Neurological Immunotoxicity from Cancer Treatment

The emergence of immune-based treatments for cancer has led to a growing field dedicated to understanding and managing iatrogenic immunotoxicities that arise from these agents. Immune-related adverse events (irAEs) can develop as isolated events or as toxicities affecting multiple body systems. In particular, this review details the neurological irAEs from immune checkpoint inhibitors (ICI) and chimeric antigen receptor (CAR) T cell immunotherapies. The recognition and treatment of neurological irAEs has variable success, depending on the severity and nature of the neurological involvement. Understanding the involved mechanisms, predicting those at higher risk for irAEs, and establishing safety parameters for resuming cancer immunotherapies after irAEs are all important fields of ongoing research.     

直接竞争酶联免疫吸附法检测食品中三聚氰胺残留

采用酶标抗原建立了高效、高灵敏的三聚氰胺直接竞争的ELISA（enzyme linked immunosorbent assay,ELISA）检测方法。方法 以三聚氰胺单克隆抗体包被作为固相抗体,辣根过氧化酶（Horseradish Peroxidase,HRP）标记的抗原与标准品（或样品）中三聚氰胺竞争结合抗体,建立了直接竞争酶联免疫检测体系。结果 以三聚氰胺为标准品建立标准曲线,得到方法的IC50为8.84μg/L,灵敏度为0.65μg/L,线性范围0.9-35μg/L;检测样品的回收率为70.0%-127.9%,与三聚氰酸交叉反应率为60%,其他结构类似物基本无交叉反应。结论 本方法灵敏度高、特异性强、检测快速, 可以满足实际样品的检测需求。     

乳腺癌疫苗中肿瘤抗原的鉴定

鉴定乳腺癌疫苗中激活小鼠抗肿瘤免疫反应的肿瘤抗原。利用蛋白质组学方法根据免疫小鼠血清中存在的特异性抗体寻找肿瘤疫苗所含的肿瘤抗原。结果表明,肿瘤疫苗免疫小鼠的抗体效价大于6400,证明免疫小鼠血清中抗体含量高,且其抗肿瘤体液免疫得到增强;SDS-PAGE结合免疫印迹结果证明,软骨多糖作用后的Ca761细胞,与免疫血清在50 kDa左右出现了特异性结合的蛋白条带;二维电泳结合免疫印迹确定了激活免疫反应的肿瘤抗原为50 kDa酸性蛋白,经质谱技术鉴定,该肿瘤抗原为α-tubulin。综上所述,Ca761细胞中的肿瘤抗原α-tubulin经软骨多糖作用后,其结构及表达量发生了变化,该变化的抗原即为肿瘤疫苗中激活小鼠抗肿瘤免疫反应的抗原,因此肿瘤疫苗免疫成功小鼠可以阻止肿瘤细胞的侵袭;α-tubulin经软骨多糖作用后其免疫原性增强,因此其可以开发成为以抗原为基础的疫苗。     

Chimeric Antigen Receptor Design and Efficacy in Ovarian Cancer Treatment

Our increased understanding of tumour biology gained over the last few years has led to the development of targeted molecular therapies, e.g., vascular endothelial growth factor A (VEGF-A) antagonists, poly[ADP-ribose] polymerase 1 (PARP1) inhibitors in hereditary breast and ovarian cancer syndrome (BRCA1 and BRCA2 mutants), increasing survival and improving the quality of life. However, the majority of ovarian cancer (OC) patients still do not have access to targeted molecular therapies that would be capable of controlling their disease, especially resistant or relapsed. Chimeric antigen receptors (CARs) are recombinant receptor constructs located on T lymphocytes or other immune cells that change its specificity and functions. Therefore, in a search for a successful solid tumour therapy using CARs the specific cell surface antigens identification is crucial. Numerous in vitro and in vivo studies, as well as studies on humans, prove that targeting overexpressed molecules, such as mucin 16 (MUC16), annexin 2 (ANXA2), receptor tyrosine-protein kinase erbB-2 (HER2/neu) causes high tumour cells toxicity and decreased tumour burden. CARs are well tolerated, side effects are minimal and they inhibit disease progression. However, as OC is heterogenic in its nature with high mutation diversity and overexpression of different receptors, there is a need to consider an individual approach to treat this type of cancer. In this publication, we would like to present the history and status of therapies involving the CAR T cells in treatment of OC tumours, suggest potential T cell-intrinsic determinants of response and resistance as well as present extrinsic factors impacting the success of this approach.     

