Perivascular Inflammatory Cell Research Articles

Pathologic Features of Anti-Ku Myositis.

Characteristics of myositis with anti-Ku antibodies are poorly understood. The purpose of this study was to elucidate the pathologic features of myositis associated with anti-Ku antibodies, compared with immune-mediated necrotizing myopathy (IMNM) with anti-signal recognition particle (SRP) and anti-3-hydroxy-3-methylglutaryl-coenzyme A reductase (HMGCR) antibodies, in muscle biopsy-oriented registration cohorts in Japan and Germany. We performed a retrospective pathology review of patients with anti-Ku myositis samples diagnosed in the Japanese and German cohorts. We evaluated histologic features and performed HLA phenotyping. Fifty biopsied muscle samples in the Japanese cohort and 10 in the German cohort were obtained. After exclusion of myositis-specific autoantibodies or other autoimmune connective tissue diseases, 26 samples (43%) of anti-Ku antibody-positive myositis were analyzed. All the samples shared some common features with IMNM, whereas they showed expression of MHC class II and clusters of perivascular inflammatory cells more frequently than the anti-SRP/HMGCR IMNM samples (71% vs 7%/16%; p < 0.005/<0.005; 64% vs 0%/0%; p < 0.005/<0.005). Anti-Ku myositis biopsies could be divided into 2 subgroups based on the extent of necrosis and regeneration. The group with more abundant necrosis and regeneration showed a higher frequency of MHC class II expression and perivascular inflammatory cell clusters. HLA phenotyping in the 44 available patients showed possible associations of HLA-DRB1*03:01, HLA-DRB1*11:01, and HLA-DQB1*03:01 (p = 0.0045, 0.019, and 0.027; odds ratio [OR] 50.2, 4.6, and 2.8; 95% CI 2.6-2942.1, 1.1-14.5, and 1.0-7.0) in the group with less conspicuous necrosis and regeneration. On the contrary, in the group of more abundant necrosis and regeneration, the allele frequencies of HLA-A*24:02, HLA-B*52:01, HLA-C*12:02, and HLA-DRB1*15:02 were lower than those of healthy controls (p = 0.0036, 0.027, 0.016, and 0.026; OR = 0.27, 0, 0, and 0; 95% CI 0.1-0.7, 0-0.8, 0-0.8, and 0-0.8). However, these HLA associations did not remain significant after statistical correction for multiple testing. While anti-Ku myositis shows necrotizing myopathy features, they can be distinguished from anti-SRP/HMGCR IMNM by their MHC class II expression and clusters of perivascular inflammatory cells. The HLA analyses suggest that anti-Ku myositis may have different subsets associated with myopathologic subgroups.

Corosolic acid attenuates platelet-derived growth factor signaling in macrophages and smooth muscle cells of pulmonary arterial hypertension

Pulmonary arterial hypertension (PAH) is a progressive and life-threatening disease that is characterized by vascular remodeling of the pulmonary artery. Pulmonary vascular remodeling is primarily caused by the excessive proliferation and migration of pulmonary arterial smooth muscle cells (PASMCs), which are facilitated by perivascular inflammatory cells including macrophages. Corosolic acid (CRA) is a natural pentacyclic triterpenoid that exerts anti-inflammatory effects. In the present study, the effects of CRA on the viability of macrophages were examined using monocrotaline (MCT)-induced PAH rats and human monocyte-derived macrophages. Although we previously reported that CRA inhibited signal transducer and activator of transcription 3 (STAT3) signaling and ameliorated pulmonary vascular remodeling in PAH, the inhibitory mechanism remains unclear. Therefore, the underlying mechanisms were investigated using PASMCs from idiopathic PAH (IPAH) patients. In MCT-PAH rats, CRA inhibited the accumulation of macrophages around remodeled pulmonary arteries. CRA reduced the viability of human monocyte-derived macrophages. In IPAH-PASMCs, CRA attenuated cell proliferation and migration facilitated by platelet-derived growth factor (PDGF)-BB released from macrophages and PASMCs. CRA also downregulated the expression of PDGF receptor β and its signaling pathways, STAT3 and nuclear factor-κB (NF-κB). In addition, CRA attenuated the phosphorylation of PDGF receptor β and STAT3 following the PDGF-BB simulation. The expression and phosphorylation levels of PDGF receptor β after the PDGF-BB stimulation were reduced by the small interfering RNA knockdown of NF-κB, but not STAT3, in IPAH-PASMCs. In conclusion, CRA attenuated the PDGF-PDGF receptor β-STAT3 and PDGF-PDGF receptor β-NF-κB signaling axis in macrophages and PASMCs, and thus, ameliorated pulmonary vascular remodeling in PAH.

Bifidobacterium animalis subsp. lactis CCFM1274 relieved allergic asthma symptoms by modifying intestinal tryptophan metabolism in mice.

