Retinoic acid-inducible gene I (RIG-I) is a cytosolic viral RNA receptor. Upon viral infection, the protein recognizes and then recruits adapter protein mitochondrial antiviral signaling (MAVS) protein, initiating the production of interferons and proinflammatory cytokines to establish an antiviral state. In the present study, we identify zinc finger protein 205 (ZNF205) which associates with RIG-I and promotes the Sendai virus (SeV)-induced antiviral innate immune response. Overexpression of ZNF205 facilitates interferon-beta (IFN-β) introduction, whereas ZNF205 deficiency restricts its introduction. Mechanistically, the C-terminal zinc finger domain of ZNF205 interacts with the N-terminal tandem caspase recruitment domains (CARDs) of RIG-I; this interaction markedly promotes K63 ubiquitin-linked polyubiquitination of RIG-I, which is crucial for RIG-I activation. Thus, our results demonstrate that ZNF205 is a positive regulator of the RIG-I-mediated innate antiviral immune signaling pathway.
Immune checkpoint inhibitors（ICIs） have improved durable clinical responses and longer survival time in patients with cancer. However, biomarkers of long-term survival are poorly understood to predict patients who will benefit from ICIs treatment. Here, by analyzing bulk RNA-seq of two ICIs-treated discovery cohorts（160patients）, we found that CHIT1 is significantly higher expressed in patients who acquire long-term overall survival（OS）. Further, we demonstrate that higher CHIT1 markedly prolonged progression-free survival（PFS） and OS in the two ICIs-treated discovery and five validation cohorts（198 patients）. Moreover, baseline of chitotriosidase greater than15 ng/ml in serum of anti-PD-1 treated patients gained longer PFS（12.4 months vs 9.5 months, P=0.039, HR=0.2,95% CI=0.04 to 1.07）. In addition, gene set enrichment analysis（GSEA） indicates that CHIT1 is positively related to hot tumor immune microenvironment（TIME） in the above cohorts and TCGA data. Together, our data suggests that CHIT1 is a novel potential biomarker for predicting the long-term OS in ICIs-treated cohorts.
目的 研究mTORC1抑制剂通过肿瘤浸润T淋巴细胞生物学活性影响KRAS基因突变型结肠癌的进展.方法 建立KRAS突变型结肠癌小鼠模型,将小鼠随机分为4组:对照组,KRAS突变组,mTORC1抑制剂组和KRAS突变+mTORC1抑制剂组;流式细胞术检测肿瘤组织中肿瘤浸润淋巴T细胞的数量;ELISA法检测小鼠血清中IL-2、IL-10、IFN-γ、TGF-β的表达情况;另外构建KRAS突变型结肠癌细胞系HT29,将细胞分为4组:HT-29组,KRAS突变组,mTORC1抑制剂组和KRAS突变+mTORC1抑制剂组;CCK-8法检测结肠癌细胞的增殖情况;流式细胞术检测其凋亡情况;Western blot法检测p53、Bcl-2、Bax、cleaved caspase3蛋白的表达水平.结果 与KRAS突变组结肠癌小鼠相比较,KRAS突变+mTORC1抑制剂组小鼠肿瘤组织中CD4+T细胞、CD8+T细胞数量增加(P＜0.01,P＜0.001),Foxp3+Treg细胞的数量减少(P＜0.000 1),小鼠血清中IL-2、IFN-γ的水平升高(P＜0.000 1,P＜0.001),IL-10、TGF-β的水平降低(均P＜0.001);另外,与KRAS突变型HT29组细胞相比,KRAS突变+mTORC1抑制剂组细胞的增殖能力减弱(P＜0.05),并且凋亡情况显著提高(P＜0.000 1),并且Bcl-2蛋白水平降低(P＜0.001),Bax、cleavedcaspase3、p53蛋白水平显著升高(P＜0.01,P＜0.001,P＜0.001).结论 mTORC1抑制剂通过影响肿瘤浸润淋巴细胞的分布情况以及通过p53/caspase相关通路抑制KRAS基因突变型结肠癌的进展.
