CN117054649A - Chronic atrophic gastritis transformation marker for stomach cancer and application thereof - Google Patents
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Abstract
本发明提供了一种慢性萎缩性胃炎向胃癌转化标志物及其应用，所述标志物为PMN‑MDSCs。本发明首次经过研究发现PMN‑MDSCs的表达量随胃“炎‑癌”进展而逐渐上调，PMN‑MDSCs含量与慢性萎缩性胃炎患者OLGA分级分期呈正相关)。进一步计算PMN‑MDSCs对于OLGAIII/IV分期的分段效能，发现ROC曲线下面积为0.9333，灵敏度为1，特异性为0.8。因此，PMN‑MDSCs可作为慢性萎缩性胃炎向胃癌转化的特异性标志物，对慢性萎缩性胃炎高癌变风险患者具有优异的诊断效能，有助于对慢性萎缩性胃炎患者进行危险分层，筛选出胃癌高风险患者，从而实现胃癌的早诊早治。
The invention provides a marker for the transformation of chronic atrophic gastritis into gastric cancer and its application. The marker is PMN-MDSCs. For the first time, the present invention found that the expression of PMN-MDSCs gradually increases with the progression of gastric "inflammation-cancer", and the content of PMN-MDSCs is positively correlated with the OLGA classification and staging of patients with chronic atrophic gastritis). Further calculation of the segmental efficacy of PMN‑MDSCs for OLGA III/IV staging revealed that the area under the ROC curve was 0.9333, the sensitivity was 1, and the specificity was 0.8. Therefore, PMN-MDSCs can be used as a specific marker for the transformation of chronic atrophic gastritis into gastric cancer, have excellent diagnostic performance for patients with chronic atrophic gastritis and high cancer risk, and are helpful for risk stratification and screening of patients with chronic atrophic gastritis. Patients at high risk of gastric cancer can be identified to achieve early diagnosis and treatment of gastric cancer.
Description
技术领域Technical field
本发明属于分子诊断技术领域，具体涉及一种慢性萎缩性胃炎向胃癌转化标志物及其应用。The invention belongs to the field of molecular diagnosis technology, and specifically relates to a marker for the transformation of chronic atrophic gastritis into gastric cancer and its application.
背景技术Background technique
我国每年新发胃癌约41万例，属于影响国民健康的重大慢性非传染性疾病，降低我国胃癌的发病率和病死率成为亟待解决的重大公共健康问题。我国胃癌以肠型为主，肠型胃癌具有相对固定的演化范式，即从慢性炎症开始，发展为慢性萎缩性胃炎(chronicatrophic gastritis，CAG)、肠上皮化生、上皮内瘤变，最终到胃癌。其中慢性萎缩性胃炎及其伴发的伴肠上皮化生、上皮内瘤变是重要的胃癌前疾病，与胃癌的发生关系密切。因此，开发能够监测慢性萎缩性胃炎“炎-癌”转化和预测胃癌风险的生物标志物对于胃癌的早诊早治有重要意义。There are approximately 410,000 new cases of gastric cancer in my country every year, which is a major chronic non-communicable disease that affects national health. Reducing the incidence and mortality of gastric cancer in my country has become a major public health issue that needs to be solved urgently. Gastric cancer in my country is dominated by intestinal type. Intestinal type gastric cancer has a relatively fixed evolution paradigm, that is, it starts from chronic inflammation, develops into chronic atrophic gastritis (CAG), intestinal metaplasia, intraepithelial neoplasia, and finally gastric cancer. . Among them, chronic atrophic gastritis and its associated intestinal metaplasia and intraepithelial neoplasia are important gastric precancerous diseases and are closely related to the occurrence of gastric cancer. Therefore, the development of biomarkers that can monitor the "inflammatory-cancer" transformation of chronic atrophic gastritis and predict the risk of gastric cancer is of great significance for the early diagnosis and treatment of gastric cancer.
