芫根调节免疫功能的小肠转录组学研究

利用生物信息学技术初步探索芫根调控肠道免疫功能的分子生物学机制。方法 选择雄性SPF级BALB/c小鼠18只，随机分为3组，每组6只。SC1组小鼠每只每次灌胃芫根提取液150 μL，SC2组小鼠每只每次灌胃绞碎芫根原浆悬液150 μL，NC组小鼠每只每次灌胃生理盐水150 μL，连续7 d每日灌胃一次。分别取每组小鼠小肠组织样品提取RNA，总RNA的质检合格后，进行转录组测序。按GO功能和KEGG信号通路对差异表达基因聚类分析揭示差异表达高度富集的免疫相关通路。从免疫角度分析芫根灌胃小鼠小肠组织的基因表达变化情况。结果 转录组测序共检测到27733条 mRNA的表达，SC1组与NC组比较，显著差异表达基因1635个，其中上调差异表达基因1236个，下调差异表达基因399个；SC2组与NC组比较，显著差异表达基因2872个，其中上调差异表达基因2233个，下调差异表达基因639个（P＜0.05）。按GO功能和KEGG信号通路聚类分析显示差异表达基因在白介素分泌、干扰素反应、T细胞受体信号通路及维生素和脂肪消化吸收等途径的富集度在两个芫根组中均居于前20位（P＜0.01），肠黏膜趋化因子CCL20和巨噬细胞极化标志物CD274等免疫蛋白编码基因差异表达显著且同属于上述多个免疫通路。结论 芫根灌胃小鼠小肠的差异表达基因在白介素分泌、干扰素反应、T细胞受体信号通路、营养物质消化吸收等途径高度富集，提示芫根对肠道免疫系统及肠黏膜功能的调节，其中CD274的表达上调和CCL20的表达下调提示诱导M1型巨噬细胞极化和T细胞活化可能是芫根调控免疫和维护肠道正常结构功能的重要途径。     

Tumor-associated high endothelial venules mediate lymphocyte entry into tumors and predict response to PD-1 plus CTLA-4 combination immunotherapy

1. Download : Download high-res image (268KB)
2. Download : Download full-size image

     

ω-3多不饱和脂肪酸对慢性移植物抗宿主病狼疮小鼠的免疫调控作用

目的探讨ω-3多不饱和脂肪酸(ω-3 poly unsaturated fatty acids,ω-3 PUFAs)对慢性移植物抗宿主病(chronic graft versus host disease,cGVH)狼疮模型小鼠的免疫调节功能影响。方法单次腹腔注射bm12小鼠淋巴细胞建立cGVH小鼠模型,造模当天分别给予90%ω-3 PUFAs和97%EPA灌胃治疗,持续14 d。通过流式细胞术检测小鼠脾脏淋巴细胞中各免疫指标,ELISA检测小鼠血清中总IgG水平。结果与正常对照组相比,cGVH组明显下调Treg亚群,明显上调T_FH细胞、生发中心B细胞和浆细胞亚群;与模型对照组相比,90%ω-3和97%EPA均明显上调Treg亚群,并明显下调T_FH细胞、生发中心B细胞和浆细胞亚群;且97%EPA组小鼠血清中总IgG水平明显下降。结论在cGVH狼疮小鼠模型中,ω-3 PUFAs通过增加Treg细胞,减少T_FH细胞、生发中心B细胞和浆细胞,抑制IgG的分泌,从而发挥免疫抑制作用,且其免疫抑制作用可能与ω-3 PUFAs含量相关。     

Newly recruited intraepithelial Ly6A+CCR9+CD4+ T cells protect against enteric viral infection

1. Download : Download high-res image (215KB)
2. Download : Download full-size image

     

The Neutrophil to Lymphocyte Ratio Predicts the Response to Neoadjuvant Chemotherapy in Luminal B Breast Cancer

Objective: Tumor response to neoadjuvant chemotherapy (NAC) in breast cancer (BC) patients is a predictor foroverall survival. The aim of our study was to determine a relationship between the neutrophil to lymphocyte ratio(NLR) prior to NAC, BC subtypes and the probability of a pathologic complete response (pCR). Materials andMethods: Medical records were collected retrospectively from Centro de Cancer at Red Salud UC-Christus. Clinicaldata collected included peripheral blood cell counts, BC subtype at diagnosis and the pathology report of surgeryafter chemotherapy. Results: A total of 88 patients were analyzed. Approximately, a 25% had a pCR, and displayed asignificant correlation between BC subtype and pCR (p= 0.0138 Chi2); this was more frequent in epidermal growthfactor receptor type 2 (HER2) enriched subtype patients (54%). Luminal B BC patients with a pCR had significantlylower NLR levels (t test, p= 0.0181). Conclusions: HER2-enriched tumors had a higher probability of pCR. In LuminalB tumors, NLR had a statistically significant relationship with the probability of pCR. In this subtype, NLR could bea useful biomarker to predict tumor response to NAC. Further studies including other clinical parameters for systemicinflammation such as platelet counts, intratumoral NLR or body mass index could help identify patients that wouldget the most benefit from NAC.     

