforming with a hollow mandrel (tube) in this study. The finite-element-analysis is carried out by ABAQUS software, the variations of (residual) contact pressure and contact area under different structures are obtained, and the torsional angle variations after assembly are measured. Further, the connection strength of the assembled camshaft under three structures is discussed. The results show that the evaluation of connection strength of the assembled camshaft is affected by many factors, including contact pressure, maximum residual contact pressure, axial and circular residual contact pressure, contact area and its rate, residual contact area percentage and torsional angle. Through the comprehensive analysis of various factors, the torsional angle of the camshaft with circular structure is the largest, i.e. poor connection strength. By contrast, the torsional strength of the camshaft with isometric-trilateral profile is the largest, namely, the best connection strength.
ABSTRACT
Meningioma is one of the most common primary neoplasms in the central nervous system, whereas there is still no specific molecularly targeted therapy that has been approved for the clinical treatment of aggressive meningiomas. There is therefore an urgent demand to decrypt the biological and molecular landscape of malignant meningioma. Here, through the in-silica prescreening and 10-year follow-up of 445 meningioma patients, we uncovered that CBX7 is progressively decreased with malignancy grade and neoplasia stage in meningioma and a high CBX7 expression level predicts a favorable prognosis in meningioma patients. CBX7 restoration significantly induces cell cycle arrest and inhibits meningioma cell proliferation. iTRAQ-based proteomics analysis indicated that CBX7 restoration triggers the metabolic shift from glycolysis to oxidative phosphorylation. The mechanistic study demonstrated that CBX7 promotes the proteasome-dependent degradation of c-MYC proteins by transcriptionally inhibiting the expression of a c-MYC deubiquitinase, USP44, which attenuates c-MYC-mediated transactivation of LDHA transcripts and further inhibits glycolysis and subsequent cellular proliferation. More importantly, the functional role of CBX7 was further confirmed in both subcutaneous and orthotopic meningioma xenografts mouse models and human meningioma patients. Together, our results shed light on the critical role of CBX7 during meningioma malignancy progression and identified the CBX7/USP44/c-MYC/LDHA axis as a promising therapeutic target against meningioma progression.
ABSTRACT
Replacement of insulin-producing pancreatic beta-cells by islet transplantation offers a functional cure for type-1 diabetes (T1D). We recently demonstrated that a clinical grade alginate micro-encapsulant incorporating the immune-repellent chemokine and pro-survival factor CXCL12 could protect and sustain the integrity and function of autologous islets in healthy non-human primates (NHPs) without systemic immune suppression. In this pilot study, we examined the impact of the CXCL12 micro encapsulant on the function and inflammatory and immune responses of xenogeneic islets transplanted into the omental tissue bilayer sac (OB; n = 4) and diabetic (n = 1) NHPs. Changes in the expression of cytokines after implantation were limited to 2-6-fold changes in blood, most of which did not persist over the first 4 weeks after implantation. Flow cytometry of PBMCs following transplantation showed minimal changes in IFNγ or TNFα expression on xenoantigen-specific CD4+ or CD8+ T cells compared to unstimulated cells, and these occurred mainly in the first 4 weeks. Microbeads were readily retrievable for assessment at day 90 and day 180 and at retrieval were without microscopic signs of degradation or foreign body responses (FBR). In vitro and immunohistochemistry studies of explanted microbeads indicated the presence of functional xenogeneic islets at day 30 post transplantation in all biopsied NHPs. These results from a small pilot study revealed that CXCL12-microencapsulated xenogeneic islets abrogate inflammatory and adaptive immune responses to the xenograft. This work paves the way toward future larger scale studies of the transplantation of alginate microbeads with CXCL12 and porcine or human stem cell-derived beta cells or allogeneic islets into diabetic NHPs without systemic immunosuppression.