目的 研究培菲康通过下调Toll样受体4(TLR4)/核转录因子-κB(NF-κB)信号通路减轻哮喘小鼠气道炎症及气道重塑的机制.方法 以卵白蛋白和氢氧化铝混悬液致敏、激发建立急性哮喘小鼠模型,随机分为4组(每组12只):模型组、培菲康低剂量(0.1g/kg)组、培菲康高剂量(0.2g/kg)组、培菲康(0.2g/kg)+脂多糖(TLR4激活剂,0.4 mg/kg)组,另取12只小鼠以等剂量生理盐水处理作为对照组以培菲康、LPS对小鼠分组进行干预后,评测各组小鼠哮喘症状评分;以瑞氏染色分类计数各组小鼠支气管肺泡灌洗液(BALF)中炎症细胞;以HE染色、Masson染色检测各组小鼠肺组织病理形态和气道重塑,并评测小鼠气管壁厚度和气道平滑肌厚度;以酶标仪检测各组小鼠BALF中炎症相关因子白细胞介素(IL)-17、肿瘤坏死因子-α(TNF-α)、IL-4及IL-10水平;以蛋白免疫印迹检测各组小鼠肺组织TLR4/NF-κB信号通路相关蛋白表达.结果 与对照组相比,模型组小鼠肺组织出现严重病理损伤和气道重塑,哮喘症状评分、气管壁厚度、气道平滑肌厚度、BALF中巨噬细胞、淋巴细胞计数和I L-17、TNF-α、IL-4水平、肺组织p-NF-κB p65/NF-κB p65及TLR4蛋白表达水平显著升高(P＜0.05),抗炎因子IL-10水平显著降低(P＜0.05);与模型组相比,培菲康低、高剂量组小鼠肺组织病理损伤和气道重塑均减轻,哮喘症状评分、气管壁厚度、气道平滑肌厚度、BALF中巨噬细胞、淋巴细胞计数和IL-17、TNF-α、IL-4水平、肺组织p-NF-κBp65/NF-κB p65及TLR4蛋白表达水平均降低(P＜0.05),抗炎因子IL-10水平均升高(P＜0.05);与培菲康低剂量组相比,培菲康高剂量组小鼠肺组织病理损伤和气道重塑减轻,哮喘症状评分、气管壁厚度、气道平滑肌厚度、BALF中巨噬细胞、淋巴细胞计数和IL-17、TNF-α、IL-4水平、肺组织p-NF-κB p65/NF-κB p65及TLR4蛋白表达水平降低(P＜0.05),抗炎因子IL-10水平升高(P＜0.05);与培菲康高剂量组相比,培菲康+LPS组小鼠肺组织病理损伤和气道重塑加重,哮喘症状评分、气管壁厚度、气道平滑肌厚度、BALF中巨噬细胞、淋巴细胞计数和IL-17、TNF-α、IL-4水平、肺组织p-NF-κB p65/NF-κB p65及TLR4蛋白表达水平升高(P＜0.05),抗炎因子IL-10水平降低(P＜0.05).结论 培菲康可通过抑制TLR4/NF-κB信号而减轻哮喘小鼠气道炎症,缓解其肺组织病理损伤和气道重塑.
