scholarly journals Passive transfer of fibromyalgia pain from patients to mice

2019 ◽  
Author(s):  
Andreas Goebel ◽  
Clive Gentry ◽  
Ulku Cuhadar ◽  
Emerson Krock ◽  
Nisha Vastani ◽  
...  
Keyword(s):  
Ex Vivo ◽  
Sensory Nerve ◽  
Immunohistochemical Analysis ◽  
Myelinated Fibre ◽  
Mechanical Pressure ◽  
Satellite Glial Cells ◽  
Chronic Pain Condition ◽  
Nociceptive Afferents ◽  
Healthy Control ◽  
Effective Treatments

SUMMARYFibromyalgia syndrome (FMS) is a chronic pain condition characterized by widespread pain and tenderness1,2. The etiology and pathophysiology of fibromyalgia are unknown and there are no effective treatments. Here we show that sensory hypersensitivity in FMS is caused by autoantibodies that act by sensitizing nociceptive sensory neurons. Administration of IgG from FMS patients increased mouse pain sensitivities to stimulation with mechanical pressure and cold. In contrast, transfer of IgG depleted samples from FMS patients or IgG from healthy control subjects had no effect on pain sensitivity. Sensory nerve fibres in ex vivo skin-nerve preparations from mice treated with FMS IgG were hypersensitive to mechanical stimulation. Immunohistochemical analysis revealed that IgG from FMS patients specifically labeled satellite glial cells and myelinated fibre tracts, as well as a small number of macrophages and endothelial cells in mouse dorsal root ganglia but not skin, muscle, spinal cord and brain. Our results demonstrate that fibromyalgia pain is caused by IgG autoantibodies that sensitize peripheral nociceptive afferents neurons and suggest that therapies that reduce patient IgG titres may be effective treatments of fibromyalgia pain.

The effect of bevacizumab on targeting of anti-epidermal growth factor receptor and anti-insulin-like growth factor 1 receptor antibodies.

2012 ◽  
Vol 30 (15_suppl) ◽  
pp. 3035-3035
Author(s):  
Sandra Heskamp ◽  
Otto C. Boerman ◽  
Janneke D.M. Molkenboer-Kuenen ◽  
Wim J.G. Oyen ◽  
Winette T.A. Van Der Graaf ◽  
...  
Keyword(s):  
Growth Factor ◽  
Ex Vivo ◽  
Growth Factor Receptor ◽  
Progression Free Survival ◽  
Immunohistochemical Analysis ◽  
Control Group ◽  
Vascular Density ◽  
Tumor Vascularity ◽  
Bevacizumab Treatment ◽  
Ex Vivo Biodistribution

3035 Background: Bevacizumab and cetuximab are approved antibodies for treatment of patients with metastasized colorectal cancer. However, the combination of bevacizumab and cetuximab does not improve progression free survival (Tol et al. NEJM 2009). This may be explained by the disruption of tumor vascularity by bevacizumab, thereby reducing targeting of other antibodies to the tumor. The aim of this study was to determine the effect of bevacizumab on the targeting of anti-EGFR and IGF-1R antibodies in tumors with SPECT/CT imaging. Methods: Mice with subcutaneous EGFR and IGF-1R-expressing SUM149 xenografts were injected intraperitoneally with a single dose of bevacizumab (10 mg/kg). After four days, mice received an intravenous injection of 17 MBq 111In-labeled cetuximab, an anti-EGFR antibody, or R1507, an anti-IGF-1R antibody. A control group was injected with labeled hLL2 anti-CD22, an irrelevant IgG . Three days after injection, SPECT/CT images were acquired and mice were dissected for ex vivo biodistribution. Tumors were analyzed immunohistochemically to determine vascular density (CD34), EGFR and IGF-1R expression. Results: SPECT imaging revealed that bevacizumab treatment reduced targeting of anti-EGFR and anti-IGF-1R antibodies by 42% and 35%, respectively. Ex vivo biodistribution showed that uptake of 111In-cetuximab in untreated tumors was 35.2 ± 1.6 %ID/g, compared with 19.7 ± 5.3 %ID/g for bevacizumab treated tumors (p = 0.009). A similar effect was observed for 111In-R1507 (control: 26.7 ± 2.8 %ID/g, bevacizumab:18.9 ± 2.8 %ID/g, p =0.009). No significant differences in tumor uptake were observed in mice that received the irrelevant IgG. Immunohistochemical analysis showed that vascular density decreased with 40%, while EGFR and IGF-1R expression was unaltered. Conclusions: Bevacizumab treatment can significantly reduce targeting of other antibodies to tumors. This underlines the importance of timing and sequencing of bevacizumab therapy in combination with other antibodies.