Subject(s)
Diabetes Mellitus , Islets of Langerhans Transplantation , Islets of Langerhans , Animals , Alginates , Chemokine CXCL12 , Graft Survival , Immunosuppression Therapy/methods , Islets of Langerhans Transplantation/methods , Pilot Projects , Primates , Swine , Transplantation, Heterologous/methodsABSTRACT
In this study, male mice were treated with fermented and unfermented Lactobacillus plantarum, Lactobacillus bulgaricus, and Lactobacillus rhamnosus black wolfberry juice (10 mL/kg/day) for 40 days, and their prophylactic effects on ulcerative colitis (UC) induced by dextran sodium sulfate were investigated. The intervention of black wolfberry juice reduced the levels of pro-inflammatory cytokines and increased the content of anti-inflammatory cytokines in the serum and colon. In addition, the pathological changes in colon tissue were alleviated, the expression of Bcl-2 protein in the colon was enhanced, and the intestinal microbiota of the mice was regulated, with an increase in Bacteroidetes and a decrease in Helicobacter. These results suggested that black wolfberry juice had an anti-UC function and Lactobacillus fermentation enhanced the anti-inflammatory effect of black wolfberry juice by modulating the intestinal microbiota.
Subject(s)
Colitis, Ulcerative , Colitis , Lycium , Probiotics , Male , Animals , Mice , Colitis, Ulcerative/chemically induced , Colitis, Ulcerative/drug therapy , Colitis, Ulcerative/prevention & control , Dextran Sulfate/adverse effects , Lactobacillus/metabolism , Colon/metabolism , Anti-Inflammatory Agents/pharmacology , Cytokines/metabolism , Probiotics/therapeutic use , Disease Models, Animal , Mice, Inbred C57BL , Colitis/chemically inducedABSTRACT
The transplanting islets to the liver approach suffers from an immediate posttransplant loss of islets of more than 50%, progressive graft dysfunction over time, and precludes recovery of grafts should there be serious complications such as the development of teratomas with grafts that are stem cell-derived islets (SC-islets). The omentum features an attractive extrahepatic alternative site for clinical islet transplantation. We explore an approach in which allogeneic islets are transplanted onto the omentum, which is bioengineered with a plasma-thrombin biodegradable matrix in three diabetic non-human primates (NHPs). Within 1 week posttransplant, each transplanted NHP achieves normoglycemia and insulin independence and remains stable until termination of the experiment. Success was achieved in each case with islets recovered from a single NHP donor. Histology demonstrates robust revascularization and reinnervation of the graft. This preclinical study can inform the development of strategies for ß cell replacement including the use of SC-islets or other types of novel cells in clinical settings.
Subject(s)
Islets of Langerhans Transplantation , Islets of Langerhans , Animals , Omentum/surgery , Islets of Langerhans/surgery , Islets of Langerhans/metabolism , Transplantation, Homologous , Islets of Langerhans Transplantation/adverse effects , Islets of Langerhans Transplantation/pathology , Primates , AllograftsABSTRACT
BACKGROUND: Pancreatic ductal adenocarcinoma (PDAC) is a rare and refractory malignancy. Early-onset pancreatic cancer (EOPC), defined as pancreatic cancer diagnosed before the age of 50 years, is very rare. Clinical presentation and oncological outcomes of EOPC are confusing according to previous studies. METHODS: We performed a retrospective, population-based study by querying the SEER database to analyze patients with PDAC from 2004 to 2018. Data on demographics, pathological characteristics, treatment patterns, and survival outcomes were compared between EOPC and pancreatic cancer in older patients. Propensity score matching (PSM) was used to minimize the potential bias of baseline characteristics between the two groups. The effect of age on changes in treatment modalities was evaluated using the Cochran-Armitage trend test. RESULTS: The entire study enrolled 42,414 patients, including 2,916 (6.9%) patients with EOPC. Patients with EOPC were more likely to be male (56.6% vs. 51.0%, P < 0.001) and more frequently to present with a larger tumor size (40 mm vs. 37 mm, P < 0.001), vascular invasion (28.6% vs. 25.9%, P = 0.022) and distant metastasis (56.2% vs. 50.8%, P < 0.001) compared with older group. However, surgical resection rates (29.3% vs. 28.3%, P = 0.284) were fairly comparable, and most clinicopathologic characteristics were similar in the patients underwent resection. Younger patients had longer 5-year overall survival (6.9% vs. 5.5%, P < 0.001) and 5-year cancer-specific survival (8.4% vs. 7.3%, P < 0.001) among the overall cohort but had comparable prognosis among patients received surgery (both P > 0.05). Similar survival outcomes were obtained after PSM. In addition, operated patients tended to receive fewer systemic treatments at an increasing age (Ptrend < 0.001). The survival analysis, which was stratified by age groups, suggested that younger patients only had a better prognosis than those over 70. CONCLUSIONS: Patients with EOPC exhibited an advanced stage and a male predilection at diagnosis in the overall cohort but broadly similar clinicopathologic characteristics in the operated patients. In the surgical cohort, although younger patients were more likely to receive systemic treatment, patients with EOPC presented comparable outcomes compared with elderly patients. We suggest that more research should be conducted to uncover the unique characteristics of EOPC for better clinical management.