目的 研究N-乙酰半胱氨酸酰胺(NACA)对脂多糖(LPS)诱导的RAW264.7巨噬细胞炎症的保护作用及作用机制,为NACA治疗脓毒症提供理论依据.方法 CCK-8法检测NACA和N-乙酰半胱氨酸(NAC)对RAW264.7细胞活力的影响;采用ELISA检测NACA和NAC对LPS诱导的RAW264.7细胞上清中TNF-α和IL-6水平;qRT-PCR检测NACA对LPS诱导的RAW264.7 细胞中 TNF-α、IL-6、IL-1β、ATF4、CHOP 的 mRNA 表达的影响;Western blot 检测 NACA 对 LPS 诱导的 RAW264.7 细胞中TLR4/NF-κB及PERK/ATF4/CHOP信号通路中关键蛋白表达的影响.结果 0.1～5 mmol/L的NACA和NAC对RAW264.7细胞活力无明显的影响(P＞0.05).1 mmol/L和5 mmol/L的NACA和NAC可显著降低LPS诱导的RAW264.7细胞上清中TNF-α和IL-6的水平(P＜0.01),且NACA的降低效果优于统计量的NAC.1 mmol/L和5 mmol/L的NACA也可显著降低LPS诱导的 RAW264.7 细胞中 TNF-α、IL-6、IL-1β、ATF4、CHOP mRNA 的表达(P＜0.05、P＜0.01).Western blot结果表明,1 mmol/L和5 mmol/L 的 NACA 可显著降低 TLR4、p-IκB、p-65、p-PERK、p-eIF2α、ATF4、Bip 和 CHOP 蛋白的表达(P＜0.05、P＜0.01).结论 NACA可能通过TLR4/NF-κB及PERK/ATF4/CHOP信号通路抑制LPS诱导的RAW264.7巨噬细胞的炎症反应,为NACA治疗脓毒症的抗炎性药物筛选提供了实验依据.
This study aimed to establish a mouse model with diphtheria toxin-induced depletion of CD4 + T cells（referred to as Cd4-DTR mouse） by CRISPR/Cas9 technology, that is, IRES-DTR segment was inserted at the stop codon of Cd4 gene by homologous recombination. The proportion and number of CD4 + T cells in the spleen and cranial dura were detected by flow cytometry. Data showed that the number and proportion of CD4 + T cells in the spleen and cranial dura decreased significantly after Cd4-DTR mice were injected with 75 mg/kg diphtheria toxin.In the spleen, CD4 + T cell ablation could last more than 1 week after a single diphtheria toxin injection. And we found that the colon pathological changes were milder in Cd4-DTR mice with acute colitis induced by 3% DSS. So,deletion of T cells may have a protective effect on DSS-induced acute colitis. Therefore, Cd4-DTR murine model has been constructed successfully, which provides a new mouse model for the study of CD4 + T cell function.
目的 探讨T辅助细胞22(Th22)在小鼠Lewis肺癌抗肿瘤免疫中的作用及其机制.方法 通过皮下接种Lewis肺癌细胞悬液建立Lewis肺癌小鼠模型,IL-22中和抗体(IL-22 nAb)组小鼠在此基础上腹腔注射IL-22 nAb(50 μg/kg).14d后处死小鼠,计算肿瘤质量和抑瘤率.流式细胞术检测小鼠Th22和调节性T细胞(Treg)的表达情况;RT-PCR法检测小鼠肿瘤组织TAP1和CTLA-4 mRNA表达;酶联免疫吸附法检测小鼠血清IL-4、IL-10、IL-22和TGF-β含量.结果 与对照组比较,模型组小鼠Th22细胞数量和IL-22水平均明显升高(P＜0.05);与模型组比较,IL-22nAb组小鼠肺癌组织的质量和Treg细胞数量明显降低(P＜0.05),肺癌组织TAP1和CTLA-4mRNA表达显著降低(P＜0.05),血清IL-4、IL-10和TGF-β表达明显降低(P＜0.05).结论 IL-22通过调控Treg细胞功能,参与肺癌免疫逃逸过程.
