Mastócitos e síndrome de ativação mastocitária: o que há de novo? / Mast cells and mast cell activation syndrome: what's new?

Os mastócitos são as principais células efetoras da resposta alérgica aguda, desempenhando também um papel importante na angiogênese, tolerância imunológica, regulação da fibrinólise, regeneração neuronal e osteoclastogênese. Localizam-se maioritariamente na pele e nas mucosas do intestino e pulmões, onde exercem uma função "sentinela". As síndromes de ativação mastocitária são caracterizadas pela ocorrência de episódios recorrentes de manifestações clínicas resultantes da libertação de mediadores mastocitários. Esta constitui-se como entidade complexa com um espectro de sintomas associados, representando um desafio diagnóstico e terapêutico. Nesta revisão, os autores pretendem apresentar uma visão geral sobre a estrutura e função dos mastócitos e sobre os critérios diagnósticos e abordagem terapêutica da síndrome de ativação mastocitária.

Mast cells are the main effector cells of acute allergic response, also playing an important role in angiogenesis, immune tolerance, regulation of fibrinolysis, neuronal regeneration, and osteoclastogenesis. They are generally located in the skin and mucous membranes of the intestines and lungs, where they perform a "sentinel" function. Mast cell activation syndrome is characterized by recurrent clinical manifestations resulting from the release of mast cell mediators. This complex entity, which involves a spectrum of associated symptoms, is a diagnostic and therapeutic challenge. In this article we overview of the structure and function of mast cells, in addition to the diagnostic criteria and therapeutic approaches to mast cell activation syndrome.

Correlation and clinical significance of VISTA expression and cervical squamous cell carcinoma / 中华肿瘤杂志

Objective: To explore the relationship between the expression of the T-cell activation suppressor-immunoglobulin variable region (VISTA) and the development of cervical squamous cell carcinoma (CSCC), and the impact on the prognosis of CSCC patients. Methods: Cervical tissue samples from 116 CSCC, including 23 cervical intraepithelial neoplasia (CIN) grade I, 23 CIN grade Ⅱ-Ⅲ, and 23 chronic cervicitis patients, were collected from the First Hospital of Soochow University between March 2014 and April 2019. The expression of VISTA in each group was detected by immunohistochemistry (IHC). Survival data of CSCC patients were obtained by follow-up. The survival analysis was performed by Kaplan-Meier method, and survival differences between groups were compared by Log rank test. Prognostic impact factors were analyzed using a multifactorial Cox proportional hazards model. Results: The positive rate of VISTA expression in CSCC group was 32.8% (38/116), and which of grade Ⅱ-Ⅲ was 17.4% (4/23). VISTA expression results showed no positive expression patients in the cervical intraepithelial neoplasia grade I and chronic cervicitis groups. The differences between the CSCC group and other groups were statistically significant (P<0.01). In 116 CSCC patients, VISTA expression was associated with International Federation of Gynecology and Obstetrics (FIGO) stage and lymph node metastasis (P<0.01). The mean survival time of patients in the VISTA positive expression group was 30.7 months, and the 3-year survival rate was 44.7% (17/38). However, the mean survival time of the patients in the VISTA negative expression group was 49.1 months, and the 3-year survival rate was 87.2% (68/78). The Cox regression model found that VISTA expression positivity (P=0.001) and FIGO stage (P=0.047) were prognostic factors for CSCC, and patients with VISTA-positive CSCC had a 4.130-fold risk of death higher than those with VISTA-negative expression. Conclusions: The VISTA protein is highly expressed in CSCC tissues, and its expression level is closely related to the occurrence and development of CSCC. The expression of VISTA can be used as an independent predictor of CSCC prognosis and can provide a strong basis for the treatment of CSCC with immune checkpoint inhibitors.

A preliminary study on the role of V-domain Ig suppressor of T cell activation in juvenile idiopathic arthritis / 中国当代儿科杂志

OBJECTIVES@#To study the expression of V-domain Ig suppressor of T cell activation (VISTA) in peripheral blood of children with juvenile idiopathic arthritis (JIA) and its role in the pathogenesis of JIA.@*METHODS@#In this prospective study, peripheral blood was collected from 47 children with different subtypes of JIA and 10 healthy children. Flow cytometry was used to measure the expression levels of VISTA, interferon-γ (IFN-γ), and tumor necrosis factor-α (TNF-α) on CD14+ mononuclear cells, CD4+ T lymphocytes, and CD8+ T lymphocytes.@*RESULTS@#The children with JIA had a significantly lower expression level of VISTA than the healthy children (P<0.05). There was a significant difference in the expression of VISTA between the children with different subtypes of JIA, with the lowest expression level in those with systemic JIA (P<0.05). There was also a significant difference in the expression of VISTA between different immune cells, with a significantly higher expression level on the surface of monocytes (P<0.05). Correlation analysis showed that VISTA was negatively correlated with the expression of IFN-γ and TNF-α on CD4+ T cells (r=-0.436 and -0.382 respectively, P<0.05), CD8+ T cells (r=-0.348 and -0.487 respectively, P<0.05), and CD14+ mononuclear cells (r=-0.582 and -0.603 respectively, P<0.05).@*CONCLUSIONS@#The insufficient expression of VISTA may be associated with the pathogenesis of JIA, and enhancing the immunomodulatory effect of VISTA might be one option for the treatment of JIA in the future.