髓系抑制性细胞(MDSCs)是一群具有强大免疫抑制作用的细胞群体，在形成免疫抑制微环境中发挥关键作用。MDSCs由髓样细胞正常分化受阻大量扩增而来。正常生理情况下，造血干细胞分化为共同髓系祖细胞，共同髓系祖细胞再分化为未熟髓系细胞(IMCs)，IMCs进一步分化为成熟的中性粒细胞、巨噬细胞或树突状细胞。当机体发生肿瘤、炎症、感染、创伤等病理情况时，IMCs的正常分化可被炎性因子或肿瘤来源的细胞因子阻滞，诱导其成为MDSCs，并在外周血、骨髓或病变部位募集、扩增、活化。MDSCs能否识别慢性萎缩性胃炎患者的胃癌风险尚未见报道。Myeloid suppressor cells (MDSCs) are a group of cells with powerful immunosuppressive effects and play a key role in forming an immunosuppressive microenvironment. MDSCs are massively expanded from myeloid cells that are blocked from normal differentiation. Under normal physiological conditions, hematopoietic stem cells differentiate into common myeloid progenitor cells, which then differentiate into immature myeloid cells (IMCs), and IMCs further differentiate into mature neutrophils, macrophages or dendritic cells. . When tumors, inflammation, infection, trauma and other pathological conditions occur in the body, the normal differentiation of IMCs can be blocked by inflammatory factors or tumor-derived cytokines, inducing them to become MDSCs and recruit and expand in peripheral blood, bone marrow or diseased sites. Increase, activate. Whether MDSCs can identify the risk of gastric cancer in patients with chronic atrophic gastritis has not been reported.
发明内容Contents of the invention
基于此，本发明的目的在于提供一种慢性萎缩性胃炎向胃癌转化标志物PMN-MDSC及其在制备慢性萎缩性胃炎向胃癌转化风险评估试剂中的应用。Based on this, the purpose of the present invention is to provide a PMN-MDSC marker for the transformation of chronic atrophic gastritis to gastric cancer and its application in preparing a risk assessment reagent for the transformation of chronic atrophic gastritis to gastric cancer.
为达到上述目的，本发明采用如下技术方案。In order to achieve the above object, the present invention adopts the following technical solutions.
人PMN-MDSC作为标志物在制备慢性萎缩性胃炎向胃癌转化风险评估试剂中的应用。Application of human PMN-MDSC as a marker in the preparation of risk assessment reagents for the transformation of chronic atrophic gastritis to gastric cancer.
本发明还提供了检测人PMN-MDSC在生物样本中含量的试剂在制备慢性萎缩性胃炎向胃癌转化风险评估产品中的应用。The present invention also provides the application of a reagent for detecting the content of human PMN-MDSC in biological samples in preparing a risk assessment product for the transformation of chronic atrophic gastritis to gastric cancer.
在一些实施例中，所述生物样本为人外周血样本。In some embodiments, the biological sample is a human peripheral blood sample.
在一些实施例中，所述产品为人流式细胞术检测试剂盒。In some embodiments, the product is a human flow cytometry detection kit.
本发明还提供了人PMN-MDSC作为标志物在筛选慢性萎缩性胃炎向胃癌转化风险评估试剂中的应用。The present invention also provides the application of human PMN-MDSC as a marker in screening reagents for risk assessment of the transformation of chronic atrophic gastritis to gastric cancer.
在一些实施例中，所述人PMN-MDSC的流式细胞术分析标记为In some embodiments, the flow cytometric analysis of human PMN-MDSC is labeled as
HLA-DR-CD11b+CD14-CD15+，HLA-DR阴性、CD11b阳性、CD14阴性、CD15阳性。HLA-DR - CD11b + CD14 - CD15 + , HLA-DR negative, CD11b positive, CD14 negative, CD15 positive.
本发明还提供了一种慢性萎缩性胃炎向胃癌转化风险评估试剂盒，所述试剂盒包括检测人PMN-MDSC在生物样本中含量的试剂。The present invention also provides a risk assessment kit for the transformation of chronic atrophic gastritis into gastric cancer, which kit includes a reagent for detecting the content of human PMN-MDSC in biological samples.
在一些实施例中，所述试剂包括以下抗体：人HLA-DR抗体、人CD11b抗体、人CD14抗体、人CD15抗体。In some embodiments, the reagents include the following antibodies: human HLA-DR antibody, human CD11b antibody, human CD14 antibody, human CD15 antibody.
在一些实施例中，所述抗体上修饰有不同荧光基团。In some embodiments, the antibodies are modified with different fluorescent groups.