This study was designed to investigate the effects of ethanol extract from Rubus rosaefolius Smith on acute gouty arthritis in rats.The model of acute gouty arthritis was established by monosodium urate in rats.The swelling degree of ankle and the scores of gait were measured;the levels of TNF-α,IL-1β and IL-6 were detected by ELISA;the mRNA levels of NF-κB P65 and its inhibitor α （IκBα） in synovial tissue of ankle were detected by qRT-PCR.Histopathological observation of synovial tissue was carried out,and NF-κB p65 in synovial tissue was detected by immunohistochemistry.Data showed that compared with the model group,middle and high doses of the ethanol extract could significantly decrease the swelling degree of ankle and the scores of gait （P＜0.05 or P＜0.01）.Middle and high doses of the ethanol extract could significantly decrease the level of TNF-α （P＜0.05 or P＜0.01）.High dose of the ethanol extract could significantly decrease the levels of IL-1β and IL-6 （P＜0.05 or P＜0.01）,and down-regulate the expression of NF-κB p65 mRNA （P＜0.05）.Middle and high doses of the ethanol extract could significantly up-regulate the expression of IκBα mRNA（P＜0.05）.The pathological results showed that the ethanol extract could alleviate the damage in synovial tissue;the results of immunohistochemistry showed that there were fewer positive staining cells,lighter staining and lower average optical density in the high dose group compared with the model group （P＜0.01）.In conclusion,the ethanol extract from Rubus rosaefolius Smith has mitigative effect on acute gouty arthritis in rats,which is probably related to its inhibitory effects on inflammatory factors and NF-κB.
Intestinal tract is an important digestive and immune organ, and its immune dysfunction is an important pathological feature of inflammatory bowel disease（IBD）. Understanding of the immunopathology of IBD has promoted the development of targeted therapy for IBD. Now antibody biologics targeting different targets have been developed, which has changed the treatment mode of IBD. Here we will review the research progress of therapeutic antibody in IBD.
It is well-recognized that gram-negative bacteria can induce and aggravate inflammatory process in chronic rhinosinusitis（CRS）. However, the pathological mechanism needs to be further confirmed. Pyroptosis has been found to be closely related to inflammatory process. This study aimed at exploring whether LPS, a key component derived from Gram-negative bacteria, could induce and aggravate pyroptosis in primary human nasal epithelial cells（HNEpCs） in CRS. Nasal mucosa tissues were collected from CRS patients and control individuals.The expression levels of pyroptosis-related proteins were detected by IHC. Normal HNEpCs（N-HNEpCs） and CRS HNEpCs（CRS-HNEpCs） were extracted from nasal mucosa tissues and cultured. Then N-HNEpCs and CRSHNEpCs were randomly divided into four groups（Control group, LPS 1 mg/L group, LPS 5 mg/L group and LPS 25mg/L group）. Cell survival rate was detected by CCK-8;EthD-I staining combined with GSDMD was used to detect pyroptosis; the expression of inflammatory mediators IL-18, IL-1β and NLRP3 inflammasome（NLRP3, ASC, cleaved Caspase-1） were detected by immunofluorescence, Western blotting and RT-qPCR. IHC showed that the expressions of NLRP3, cleaved Caspase-1, IL-18 and IL-1β were significantly increased（P＜0.01） in CRS patients. In vitro, LPS stimulation of different concentrations could decrease the survival rate of N-HNEpCs and CRS-HNEpCs（P＜0.05）, while elevate the number of EthD-I staining positive cells and the expression of GSDMD（P＜0.05） in a dose-dependent manner.Immunofluorescence, Western blot and RT-qPCR results showed that 5 mg/L LPS stimulation could significantly up-regulate the expression of inflammatory mediators IL-18, IL-1β and NLRP3 inflammasome in N-HNEpCs and CRS-HNEpCs（P＜0.05）. In conclusion, LPS can induce pyroptosis in N-HNEpCs by activating NLRP3inflammasome, and can further promote the progression of pyroptosis in CRS-HNEpCs, suggesting that Gramnegative bacteria may cause and aggravate CRS lesions by inducing nasal epithelial cells pyroptosis.