Knockout of BRCC3 aggravates acute GVHD in allogeneic hematopoietic stem cell transplant recipient mice / 中华放射医学与防护杂志

Objective:To investigate the effect and underlying mechanism of BRCC3 knockout on acute GVHD(aGVHD) of mice.Methods:A total of 12 recipient C57BL/6J mice were divided into two groups, including 6 wild type(WT) and BRCC3 -/-(KO). The recipients were exposed to 4.5 Gy + 4.5 Gy 60Co γ-rays in total body irradiation (TBI) at 30 min intervals. At 6 h post-irradiation, 1×10 7bone marrow cells and 8×10 6 splenocytes from BALB/c mice were infused into C57BL/6J mouse via tail vein to develop aGVHD mouse model. BRCC3 was specifically knocked out in aGVHD mouse model. The organ damage was examined through histopathology. The levels of serum cytokines were measured by enzyme-linked immuno sorbent assay (ELISA) and cytometric bead array (CBA), respectively. Spleen, liver and small intestine lymphocytes were isolated at 9 d post-transplantation, and the infiltration and activation of T cells in the target organs were assayed using flow cytometry. Results:The absence of BRCC3 in recipient mice significantly shortened survival ( P<0.05) with increased liver injury of aGVHD mice. In BRCC3 -/-recipient mice, the proportions of CD8+ T cells and CD8+ CD25+ T cells were significantly higher than those in the spleen( t=6.53, 5.52, P<0.05), and the proportions of CD8+ T cells and CD8+ CD25+ T cells were significantly increased in the liver ( t=3.74, 3.19, P<0.05). Similarly, the proportions of CD8+ T cells, CD8+ CD25+ T cells and CD8+ CD69+ T cells were significantly elevated in the small intestine ( t=3.52, 4.06, 3.29, P<0.05). Conclusions:BRCC3 deletion increased the proliferation and activation of donor CD8+ T cells and aggravated aGVHD, which might provide a new prevention and treatment target for aGVHD.

Mechanism of Duanteng Yimu Decoction on Human Umbilical Vein Endothelial Cell Model / 中国实验方剂学杂志

Objective:To explore the effect of Duanteng Yimu decoction （DTYM） on the activation of the human umbilical vein endothelial cell （HUVEC） model and the effect on related activated proteins and vascular endothelial growth factor （VEGF） signaling pathway. Method:After DTYM （200， 400 g·mL^-1） treatment of HUVEC induced by VEGF and tumor necrosis factor-<italic>α</italic> （TNF-<italic>α</italic>）， cell proliferation， migration， and tubulogenesis were detected by cell counting kit-8 （CCK-8） assay， 5-ethynyl-2'-deoxyuridine （EdU） assay， transwell migration assay， phalloidin staining， and matrix gel card method. The mRNA expression of adhesion factors， including E-selectin， intercellular adhesion molecule-1 （ICAM-1）， and vascular cell adhesion molecule-1 （VCAM-1） was detected by real-time fluorescence quantitative polymerase chain reaction （Real-time PCR）. The expression of von Willebrand factor （VWF）， platelet-endothelial cell adhesion molecule-31 （CD31）， angiogenic factor cysteine-rich-61 （CYR61）， angiopoietin-1 （ANG-1）， VEGF， and VEGF receptor-2 （VEGFR2） was detected by Western blot. Immunofluorescence was used to determine CD31 expression. Result:Compared with the normal group， the model group showed potentiated proliferation， migration， and tubulogenesis of HUVEC （<italic>P</italic><0.05， <italic>P</italic><0.01）， elevated mRNA expression of E-selectin， ICAM-1， and VCAM-1 （<italic>P</italic><0.01）， up-regulated protein expression of VWF， CD31， ANG-1， CYR61， VEGF-<italic>α</italic>， and phospho （p）-VEGFR2 （<italic>P</italic><0.05，<italic> P</italic><0.01）， and increased CD31 immunofluorescence intensity （<italic>P</italic><0.01）. Compared with the model group， the DTYM groups displayed blunted proliferation， migration， and tubulogenesis of HUVEC （<italic>P</italic><0.05，<italic> P</italic><0.01）， decreased mRNA expression of E-selectin， ICAM-1， and VCAM-1 （<italic>P</italic><0.05， <italic>P</italic><0.01）， down-regulated protein expression of VWF， CD31， ANG-1， CYR61， VEGF-<italic>α</italic>， and p-VEGFR2 （<italic>P</italic><0.05， <italic>P</italic><0.01）， and weakened CD31 immunofluorescence intensity （<italic>P</italic><0.01）. Conclusion:DTYM inhibits HUVEC proliferation， migration， adhesion， and tubulogenesis， which is associated with the regulation of CD31， VWF， CYR61， and ANG-1 expression in HUVEC and the VEGF signaling pathway.