ABSTRACT
Cardiovascular diseases (CVDs) remain the world's leading cause of death despite the best available healthcare and therapy. Emerging as a key mediator of intercellular and inter-organ communication in CVD pathogenesis, extracellular vesicles (EVs) are a heterogeneous group of membrane-enclosed nano-sized vesicles released by virtually all cells, of which their RNA cargo, especially non-coding RNAs (ncRNA), has been increasingly recognized as a promising diagnostic and therapeutic target. Recent evidence shows that ncRNAs, such as small ncRNAs, circular RNAs, and long ncRNAs, can be selectively sorted into EVs or other non-vesicular carriers and modulate various biological processes in recipient cells. In this review, we summarize recent advances in the literature regarding the origin, extracellular carrier, and functional mechanisms of extracellular ncRNAs with a focus on small ncRNAs, circular RNAs, and long ncRNAs. The pathophysiological roles of extracellular ncRNAs in various CVDs, including atherosclerosis, ischemic heart diseases, hypertension, cardiac hypertrophy, and heart failure, are extensively discussed. We also provide an update on recent developments and challenges in using extracellular ncRNAs as biomarkers or therapeutical targets in these CVDs.
ABSTRACT
Reverse osmosis (RO) membrane has been widely used in various water treatment fields as an efficient desalination technology, but serious biofouling problem arises in the actual application process. Curcumin is known as a natural compound that can reduce biofouling by inhibiting the growth of microorganisms based on quorum sensing. Dopamine, a molecule with excellent adhesion and functionalization on the material's surface, has high research value for applying a curcumin coating to the membrane surface. Curcumin degrades under alkaline conditions, whereas dopamine must polymerize under alkaline conditions. Simultaneously, a coating may adversely affect curcumin. Therefore, a two-step coating process was considered by self-polymerizing dopamine on the thin-film composite membrane surface and then dip-coating curcumin attached to the polydopamine layer. Furthermore, the effect of time and concentration on the surface modification before and after membrane modification was investigated. The highest permeability of 1.39 L/m2/hr/bar was achieved with the modified membranes. The number of gram-positive bacteria decreased from 6.71 × 106 to 9.67 × 105 CFU/mL. This result is meaningful for antifouling through modification of the membrane surface. Use of curcumin can be applied to reduce biofouling and extend the lifetime of the membrane without pretreatment or membrane cleaning.