在一些实施例中，所述试剂盒为人流式细胞术检测试剂盒。In some embodiments, the kit is a human flow cytometry detection kit.
本发明首次经过研究发现慢性浅表性胃炎、慢性萎缩性胃炎和胃癌患者的血液样本中PMN-MDSCs的表达量随胃“炎-癌”进展而逐渐上调，差异具有统计学意义。PMN-MDSCs含量与慢性萎缩性胃炎患者OLGA分级分期呈正相关(r＝0.6427,p＝0.0072)。进一步计算PMN-MDSCs对于OLGAIII/IV分期的分段效能，发现ROC曲线下面积为0.9333，灵敏度为1，特异性为0.8，p＝0.0048。因此，PMN-MDSCs可作为慢性萎缩性胃炎向胃癌转化的特异性标志物，对慢性萎缩性胃炎高癌变风险患者具有优异的诊断效能，有助于对慢性萎缩性胃炎患者进行危险分层，筛选出胃癌高风险患者，从而实现胃癌的早诊早治。For the first time, the present invention found through research that the expression of PMN-MDSCs in blood samples of patients with chronic superficial gastritis, chronic atrophic gastritis and gastric cancer gradually increases with the progression of gastric "itis-cancer", and the difference is statistically significant. There was a positive correlation between PMN-MDSCs content and OLGA classification and stage in patients with chronic atrophic gastritis (r=0.6427, p=0.0072). Further calculation of the segmental efficacy of PMN-MDSCs for OLGA III/IV staging revealed that the area under the ROC curve was 0.9333, the sensitivity was 1, the specificity was 0.8, p=0.0048. Therefore, PMN-MDSCs can be used as a specific marker for the transformation of chronic atrophic gastritis into gastric cancer, have excellent diagnostic performance for patients with chronic atrophic gastritis and high cancer risk, and are helpful for risk stratification and screening of patients with chronic atrophic gastritis. Patients at high risk of gastric cancer can be identified to achieve early diagnosis and treatment of gastric cancer.
附图说明Description of the drawings
图1为典型的流式细胞术检测分析结果。Figure 1 shows typical flow cytometry analysis results.
图2为各组患者生物样本中PMN-MDSCs含量的统计分析结果。Figure 2 shows the statistical analysis results of PMN-MDSCs content in biological samples of patients in each group.
图3为PMN-MDSCs与OLGA分级分期的相关性分析结果。Figure 3 shows the correlation analysis results between PMN-MDSCs and OLGA classification and staging.
图4为PMN-MDSCs用于评估胃癌风险的ROC曲线分析结果。Figure 4 shows the ROC curve analysis results of PMN-MDSCs for assessing the risk of gastric cancer.
具体实施方式Detailed ways
本发明下列实施例中未注明具体条件的实验方法，通常按照常规条件，或按照制造厂商所建议的条件。实施例中所用到的各种常用化学试剂，均为市售产品。Experimental methods without specifying specific conditions in the following examples of the present invention usually follow conventional conditions or conditions recommended by the manufacturer. Various commonly used chemical reagents used in the examples are all commercially available products.
除非另有定义，本发明所使用的所有的技术和科学术语与属于本发明的技术领域的技术人员通常理解的含义相同。本发明的说明书中所使用的术语只是为了描述具体的实施例的目的，不用于限制本发明。Unless otherwise defined, all technical and scientific terms used in the present invention have the same meanings commonly understood by those skilled in the technical field belonging to the present invention. The terms used in the description of the present invention are only for the purpose of describing specific embodiments and are not used to limit the present invention.
本发明的术语“包括”和“具有”以及它们任何变形，意图在于覆盖不排他的包含。例如包含了一系列步骤的过程、方法、装置、产品或设备没有限定于已列出的步骤或模块，而是可选地还包括没有列出的步骤，或可选地还包括对于这些过程、方法、产品或设备固有的其它步骤。The terms "including" and "having" and any variations thereof herein are intended to cover a non-exclusive inclusion. For example, a process, method, device, product or equipment that includes a series of steps is not limited to the listed steps or modules, but optionally also includes unlisted steps, or optionally also includes steps for these processes, Other steps inherent to the method, product, or device.