Effects of length and chemical modification on the activation of vascular endothelial cells induced by multi walled carbon nanotubes / 北京大学学报(医学版)

OBJECTIVE@#To investigate the effects of multi-walled carbon nanotubes (MWCNTs) with different length or chemical modification on endothelial cell activation and to explore the role of nucleotide-binding oligomerization domain-like receptor family pyrin domain containing 3 (NLRP3) inflammasome.@*METHODS@#MWCNTs were characterized by dynamic light scattering (DLS) after being suspended in culture medium. The immortalized mouse cerebral microvascular endothelial cell line b.End3 was treated with short MWCNTs (S-MWCNT, 0.5 to 2 μm), long MWCNTs (L-MWCNT, 10 to 30 μm) and the above long MWCNTs functionalized by carboxyl-(L-MWCNT-COOH), amino-(L-MWCNT-NH2) or hydroxyl-(L-MWCNT-OH) modification. Cytotoxicity of MWCNTs in b.End3 cells was determined by cell counting kit-8 (CCK-8) assay and lactate dehydrogenase (LDH) release assay, and non-toxic low dose was selected for subsequent experiments. Effects of all types of MWCNTs on the endothelial activation of b.End3 were determined by the measurement of vascular cell adhesion molecule-1 (VCAM-1) concentration in cell supernatant and adhesion assay of human monocytic cell line THP-1 to b.End3.To further elucidate the mechanism involved, the protein expressions of nucleotide-binding oligomerization domain-like receptor family pyrin domain containing 3(NLRP3) in cells treated with S-MWCNT, L-MWCNT and L-MWCNT-COOH were measured by Western blot.@*RESULTS@#At a higher concentration (125 μg/cm2) and treated for 24 h, all types of MWCNTs significantly inhibited viability of b.End3 cells. At a sub-toxic concentration (6.25 μg/cm2), all types of MWCNTs treated for 12 h significantly induced the activation of b.End3 cells, as evidenced by the elevated VCAM-1 release and THP-1 adhesion. Compared with S-MWCNT, L-MWCNT significantly promoted endothelial cell activation. L-MWCNT and L-MWCNT-COOH activated b.End3 cells to a similar extent. Furthermore, treatment with S-MWCNT, L-MWCNT and L-MWCNT-COOH increased NLRP3 expression in a time-dependent manner at 6.25 μg/cm2. Compared with S-MWCNT, cells treated with L-MWCNT for 4 h and 12 h exhibited significantly increased protein expressions of NLRP3. However, no significant differences were detected in the level of NLRP3 protein in cells treated with L-MWCNT and L-MWCNT-COOH.@*CONCLUSION@#Compared with the surface chemical modification, length changes of MWCNTs exerted more influence on endothelial cell activation, which may be related to the activation of NLRP3 inflammasome. Our study contributes further understanding of the impact of MWCNTs on endothelial cells, which may have implications for the improvement of safety evaluation of MWCNTs.

The clinical significance of the transforming growth factor-β receptor I expression in na?ve CD4 + T cells from patients with systemic lupus erythematosus / 中华风湿病学杂志