Subject(s)
Biofouling , Curcumin , Water Purification , Biofouling/prevention & control , Curcumin/pharmacology , Osmosis , Membranes, Artificial , DopamineABSTRACT
Objective:To analyze the relationship between killer cell immunoglobulin-like receptor ( KIR) genes and immune reconstitution failure in human immunodeficiency virus infection/acquired immunodeficiency syndrome (HIV/AIDS) patients after anti-retroviral therapy (ART). Methods:HIV/AIDS patients receiving ART for ≥1 year who attended the AIDS outpatient clinics of Wuming Hospital of Guangxi Medical University and People′s Hospital of Mashan from May 2007 to December 2019 were included. Patients were divided into immune reconstitution failure group and full immune reconstitution group. Polymerase chain reaction with sequence specific primers (PCR-SSP) was used to detect KIR genotypes in all subjects, and the genotype frequency (PF) of 16 KIR genotypes was calculated. Statistical analysis was conducted using chi-square test. Multivariate logistic regression was used to analyze the relationship between KIR genotypes and immune reconstitution failure.Results:There were 102 patients with HIV/AIDS, including 44 immunological non-responders and 58 immunological responders. The PF of KIR2 DL5 in immune reconstitution failure group was 59.09%(26/44), which was higher than 36.21%(21/58) in full immune reconstitution group, and the difference was statistically significant ( χ2=5.27, P=0.022). Multivariate logistics regression analysis showed that KIR2 DL5 was associated with immune reconstitution failure when adjusted for age and baseline CD4 + T cell count. Positive expression of KIR2 DL5 may be a risk factor for immune reconstitution failure (adjusted odds ratio (a OR)=2.431, 95% confidence interval 1.012 to 5.844, P=0.047). Conclusions:Positive expression of KIR2 DL5 may be related to immune reconstitution failure in HIV/AIDS patients after ART.
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Objective To explore the characteristics of opportunistic infections(OIs)in HIV-infected indi-viduals with suboptimal immune reconstitution after ART treatment so as to provide a reference for preventing and managing HIV infections.Methods The clinical data including opportunistic infections specifically were acquired from 112 HIV-infected individuals with suboptimal immune reconstitution from the outpatient department of Wuming Hospital,Guangxi Medical University.The impact of baseline CD4+T lymphocyte counts on the incidence,type,and mixed infection rates of the opportunistic infections were analyzed.Results The opportunistic infection rate among the 112 HIV-infected individuals with suboptimal immune reconstitution was 42.86%,among which fungal infections were the most commonly seen.The opportunistic infection rate of the patients with a baseline of CD4+T lymphocyte counts≤50/μL was significantly higher than that of the patients with a baseline of CD4+T lymphocyte counts>50/μL,and there was no significant difference in the type of opportunistic infections as well as the rate of mixed infections.Conclusion HIV-infected people with suboptimal immune reconstitution in Guangxi are susceptible to HIV OIs.Among them,the group with a baseline CD4+ T lymphocyte counts≤50/μL has a higher rate of OIs,mainly fungal infections.
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Cardiovascular disease (CVD) is a leading cause of morbidity and mortality worldwide that bears an enormous healthcare burden and aging is a major contributing factor to CVDs. Functional gene expression network during aging is regulated by mRNAs transcriptionally and by non-coding RNAs epi-transcriptionally. RNA modifications alter the stability and function of both mRNAs and non-coding RNAs and are involved in differentiation, development, and diseases. Here we review major chemical RNA modifications on mRNAs and non-coding RNAs, including N6-adenosine methylation, N1-adenosine methylation, 5-methylcytidine, pseudouridylation, 2' -O-ribose-methylation, and N7-methylguanosine, in the aging process with an emphasis on cardiovascular aging. We also summarize the currently available methods to detect RNA modifications and the bioinformatic tools to study RNA modifications. More importantly, we discussed the specific implication of the RNA modifications on mRNAs and non-coding RNAs in the pathogenesis of aging-associated CVDs, including atherosclerosis, hypertension, coronary heart diseases, congestive heart failure, atrial fibrillation, peripheral artery disease, venous insufficiency, and stroke.