人外周血TIM3 基因敲除T淋巴细胞的制备及其抗肿瘤作用

[摘要] 目的: 通过CRISPR-Cas9 基因编辑技术及质粒电转染技术制备T 细胞免疫球蛋白黏液素3（TIM3）基因敲除的人外周血T淋巴细胞，探讨敲除TIM3 基因后能否增强T细胞免疫功能及其抗肿瘤作用。方法: 通过电转染法将hTIM3 sgRNA/Cas9 双质粒共转染EBV阳性胃癌患者外周血T淋巴细胞，电转24 h 后流式细胞术检测转染效率并利用荧光显微镜观察；体外培养过程中观察基因敲除后T细胞增殖活性，流式细胞术验证TIM3 基因敲除效率以及细胞表型的变化。用肿瘤抗原肽活化T细胞检测TIM3 基因敲除后T 细胞分泌细胞因子水平及体外杀伤胃癌AGS-EBV 细胞的能力。结果:电转染法能够有效地将hTIM3 sgRNA/Cas9 双质粒敲除体系转入人外周血T淋巴细胞，其转染效率平均达（41.5±3.6）%，基因敲除效率波动在40.0%～50.0%（均P<0.01）；经抗原活化后的基因敲除组T细胞的增殖活性和免疫表型未见明显变化，表面活化分子中仅见HLA-DR较对照组明显提高（P<0.05）；TIM3 基因敲除T细胞分泌TNF-α、IFN-γ 的水平显著增高（P<0.01 或P<0.05），体外杀伤胃癌AGS-EBV 细胞的能力也明显增强（均P<0.05）。结论: 用CRISPR-Cas9 基因编辑及质粒电转染技术制备人外周血TIM3 基因敲除T淋巴细胞的方法简易可行，在体外的扩增活化中保持TIM3 基因水平下调并具有更高的免疫应答水平以及更强的抗肿瘤作用。这一新技术的研发为基因工程化细胞免疫治疗提供了新的思路。     

D-二聚体和中性粒细胞/淋巴细胞比值评估卵巢癌预后的临床价值

目的:研究D-二聚体（D-dimer,D-D）和中性粒细胞/淋巴细胞比值（neutrophil to lymphocyte ratio,NLR）与卵巢癌临床病理特征及预后的关系。方法:回顾性分析2012年1月至2015年12月于我院妇科行手术治疗的卵巢恶性肿瘤患者387例和卵巢良性肿瘤患者250例临床资料。比较血清D-D和外周血NLR在卵巢良、恶性肿瘤中的表达水平；确定D-D和NLR临界值，D-D+NLR=0（D-D≤0.555 mg/L和NLR≤2.792）,D-D+NLR=1（D-D>0.555 mg/L或NLR>2.792），D-D+NLR=2（D-D>0.555 mg/L和NLR>2.792），分析两者联合的评分系统与卵巢癌临床病理特征和预后的关系。结果:血清D-D和外周血NLR在卵巢良、恶性肿瘤患者中的表达水平有统计学差异（P＜0.001）。D-D高水平组与低水平组相比，患者的分期、分级、淋巴结转移、腹水、CA125水平、残余瘤大小有统计学差异（P＜0.05）。NLR高水平组与低水平组相比，患者的年龄、分期、淋巴结转移、腹水、CA125水平、残余瘤大小有统计学差异（P＜0.05）。D-D+NLR为0、1、2分的平均总生存期（OS）分别为70个月、58个月、40个月。D-D+NLR评分是影响OS的独立预后因素。结论:术前血清D-D和外周血NLR与卵巢癌临床病理特征和OS相关，D-D+NLR评分可以作为评估卵巢癌预后的指标。     

Chronological Changes in Neutrophil/lymphocyte Ratio in Advanced Gastric Cancer Patients Treated with Nivolumab: a Report of Nine Cases

Background: Nivolumab has been approved for use in advanced gastric cancer (GC) after third-line chemotherapy in Japan. However, it remains difficult to predict favorable nivolumab response before treatment. Methods: We evaluated the clinical course with a focus on the chronological changes in neutrophil/lymphocyte ratio (NLR) throughout the chemotherapy and assessed the relationship between nivolumab response and chronological changes in NLR before nivolumab administration. Results: We experienced nine cases who received nivolumab monotherapy for unresectable advanced or postoperative recurrent GC. Nivolumab was used as third-line chemotherapy in all patients, and partial response (PR) and stable disease (SD) were observed in two patients each. Nivolumab treatment resulted in progressive disease (PD) in five patients. In patients with PR or SD, changes in the NLR tended to correspond to the response of target metastatic lymph nodes to first- and second-line chemotherapy. In the four cases with PR or SD following nivolumab, ∆NLRresponses that was the difference in the degree of decline during the most effective pretreatment chemotherapy were 1.39, 0.73, 1.62, and 1.22. However, the patients with PD showed lower ∆NLRresponses, at 0.66, 0.66, 0.25, 0.13, and -0.05 in the five cases. Mean ∆NLRresponses in the patients with PR or SD and patients with PD were 1.17 and 0.33, respectively (P = 0.0008). Conclusions: We experienced nine GC cases treated with nivolumab and assessed the association between chronological NLR changes throughout chemotherapy and tumor response to nivolumab. Changes in NLR during pretreatment chemotherapy might predict tumor response to nivolumab monotherapy in patients with advanced GC.     

支原体肺炎患儿免疫功能变化的检测价值

目的分析免疫功能变化在支原体肺炎感染患儿中的检测价值。方法在2016年8月-2018年8月在医院接受诊治的114例支原体肺炎患儿与114例健康体检者作为本次研究。按根据是否属于支原体肺炎划分为试验组与常规组。试验组与常规组均接受免疫功能变化检测,总结并对比两组补体、免疫球蛋白水平、红细胞免疫水平以及T淋巴细胞亚群水平。结果试验组患儿的各项补体、免疫球蛋白水平相对于常规组而言均有明显数据差异(P<0.05);试验组患儿的各项细胞免疫水平相对于常规组而言均有明显数据差异(P<0.05);试验组患儿的各项T淋巴细胞亚群水平相对于常规组而言均有明显数据差异(P<0.05)。结论免疫功能变化在支原体肺炎感染患儿中的检测价值突出,可以作为患儿症状的检测与症状严重程度的判断依据。