Objective:To investigate the clinical significance of the transforming growth factor-β receptor I (TGF-βRⅠ) expression in na?ve CD4 + T cells from patients with systemic lupus erythematosus (SLE). Methods:Na?ve CD4 + T cells were purified using magnetic microbeads from peripheral blood mononuclear cells of SLE patients and healthy controls. Real-time quantitative PCR was used to detect TGF-βRⅠ mRNA level, and flow cytometry was used to detect the percentage of CD69 +CD4 + T cells. Data were analyzed by t test and Pearson correlation analysis. Results:The level of TGF-βR Ⅰ mRNA in na?ve CD4 + T cells from SLE patients was significantly lower than that in healthy controls [(0.674±0.873) vs (1.445±1.112), t=2.301, P<0.05]. The TGF-βR Ⅰ mRNA level was negatively correlated with systemic lupus erythematosus disease activity index (SLEDAI) ( r=-0.376, P<0.05), erythrocyte sedimentation rate (ESR) ( r=-0.376, P<0.05), serum creatinine ( r=-0.323, P<0.05) and 24 h urine protein ( r=-0.331, P<0.05), and positively correlated with serum com-plement C3 ( r=0.528, P<0.01). The level of TGF-βRⅠ mRNA level in na?ve CD4 + T cells in SLE patients with renal involvement was lower than that in SLE patients without renal involvement [(0.525±0.536) vs (1.071±1.007), t=2.198, P<0.05]. The TGF-βR Ⅰ mRNA level in the na?ve CD4 + T cells in anti-dsDNA antibody positive group was lower than that in the anti-dsDNA antibody negative group [(0.344±0.315) vs (0.958±1.076), t=2.277, P<0.05]. The expression of TGF-βRⅠ mRNA in na?ve CD4 + T cells from SLE patients was reduced after 24 h stimulation with anti-CD3/CD28 beads [(0.047±0.013) vs (1.008±0.129), t=14.38, P<0.01], which was partially reversed by dexamethasone treatment [(0.240±0.042) vs (0.047±0.013), t=7.845, P<0.01]. Meanwhile, dexamethasone significantly decreased the expression of CD69 in CD4 + T cells [(15.0±2.1)% vs(34.9±2.0)%, t=32.57, P<0.01]. Conclusion:The abnormally low expression of TGF-βRⅠ in na?ve CD4 + T cells may be involved in the pathogenesis of SLE. Glucocorticoid treatment can upregulate the expression of TGF-βRI and inhibit the activation of T cells, This suggests suggesting that TGF-βRⅠ may be a potential target for SLE treatment.

miR-214 promotes the progression of liver fibrosis by activating hepatic stellate cells / 上海交通大学学报（医学版）

Objective • To explore the biological role of miR-214 in the activation of hepatic stellate cells (HSCs) and the procession of liver fibrosis and its possible mechanism. Methods • Quantitative real-time PCR (qPCR) was used to detect the expression of miR-214 in the activation of HSCs and the progression of liver fibrosis in rats induced by CCl4 (liver fibrosis model). HSC-T6 cells were treated with lentivirus infection and divided into miR-214 overexpression group, miR-214 knockout group and negative control lentivirus group (mock group). qPCR and Western blotting were used to detect the gene and protein expression levels of collagen type 1 (COL1) and α-smooth muscle actin (α-SMA) in the three groups, respectively. Transwell assay and flow cytometry were used to detect the migration and apoptosis of HSCs in the three groups, respectively. Double luciferase reporter gene assay was used to detect weather Hif1an was the target gene of miR-214, and then qPCR and Western blotting were used to detect the expression levels of HIF1AN in the three groups, the activation of HSCs and the liver fibrosis model. Results • miR-214 was upregulated during HSCs activation and the progression of liver fibrosis (both P<0.05). Compared with the mock group, the gene and protein expressions of COL1 and α-SMA in the miR-214 overexpression group were increased, and HSCs migration ability was increased and apoptosis rate was decreased (all P<0.05); the expressions of COL1 and α-SMA in the miR- 214 knockdown group were decreased, and HSCs migration ability was decreased (all P<0.05). Double luciferase reporter gene assay showed that Hif1an was the target gene of miR-214. Compared with the mock group, the gene and protein expressions of HIF1AN in the miR-214 overexpression group were decreased (both P<0.05), and those in the miR-214 knockdown group were increased (both P<0.05). The expression of HIF1AN was decreased during HSCs activation and the progression of liver fibrosis (all P<0.05). Conclusion • miR-214 may promote the migration and activation of HSCs by targeting Hif1an, and then promote the progression of liver fibrosis, suggesting that miR-214 may be a new marker and potential target for the treatment of liver fibrosis.

Uma visão atualizada da anafilaxia idiopática: a ausência de evidência não é a evidência de ausência / An updated view of idiopathic anaphylaxis: absence of evidence is not evidence of absence