Subject(s)
Cardiovascular Diseases , RNA, Long Noncoding , Humans , Cardiovascular Diseases/genetics , Ribose , Aging/genetics , RNA, Messenger , RNA , Adenosine/metabolism , RNA, Long Noncoding/geneticsABSTRACT
BACKGROUND: The treatment of diabetes by islet cell transplantation has become an accepted therapy, with transplantation of xenogeneic islet cells an attractive alternative to the problem. Previous studies in mice have demonstrated that anti-CD45RB induce immune tolerance in human pancreatic islet cells. The current study was to define the mechanism of action of anti-CD45RB induced nonspecific immune tolerance to heteroantigens. METHODS: A total of 1500 IEQ human islets were transplanted to diabetic B6µMT-/- mice, B6 mice, and µMT-/- diabetic mice undergoing thymectomy. These mice were treated short-term with doses of anti-CD45RB. CD4+Foxp3+Tregs were detected in the blood, peripheral lymphatic organs by flow cytometry, and immunohistochemistry. In addition, anti-CD25 mAb was administered to tolerant human islet cells B6µMT-/-mice. Mice then were transplanted with other human islet cells and received CD4+CD25+Tregs isolated from tolerant human islets mice to observe islet destruction. RESULTS: Anti-CD45RB treatment-induced tolerance to islets in both immunocompetent and B-cell-deficient mice (µMT-/- mice) by processes that were dependent on CD25+ Tregs, but not B cells. Anti-CD45RB treatment increased the number of CD4+Foxp3+Tregs cells. Anti-CD45RB treatment-induced immune tolerance that was antigen nonspecific, with Tregs playing an important role. Anti-CD45RB treatment-induced tolerance generated Tregs that could be transferred to another individual to manifest nonspecific immune tolerance. CONCLUSION: The results of the experiment suggest that anti-CD45RB induced tolerance to human islet xenografts is mediated by the proliferation of Tregs. These tolerogenic Tregs can be transferred to other mice and induce nonspecific immune tolerance.
Subject(s)
Diabetes Mellitus, Experimental , Islets of Langerhans Transplantation , Humans , Mice , Animals , T-Lymphocytes, Regulatory , Transplantation Tolerance , Transplantation, Heterologous/methods , Graft Survival , Islets of Langerhans Transplantation/methods , Immune Tolerance , Mice, Inbred C57BLABSTRACT
B lymphocytes have long been recognized for their critical contributions to adaptive immunity, providing defense against pathogens through cognate antigen presentation to T cells and Ab production. More recently appreciated is that B cells are also integral in securing self-tolerance; this has led to interest in their therapeutic application to downregulate unwanted immune responses, such as transplant rejection. In this study, we found that PMA- and ionomycin-activated mouse B cells acquire regulatory properties following stimulation through TLR4/TLR9 receptors (Bregs-TLR). Bregs-TLR efficiently inhibited T cell proliferation in vitro and prevented allograft rejection. Unlike most reported Breg activities, the inhibition of alloimmune responses by Bregs-TLR relied on the expression of TGF-ß and not IL-10. In vivo, Bregs-TLR interrupted donor-specific T cell expansion and induced Tregs in a TGF-ß-dependent manner. RNA-Seq analyses corroborated the involvement of TGF-ß pathways in Breg-TLR function, identified potential gene pathways implicated in preventing graft rejection, and suggested targets to foster Breg regulation.
Subject(s)
B-Lymphocytes, Regulatory , Allografts , Animals , B-Lymphocytes, Regulatory/metabolism , Lymphocyte Activation , Mice , Signal Transduction , Toll-Like Receptor 4/metabolism , Transforming Growth Factor beta/metabolismABSTRACT
Islet transplantation to treat insulin-dependent diabetes is greatly limited by the need for maintenance immunosuppression. We report a strategy through which cotransplantation of allogeneic islets and streptavidin (SA)-FasL-presenting microgels to the omentum under transient rapamycin monotherapy resulted in robust glycemic control, sustained C-peptide levels, and graft survival in diabetic nonhuman primates for >6 months. Surgical extraction of the graft resulted in prompt hyperglycemia. In contrast, animals receiving microgels without SA-FasL under the same rapamycin regimen rejected islet grafts acutely. Graft survival was associated with increased number of FoxP3+ cells in the graft site with no significant changes in T cell systemic frequencies or responses to donor and third-party antigens, indicating localized tolerance. Recipients of SA-FasL microgels exhibited normal liver and kidney metabolic function, demonstrating safety. This localized immunomodulatory strategy succeeded with unmodified islets and does not require long-term immunosuppression, showing translational potential in ß cell replacement for treating type 1 diabetes.