Approximately two-thirds of patients with asthma, a common inflammatory airway disease, are thought to present with allergies. Probiotics and tryptophan metabolites are becoming increasingly important in treating allergic asthma. This study aimed to identify potential probiotic strains and tryptophan metabolites that could alleviate asthma symptoms. Based on in vitro fermentation experiments, we evaluated variations in probiotic capacity to metabolize tryptophan. Of the eight tested strains, Bifidobacterium animalis subsp. lactis CCFM1274 produced relatively high levels of indole-3-carboxaldehyde (I3C). A mouse model of allergic asthma was established by oral administration of ovalbumin (OVA) and was subjected to oral administration of probiotics. The results demonstrated that treatment with CCFM1274 reduced the tendency for body weight loss and mortality in OVA-induced asthmatic mice. Ingestion of CCFM1274 improved the infiltration of perivascular and peribronchial inflammatory cells in the lung sections stained with hematoxylin and eosin (H&E). This outcome was accompanied by a reduction in the serum levels of OVA-specific immunoglobulin E (OVA-sIgE) and in the levels of IL-10 and IL-17 in the bronchoalveolar lavage fluid (BALF). The linear discriminant analysis effect size (LEfSe) of the gut microbiota showed that CCFM1274 increased the relative abundance of Bifidobacterium. In conclusion, CCFM1274 remodeled intestinal tryptophan metabolism in mice and contributed to the improvement of allergic asthma.

Collagen-Filler Injection Associated Dermopathy and COVID-19

COVID-19 and cosmetic skin-fillers are two prevalent topics of today’s medicine, yet their interaction is not sufficiently studied. This article is based on a clinico-morphological case where the patient, a 37-year-old female, visited the clinic with complaints of painless palpable subcutaneous pathologic nodular lesions at the site of collagen cosmetic filler injection after severe acute respiratory syndrome coronavirus 2 infection. In order to verify the pathological processes of the lesions, punch biopsy of the affected skin was taken, and histological, histochemical, and immunohistochemical studies were conducted. Atrophy, acanthosis, parakeratosis with vacuolisation of nuclei of the epidermis; sclerosis and abnormal deposition of collagen fibres in the subepithelial layer of dermis; and vasculitis with endothelial hypertrophy and lymphoid perivascular infiltration (CD3 lymphocytes and CD68 macrophages) were found. Spike and nuclear capsid proteins of severe acute respiratory syndrome coronavirus 2 were localised in cells of perivascular inflammatory infiltrates, endothelial cells, and epithelium of glands and epidermis of the skin. The association between the dermatopathy in COVID-19 virus infection and cosmetic fillers were established. The authors discuss and hypothesise possible autoimmune processes that lead to autoimmune vasculitis.

Comparison of ultrasound signs, computed tomography data and morphological examination of the lungs in patients with coronavirus infection: post hoc analysis

INTRODUCTION: Ultrasound examination (US) of the lungs has shown high efficiency in the diagnosis of COVID-19 pneumonia. The aim of the research was studying the correspondence of computed tomography (CT) US signs of the lungs and morphological data in patients with COVID-19 pneumonia. MATERIALS AND METHODS: The post hoc analysis included 388 patients who simultaneously underwent ultrasound and CT of the lungs. Lung ultrasound was performed according to the 16-zone “Russian Protocol”. Morphological data were obtained from the results of pathoanatomic examination of deceased patients. RESULTS: The comparison of signs detected by CT and ultrasound of the lungs was performed during a multidimensional correspondence analysis. The analysis was carried out using a three-dimensional solution that explained 64.9 % of inertia (p &lt; 0.001). CT signs of “ground glass opacity” (100 %) corresponded to the B-line at ultrasound (100 %), CT of the consolidation sign (44.8 %) — ultrasound signs of consolidation (46.9 %), aerobronchogram of CT (34 %) — aerobronchogram of ultrasound (36.9 %), free liquid CT (11.1 %) — free liquid Ultrasound (13.9 %). CT signs of reticular changes (29.6 %) and “cobblestone pavement” (12.4 %) corresponded to various combinations of ultrasound signs of subpleural consolidation and B-lines. The B-lines were caused by the exudation of fluid and protein molecules into the intraalveolar space against the background of massive death of alveolocytes and formed by the development of intraalveolar edema and the formation of hyaline membranes. The ultrasonic sign of consolidation appeared in the airless zone of the lungs. Subpleural consolidation are caused by thickening and inflammatory infiltration of the pleura, diffuse alveolar damage, with intraalveolar edema, death and decay of alveolocytes, perivascular inflammatory cell reaction/ During treatment for more than 7 days, consolidation in the lung tissue developed due to the disorganization of the organ structure due to the progression of fibrosis. CONCLUSION: Multivariate correspondence analysis showed correlation of CT signs and US signs of the lungs. Morphological analysis showed polymorphism of histological data that caused the formation of ultrasound signs.

A fatal case of fulminant group: a streptococcal infection in a neonate

BackgroundFulminant hemolytic streptococcal infection is a condition of sudden onset and rapidly progressing septic shock caused by Streptococcus pyogenes. It causes beta (complete) hemolysis. Although type A Streptococcus occurs more frequently, all streptococci that cause beta-hemolysis are eligible as causes. This report describes a rare autopsy case of fulminant group A streptococcal infection in a neonate.Case presentationA 16-day-old girl, 3300 g, born by spontaneous delivery at 41 weeks 1 day, experienced a 1-day history of low-grade fever, malaise, and a few hours of cyanosis and anuria, prompting her parents to bring her to the hospital. Her eldest brother, who lived with her, had been infected with Streptococcus approximately one month earlier, and had been treated with ten days of antibiotics. The infant died three hours after presentation. Autopsy findings indicated her to be 52 cm in length, weighing 3585 g, with medium build and normal nutrition. Her lungs were slightly oligemic with decreased volumes. The liver and kidneys were mildly enlarged. The spleen was markedly enlarged. The adrenal glands showed diffuse cortical hemorrhage (Fig. 1). There was some thymic atrophy (thymus weight 7.4 g, < 1 SD below the mean). Histopathological findings included chronic and neutrophilic infiltration of the tonsils and multiple septic emboli containing cocci in the lungs. Perivascular inflammatory cell infiltrates were observed in the lungs, myocardium, kidneys, adrenal glands, brain, meninges, and liver, with micro-necrotic changes in the kidneys and liver. Fibrin thrombi were observed in multiple renal glomeruli. Hemophagocytosis by macrophages was observed in the spleen, liver, lymph nodes, bone marrow, and adrenal glands. Immunostaining showed positive results for group A Streptococcus antibody in the tonsils, with bacterial masses. Blood cultures were positive for group A Streptococcus pyogenes.ConclusionsBecause of the lower respiratory tract infection and suspected sibling transmission, in addition to the adrenal hemorrhages, a diagnosis of fulminant sepsis with group A Streptococcus pyogenes with associated Waterhouse–Friderichsen syndrome was made.