TRPA1 in bradykinin-induced mechanical hypersensitivity of vagal C fibers in guinea pig esophagus

2009 ◽  
Vol 296 (2) ◽  
pp. G255-G265 ◽  
Author(s):  
Shaoyong Yu ◽  
Ann Ouyang
Keyword(s):  
Guinea Pig ◽  
Action Potentials ◽  
Transient Receptor Potential ◽  
Ex Vivo ◽  
Sensory Nerve ◽  
Sensory Nerves ◽  
Peripheral Sensitization ◽  
Nerve Terminals ◽  
Mechanical Hypersensitivity ◽  
C Fibers

Bradykinin (BK) activates sensory nerves and causes hyperalgesia. Transient receptor potential A1 (TRPA1) is expressed in sensory nerves and mediates cold, mechanical, and chemical nociception. TRPA1 can be activated by BK. TRPA1 knockout mice show impaired responses to BK and mechanical nociception. However, direct evidence from sensory nerve terminals is lacking. This study aims to determine the role of TRPA1 in BK-induced visceral mechanical hypersensitivity. Extracellular recordings of action potentials from vagal nodose and jugular neurons are performed in an ex vivo guinea pig esophageal-vagal preparation. Peak frequencies of action potentials of afferent nerves evoked by esophageal distension and chemical perfusion are recorded and compared. BK activates most nodose and all jugular C fibers. This activation is repeatable and associated with a significant increase in response to esophageal distension, which can be prevented by the B2 receptor antagonist WIN64338. TRPA1 agonist allyl isothiocyanate (AITC) activates most BK-positive nodose and jugular C fibers. This is associated with a transient loss of response to mechanical distensions and desensitization to a second AITC perfusion. Desensitization with AITC and pretreatment with TRPA1 inhibitor HC-030031 both inhibit BK-induced mechanical hypersensitivity but do not affect BK-evoked activation in nodose and jugular C fibers. In contrast, esophageal vagal afferent Aδ fibers do not respond to BK or AITC and fail to show mechanical hypersensitivity after BK perfusion. This provides the first evidence directly from visceral sensory afferent nerve terminals that TRPA1 mediates BK-induced mechanical hypersensitivity. This reveals a novel mechanism of visceral peripheral sensitization.

scholarly journals Joint ex vivo MRI and histology detect iron-rich cortical gliosis in Tau and TDP-43 proteinopathies

2021 ◽  
Author(s):  
M. Dylan Tisdall ◽  
Daniel T Ohm ◽  
Rebecca Lobrovich ◽  
Sandhitsu R Das ◽  
Gabor Mizsei ◽  
...  
Keyword(s):  
Clinical Symptoms ◽  
Ex Vivo ◽  
Cortical Layer ◽  
Heme Iron ◽  
Therapeutic Trials ◽  
Hyperintense Signal ◽  
Mri Features ◽  
Healthy Control ◽  
Hypointense Signal