目的 通过评估系统性红斑狼疮(systemic lupus erythematosus,SLE)患者,尤其是合并感染的SLE患者,淋巴细胞亚群和细胞因子的变化,以期待找到SLE患者的免疫特征.方法 回顾性分析2019年5月至2022年6月共150例SLE患者(其中58例合并感染)的基本信息、临床表现和实验数据.同时采用流式细胞术检测22例健康对照者外周血淋巴细胞亚群和细胞因子水平.结果 与健康对照组相比,SLE患者中CD19+B细胞的绝对细胞数目、百分比显著增高,而CD16+CD56+NK细胞的绝对数、百分比显著下降(均P＜0.01);白介素-6、白介素-10、肿瘤坏死因子-α显著增高(均P＜0.05),而干扰素-γ明显降低(P＜0.05).与非感染SLE患者相比,感染性SLE患者发热(27.6％vs 12％)、浆膜炎(39.7％vs19.6％)的发生率明显增高(均P＜0.05),皮疹(3.4％vs30.4％)、关节炎(8.6％vs34.8％)发生率降低;红细胞沉降率、C反应蛋白、抗双链DNA抗体水平显著升高(均P＜0.05);而血红蛋白水平显著降低(均P＜0.05);系统性红斑狼疮疾病活动指数无明显差异.本研究还发现,与非感染性SLE患者相比,感染性SLE患者外周血中CD3+T细胞绝对数显著下调[1 040.5(655.3～1 436.8)vs 716.5(406.5～1 115.8)]、CD4*T细胞绝对数下调[366(239.8～525.5)vs 234.5(162.75～373.3),均P＜0.01].进一步的相关性分析结果表明,SLE患者外周血CD3+T细胞的绝对数量与红细胞沉降率(rs=-0.392,P＜0.001)和C反应蛋白(rs=-0.178,P=0.026)呈负相关.结论 SLE患者外周淋巴细胞亚群和细胞因子分布异常,下降的CD3+T细胞与红细胞沉降率和C反应蛋白成负相关,感染性SLE患者具有更低的CD3+和CD4+T淋巴细胞.因此,对SLE患者监测淋巴细胞亚群及细胞因子,对判断是否感染及指导临床治疗具有重要意义.
B cells are derived from the bone marrow（BM） and maturated in peripheral organs, which is important for humoral immunity. Upon stimulation with antigen, B cells differentiate into plasma cells to produce specific antibodies or differentiate into memory B cells. Mitochondria are the energy production and metabolism center of cells and responsible for providing sufficient energy to cells. When the function of mitochondria is damaged, the energy supply is affected, which changes the fate of cells. Conversely, different states of the cell will also affect the function of mitochondria. Recent studies have shown that mitochondria function play an important role in the development and activation of B cells to adapt to phenotypic and environmental changes encountered during the cell life cycle. This article mainly reviews the role of mitochondria in the development, differentiation and activation of B cells, as well as the changes that occur at different stages and states of B cells, which may provide a new idea to explore the mechanism of mitochondrial diseases and a new theoretical basis for the treatment of B-cell related diseases.
This study was designed to investigate the effect of interleukin-12（IL-12） on the biological behavior and immune escape factor secretion of non-small cell lung cancer（NSCLC） cells. The expression levels of IL-12 and STAT3 mRNA in 126 patients with NSCLC were detected by real-time quantitative PCR. A human lung adenocarcinoma A549 cell model was constructed and divided into blank control group, adenovirus control group,IL-12 adenovirus group, plasmid control group. STAT3 plasmid group, and IL-12 + STAT3 co-transfection groups.The proliferation and apoptosis of A549 cells in each group were compared; the secretion of IL-2, INF-γ and TNF-α in the cell culture medium were detected by ELISA, and the expression levels of IL-12, STAT3 and pSTAT3protein were detected by Western blotting. Data showed that the expression of IL-12 mRNA was decreased and the expression of STAT3 mRNA was increased in lung cancer tissues compared with adjacent non-cancerous tissues.IL-12 adenovirus group showed decreased cancer cell proliferation and viability, and increased apoptosis compared with each control group, while the results of cell proliferation and apoptosis in STAT3 plasmid group were opposite.Cell proliferation, viability and apoptosis in IL-12 +STAT3 co-transfection groups were intermediate between those in IL-12 adenovirus group and STAT3plasmid group. In addition, the contents of IL-12, TNF-α and IFN-γ in the cell culture medium of the IL-12 adenovirus group were significantly higher than those of the STAT3 plasmid group, while the expression levels of STAT3 and p-STAT3 protein in the STAT3 plasmid group were significantly higher than those in the IL-12adenovirus group. In conclusion, IL-12 can inhibit the proliferation, decrease the viability and induce the apoptosis of NSCLC cells, which may be achieved by inhibiting STAT3 pathway and immune escape.