Cancer-associated Regional Ischemic Myopathy: A Rare Immune Myopathy (P9-8.008)

<h3>Objective:</h3> To describe clinical and pathological features of a rare immune myopathy featured by regional ischemia in the setting of cancer. <h3>Background:</h3> Regional ischemic immune myopathy (RIIM) is a rare disorder attributed to myovasculopathy and considered paraneoplastic. It has a high mortality rate. Contrary to dermatomyositis, where microvasculopathy leads to perifascicular pathology and sometimes muscle ischemia, RIIM is characterized by regional necrosis in border zones between perimysial vessels. <h3>Design/Methods:</h3> Review of patient’s clinical and laboratory findings. <h3>Results:</h3> An 81-year-old man manifested subacute proximal limb weakness, dysphagia, and 25-pound weight loss. He had stage IV prostate cancer, diagnosed 3 months prior to weakness-onset. Examination revealed mild facial weakness, flaccid dysarthria, moderate-to-severe neck and limb girdle weakness and atrophy. Videofluoroscopic swallow study showed severe oropharyngeal dysphagia with aspiration. CK was 689 U/L (normal 39–108 U/L) at presentation but 6,145 U/L six weeks earlier. Myositis-associated and HMGCR antibodies were negative. EMG showed proximal-predominant myopathic changes with fibrillation potentials. Muscle biopsy revealed several well-demarcated areas of ischemia involving adjacent fascicles. Necrotic fibers were in a similar early stage of necrosis with no macrophagic infiltration and diffuse sarcoplasmic C5b-9 deposition. A few small collections of perimysial perivascular inflammatory cells were present in the areas of the specimen not affected by ischemia. There was patchy capillary depletion and dilation; some capillaries in areas not affected by ischemia showed complement deposition. Myxovirus resistance protein A (MxA) expression occurred diffusely, especially in necrotic fibers, but without perifascicular predominance. Patient received IVIG 2g/Kg and prednisone 40 mg daily. The limb weakness stabilized while the dysphagia progressed. He expired 3 weeks later. <h3>Conclusions:</h3> RIIM is a rare myovasculopathy leading to muscle weakness in the setting of cancer. In keeping with published data, it has a poor prognosis. The identification of RIIM myopathological changes should trigger the search for underlying malignancy in patients without known cancer. <b>Disclosure:</b> Dr. Granger has nothing to disclose. Dr. Soontrapa has nothing to disclose. Dr. Klein has a non-compensated relationship as a Klein with Neurology Journal that is relevant to AAN interests or activities. Dr. Milone has received personal compensation in the range of $500-$4,999 for serving on a Scientific Advisory or Data Safety Monitoring board for Argenx. Dr. Milone has received personal compensation in the range of $500-$4,999 for serving as an Editor, Associate Editor, or Editorial Advisory Board Member for Neurology Genetics, AAN. The institution of Dr. Milone has received research support from Mayo Clinic, CCaTS-CBD. The institution of Dr. Milone has received research support from Mayo Clinic, SGP Award. The institution of Dr. Milone has received research support from MDA for Care Center grant. The institution of Dr. Milone has received research support from Regenerative medicine Minnesota.

Effect of Calycosin on Airway Inflammation and Airway Remodeling in Allergic Asthma Mouse Model

Background: This study aimed to construct a TDI-induced mouse model of asthma, and evaluate the potential effects and possible molecular mechanisms of Calycosin on airway inflammation and airway remodeling in mouse model. Material and methods: ELISA method was applied to detect the total serum IgE level and the inflammatory cytokine level in the bronchoalveolar lavage fluid. The total number of cells and the proportion of inflammatory cells in BALF were evaluated under an optical microscope. HE was employed to assess and score the infiltration of peritracheal and perivascular inflammatory cells in lung tissue, and PAS staining was used to assess the proportion of goblet cells in the airway epithelium and the thickness of airway epithelial reticular basement membrane in each group of mice. WB was used to detect the expressions of HMGB1 and a-SMA in cells. Immunofluorescence staining was used to detect the expressions of HMGB1 and a-SMA in 16HBEs. Results: The airway hyperresponsiveness of the Calycosin TDI asthma mice decreased, the inflammatory factors in BALF and the total serum IgE levels decreased, the airway epithelial goblet cell metaplasia and the thickness of the airway epithelial reticular basement membrane were improved, thus reducing the up-regulation of HMGB1 and a-SMA expression of 16HBES induced by TDI-HSA. Conclusion: In our study, in the TDI-induced mouse model of asthma, the administration of drug to inhibit the activation of AKT can reduce airway inflammation and airway remodeling. These findings have enriched the current understanding of Calycosin and provided a basis for future research. However, there are also some limitations: How does TDI activate the AKT signaling pathway? After the activation of the AKT pathway, the mechanism by which the expressions of HMGB1, α-SMA and Collagen-I were up-regulated has not been fully elucidated.