Frontotemporal lobar degeneration (FTLD) is a heterogeneous spectrum of age-associated neurodegenerative diseases that include two main pathologic categories of tau (FTLD-Tau) and TDP-43 (FTLD-TDP) proteinopathies. These distinct proteinopathies are often clinically indistinguishable during life, posing a major obstacle for diagnosis and emerging therapeutic trials tailored to disease-specific mechanisms. Moreover, MRI-derived measures have had limited success to date discriminating between FTLD-Tau or FTLD-TDP. T2*-weighted (T2*w) ex vivo MRI has previously been shown to be sensitive to non-heme iron in healthy intracortical lamination and myelin, and to pathological iron deposits in amyloid-beta plaques and activated microglia in Alzheimer's disease (AD). However, an integrated, ex vivo MRI and histopathology approach is understudied in FTLD. We apply joint, whole-hemisphere ex vivo MRI at 7T and histopathology to the study autopsy-confirmed FTLD-Tau (n=3) and FTLD-TDP (n=2), relative to an AD disease-control brain with antemortem clinical symptoms of frontotemporal dementia and an age-matched healthy control. We detect distinct laminar patterns of novel iron-laden glial pathology in both FTLD-Tau and FTLD-TDP brains. We find iron-positive ameboid and hypertrophic microglia and astrocytes largely in deeper GM and adjacent WM in FTLD-Tau. In contrast, FTLD-TDP presents prominent superficial cortical layer iron reactivity in astrocytic processes enveloping small blood vessels with limited involvement of adjacent WM, as well as more diffuse distribution of punctate iron-rich dystrophic microglial processes across all GM lamina. This integrated MRI/histopathology approach reveals ex vivo MRI features that are consistent with these pathological observations distinguishing FTLD-Tau and FTLD-TDP, including prominent irregular hypointense signal in deeper cortex in FTLD-Tau whereas FTLD-TDP showed upper cortical layer hypointense bands and diffuse cortical speckling. Moreover, differences in adjacent WM degeneration and iron-rich gliosis on histology between FTLD-Tau and FTLD-TDP were also readily apparent on MRI as hyperintense signal and irregular areas of hypointensity, respectively that were more prominent in FTLD-Tau compared to FTLD-TDP. These unique histopathological and radiographic features were distinct from HC and AD brains, suggesting that iron-sensitive T2*w MRI, adapted to in vivo application at sufficient resolution, may offer an opportunity to improve antemortem diagnosis of FTLD proteinopathies using tissue-validated methods.

scholarly journals Effect of TGF-β1 on eosinophils to induce cysteinyl leukotriene E4 production in aspirin-exacerbated respiratory disease

PLoS ONE ◽  
2021 ◽  
Vol 16 (8) ◽  
pp. e0256237
Author(s):  
Youngwoo Choi ◽  
Soyoon Sim ◽  
Dong-Hyun Lee ◽  
Hee-Ra Lee ◽  
Ga-Young Ban ◽  
...  
Keyword(s):  
Respiratory Disease ◽  
Transforming Growth Factor Beta ◽  
Transforming Growth Factor ◽  
Ex Vivo ◽  
Cysteinyl Leukotriene ◽  
Aspirin Exacerbated Respiratory Disease ◽  
Healthy Control ◽  
Tgf Β1 ◽  
Eosinophil Degranulation

Cysteinyl leukotriene (cysLT) overproduction and eosinophil activation are hallmarks of aspirin-exacerbated respiratory disease (AERD). However, pathogenic mechanisms of AERD remain to be clarified. Here, we aimed to find the significance of transforming growth factor beta 1 (TGF-β1) in association with cysteinyl leukotriene E4 (LTE4) production, leading to eosinophil degranulation. To evaluate levels of serum TGF-β1, first cohort enrolled AERD (n = 336), ATA (n = 442) patients and healthy control subjects (HCs, n = 253). In addition, second cohort recruited AERD (n = 34) and ATA (n = 25) patients to investigate a relation between levels of serum TGF-β1 and urinary LTE4. The function of TGF-β1 in LTE4 production was further demonstrated by ex vivo (human peripheral eosinophils) or in vivo (BALB/c mice) experiment. As a result, the levels of serum TGF-β1 were significantly higher in AERD patients than in ATA patients or HCs (P = .001; respectively). Moreover, levels of serum TGF-β1 and urinary LTE4 had a positive correlation (r = 0.273, P = .037). In the presence of TGF-β1, leukotriene C4 synthase (LTC4S) expression was enhanced in peripheral eosinophils to produce LTE4, which sequentially induced eosinophil degranulation via the p38 pathway. When mice were treated with TGF-β1, significantly induced eosinophilia with increased LTE4 production in the lung tissues were noted. These findings suggest that higher levels of TGF-β1 in AERD patients may contribute to LTE4 production via enhancing LTC4S expression which induces eosinophil degranulation, accelerating airway inflammation.