A anafilaxia idiopática não apresenta etiologia conhecida. A sua prevalência é estimada entre 10-35% de todas as modalidades de anafilaxia. A sintomatologia apresentada é a mesma de qualquer outra anafilaxia: urticária, angioedema, ruborização, prurido, hipotensão arterial, taquicardia, manifestações gastrointestinais (disfagia, náusea, vômitos, cólicas abdominais, diarreia), asma, edema laríngeo, tontura e síncope. A mortalidade é rara. Não há transmissão genética, mas 40% dos pacientes são atópicos. É mais frequente nos adultos do que nas crianças, e principalmente em mulheres. É um diagnóstico de exclusão. Ocorre ativação mastocitária com desgranulação citoplasmática dos mediadores de anafilaxia (triptase, histamina, entre outros). É uma anafilaxia com boa resposta aos corticoides, e, portanto, caso não haja resposta adequada a doses eficazes de prednisona/prednisolona, o seu diagnóstico deve ser revisto. O diagnóstico diferencial da anafilaxia idiopática inclui: a mastocitose sistêmica indolente, síndromes de ativação mastocitária monoclonais, alergia à galactose-alfa-1,3 galactose, anafilaxia induzida por exercícios (com e sem dependência alimentar e medicamentosa), angioedema hereditário (congênito e adquirido), feocromocitoma, síndrome carcinoide, anafilaxia oral acarina, alergia ao Anisakis simplex, disfunção das cordas vocais, síndrome escombroide, alergia ao sêmen, alergia ao látex, manifestações psicossomáticas (síndrome do pânico, globus hystericus e a síndrome de Münchausen), bem como as tradicionais e mais frequentes modalidades de anafilaxia (alergia a alimentos, medicamentos e insetos). O tratamento na crise aguda da anafilaxia idiopática é o mesmo do que nas demais anafilaxias, incluindo a administração intramuscular imediata de epinefrina. Deve haver uma generosa e prolongada prescrição de corticoterapia oral, e também a instituição de medicação preventiva (anti-histamínicos anti- H1 e anti-H2, cetotifeno, albuterol oral, montelucaste, cromoglicato de sódio, e por último o omalizumabe). Os pacientes devem portar epinefrina autoinjetora e ser instruídos sobre como agir em caso de um episódio anafilático. Eles respondem bem à administração de epinefrina. A corticoterapia oral, por 4-6 semanas, pode induzir uma remissão completa.

Idiopathic anaphylaxis is a condition of unknown etiology. Its prevalence ranges from 10 to 35% of all cases of anaphylaxis. Clinical symptoms and signs are those of classic anaphylaxis, including urticaria, angioedema, flushing, itching, hypotension, tachycardia, gastrointestinal manifestations (dysphagia, nausea, vomiting, abdominal cramps, and diarrhea), asthma, laryngeal edema, dizziness, and syncope. Mortality is rare. There is no genetic transmission, but about 40% of patients are atopic. It is more common in adults than in children, affecting mainly women. It is considered a diagnosis of exclusion of other known forms of anaphylaxis. Mast cell activation occurs with cytoplasmatic degranulation of mediators of anaphylaxis (tryptase and histamine, among others). Because idiopathic anaphylaxis is a steroid-responsive condition, if it is not controlled with adequate doses of prednisone/prednisolone, the diagnosis should be challenged. The differential diagnosis of idiopathic anaphylaxis includes indolent systemic mastocytosis, clonal mast cell activation syndromes, galactose-alpha-1,3- galactose allergy, exercise-induced anaphylaxis (both food- and drug-dependent and -independent), hereditary angioedema (congenital and acquired), pheochromocytoma, carcinoid syndrome, oral mite anaphylaxis, Anisakis simplex allergy, vocal cord dysfunction, scombroid poisoning, semen allergy, latex allergy, psychosomatic conditions (panic attacks, globus hystericus, and Münchausen syndrome), and the classic forms of anaphylaxis (food, drug, and insect allergies). Treatment of acute idiopathic anaphylaxis is the same as in the other forms of anaphylaxis, including intramuscular epinephrine, but with prolonged oral corticosteroid therapy. It might also include other oral preventive medications (H1 and H2 antihistamines, ketotifen, oral albuterol, montelukast, sodium cromoglycate, and recently omalizumab). Patients should have an epinephrine auto-injector and be instructed on self-management of anaphylaxis. Good response to epinephrine is observed, and oral corticosteroid therapy for 4-6 weeks can induce complete remission.

Effects of thyroid hormones on cardiac function in mice by inducing expression of B-cell activating factor in cardiomyocytes / 医学研究生学报