Perivascular inflammatory cells and their association with pulmonary arterial remodelling in dogs with pulmonary hypertension due to myxomatous mitral valve disease.

Pulmonary hypertension (PH), an increase in pulmonary arterial pressure (PAP), may occur in dogs affected with myxomatous mitral valve disease (MMVD). Recent studies suggest that an accumulation of perivascular inflammatory cells may be involved with medial thickening which is a sign of the pulmonary artery remodelling in PH. The aim of this study was to characterise perivascular inflammatory cells in the surrounding pulmonary arteries of dogs with PH due to MMVD compared to MMVD dogs and healthy control dogs. Nineteen lung samples were collected from cadavers of small-breed dogs (control n = 5; MMVD n = 7; MMVD + PH n = 7). Toluidine blue stain and multiple IHC targeting α-SMA, vWF, CD20, CD68 and CD3 was performed to examine intimal and medial thickening, assess muscularisation of the small pulmonary arteries and characterise perivascular leucocytes. Medial thickening without intimal thickening of pulmonary arteries and muscularisation of normally non-muscularised small pulmonary arteries was observed in the MMVD and MMVD + PH groups compared with the control group. The perivascular numbers of B lymphocytes, T lymphocytes and macrophages was significantly increased in the MMVD + PH group compared with the MMVD and control groups. In contrast, the perivascular number of mast cells was significantly higher in the MMVD group compared with the MMVD + PH and control groups. This study suggested that pulmonary artery remodelling as medial thickening and muscularisation of the normally non-muscular small pulmonary arteries is accompanied by the accumulation of perivascular inflammatory cells.

Dermal Hypersensitivity Reaction to Semaglutide: Two Case Reports.

Glucagon-like peptide-1 (GLP-1) receptor agonists are a class of antidiabetic agents that work by inducing insulin secretion and inhibiting release of glucagon in a glucose-dependent manner. They are especially promising given their long duration of action, decreased risk of hypoglycemia, and added benefit of weight loss. Semaglutide is a GLP-1 receptor agonist that has been approved for type II diabetes and chronic weight management in obese adults. Cases of hypersensitivity reactions have been previously reported in patients taking GLP-1 receptor agonists dulaglutide and liraglutide. However, to our knowledge, there have been no reports of hypersensitivity reactions to semaglutide. Here, we present two cases of dermal hypersensitivity reactions in patients taking semaglutide for type II diabetes. In the first case, a 75-year-old woman who had been taking semaglutide for 10 months presented with an eruption on her legs, back, and chest for 3 months duration. Histology showed a subepidermal blister with eosinophils, suggestive of a drug hypersensitivity reaction. In the second case, a 74-year-old white man who had been taking semaglutide for 1 month presented with an eruption on the bilateral flanks and lower abdomen for 3 weeks duration. Histology revealed perivascular inflammatory cell infiltrate with eosinophils, also suggestive of a drug hypersensitivity reaction. Both patients began experiencing resolution of their symptoms within 1 month of discontinuing semaglutide. J Drugs Dermatol. 2023;22(4): doi:10.36849/JDD.6550 Citation: Ouellette S, Frias G, Shah R, et al. Dermal hypersensitivity reaction to semaglutide: Two case reports. J Drugs Dermatol. 2023;22(4):413-415. doi:10.36849/JDD.6550.

PS-B01-7: VASCULAR SMOOTH MUSCLE CELL NOX5 REGULATES FIBROBLAST PHENOTYPIC SWITCH AND CARDIOVASCULAR FIBROSIS

Objective: We previously observed that mice expressing human Nox5 (hNox5) in VSMC exhibit vascular dysfunction and cardio-renal fibrosis, where mechanisms are unknown. Here we postulated that VSMC-Nox5 promotes fibroblast phenotypic changes leading to myofibroblast differentiation and pro-fibrotic responses. Design and Methods: Fibroblasts were cultured from wildtype (WT) and hNOX5 mice (Nox5 + SM22 +). Mice (20 weeks old) were infused with Ang II (600 ng/Kg/day) for 28 days and renal fibrosis/inflammation studied. Markers of myofibroblasts, pro-fibrotic and inflammatory phenotypes were assessed by qPCR and immunoblotting. Results: Fibroblasts from Nox5 + SM22 + mice exhibited increased mRNA of pro-fibrotic markers, such as Col1A1 (2-ddC:1.74 ± 0.16 vs. WT 0.67 ± 0.11), Col3A1 (2-ddC:1.74 ± 0.18 vs. WT 0.96 ± 0.24) and TIMP3 (2-ddC:2.65 ± 0.25 vs. WT 0.38 ± 0.07), p &lt; 0.05. Myofibroblast phenotype (SMC) markers mRNA, aSMA (2-ddC:1.54 ± 0.05 vs. WT 0.78 ± 0.17) and Myocd (2-ddC:1.36 ± 0.17 vs. WT 0.39 ± 0.22) were also increased, p &lt; 0.05. mRNA expression of CD36 (2-ddC:1.37 ± 0.07 vs. WT 86 ± 0.24), TNFa (2-ddC:1.32 ± 0.2 vs. WT 0.71 ± 0.17) and TNFR1 (2-ddC:1.26 ± 0.04 vs. WT 1.02 ± 0.10) were increased, while CD68 expression was decreased (2-ddC:0.82 ± 0.11 vs. WT 1.36 ± 0.18) in fibroblasts from Nox5+SM22+ mice (p &lt; 0.05). In Nox5 mice fibroblasts, ROS production and TGFb protein expression (AU:1.8 ± 0.05 vs. WT 1.4 ± 0.06), as well as TGFbR2 gene expression (2-ddC:2.04 ± 0.17 vs. WT 0.57 ± 0.12), were increased (p &lt; 0.05). Kidneys from Ang II-infused Nox5+SM22+ mice exhibited significant perivascular fibrosis and inflammatory cell infiltration compared to WT. Kidney expression of vimentin (AU:1.01 ± 0.05 vs WT 0.85 ± 0.03) and aSMA (AU:0.44 ± 0.03 vs WT 0.33 ± 0.01), markers of myofibroblast differentiation, were increased in Nox5 mice (p &lt; 0.05). Gene expression of DNMT3a and TET2, DNA methylation regulatory enzymes, were also increased in fibroblasts from Nox5+SM22+ mice, p &lt; 0.05. Conclusion: VSMC-Nox5 regulates fibroblast phenotypic switch leading to fibrosis; a process that may involve increased production of ROS and TGFb, and activation of DNA methylation pathways.