scholarly journals The role of the Oncostatin M/ OSM receptor β axis in activating dermal microvascular endothelial cells in Systemic Sclerosis

2020 ◽  
Author(s):  
Grace Marden ◽  
Qianqian Wan ◽  
James Wilks ◽  
Katherine Nevin ◽  
Maria Feeney ◽  
...  
Keyword(s):  
Endothelial Cells ◽  
Cell Migration ◽  
Ex Vivo ◽  
Oncostatin M ◽  
Microvascular Endothelial Cells ◽  
Mesenchymal Transition ◽  
Healthy Control ◽  
Microvascular Endothelial ◽  
Endothelial To Mesenchymal Transition

Abstract Background Scleroderma (SSc) is a rare autoimmune disease characterized by vascular impairment and progressive fibrosis of the skin and other organs. Oncostatin M, a member of the IL-6 family, is elevated in SSc serum and was recognized as a significant player in various stages of fibrosis. The goal of this study was to assess the contribution of the OSM/OSMRβ pathway to endothelial cell (EC) injury and activation in SSc. Methods IHC and IF were used to assess the distribution of OSM and OSMRβ in SSc (n = 14) and healthy control (n = 7) skin biopsies. Cell culture experiments were performed in human dermal microvascular endothelial cells (HDMECs) and included mRNA and protein analysis, and cell migration and proliferation assays. Ex vivo skin organoid culture was used to evaluate the effect of OSM on perivascular fibrosis. Results OSMRβ protein was elevated in dermal ECs and in fibroblasts of SSc patients. Treatments of HDMECs with OSM or IL-6 + sIL-6R have demonstrated that both cytokines similarly stimulated proinflammatory genes and genes related to endothelial-to mesenchymal transition ((EndMT). OSM was more effective than IL-6 + sIL-6R in inducing cell migration, while both treatments similarly induced cell proliferation. The effects of OSM were mediated via OSMRβ and STAT3, while the LIFR did not contribute to these responses. Both, OSM and IL-6 + sIL-6R induced profibrotic gene expression in HDMECs, as well as expansion of the perivascular PDGFRβ+ cells in the ex vivo human skin culture system. Additional studies in HDMECs showed that siRNA-mediated downregulation of FLI1 and its close homolog ERG resulted in increased expression of OSMRβ in HDMECs. Conclusions This work provides new insights into the role of the OSM/OSMRβ axis in activation/injury of dermal ECs and supports the involvement of this pathway in SSc vascular disease.

scholarly journals A234 EVALUATING THE EFFECT OF STOOL SUPERNATANTS FROM IBS PATIENTS AND HEALTHY CONTROLS ON THE EXCITABILITY OF DRG NEURONS

2021 ◽  
Vol 4 (Supplement_1) ◽  
pp. 283-284
Author(s):  
E Neary ◽  
N N Jiménez-Vargas ◽  
S Osman ◽  
D E Reed ◽  
S Vanner ◽  
...  
Keyword(s):  
Neuronal Excitability ◽  
Ex Vivo ◽  
Resting Membrane Potential ◽  
Krebs Solution ◽  
Drg Neurons ◽  
Healthy Control ◽  
Underlying Mechanisms ◽  
Irritable Bowel ◽  
And Control ◽  
Perforated Patch Clamp