This study was designed to detect blood cell-derived inflammatory indexes in patients with psoriasis vulgaris （PsV） and evaluate their correlation with the severity and histopathology changes of PsV.The clinical data of 99 patients with PsV （PsV group） and 99 surgical inpatients with no history of psoriasis or inflammatory disease （control group） were collected retrospectively in our hospital,and the blood cell-derived inflammatory indexes were compared between the two groups.According to the PASI score,the PsV group was divided into mild （PASI＜10） and moderate-to-severe （PASI≥10） subgroups,and the blood cell-derived inflammatory indexes were compared in the two subgroups.Spearman correlation analysis was carried out to evaluate the correlation between blood cell-derived inflammatory indexes and with PASI score in PsV patients;receiver operating characteristic （ROC） curve was used to evaluate the predictive value of blood cell-derived inflammatory indexes for the severity of PsV.The clinical data of 41 PsV patients underwent skin biopsy and blood routine examination were collected,and spearman correlation analysis was used to evaluate the relationship of blood cells and their derived inflammatory indexes with the morphometric results of skin biopsy,including maximum epidermal thickness （ETmax） and minimum epidermal thickness of nipple （SPETmin）.Data showed that the NLR,MLR,SIRI,SII,AISI,d NLR and NLPR values of the PsV group were significantly higher than those of the control group （all P＜0.05）;the PLR,MLR,SIRI,SII,AISI and d NLR values of the moderate-to-severe group were significantly higher than those of the mild psoriasis group （all P＜0.05）;the PLR,MLR,SIRI,SII,AISI and d NLR were positively correlated with PASI score （r=0.23,0.27,0.25,0.25,0.31,0.30;all P＜0.05）;ROC curve showed that the best sensitivity of PLR,SII and AISI to predict the severity of PsV were 62.9%,40.5%and 56.8%,while the best specificity were 70.3%,85.5%and 77.4%.Among them,AISI had the best value in predicting the severity of PsV,with area under curve （AUC） of 0.711[95%CI （0.606,0.815）].Furthermore,NEU was positively correlated with ETmax （r=0.40,P＜0.01）.In conclusion,the PLR,SII and AISI values are positively correlated with the severity of PsV,and have a certain value for evaluating the severity of PsV;the increase of NEU in peripheral blood may be related to the maximum epidermal thickening of skin biopsy histomorphometry in PsV patients.
目的 研究lncRNA PCGEM1/miR-155-5p轴对LPS诱导的气管平滑肌细胞增殖和凋亡及炎症反应的影响.方法 采用100μg/ml LPS刺激支气管平滑肌细胞BSMC 12 h以诱导细胞损伤.实时荧光定量PCR(RT-qPCR)检测细胞lncRNA PCGEM1和miR-155-5p表达水平;细胞计数试剂盒(CCK-8)法检测细胞增殖;流式细胞术检测细胞凋亡;蛋白质印迹(Western blot)法检测细胞CyclinD1、Cleaved-caspase-3蛋白表达;双荧光素酶报告实验检测lncRNA PCGEM1和miR-155-5p靶向关系.结果 与NC组比较,LPS组细胞lncRNA PCGEM1表达水平显著降低,miR-155-5p表达水平显著升高(P＜0.05).与pcDNA+LPS组比较,pcDNA-lncRNA PCGEM1+LPS组lncRNA PCGEM1表达水平显著升高,吸光度值、CyclinD1蛋白表达显著降低,细胞凋亡率、Cleaved-caspase-3 蛋白表达显著升高,TNF-α、IL-6、IL-33 水平显著降低(P＜0.05).与 anti-miR-NC+LPS 组比较,anti-miR-155-5p+LPS组miR-155-5p表达水平显著降低,吸光度值、CyclinD1蛋白表达显著降低,细胞凋亡率、Cleaved-caspase-3蛋白表达显著升高,TNF-α、IL-6、IL-33 水平显著降低(P＜0.05).与 miR-NC+pcDNA-lncRNA PCGEM1+LPS组比较,miR-155-5p+pcDNA-lncRNAPCGEM1+LPS组细胞miR-155-5p表达水平显著升高,吸光度值、CyclinD1蛋白表达显著升高,细胞凋亡率、Cleaved-caspase-3蛋白表达显著降低,TNF-α、IL-6、IL-33水平显著升高(P＜0.05).结论 lncRNA PCGEM1 下调miR-155-5p降低LPS诱导的气管平滑肌细胞凋亡及炎症反应,抑制增殖.