Fingolimod attenuates ovalbumin-induced airway inflammation via inhibiting MAPK/ERK signaling in mice.

The current study was designed to investigate the potential anti-inflammatory and antioxidant effects of fingolimod against Ovalbumin (Ova)-induced allergic airway inflammation compared to dexamethasone. Fingolimod was given (0.5 mg/kg/day, p.o.) for sensitized mice 1 hbefore Ova challenge from Days 19 to 24. Fingolimod significantly inhibited Ova-induced elevation of inflammatory cells and eosinophils numbers in bronchoalveolar lavage fluid(BALF) and reduced concentrations of immunoglobulin E in serum and of sphingosine-1-phosphate, interleukin(IL)-4, and IL-13 in BALF. Fingolimod inhibited microvascular leakage and edema as reflected by the decreased lung/body weight index. These findings were supported by histopathological examination results showing that fingolimod substantially decreased perivascular edema and inflammatory cell infiltration. Fingolimod also attenuated Ova-induced oxidative stress as evidenced by decreased malondialdehyde concentration along with increasing concentrations of reduced glutathione and superoxide dismutase in lung tissues. Fingolimod also significantly decreased monocyte chemoattractant protein-1(MCP-1), p-ERK,and p-P38 in lung tissues of Ova-challenged mice. In conclusion, the current study demonstrated the anti-inflammatory and antioxidant effects of fingolimod in allergic airway inflammation that might be associated with the downregulation of mitogen activated kinases signaling to decrease T helper 2 cytokine secretion (IL-4 and IL-13) and MCP-1 expression, along with the inhibition of oxidative stress.

Obstacles in Diagnosing and Treating a Case of Primary Cutaneous CD8-Positive Aggressive Epidermotropic T-Cell Lymphoma; A Case Report

Cutaneous T-cell lymphomas (CTCL) have a wide spectrum of clinical, histological, immunophenotypic features and genetic alterations which make their diagnosis challenging. Due to the rarity and poor characterization of Primary cutaneous CD8-positive aggressive epidermotropic T-cell lymphoma ((PCAECyTCL), it remains a provisional entity in WHO classification (2018) (1). The constant immunohistochemical finding in PCEACyTCL is CD8 +/ CD4 - expression that is non-specific. So, it needs a consensus on the correlation of clinical & lab for accurate diagnosis (2).In addition to the aggressive presentation of the disease and the associated burden of infection make treatment decisions very challenging. Herein, we are presenting our experience in treating an aggressive case of PCEACyTCL, and how we dealt with the disease and its complications. a 58-year-old male, with a history of Psoriasis, started with him 30 years and he was on topical treatment for it. Referred to us after 3 months journey in the Dermatological department with rapidly progressive maculopapular lesions & tumors, showing central ulcerations, necrosis, and complicated by purulent bacterial infection involving his face, scalp, neck, all 4 limbs, whole trunk, perineum with the oral cavity and oropharynx. There was also generalized lymphadenopathy (Figure 1). His lab results were within the normal range with no evidence of any abnormal infiltration by lymphocytes in peripheral blood and Bone marrow examination. Punch skin biopsy showed that the dermis was the seat of dense localized upper & mid dermal wedge-shaped angiocentric & perivascular inflammatory cell infiltrate composed mainly of activated lymphocytes mixed with histiocytes, plasma cells & neutrophils with atypical, transformed lymphocytes. The infiltrate is encroaching upon the epidermis with degeneration of the basal cell layer & epidermotropism. The Immunohistochemistry revealed diffuse positivity of CD3, weak positivity of CD4, and strong positivity of CD8 with negativity for CD7, CD20, CD30, and ALK in the atypical, transformed T-lymphocytes (Figure 2). The patient was diagnosed to have pcAECyTCL. The staging was done using Contrast CT of the chest, abdomen, and pelvis and showed generalized lymphadenopathy with pulmonary nodules, so he was staged (IVB). After mild improvement of the patient's general condition by anti-biotics, painkillers, and supportive treatment, he started CHOP/Etoposide (doxorubicin 50 mg/m2, cyclophosphamide 750 mg/m2, and vincristine 1.4 mg/m2 by intravenous infusion on day 1, etoposide 100 mg/m2 intravenously days 1-3) to be repeated/3 weeks, with G-CSF as our patient showed aggressive myelosuppression after cycles. There was a very promising improvement in his skin lesions (Figure 3), oral ulcers, and general condition. After 3 cycles of chemotherapy, a PET scan showed: Non metabolically active lymphadenopathy. Metabolically active diffuse skin lesions forming irregular skin thickening over the trunk & limbs (SUV max 5). So, he continued further 3 cycles with preparation for an ASCT. Unfortunately, before stem cell transplantation, the skin lesions progressed again with the same aggression. So, he was given 1 cycle of GEMOX (Gemcitabine1 gm/m2 and Oxaliplatin 100 mg/m2 to be repeated/14 days) then he got COVID-19 infection and is still under treatment for it. In our patient, psoriasis can be considered a susceptible predisposing factor for pcAECyTCL according to the proposed theory by Cai, Fleming and Yan, and others (3,4). One of the obvious problems of pcAECyTCL is the exhibition of T-helper-1 cytokine profile, this makes many treatment options inappropriate as Interferons (5). Theoretically, multi-agent chemotherapies appear to be better options, but they carry the risk of toxicity and myelosuppression, and their response is unsatisfactory till now. There are emerging novel therapies but still not applied to this type of lymphoma due to its rarity (6, 7, 8). Bastidas et al. was suggesting that Ruxolitinib may play a future role in its treatment, but this is still in need of future research (9). The CHOP/etoposide protocol was promising in our patient, but it didn't give a durable response. The plan after improvement from COVID-19 infection is to give a targeted therapy as Brentuximab if available or an aggressive protocol as Hyper-CVAD followed by ASCT hoping for CR, longer OS, and better quality of life. Figure 1View largeDownload PPTFigure 1View largeDownload PPT Close modal