Abstract Background Abdominal pain is commonly described in chronic disorders such as irritable bowel syndrome (IBS), but the underlying mechanisms are currently unclear. The stool metabolomic and microbiota profiles of IBS and healthy patients have shown distinct differences. Additionally, IBS stool supernatants have previously been demonstrated to induce hypersensitivity of nociceptive nerves in the ex vivo mouse colon, suggesting that mediators in the stool can sensitize nociceptors. However, the effects of healthy control (HC) or IBS patient stool supernatants on the excitability of DRG neurons have not been clarified. Aims To evaluate the effect of HC and IBS supernatant on DRG neurons. Methods HC (n=8 patients) or IBS (n=10 patients) stool was collected, dissolved and homogenized with bicarbonate-buffered Krebs solution at 37°C in a 1/10 dilution. DRG neurons from C57BL/6 mice were dissociated and incubated overnight with HC or IBS supernatant in a Krebs dissolution. Changes in DRG neuronal excitability were recorded using perforated patch-clamp techniques to measure the rheobase (amount of current needed to elicit an action potential). The effect of the IBS and HC stool supernatants on the resting membrane potential (RMP) was also recorded. Results Overnight incubations with supernatant of HC stool diluted in Krebs solution (n=28 neurons) did not significantly decrease the rheobase compared to control neurons (n=22) (62.7 ± 3.9 pA vs 64.2 ± 2.7 pA). In a parallel experiment, we evaluated the effect of IBS stool supernatants diluted in Krebs (n=52 neurons) and found that they significantly decreased the rheobase compared to the supernatant of HC diluted in Krebs and control neurons (52.3 ± 2.3; p<0.05). The data were analyzed with a one-way ANOVA and Tukey’s test. Incubations with IBS supernatant decreased the RMP compared to HC supernatant (-42.6 ± 0.6 mV vs. -46.0 ± 0.9 mV; p<0.01), which was calculated with an unpaired t-test. Conclusions These findings suggest that mediators in IBS stool increase the excitability of DRG neurons compared to HC stool supernatant, and thus may contribute to pain signaling in IBS patients. Funding Agencies CIHR

scholarly journals Synthesize of a novel 89Zr labeled HER2 affibody and its application study in tumor PET imaging

2020 ◽  
Author(s):  
Yuping Xu ◽  
Lizhen Wang ◽  
Donghui Pan ◽  
Junjie Yan ◽  
Xinyu Wang ◽  
...  
Keyword(s):  
Pet Imaging ◽  
Ex Vivo ◽  
Growth Factor Receptor ◽  
Immunohistochemical Analysis ◽  
In Vivo Studies ◽  
Her2 Overexpression ◽  
Potential Candidate ◽  
Synthesis Time ◽  
Ex Vivo Autoradiography

Abstract Background: Human epidermal growth factor receptor-2 (HER2) is an important biomarker for tumor diagnosis and therapy. Affibody is an ideal vector for preparing HER2 specific probes due to the advantages such as high affinity and rapid blood clearance etc. 89Zr is a novel PET imaging isotope with long half-lives and suitable for tracking biological processes for longer periods. In this study, a novel 89Zr labeled HER2 affibody, 89Zr-DFO-MAL-Cys-MZHER2, was synthesized and its imaging properties were also evaluated. Results: The precursors, DFO-MAL-Cys-MZHER2, were obtained with the yields of nearly 50%. The yields of 89Zr -DFO-MAL-Cys-MZHER2 were 90.2±1.9% and the radiopurities were more than 95%. The total synthesis time was only 30 minutes. The probes were stable in PBS and serum. The tracer accumulated in HER2 overexpression human ovarian cancer SKOV-3 cells. In vivo studies in mice bearing tumors showed that the probe highly retained in SKOV-3 xenografts even for 48 hours. The tumors were visualized with good contrast to normal tissue. ROI analysis revealed that the average uptakes in the tumor were greater than 5 %ID/g. On the contrary, the counterparts of MCF-7 tumors kept low levels of （~1%ID/g）. The outcome was consistent with the immunohistochemical analysis and ex vivo autoradiography. The probe quickly cleared from the blood and normal organs, and mainly excreted through the urinary system. Conclusion: The novel HER2 affibody for PET imaging was easily prepared with satisfactory labeling yield and radiochemical purity. 89Zr-DFO-MAL-MZHER is a potential candidate for monitoring HER2 expression and may play specific roles in clinical cancer theranostics.

scholarly journals Synthesis of a novel 89Zr labeled HER2 affibody and its application study in tumor PET imaging

2020 ◽  
Author(s):  
Yuping Xu ◽  
Lizhen Wang ◽  
Donghui Pan ◽  
Junjie Yan ◽  
Xinyu Wang ◽  
...  
Keyword(s):  
Pet Imaging ◽  
Radiochemical Purity ◽  
Ex Vivo ◽  
Radiochemical Yield ◽  
Immunohistochemical Analysis ◽  
In Vivo Studies ◽  
Potential Candidate ◽  
Synthesis Time ◽  
Normal Tissues