Neuroblastoma is one of the most common extracranial solid tumors in children, accounting for 15% of pediatric cancer-related deaths. CDK2 silencing leads to apoptosis in MYCN positive neuroblastoma, but not in MYCN negative neuroblastoma. However, little is known about whether cyclin dependent kinase 2（CDK2） inhibits MYCN negative neuroblastoma tumorigenesis and its effect on tumor immune microenvironment. In this study, we evaluated the effect of CDK2 inhibitor PF-07104091 or CDK2 stable knockdown on the tumor growth of subcutaneous MYCN negative neuroblastoma cells SH-EP in C57BL/6N WT mice, so as to explore whether CDK2inhibitors or knockdown of CDK2 affect the immune microenvironment of MYCN negative neuroblastoma. Data showed that both PF-07104091 and CDK2 knockdown significantly inhibited the growth of MYCN negative neuroblastoma and increased murine overall survival of mice; CDK2 inhibitors or CDK2 knockdown did not affect the proliferation or apoptosis of MYCN negative neuroblastoma cells. PF-07104091 and CDK2 knockdown groups demonstrated significant increase in the percentages of CD3 + T cells, CD4 + T cells, CD8 + T cells, and DC cells. After CDK2 inhibitors or CDK2 knockdown treatment, the level of IFN-γ in MYCN negative neuroblastoma increased,and the expressions of the key genes of IFN-γ pathway IFNB1, STAT1, TLR3, DDX58, CCL17, TAP1, TX2 were up-regulated. Intraperitoneal injection of CDK2 inhibitor PF-07104091 did not affect the growth rate and survival IFNGR1with or without CDK2 knockdown had similar tumor growth rates and survival rates in IFNGR1 -/- mice.Therefore, CDK2 affects the ratio of CD3 + T cells, CD4 + T cells, CD8 + T cells, and DC cells in the tumor immune microenvironment through the IFN-γ pathway.
Even though smallpox has been eradicated, the threat of accidental or intentional release has highlighted the fact that there is little consensus about correlates of protective immunity or immunity against reinfection with the causative poxvirus, variola virus（VARV）. As the existing vaccine for smallpox has unacceptable rates of side effects and complications, new vaccines are urgently needed. Surrogate animal models of VARV infection in humans, including vaccinia virus（VACV） and ectromelia virus（ECTV） infection in mice, monkeypox virus（MPXV） infection in macaques, have been used as tools to dissect the immune response to poxviruses.Mousepox, caused by ECTV, a natural mouse pathogen, is arguably the best surrogate small-animal model, as it shares many aspects of virus biology, pathology and clinical features with smallpox in humans. The requirements for recovery from a primary ECTV infection have been well characterized, which include type I and II interferons,natural killer cells, CD4T cells, CD8T cell effector function and antibody. From a vaccine standpoint, it is also imperative that the requirement for recovery from secondary infection. We have investigated host immune parameters in immune responses to a secondary ECTV infection, and have identified that interferon and CD8T cell effector functions are not essential; however, T-and B-cell interaction and antibody are absolutely critical for recovery from a secondary challenge. The central role of antibody has been also identified in the secondary response to other poxviruses. These findings have important clinical implications and would greatly assist the design of therapeutic interventions and new vaccines for smallpox.