Abstract 14566: The Role of Eosinophils in Pulmonary Hypertension

Introduction: Pulmonary hypertension (PH) is a severe life-threatening disease with insidious onset and high mortality, featured by pulmonary vascular remodeling and perivascular inflammatory cell infiltration. However, the precise immune mechanisms underlying PH remain unclear. Although an enhanced Th2 immune response in PH has been reported for years, the role of eosinophils (EOS), one of the Th2 effector cells, in PH is seldom reported. Investigating the roles and molecular mechanisms of EOS may present a novel paradigm of PH development. Hypothesis: EOS protect against PH through lipid metabolites. Methods: EOS infiltration and chemotaxis were investigated in peripheral blood and lung tissues from PH patients and sugen/hypoxia-induced PH mice. ΔdblGATA mice and anti-IL5 antibody (TRFK5)-treated mice were applied to investigate the role of EOS in PH development. Flow cytometry-based assays were used to quantify infiltration of different immune cells. The culture supernatants and lysates of EOS were collected to explore the mechanisms in co-culture cell experiments. Results: EOS accumulated in lung tissues from PH patients and mice, and the infiltration of EOS into pulmonary tissues during PH may be promoted by fibroblast-derived CCL11. Furthermore, EOS deficiency increased neutrophil (p=0.0038) and monocyte/macrophage (p=0.0344) infiltration and promoted pulmonary arterial smooth muscle cell proliferation, leading to aggravated PH in mice. Mechanistically, arachidonate 15-lipoxygenase (ALOX15), which catalyzes the production of pro-resolving lipid mediators, is specifically expressed by EOS. Conclusions: EOS play a beneficial role during PH development via ALOX15 and its downstream lipid mediators, suppressing perivascular inflammation and maintaining smooth muscle cell homeostasis.

The role of calcium-sensitive receptor in ovalbumin-induced airway inflammation and hyperresponsiveness in juvenile mice with asthma.

The role of the calcium-sensitive receptor (CaSR) was assessed in a juvenile mouse model of asthma induced by ovalbumin (OVA). The experiment was divided into normal control, OVA, and OVA +2.5/5mg/kg NPS2143 (a CaSR antagonist) groups. OVA induction was performed in all groups except the normal control, followed by assessing airway hyperresponsiveness (AHR) and lung pathological changes. Serum OVA-specific IgE and IgG1 were detected with an enzyme-linked immunosorbent assay (ELISA), and inflammatory cells were counted in bronchoalveolar lavage fluid (BALF). Real-time quantitative polymerase chain reaction, ELISA, and western blotting were performed to detect gene and protein expression. NPS2143 improved the OVA-induced AHR in mice, and AHR was higher in the OVA +2.5mg/kg NPS2143 group than in the OVA +5mg/kg NPS2143 group. Furthermore, NPS2143 reduced the production of OVA-specific IgE and IgG1 in serum and the number of eosinophils and lymphocytes in BALF in OVA mice with reduced CaSR expression in lung tissues. Besides, OVA-induced mice exhibited peribronchial and perivascular inflammatory cell infiltration, which was accompanied by severe goblet cell hyperplasia/hyperplasia and airway mucus hypersecretion. Furthermore, these mice exhibited increased levels of Interleukin (IL)-5, IL-13, MCP-1, and eotaxin, which were alleviated by NPS2143. The 5mg/kg NPS2143 showed more effective than the 2.5mg/kg treatment. CaSR expression was elevated in the lung tissues of OVA-induced asthmatic juvenile mice, whereas the CaSR antagonist NPS2143 reduced AHR and attenuated the inflammatory response in OVA-induced juvenile mice, possibly exerting therapeutic effects on childhood asthma.