Abstract Background: Human epidermal growth factor receptor-2 (HER2) is an essential biomarker for tumor treatment. Affibody is an ideal vector for preparing HER2 specific probes because of high affinity and rapid clearance from normal tissues etc. Zirconium-89 is a PET imaging isotope with a long half-life and suitable for monitoring biological processes for more extended periods. In this study, a novel 89Zr-labeled HER2 affibody, [89Zr]Zr-DFO-MAL-Cys-MZHER2, was synthesized, and its imaging characters were also assessed. Results: The precursor, DFO-MAL-Cys-MZHER2, was obtained with a yield of nearly 50%. The radiochemical yield of [89Zr]Zr -DFO-MAL-Cys-MZHER2 was 90.2±1.9% , and the radiochemical purity was higher than 95%. The total synthesis time was only 30 minutes. The probe was stable in PBS and serum. The tracer accumulated in HER2 overexpressing human ovarian cancer SKOV-3 cells. In vivo studies in mice bearing tumors showed that the probe highly retained in SKOV-3 xenografts even for 48 hours. The tumors were visualized with good contrast to normal tissues. ROI analysis revealed that the average uptake values in the tumor were greater than 5%IA/g during 48 hours postinjection. On the contrary, the counterparts of MCF-7 tumors kept low levels（~1%IA/g）. The outcome was consistent with the immunohistochemical analysis and ex vivo autoradiography. The probe quickly cleared from the normal organs except kidneys and mainly excreted through the urinary system. Conclusion: The novel HER2 affibody for PET imaging was easily prepared with satisfactory labeling yield and radiochemical purity. [89Zr]Zr-DFO-MAL-Cys-MZHER2 is a potential candidate for detecting HER2 expression. It may play specific roles in clinical cancer theranostics.

scholarly journals Effect of synthetic cationic protein on mechanoexcitability of vagal afferent nerve subtypes in guinea pig esophagus

2011 ◽  
Vol 301 (6) ◽  
pp. G1052-G1058 ◽  
Author(s):  
Shaoyong Yu ◽  
Ann Ouyang
Keyword(s):  
Action Potentials ◽  
Ex Vivo ◽  
Sensory Nerve ◽  
Vagal Afferents ◽  
Spontaneous Discharge ◽  
Cationic Protein ◽  
C Fibers ◽  
Potentiation Effect ◽  
Before And After ◽  
Evoke Action Potential

Eosinophilic esophagitis is characterized by increased infiltration and degranulation of eosinophils in the esophagus. Whether eosinophil-derived cationic proteins regulate esophageal sensory nerve function is still unknown. Using synthetic cationic protein to investigate such effect, we performed extracellular recordings from vagal nodose or jugular neurons in ex vivo esophageal-vagal preparations with intact nerve endings in the esophagus. Nerve excitabilities were determined by comparing action potentials evoked by esophageal distensions before and after perfusion of synthetic cationic protein poly-l-lysine (PLL) with or without pretreatment with poly-l-glutamic acid (PLGA), which neutralized cationic charges of PLL. Perfusion with PLL did not evoke action potentials in esophageal nodose C fibers but increased their responses to esophageal distension. This potentiation effect lasted for 30 min after washing out of PLL. Pretreatment with PLGA significantly inhibited PLL-induced mechanohyperexcitability of esophageal nodose C fibers. In esophageal nodose Aδ fibers, perfusion with PLL did not evoke action potentials. In contrast to nodose C fibers, both the spontaneous discharges and the responses to esophageal distension in nodose Aδ fibers were decreased by perfusion with PLL, which can be restored after washing out PLL for 30–60 min. Pretreatment with PLGA attenuated PLL-induced decrease in spontaneous discharge and mechanoexcitability of esophageal nodose Aδ fibers. In esophageal jugular C fibers, PLL neither evoked action potentials nor changed their responses to esophageal distension. Collectively, these data demonstrated that synthetic cationic protein did not evoke action potential discharges of esophageal vagal afferents but had distinctive sensitization effects on their responses to esophageal distension.

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