Objective　High levels of triiodothyronine (T3) can lead to hyperthyroid heart disease, but its mechanism is unclear. This study aims to investigate the effects of T3 on the expression of B-cell activating factor (BAFF) in cardiomyocytes and to explore its possible role in the pathogenesis of hyperthyroid heart disease.　Methods　Sixty healthy C57BL/6J mice were selected and randomly divided into two groups: the experimental group and the control group. The experimental group received intraperitoneal injection of T3 at 5 μg/ml, one time/d, for 42 consecutive days. The concentrations of serum T3 and tetraiodothyronine (T4) were detected by radioimmunoassay; ELISA was used to determine BAFF expression in peripheral blood, and the cardiac index and the transverse diameter of myocardial cells in each group were determined. Immunohistochemistry and Western blot were used to detect the expression of BAFF protein in myocardium and of myocardial tumor necrosis factor-α (TNF-α) protein; the expression of BAFF mRNA in myocardium was detected by Real-Time PCR; flow cytometry (FCM) was used to detect changes in the proportion of B-cells in the heart.　Results　Compared with the control group, the serum T3 concentration, cardiac index, BAFF and myocardial cell transverse diameter of the experimental group significantly increased (P<0.05), and the T4 concentration decreased (P<0.05). Under the light microscope, the cardiomyocytes of the control group were normal, while those of the experimental group were hypertrophied and disordered in structure. Compared with the control group (0.765±0.164), BAFF protein expression significantly increased in the experimental group (1.865±0.290) (P<0.05). Compared with the control group (0.537±0.089), the expression of TNF-α protein significantly increased in the experimental group (0.737±0.065) (P<0.05). Correlation analysis of T3 with BAFF gene expression in cardiomyocytes and BAFF level in peripheral blood showed that T3 was positively correlated with both the former with a correlation coefficient of 0.637 (P<0.01) and the latter with 0.778 (P<0.01). For FCM, compared with the control group [(12.40±1.09)%], the proportion of myocardial B-cells increased in the experimental group [(16.12±0.631)%] (P<0.05).　Conclusion　High concentration of T3 can promote the expression of BAFF in myocardial cells and lead to the activation of B-cells, thus increasing the inflammatory response and leading to myocardial hypertrophy.

Expression profile analysis of long noncoding RNAs and mRNAs in the human colon cancer cells regulated by VTCN1 / 上海交通大学学报（医学版）

Objective • To investigate the effect of V-set domain containing T cell activation inhibitor 1 (VTCN1) on long noncoding RNAs (lncRNAs) and mRNAs expression in colon cancer cells. Methods • VTCN1 was overexpressed by lentivirus plasmid in colon cancer cell line SW1116. RNA was extracted and sequenced. The differentially expressed lncRNAs and mRNAs were compared with the negative control group. The accuracy of RNA sequencing was verified by real-time quantitative PCR (qRT-PCR) using three differentially expressed lncRNAs and two mRNAs. BLAST2GO and Kyoto Encyclopedia of Genes and Genomes (KEGG) were used to analyze and predict the functions of these differentially expressed lncRNAs and mRNAs. The online platform GEPIA18 (Gene Expression Profiling Interactive Analysis) was used to analyze the correlation between differentially expressed lncRNAs and survival of patients with colorectal cancer. Results • A total of 167 differential genes, i.e., 39 lncRNAs and 128 mRNAs, were identified by RNA sequencing in VTCN1 overexpressed SW1116 cells. The results of qRT-PCR were consistent with those of RNA sequencing. Bioinformatics analysis showed that these different genes regulated by VTCN1 may be involved in endoplasmic reticulum protein processing and RNA monitoring signaling pathways. In addition, three lncRNAs (DNA JC9-AS1, HCG27, and RP11-339B21.13), which were significantly up-regulated in colorectal cancer cells overexpressing VTCN1, were also independent predictors of overall survival of colorectal cancer patients. Conclusion • In colon cancer cells, VTCN1 regulates several downstream lncRNAs and mRNAs, which may be involved in endoplasmic reticulum protein processing and mRNA monitoring signaling pathways.

Single-cell Analysis of CAR-T Cell Activation Reveals A Mixed T1/T2 Response Independent of Differentiation / 基因组蛋白质组与生物信息学报·英文版

The activation mechanism of chimeric antigen receptor (CAR)-engineered T cells may differ substantially from T cells carrying native T cell receptor, but this difference remains poorly understood. We present the first comprehensive portrait of single-cell level transcriptional and cytokine signatures of anti-CD19/4-1BB/CD28/CD3ζ CAR-T cells upon antigen-specific stimulation. Both CD4 helper T (T) cells and CD8 cytotoxic CAR-T cells are equally effective in directly killing target tumor cells and their cytotoxic activity is associated with the elevation of a range of T1 and T2 signature cytokines, e.g., interferon γ, tumor necrotic factor α, interleukin 5 (IL5), and IL13, as confirmed by the expression of master transcription factor genes TBX21 and GATA3. However, rather than conforming to stringent T1 or T2 subtypes, single-cell analysis reveals that the predominant response is a highly mixed T1/T2 function in the same cell. The regulatory T cell activity, although observed in a small fraction of activated cells, emerges from this hybrid T1/T2 population. Granulocyte-macrophage colony stimulating factor (GM-CSF) is produced from the majority of cells regardless of the polarization states, further contrasting CAR-T to classic T cells. Surprisingly, the cytokine response is minimally associated with differentiation status, although all major differentiation subsets such as naïve, central memory, effector memory, and effector are detected. All these suggest that the activation of CAR-engineered T cells is a canonical process that leads to a highly mixed response combining both type 1 and type 2 cytokines together with GM-CSF, supporting the notion that polyfunctional CAR-T cells correlate with objective response of patients in clinical trials. This work provides new insights into the mechanism of CAR activation and implies the necessity for cellular function assays to characterize the quality of CAR-T infusion products and monitor therapeutic responses in patients.