Abstract 127: Nox5 Expression In A Vascular Smooth Muscle Cell-specific Manner Induces Fibroblast To Myofibroblast Differentiation

Mice expressing human Nox5 (hNox5) in VSMC exhibit cardfiovascular fibrosis, where mechanisms are unknown. We postulated that VSMC-Nox5 promotes fibroblast to myofibroblast differentiation that contributes to fibrosis. Fibroblasts were cultured from wildtype (WT) and hNOX5 mice. Mice (20 weeks old) were infused with Ang II (600 ng/Kg/day) for 28 days and renal fibrosis/inflammation studied. Markers of myofibroblasts (αSMA), and pro-fibrotic and inflammatory signaling molecules were assessed by qPCR and immunoblotting. Inflammatory infiltrate was assessed by FACS. Fibroblasts from Nox5 mice exhibited increased mRNA of markers of myofibroblast αSMA (2 -ddC :1.54±0.05 vs. WT 0.78±0.17) and Myocd (2 -ddC :1.36±0.17 vs. WT 0.39±0.22), as well as, pro-fibrotic markers, Col1A1 (2 -ddC :1.74±0.16 vs. WT 0.67±0.11), Col3A1 (2 -ddC :1.74±0.18 vs. WT 0.96±0.24) and TIMP3 (2 -ddC :2.65±0.25 vs. WT 0.38±0.07), p&lt;0.05. mRNA expression of CD36 (2 -ddC :1.37±0.07 vs. WT 86±0.24), TNFα (2 -ddC :1.32±0.2 vs. WT 0.71±0.17) and TNFR1 (2 -ddC :1.26±0.04 vs. WT 1.02±0.10) were increased, while CD68 expression was decreased (2 -ddC :0.82±0.11 vs. WT 1.36±0.18) in fibroblasts from Nox5 mice (p&lt;0.05). In Nox5 mice fibroblasts, ROS production and TGFβ protein expression (AU:1.8±0.05 vs. WT 1.4±0.06), as well as TGFβR2 gene expression (2 -ddC :2.04±0.17 vs. WT 0.57±0.12), were increased (p&lt;0.05). mRNA of DNMT3a and TET2, DNA methylation regulatory enzymes, were also increased in fibroblasts from Nox5 mice, p&lt;0.05. Kidneys from Ang II-infused Nox5 mice exhibited significant perivascular fibrosis and inflammatory cell infiltration compared to WT, as well as increased protein expression of TGFβ (AU: 3.59±0.8 vs. WT 1.54±0.2) and IL-11 (AU: 0.64±0.08 vs. WT 0.39±0.04), p&lt;0.05; where levels of macrophage F4/80+ cells (%:24±2 vs WT 18±1, p&lt;0.05) and levels of cytotoxic CD8+ T cells were increased, p&lt;0.05. Kidney expression of vimentin (AU:1.01±0.05 vs WT 0.85±0.03) and αSMA (AU:0.44±0.03 vs WT 0.33±0.01), were increased in Nox5 mice (p&lt;0.05). Nox5 regulates fibrosis by inducing fibroblast-to-myofibroblast differentiation, possibly through increased ROS. These processes may be important in Nox5-assocated cardiovascular-renal fibrosis.

Examination of nailfold videocapillaroscopy findings in ANCA-associated vasculitis.

The objective of this study was to evaluate nailfold videocapillaroscopy (NVC) as a useful tool for assessing the disease activity of ANCA-associated vasculitis (AAV). This study enrolled 51 patients with AAV and 21 healthy controls. We scored NVC findings semiquantitatively, and compared them between AAV patients and controls. We examined the association of NVC findings with disease activity indicators, histopathological findings of skin biopsies, and high-resolution CT (HRCT) scores in AAV. Additionally, we repeatedly rated the NVC findings 3 months after immunosuppressive therapy. Of the 51 enrolled patients, 36 (70.6%) showed a microangiopathy pattern and 4 (7.8%) showed a scleroderma pattern in AAV. The scores for microhaemorrhage, capillary loss, neoangiogenesis, and tortuosity were significantly higher in the AAV group than in the control group. NVC abnormalities correlated with the severity of skin, lung and kidney involvement. The scores of giant capillaries significantly correlated with the total BVAS and the chest BVAS; the scores of capillary loss correlated with the chest BVAS and the renal BVAS. The scores of microhaemorrhage significantly correlated with perivascular inflammatory cell infiltrations in the upper dermis of the purpura and tended to correlate with the total ground-glass opacity and consolidation scores on HRCT. In addition, capillary loss scores had a significant positive correlation with serum creatinine levels. Additionally, the microhaemorrhage scores were significantly reduced after 3 months of immunosuppressive therapy. In AAV patients, NVC abnormalities are significantly associated with disease severity. This result suggests that NVC is a useful tool for assessing the disease activity and treatment response in AAV.