在本发明中提及的“多个”是指两个或两个以上。“和/或”，描述关联对象的关联关系，表示可以存在三种关系，例如，A和/或B，可以表示：单独存在A，同时存在A和B，单独存在B这三种情况。字符“/”一般表示前后关联对象是一种“或”的关系。The "plurality" mentioned in the present invention means two or more. "And/or" describes the relationship between related objects, indicating that there can be three relationships. For example, A and/or B can mean: A exists alone, A and B exist simultaneously, and B exists alone. The character "/" generally indicates that the related objects are in an "or" relationship.
2005年国际萎缩研究小组提出胃黏膜炎性反应、萎缩程度和范围的分级、分期标准(即慢性胃炎OLGA分级分期系统)。该系统根据胃黏膜萎缩范围和程度评估癌变风险。In 2005, the International Atrophy Research Group proposed grading and staging standards for gastric mucosal inflammatory reaction, degree and extent of atrophy (i.e., the OLGA grading and staging system for chronic gastritis). This system assesses the risk of cancer based on the extent and degree of gastric mucosal atrophy.
OLGA分期如下：OLGA’s staging is as follows:
其中，高危等级OLGA分期(Ⅲ/Ⅳ期)与胃癌高危密切相关，Ⅲ期和Ⅳ期属于胃癌高风险患者。Among them, the high-risk OLGA stage (stage III/IV) is closely related to the high risk of gastric cancer, and stages III and IV belong to patients with high risk of gastric cancer.
下面结合具体实施例进行说明。Description will be made below with reference to specific embodiments.
实施例1Example 1
利用流式细胞术检测15例慢性浅表性胃炎(CNAG)、16例慢性萎缩性胃炎(CAG)和7例胃癌(GC)患者的外周血样本中PMN-MDSCs的含量。所有纳入检测的患者为经临床确诊的患者，与患者签署知情同意书后，静脉EDTA抗凝收集患者外周血。Flow cytometry was used to detect the content of PMN-MDSCs in peripheral blood samples of 15 patients with chronic superficial gastritis (CNAG), 16 patients with chronic atrophic gastritis (CAG), and 7 gastric cancer (GC). All patients included in the test were clinically diagnosed. After signing an informed consent form with the patients, the patients' peripheral blood was collected through intravenous EDTA anticoagulation.
利用淋巴细胞分离液(天津灏洋)分离所有外周血样本中的单个核淋巴细胞(PBMC)，操作步骤严格按照试剂盒说明书进行。将分离获得的PBMC按如下步骤进行染色：Lymphocyte separation solution (Tianjin Haoyang) was used to isolate mononuclear lymphocytes (PBMC) in all peripheral blood samples. The operating steps were strictly followed the instructions of the kit. The separated PBMCs were stained as follows:
(1)每个样本对应取1*106个PBMC加入流式管中，然后加入5ml PBS缓冲液混匀，2000rpm/min离心5min，弃上清，收集细胞沉淀；(1) For each sample, add 1*10 6 PBMC into the flow tube, then add 5 ml PBS buffer and mix well, centrifuge at 2000 rpm/min for 5 minutes, discard the supernatant, and collect the cell pellet;
(2)向所述细胞沉淀中加入100μL流式抗体稀释液，所述流式抗体稀释液包含人HLA-DR--APC-cy7抗体、人CD11b--BUV396抗体、人CD14--APC抗体、人CD15--BV605体，按1：200的比例将全部种类的抗体加入PBS缓冲液中进行稀释，得到所述流式抗体稀释液；(2) Add 100 μL of flow antibody diluent to the cell pellet. The flow antibody diluent contains human HLA-DR--APC-cy7 antibody, human CD11b--BUV396 antibody, human CD14--APC antibody, For human CD15--BV605 antibodies, add all types of antibodies into PBS buffer at a ratio of 1:200 for dilution to obtain the flow antibody diluent;
(3)将流式管置于涡旋振荡器上振荡混匀，然后于4℃条件下避光染色30min；(3) Place the flow tube on a vortex oscillator to oscillate and mix, and then stain at 4°C in the dark for 30 minutes;
(4)向流式管中加入预冷的PBS缓冲液，然后于2000rpm/min离心5min，弃上清；加入300μL预冷PBS缓冲液重悬细胞，然后使用流式分析仪(BD Canto II，USA)上机检测，获取数据，并用FlowJo10软件进行分析。(4) Add pre-cooled PBS buffer to the flow tube, then centrifuge at 2000 rpm/min for 5 minutes, discard the supernatant; add 300 μL of pre-cooled PBS buffer to resuspend the cells, and then use a flow cytometer (BD Canto II, USA) was tested on the machine, the data was obtained, and analyzed using FlowJo10 software.