Deubiquitinases as pivotal regulators of T cell functions / 医学前沿

T cells efficiently respond to foreign antigens to mediate immune responses against infections but are tolerant to self-tissues. Defect in T cell activation is associated with severe immune deficiencies, whereas aberrant T cell activation contributes to the pathogenesis of diverse autoimmune and inflammatory diseases. An emerging mechanism that regulates T cell activation and tolerance is ubiquitination, a reversible process of protein modification that is counter-regulated by ubiquitinating enzymes and deubiquitinases (DUBs). DUBs are isopeptidases that cleave polyubiquitin chains and remove ubiquitin from target proteins, thereby controlling the magnitude and duration of ubiquitin signaling. It is now well recognized that DUBs are crucial regulators of T cell responses and serve as potential therapeutic targets for manipulating immune responses in the treatment of immunological disorders and cancer. This review will discuss the recent progresses regarding the functions of DUBs in T cells.

Piperacillin-Tazobactam-Induced Linear IgA Bullous Dermatosis Supported by a T-Cell Activation Assay

Linear immunoglobulin (Ig) A bullous dermatosis (LABD) is a rare subepidermal autoimmune blistering disease characterized by linear IgA deposits at the basement membrane zone visualized with direct immunofluorescence (DIF). Most cases of LABD are idiopathic, but some are drug-induced with vancomycin being the most common causative agent. We herein report a patient presenting with blisters and erosive lesions, primarily in the intertriginous and flexor areas, consistent with a diagnosis of piperacillin-tazobactam-induced LABD based on the patient's clinical course and histopathology, DIF, and in vitro T-cell activation assay (TAA) findings. Only one case of piperacillin-tazobactam-induced LABD has been previously reported. In addition to its rarity, our case was also unique in that the skin lesions occurred in the intertriginous and flexor areas, uncommon locations for typical adult patients with LABD, and TAA strongly suggested an association with the causative drug.

Mechanism of TUDCA for retinitis pigmentosa / 国际眼科杂志(Guoji Yanke Zazhi)

@#Tauroursodeoxycholic acid(TUDCA)is formed by taurine conjugated of ursodeoxycholic acid(UDCA). It has the role of neurotrophic factor in anti-inflammatory, anti-apoptosis and reducing the activation of microglial cells. These effects may be one of the most critical of all pathological stages of retinitis pigmentosa. Preclinical trials have shown that TUDCA had potential therapeutic value for retinal degeneration disease. This article discusses how TUDCA can slow down the process of retinal degeneration.

Activation of CD8+T cells regulated by γ-aminobutyric acid and its receptors / 器官移植

Objective To evaluate the effect of γ-aminobutyric acid (GABA) and its receptors upon the proliferation of CD8+T cells.Methods The splenic CD8+T cells of Balb/c mice were obtained by CD8+f cell magnetic bead sorting kit.Under the effect of CD3/CD28-activated magnetic bead,CD8+T cells were stimulated by different concentrations of GABA.5-bromo-2-deoxyuridine (BrdU) labeling and flow cytometry were performed to detect the proliferation of CD8+T cells.The expression levels of GABA-A and GABA-B receptor before and after CD8+T cell activation were compared by fluorescent quantitative real-time polymerase chain reaction (PCR).Result Flow cytometry result revealed that GABA could inhibit the proliferation of activated CD8+T cells,manifested as significant decrease in the quantity of CD152+CD8+T cells.Fluorescent quantitative real-time PCR demonstrated that GABA-A receptor subtypes α2,α6 and GABA-B receptor subtype 1a were expressed only when the CD8+T cells were activated.After CD8+T cell activation,the quantity of GABA-A receptor subtypes α3,αs,β2,β3,γ1,γ2 and θ were significantly increased,whereas the quantity of GABA-B2R and GABA-B1b did not significantly differ before and after CD8+T cell activation.Conclusions GABA can suppress the proliferation of activated CD8+T cells.The activation of CD8+T cells is regulated by GABA receptors.However,the underlying mechanism remains to be elucidated.