Subject(s)
Diabetes Mellitus, Type 1 , Islets of Langerhans Transplantation , Microgels , Allografts/metabolism , Animals , Diabetes Mellitus, Type 1/therapy , Islets of Langerhans Transplantation/methods , Primates , Sirolimus , StreptavidinABSTRACT
Clinical islet transplantation has relied almost exclusively on intraportal administration of pancreatic islets, as it has been the only consistent approach to achieve robust graft function in human recipients. However, this approach suffers from significant loss of islet mass from a potent immediate blood-mediated inflammatory response (IBMIR) and a hypoxic environment. To avoid these negative aspects of the portal site, we explored an alternative approach in which allogeneic islets were transplanted into the intrapleural space of a non-human primate (NHP), treated with an immunosuppression regimen previously reported to secure routine survival and tolerance to allogeneic islets in NHP. Robust glycemic control and graft survival were achieved for the planned study period of >90 days. Our observations suggest the intrapleural space provides an attractive locale for islet transplantation due to its higher oxygen tension, ability to accommodate large transplant tissue volumes, and a lack of IBMIR-mediated islet damage. Our preliminary results reveal the promise of the intrapleural space as an alternative site for clinical islet transplantation in the treatment of type 1 diabetes.
Subject(s)
Diabetes Mellitus, Type 1 , Hematopoietic Stem Cell Transplantation , Islets of Langerhans Transplantation , Islets of Langerhans , Animals , Diabetes Mellitus, Type 1/surgery , Glycemic Control , Graft Survival , Islets of Langerhans Transplantation/methods , PrimatesABSTRACT
Pulmonary hypertension (PH) is a kind of disease characterized by progressive increase of pulmonary vascular resistance and occlusive vascular remodeling. Hypoxic inductive factor-2α (HIF-2α) plays an important role in the abnormal proliferation of pulmonary vascular cells and pulmonary vascular remodeling. This review focuses on the role of HIF-2α in pulmonary hypertension at the cellular and the global level, and candidates targeting HIF-2α for the treatment of pulmonary hypertension, in order to better understand the pathogenesis of PH and find effective treatments.
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Objective To summarize the clinical features of patients with Wilson's disease (WD). Methods A retrospective analysis was performed for the clinical data of 83 patients with WD who were admitted to The Third Hospital of Hebei Medical University from April 2013 to August 2021, including clinical manifestations, Imaging examinations, laboratory examinations, liver histopathological examinations, and ATP7B gene testing results. The patients were divided into groups based on different clinical types. A one-way analysis of variance was used for comparison between groups. Results The youngest age was 3 years for the 83 patients with WD, among whom 39 (46.99%) had an age of ≤18 years, with a mean age of 21.16±14.87 years for all 83 patients. Of all patients, 63.86% had liver-type WD, 31 patients (37.35%) had developed liver cirrhosis at the time of consultation, and 5 patients (6.2%) attended the hospital due to acute or acute-on-chronic liver failure. Of all patients, 62(74.69%) were positive for corneal K-F ring, and the positive rate of K-F ring was 66.04% in the patients with liver-type WD. Among the 83 patients, 79(95.18%) had a reduction in blood ceruloplasmin, and 73(87.95%) had an increase in 24-hour urine copper. The liver histopathological results of 25 patients showed varying degrees of inflammation, fibrosis, steatosis, and copper particle deposition in liver tissue. The ATP7B gene testing results of 25 patients showed that c.2333G > T/p.R778L of exon 8 was the most common mutation site. Conclusion Most patients with WD have the manifestation of liver diseases, and the examinations of corneal K-F ring, serum ceruloplasmin, and 24-h urine copper have their own limitations. Liver pathology and ATP7B gene testing can be performed when it is unable to make a confirmed diagnosis.