统计分析：采用IBM SPSS统计学软件中的秩和检验进行组间比较，相关性分析探讨PMN-MDSCs与OLGA分期相关性，ROC曲线检测PMN-MDSCs对胃癌高风险患者的诊断效能。以P＜0.05视为差异具有统计学意义。Statistical analysis: The rank sum test in IBM SPSS statistical software was used for comparison between groups, correlation analysis was used to explore the correlation between PMN-MDSCs and OLGA staging, and the ROC curve was used to detect the diagnostic performance of PMN-MDSCs in patients with high risk of gastric cancer. P<0.05 was considered as a statistically significant difference.
图1为典型的流式细胞术检测分析结果。如图2所示，GC患者外周血中PMN-MDSCs的含量明显高于CAG患者和CNAG患者；且CAG患者外周血中PMN-MDSCs的含量明显高于CNAG患者。说明慢性浅表性胃炎、慢性萎缩性胃炎和胃癌患者的血液样本中PMN-MDSCs的表达量随胃“炎-癌”进展而逐渐上调，提示PMN-MDSCs可作为胃“炎-癌”风险评估标志物。Figure 1 shows typical flow cytometry analysis results. As shown in Figure 2, the content of PMN-MDSCs in the peripheral blood of GC patients was significantly higher than that of CAG patients and CNAG patients; and the content of PMN-MDSCs in the peripheral blood of CAG patients was significantly higher than that of CNAG patients. This shows that the expression of PMN-MDSCs in blood samples of patients with chronic superficial gastritis, chronic atrophic gastritis and gastric cancer gradually increases with the progression of gastric "inflammation-cancer", suggesting that PMN-MDSCs can be used as a risk assessment for gastric "inflammation-cancer" landmark.
进一步地，为了评价PMN-MDSCs作为胃“炎-癌”风险评估标志物的效能，将CAG患者根据OLGA分级分期分为低风险患者(Ⅰ期和Ⅱ期)组(10例)与高风险患者(Ⅲ期和Ⅳ期)组(6例)，分析PMN-MDSCs与OLGA分级分期之间的相关性。如图3所示，PMN-MDSCs与OLGA分级分期呈正相关(r＝0.6427,p＝0.0072)。ROC曲线分析结果显示，PMN-MDSCs在用于评估胃癌风险(Ⅲ期和Ⅳ期为高风险)时，曲线下面积为0.9333，灵敏度为1，特异性为0.8，p＝0.0048，诊断性能较好，可作为胃“炎-癌”风险评估特异性标志物。Furthermore, in order to evaluate the effectiveness of PMN-MDSCs as a marker for gastric "inflammatory-cancer" risk assessment, CAG patients were divided into low-risk patient (stage I and II) groups (10 cases) and high-risk patients according to OLGA classification and staging. (Stage III and IV) group (6 cases), the correlation between PMN-MDSCs and OLGA classification and staging was analyzed. As shown in Figure 3, PMN-MDSCs were positively correlated with OLGA grading and staging (r=0.6427, p=0.0072). ROC curve analysis results show that when PMN-MDSCs are used to assess the risk of gastric cancer (stages III and IV are high risk), the area under the curve is 0.9333, the sensitivity is 1, the specificity is 0.8, p=0.0048, and the diagnostic performance is good. , can be used as a specific marker for gastric "itis-cancer" risk assessment.
综上所述，本发明发现PMN-MDSCs在慢性浅表性胃炎、慢性萎缩性胃炎和胃癌患者的血液样本中的表达量随胃“炎-癌”进展而逐渐上调，与OLGA分级分期呈正相关，可作为胃“炎-癌”风险评估特异性标志物，诊断性能较好。In summary, the present invention found that the expression of PMN-MDSCs in the blood samples of patients with chronic superficial gastritis, chronic atrophic gastritis and gastric cancer gradually increases with the progression of gastric "itis-cancer" and is positively correlated with OLGA grading and staging. , can be used as a specific marker for gastric "itis-cancer" risk assessment, with good diagnostic performance.