High Intensity Interval Training Increases Natural Killer Cell Number and Function in Obese Breast Cancer-challenged Mice and Obese Women

High intensity interval training (HIIT) boosts natural killer (NK) cell number and activity in normal weight breast cancer patients; however, whether this occurs in obese individuals is not well established. The goal of this study was to determine whether HIIT effectively boosts NK cells as a therapeutic strategy against breast cancer in an obese mouse model and in overweight/obese women. Diet induced female C57Bl/6 obese mice were assigned to undergo HIIT for four weeks or remain sedentary. Female participants were subjected to a six weeks HIIT protocol. HIIT mice acclimatized to treadmill running were subsequently injected with 5 × 105 polyoma middle T (MT) breast cancer cells intravenously. NK cell number and activation were monitored using flow cytometry, and tumor burden or lipid content evaluated from histological lung and liver tissues, respectively. In both mice and humans, circulating NK cell number and activation (CD3−NK1.1+CD27+ and CD3−CD56+, respectively) markedly increased immediately after HIIT. HIIT obese mice had reduced lung tumor burden compared to controls following MT challenge, and had diminished hepatic lipid deposition despite minimal body weight loss. Our findings demonstrate that HIIT can benefit obese individuals by enhancing NK cell number and activity, reducing tumor burden, and enhancing metabolic health.

Prostaglandin E2 induced myofibrolast phenotype inversion and interleukin 8 expression in prostate stromal cells / 国际检验医学杂志

Objective To investigate the effects of prostaglandin E2(PGE2) in prostate mesenchymal cell activation .Methods To culture human prostate stromal cells WPMY‐1 in vitro ,implement immunofluorescence staining after treating with DMSO and 10-9 mol/L PGE2 and detect the change of the myofibroblast phenotype .The expression of cellular inflammatory factor interleukin 8(IL‐8) was detected by real‐time quantitative PCR and the concentration of IL‐8 was detected .Results PGE2 significantly in‐creased the cellular proportion of colocalization with alpha SMA and Vimentin in WPMY‐1 cells .PGE2 promoted the expression of IL‐8 in WPMY‐1 cells .Conclusion PGE2 can increase myofibroblast phenotype in human prostate mesenchymal cells WPMY‐1 in vitro culture ,promotes the IL‐8 expression ,which has an important role in the occurrence and development of prostate cancer .

Vitamin C is taken up by human T cells via sodium-dependent vitamin C transporter 2 (SVCT2) and exerts inhibitory effects on the activation of these cells in vitro / 대한해부학회지

Vitamin C is an essential micronutrient that affects immune responses. T cells are one of the main players in acquired immunity and have been reported to be influenced by in vivo vitamin C supplementation. Yet, the way by which T cells uptake vitamin C and what direct effects vitamin C exerts on the cells are not known. To elucidate, we isolated human peripheral blood T cells and analyzed the expression of sodium-dependent vitamin C transporters (SVCT). T cells were activated in vitro in the absence or presence of vitamin C, before or after activation. As results, human T cells expressed SVCT2, but not SVCT1, and the expression level increased following activation. Vitamin C added in the culture media generally did not affect T-cell behaviors following activation, such as proliferation, apoptosis, expression of CD25 and CD69, and interleukin 2 secretion, regardless whether it was added before or after activation. However, exceptionally, high concentration vitamin C, when it was added before activation, but not after activation, did exert toxic effects on cell activation with respect to the above-mentioned parameters. In conclusion, we showed the expression of SVCT2 in human T cells for the first time. Vitamin C exerted toxic effects, at least in vitro, when the concentration was high and when it was given before activation. These toxic effects are not thought to be via anti-oxidant effects of vitamin C.

T cell receptor signaling pathway is overexpressed in CD4 + T cells from HAM/TSP individuals

ABSTRACT Human T-lymphotropic virus type 1 (HTLV-1) is a human retrovirus related to the chronic neuroinflammatory disease HTLV-1-associated myelopathy/tropical spastic paraparesis (HAM/TSP). CD4+ T cells activation appears to play a key role on HTLV-1 infection. Here we investigated the expression of genes associated to T cell activation CD3e molecule, epsilon (CD3?), lymphocyte-specific protein tyrosine kinase (LCK), vav 1 guanine nucleotide exchange factor (VAV1), and zeta-chain (TCR) associated protein kinase 70 kDa (ZAP70) on T lymphocytes of HTLV-1-infected individuals and compared to healthy uninfected individuals (CT). We observed that CD3?, LCK, ZAP70, and VAV1 gene expression were increased in CD4+ T cells from HAM/TSP group compared to HTLV-1 asymptomatic patients (HAC). Moreover, ZAP70 and VAV1 were also upregulated in HAM/TSP compared to CT group. We detected a positive correlation among all these genes. We also observed that CD3?, LCK, and VAV1 genes had a positive correlation with the proviral load (PVL) and Tax expression. These results suggest that PVL and Tax protein could drive CD3?, LCK, and VAV1 gene expression in CD4+ T cells, and these genes function on a synchronized way on the CD4+ T cell activation. The elucidation of the mechanisms underlying T cell receptor signaling pathway is of considerable interest and might lead to new insights into the mechanism of HAM/TSP.