Early Lung Cancer Diagnosis by Biosensors

Lung cancer causes an extreme threat to human health, and the mortality rate due to lung cancer has not decreased during the last decade. Prognosis or early diagnosis could help reduce the mortality rate. If microRNA and tumor-associated antigens (TAAs), as well as the corresponding autoantibodies, can be detected prior to clinical diagnosis, such high sensitivity of biosensors makes the early diagnosis and prognosis of cancer realizable. This review provides an overview of tumor-associated biomarker identifying methods and the biosensor technology available today. Laboratorial researches utilizing biosensors for early lung cancer diagnosis will be highlighted.     

shRNA沉默P115基因对胃癌细胞增殖相关蛋白表达的影响及机制

目的探讨高尔基体囊泡转运蛋白(colgi-vesicular transport protein,P115)基因沉默对胃癌细胞株BGC-823中增殖相关因子细胞周期素D1(cyclinD1)、微小染色体维持缺陷蛋白2(mini chromosome maintenance protein 2,Mcm2)和增殖细胞核抗原(proliferating Cell Nuclear Antigen,PCNA)表达的影响及其可能的机制。方法采用脂质体介导法将重组表达质粒P115-shRNA2(P115-shRNA2组)和阴性对照质粒shNC(阴性对照组)转染至高表达P115的胃癌细胞株BGC-823中,并设未转染组,经G418筛选出稳定转染细胞,实时定量PCR(Real-time PCR)法检测细胞中P115、巨噬细胞移动抑制因子(macrophage migration inhibitory factor,MIF)、cyclinD1、Mcm2和PCNA基因mRNA的转录水平;Western blot法检测细胞中P115、MIF、pERK1/2、cyclinD1、Mcm2和PCNA蛋白表达水平;免疫共沉淀法检测BGC-823细胞中P115与MIF间的相互作用;ELISA法检测细胞上清液中MIF的分泌水平。结果与未转染组和阴性对照组相比,P115-shRNA2组细胞中P115、MIF、cyclinD1、Mcm2和PCNA的mRNA和蛋白表达水平均明显降低,ERK1/2的磷酸化水平明显下调,BGC-823细胞培养上清中MIF的含量明显降低;P115蛋白可在抗MIF的免疫沉淀物中检出,P115和MIF在BGC-823细胞中存在特异性的相互作用。结论 P115基因沉默下调胃癌细胞中cyclinD1、Mcm2和PCNA的表达,其分子机制可能与P115通过调控MIF的表达和分泌,进而启动下游的ERK1/2信号通路有关。P115可作为研究胃癌细胞增殖分子机制的新靶点。     

前列腺抗原酶促化学发光免疫分析方法学的建立

应用鲁米诺-过氧化氢发光体系建立定量测定血清前列腺特异性抗原含量的酶促化学发光免疫分析方法。该方法测量范围为1.5～80μg/L,灵敏度为0.12μg/L,批内变异＜5%,批间变异＜15%。回收率为83.8%～118.7%,稀释实验测定值与稀释度呈线性相关,相关系数为0.999。本方法与免疫放射分析方法的相关方程y=1.07x+0.68,相关系数r=0.97。     

百菌清人工抗原的制备与鉴定

以杀菌剂百菌清为起始反应物,通过两步化学反应合成百菌清的衍生物(半抗原);采用活化酯法与混合酸酐法分别将半抗原与载体蛋白(牛血清蛋白、卵清蛋白)进行偶联,制备百菌清的人工抗原,同时通过免疫新西兰大白兔获得抗血清,用酶联免疫吸附法(ELISA)测定其效价。利用红外光谱(IR)、质谱(MS)、核磁共振波谱(NMR)对百菌清半抗原进行表征,并用紫外扫描的方法对人工抗原进行鉴定。结果表明,成功制备出百菌清半抗原及人工抗原,并且半抗原与载体蛋白的偶联比分别为27.86:1 和12.32:1,抗血清效价约为1:1.28 × 104。     

低剂量辐射（LDR）对淋巴细胞的效应

LDR能刺激成人、儿童和婴幼儿的淋巴细胞转化,对营养不良儿童的刺激效应较低,对脐血则刺激效应较明显。LDR对成人CD4细胞的刺激效应强于CD8细胞,而对同样的刺激效应,NK细胞则需要较大剂量。对淋巴细胞NK活性也有同样规律。肿瘤病人的NK活性的刺激效应低于正常人。LDR增加了小鼠的血和脾内的T辅助／诱导细胞的比例和肿瘤内T细胞毒性细胞。低剂理照射的淋巴细胞外液也能刺激淋巴细胞转化。预先LDR可以减轻随后大剂量照射引起的生物大分子、膜抗原和染色体的损伤。LDR可诱导鼠脾淋巴细胞产生新蛋白,它能增强人和动物的免疫功能。     

脱氧雪腐镰刀菌烯醇人工抗原的研制

本研究利用丁基硼酸封闭DON的7,15-羟基,戊二酸酐酰化3-羟基合成3-HG-DON,通过活泼酯法将3-HG-DON偶联到载体蛋白OVA,制备检测抗原。采用CDI法将DON分子与载体蛋白MBSA偶联制备的免疫抗原免疫3只BALB/c小鼠,经4次免疫后,1号小鼠血清抗体效价达1∶25600,且小鼠抗DON多克隆抗体与DON-HG-OVA的结合能被DON阻断。