以上所述实施例的各技术特征可以进行任意的组合，为使描述简洁，未对以上实施例中的各个技术特征所有可能的组合都进行描述，然而，只要这些技术特征的组合不存在矛盾，都应当认为是本说明书记载的范围。The technical features of the above embodiments can be combined in any way. To simplify the description, not all possible combinations of the technical features in the above embodiments are described. However, as long as there is no contradiction in the combination of these technical features, All should be considered to be within the scope of this manual.
以上所述实施例仅表达了本发明的几种实施方式，其描述较为具体和详细，但并不能因此而理解为对本发明专利范围的限制。应当指出的是，对于本领域的普通技术人员来说，在不脱离本发明构思的前提下，还可以做出若干变形和改进，这些都属于本发明的保护范围。因此，本发明专利的保护范围应以所附权利要求为准。The above-mentioned embodiments only express several implementation modes of the present invention, and their descriptions are relatively specific and detailed, but they should not be construed as limiting the patent scope of the present invention. It should be noted that, for those of ordinary skill in the art, several modifications and improvements can be made without departing from the concept of the present invention, and these all belong to the protection scope of the present invention. Therefore, the scope of protection of the patent of the present invention should be determined by the appended claims.
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Citations (18)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102037015A (en) * | 2007-09-21 | 2011-04-27 | 健泰科生物技术公司 | Inhibition of angiogenesis |
| US20120070461A1 (en) * | 2010-09-21 | 2012-03-22 | Immatics Biotechnologies Gmbh | Use of myeloid cell biomarkers for the diagnosis of cancer |
| CN104024276A (en) * | 2012-06-21 | 2014-09-03 | 卡姆普根有限公司 | LSR antibodies and their use for cancer therapy |
| US20150150940A1 (en) * | 2012-05-21 | 2015-06-04 | The Trustees Of Columbia University In The City Of New York | Trefoil family factor proteins and uses thereof |
| US20170247464A1 (en) * | 2014-10-24 | 2017-08-31 | Ose Immunotherapeutics | Method and Compositions for Inducing Differentiation of Myeloid Derived Suppressor Cell to Treat Cancer and Infectious Diseases |
| CN107271675A (en) * | 2017-03-24 | 2017-10-20 | 郑猛 | Anti-human ADRB3 monoclonal antibodies and its application in medical diagnosis on disease and treatment |
| WO2017212018A1 (en) * | 2016-06-10 | 2017-12-14 | INSERM (Institut National de la Santé et de la Recherche Médicale) | Methods and pharmaceutical compositions for the treatment of autoimmune inflammatory diseases |
| CN107533065A (en) * | 2015-03-13 | 2018-01-02 | 布莱恩·J·赫尔尼奇 | Methods for Monitoring CD4+ Type 1 T Helper Cell Responses in Cancer and Immune Recovery |
| US20180164313A1 (en) * | 2015-06-01 | 2018-06-14 | The Wistar Institute Of Anatomy And Biology | Methods for monitoring polymorphonuclear myeloid derived suppressor cells |
| US20180252721A1 (en) * | 2015-09-02 | 2018-09-06 | Syndax Pharmaceuticals, Inc. | Selection of patients for combination therapy |
| CN108982857A (en) * | 2018-09-16 | 2018-12-11 | 上海中医药大学附属龙华医院 | The detection method and its application of a kind of Non-small cell lung carcinoma peripheral blood marrow source property inhibition cell |
| CN109517897A (en) * | 2011-04-28 | 2019-03-26 | 南加利福尼亚大学 | Mankind's marrow source inhibitory cells cancer markers |
| US20210077582A1 (en) * | 2019-09-13 | 2021-03-18 | Duke University | Compositions and methods for increasing the efficacy of anti-pd-1 antibody immunotherapy |
| CN114814209A (en) * | 2022-04-29 | 2022-07-29 | 中山大学附属第一医院 | An early warning detection marker PMN-MDSC for early diagnosis of BPD in premature infants |