Wenyang Huazhuo Tongluo formula alleviates pulmonary vascular injury and downregulates HIF-1α in bleomycin-induced systemic sclerosis mouse model

BackgroundVascular damage, autoimmune abnormalities, and fibrosis are the three pathological features of systemic sclerosis (SSc).However, pulmonary vascular damage is the main factor affecting the progression and prognosis of SSc. The main purpose of this study was to explore the molecular mechanism of Wenyang Huazhuo Tongluo Formula in alleviating pulmonary vascular injury in bleomycin-induced SSc mouse model.MethodsMasson staining and H&E staining were used to analyze the degree of pulmonary vascular fibrosis and the infiltration of leukocyte cells in lung tissue ofbleomycin-induced SSc mouse models treated with saline (BLM group), Wenyang Huazhuo Tongluo Formula (WYHZTL group) and HIF-1α inhibitor KC7F2 (KC7F2 group). Blood vessel exudation was determined by analyzing the cell number and albumin concentration in bronchoalveolar lavage fluid using a cell counter and ELISA assay, respectively. The degree of vascular injury was assessed by measuring the expression levels of vWF, E-selectin, ICAM-1, VCAM-1, VE-cadherin and claudin-5 in serum and pulmonary vascular endothelial cells using ELISA and immunofluorescence staining. Finally, the effect of Wenyang Huazhuo Tongluo Formula on the expression of HIF-1α was detected using immunofluorescence staining.ResultsWenyang Huazhuo Tongluo Formula and KC7F2 significantly inhibited bleomycin-induced pulmonary vascular fibrosis and the level of perivascular inflammatory cell infiltration. The number of cells and the concentration of albumin were significantly reduced in the bronchoalveolar lavage fluid of the WYHZTL group and KC7F2 group compared with the BLM group. In addition, treatment with Wenyang Huazhuo Tongluo Formula and KC7F2 significantly downregulated the expression levels of vWF, E-selectin, ICAM-1, VCAM-1 and HIF-1α, but upregulated the expression of VE-cadherin and claudin-5 in serum and pulmonary vascular endothelial cells, compared with treatment with saline.ConclusionsThis study reveals that Wenyang Huazhuo Tongluo Formula plays a new role in the treatment of SSc by alleviating pulmonary vascular damage. Furthermore, we found that Wenyang Huazhuo Tongluo Formula alleviates pulmonary vascular injury and inhibits HIF-1α expression.

Effects of different probiotic strains B. lactis, L. rhamnosus and L. reuteri on brain-intestinal axis immunomodulation in an endotoxin-induced inflammation.

The study evaluated the effects of supplementation with three different probiotic strains Bifidobacterium lactis (LACT GB™), Lactobacillus rhamnosus (RHAM GB™) and Lactobacillus reuteri (REUT GB™) on brain-intestinal immunomodulation in an animal model of LPS-induced inflammation. Fifty mice Balb/C were distributed into five groups: control; lipopolysaccharide (LPS); LPS + B. lactis (LACT GB™); LPS + L. rhamnosus (RHAM GB™); and LPS + L. reuteri (REUT GB™). The animals were supplemented with their respective probiotic microorganisms daily, for 30days, at a concentration of 1 × 109CFU/animal/day. After 30days of supplementation, animals received the inflammatory insult by LPS (15mg/kg). Behavioral tests, oxidative stress and inflammation were performed, as well as gut and brain histology. In the behavioral test, LPS + B. lactis group was less anxious than the other groups. Serum interleukin IL-1β and IL-6 levels increased in all groups that received the LPS insult, and there was a reduction in inflammation in the supplemented groups when compared to the LPS group in brain and gut. There is a reduction in myeloperoxidase activity and oxidative stress in groups supplemented with probiotics. Inintestine histological analysisoccurs damage tothe tissue integrity in the LPS group, in the other hand,occurs preservation of integrity in the probioticsupplemented animals. In the brain, infiltrates of perivascular inflammatory cells can be seen in the LPS group. The three probiotic studies showed efficient immunomodulating activity and ensured integrity of the intestinal barrier function, even after the severe insult by LPS. These results show the important role of probiotics in the gut-brain axis. Graphical abstract illustratively represents the gut-brain axis and how different probiotic strains influence the immunomodulatory response releasing different pro- and anti-inflammatory cytokines, and their role in the balance of dysbiosis.

Bullous systemic lupus erythematosus with nephrotic syndrome in an adolescent girl: a case report

Bullous systemic lupus erythematosus is an uncommon autoimmune subepidermal vesiculobullous disease occurring in less than 5% of patients with systemic lupus erythematosus. The diagnosis of bullous systemic lupus erythematosus is based on clinical picture, histopathological findings, direct immunofluorescence and other immunologic tests. Importantly, establishing the diagnosis of bullous systemic lupus erythematosus requires the occurrence of systemic lupus erythematosus according to the American College of Rheumatology criteria. We report a case of bullous systemic lupus erythematosus in a 16-year-old Vietnamese girl, who demonstrated an acquired vesiculobullous eruption associated with nephrotic syndrome and fulfilled the European League Against Rheumatism/ American College of Rheumatology 2019 classification criteria of systemic lupus erythematosus. Clinical examination showed numerous tense, clear fluid-filled vesicles and bullae affecting the whole body with the predilection for the face, neck, chest, abdomen and bilateral inner thigh areas. Erosions were observed on the tongue and the buccal mucosa. Histopathologic examination showed subepidermal vesicle containing abundant neutrophils and perivascular mixed inflammatory cell infiltration. Microabscesses at tips of dermal papillae and the features of a leukocytoclastic vasculitis were not seen. Direct immunofluorescence of perilesional skin biopsies demonstrated linear deposition of IgG, IgA, IgM and C3 in the basement membrane zone. Linear immunoglobulin deposition displayed a u-serrated pattern with the predominant staining intensity of IgG. Serologic tests showed positive antinuclear antibody. Complement components assay revealed low levels of C3 (66.5 mg/dL) and C4 (3,9mg/dL). Proteinuria level was increased to 3.5g/24 hours. The final diagnosis of bullous systemic lupus erythematosus with nephrotic syndrome was made. The patient was treated with a high-dose oral glucocorticoid scheme and showed significant clinical improvement.