Based on network pharmacology and in vitro experimental verification, this study was designed to explore the potential mechanism of polydatin（PD） against bacterial pneumonia from the point of its polarization modulation on macrophages. Drug targets were predicted by SwissTargetPrediction, SEA, TCMSP and PharmMapper databases; disease targets were obtained in GeneCards and DisGeNET databases; GO and KEGG enrichment analysis were established by DAVID databases; protein interaction networks were constructed using STRING databases. And in vitro cell experiments were carried out to further verify the results of network pharmacology. The RAW264.7 cells were divided into three groups: model control group（model, M） stimulated with LPS, PD experimental group treated with PD, and blank control group（Control, C）. CCK8 was used to detect the effect of different concentrations of PD on cell viability; ELISA was used to detect the secretion levels of NF-κB and IL-1β, the terminal effector molecules of SIRT1/HMGB1 signaling pathway, and the macrophage polarization factors MCP-1 and Arg1 in the cell supernatant; qPCR was used to detect the mRNA expression of SIRT1/HMGB1 signaling pathway related molecules SIRT1, HMGB1, TLR4 and TRAF6 in cell pellets. According to the results of network pharmacological analysis, there are 61 intersection targets were screened out, in which SIRT1, HMGB1, TLR4, TRAF6 and NF-κB were identified as subsequent experimental targets by combining the results of GO and KEGG enrichment analysis and protein interaction network construction.The results of cell viability experiments confirmed that the secretion levels of NF-κB, IL-1β and MCP-1 in the PD group were significantly lower, while Arg1 was significantly higher than those in the M group; the mRNA expression of SIRT1 in PD group was significantly up-regulated, but the mRNA expressions of HMGB1, TLR4 and TRAF6were significantly down-regulated. In summary, polydatin can regulate the expression of signaling pathway molecules such as TLR4 and NF-κB and polarizing factors MCP-1 and Arg1 through SIRT1/HMGB1, and modulate macrophage polarization, thus may be a potential drug for the treatment of bacterial pneumonia.
This study was designed to investigate the effects of HOXA9 expression on the proliferation,migration and invasion of mouse bladder cancer cell BTT739. Before the cell experiment, we detected the expression of HOXA9 gene in the samples of bladder urothelial cancer and adjacent normal tissues from surgical operation by immunohistochemistry. Then in later experiments, the expression of HOXA9 gene in BTT739 cells was knockdown by sh-RNA and the proliferation, apoptosis, migration and invasion were detected by clone formation, flow cytometry, CCK8, Transwell and scratch test, respectively. The expression of epithelial to mesenchymal transition（EMT） characteristic factors, including E-cadherin, Vimentin and Snail, were measured by Western blotting. The results showed that the expression of HOXA9 gene in bladder cancer tissues was much higher than that in the adjacent normal tissues. Both the mRNA and protein expression of HOXA9 gene in BTT739 cells were successfully knocked down, which led to a significant reduction in cells proliferation, migration and invasion, a significant increase in the early apoptosis percentage and the proportion of S phase, while a significant decrease in the proportion of G1 and G2 phase. Meanwhile, EMT characteristic factor E-cadherin expression was enhanced while the expression of Vimentin and Snail were decreased. In conclusion, HOXA9 gene may be an oncogene with the function of promoting BTT739 cells proliferation, migration and invasion, and the mechanism of this process involves EMT.Therefore, HOXA9 is expect to become a potential target for immunotherapy of bladder cancer.
Adhesion-mediated colonization is closely associated with persistent Helicobacter pylori（H. pylori）infection. Various adhesion proteins on the surface of H. pylori exhibit specific tropism towards human gastric mucosal epithelial cells by recognizing specific glycan structures on the host cell surface. In recent years, the functions and structures of some adhesion proteins have been elucidated: different adhesion proteins recognize different gastric epithelial cell ligands and are associated with immunopathological damage such as chronic inflammation and intestinal epithelial chemotaxis due to infection, in addition to their involvement in bacterial adhesion. This review summarizes the progress of research on three important adhesion proteins, HpaA, BabA and SabA, and their application as potential targets in the development of vaccines against of H. pylori.