| WO2023012844A1 (en) * | 2021-08-02 | 2023-02-09 | Ospedale Pediatrico Bambino Gesu' | Antigen cd111 as a new specific diagnostic and therapeutic marker for polymorphonuclear myeloid-derived suppressor cells (pmn-mdscs) |
| CN116240277A (en) * | 2022-07-11 | 2023-06-09 | 复旦大学附属儿科医院 | Malat 1-mediated polymorphonuclear myeloid-derived suppressor cell production and its use in sepsis |
| CN116348495A (en) * | 2020-09-18 | 2023-06-27 | 瑞泽恩制药公司 | Antigen binding molecules that bind CD38 and/or CD28 and uses thereof |
| CN116380755A (en) * | 2023-03-20 | 2023-07-04 | 广州市第一人民医院（广州消化疾病中心、广州医科大学附属市一人民医院、华南理工大学附属第二医院） | Application and diagnostic kit of CD127+PMN-MDSCs in the diagnosis of bronchopulmonary dysplasia |
-
2023
- 2023-08-04 CN CN202310977830.9A patent/CN117054649A/en active Pending
Patent Citations (19)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102037015A (en) * | 2007-09-21 | 2011-04-27 | 健泰科生物技术公司 | Inhibition of angiogenesis |
| US20120070461A1 (en) * | 2010-09-21 | 2012-03-22 | Immatics Biotechnologies Gmbh | Use of myeloid cell biomarkers for the diagnosis of cancer |
| CN103180739A (en) * | 2010-09-21 | 2013-06-26 | 伊玛提克斯生物技术有限公司 | Use of myeloid cell biomarkers for the diagnosis of cancer |
| CN109517897A (en) * | 2011-04-28 | 2019-03-26 | 南加利福尼亚大学 | Mankind's marrow source inhibitory cells cancer markers |
| US20150150940A1 (en) * | 2012-05-21 | 2015-06-04 | The Trustees Of Columbia University In The City Of New York | Trefoil family factor proteins and uses thereof |
| CN104024276A (en) * | 2012-06-21 | 2014-09-03 | 卡姆普根有限公司 | LSR antibodies and their use for cancer therapy |
| US20170247464A1 (en) * | 2014-10-24 | 2017-08-31 | Ose Immunotherapeutics | Method and Compositions for Inducing Differentiation of Myeloid Derived Suppressor Cell to Treat Cancer and Infectious Diseases |
| CN107533065A (en) * | 2015-03-13 | 2018-01-02 | 布莱恩·J·赫尔尼奇 | Methods for Monitoring CD4+ Type 1 T Helper Cell Responses in Cancer and Immune Recovery |
| US20180164313A1 (en) * | 2015-06-01 | 2018-06-14 | The Wistar Institute Of Anatomy And Biology | Methods for monitoring polymorphonuclear myeloid derived suppressor cells |
| US20180252721A1 (en) * | 2015-09-02 | 2018-09-06 | Syndax Pharmaceuticals, Inc. | Selection of patients for combination therapy |
| WO2017212018A1 (en) * | 2016-06-10 | 2017-12-14 | INSERM (Institut National de la Santé et de la Recherche Médicale) | Methods and pharmaceutical compositions for the treatment of autoimmune inflammatory diseases |
| CN107271675A (en) * | 2017-03-24 | 2017-10-20 | 郑猛 | Anti-human ADRB3 monoclonal antibodies and its application in medical diagnosis on disease and treatment |
| CN108982857A (en) * | 2018-09-16 | 2018-12-11 | 上海中医药大学附属龙华医院 | The detection method and its application of a kind of Non-small cell lung carcinoma peripheral blood marrow source property inhibition cell |
| US20210077582A1 (en) * | 2019-09-13 | 2021-03-18 | Duke University | Compositions and methods for increasing the efficacy of anti-pd-1 antibody immunotherapy |
| CN116348495A (en) * | 2020-09-18 | 2023-06-27 | 瑞泽恩制药公司 | Antigen binding molecules that bind CD38 and/or CD28 and uses thereof |
| WO2023012844A1 (en) * | 2021-08-02 | 2023-02-09 | Ospedale Pediatrico Bambino Gesu' | Antigen cd111 as a new specific diagnostic and therapeutic marker for polymorphonuclear myeloid-derived suppressor cells (pmn-mdscs) |
| CN114814209A (en) * | 2022-04-29 | 2022-07-29 | 中山大学附属第一医院 | An early warning detection marker PMN-MDSC for early diagnosis of BPD in premature infants |
| CN116240277A (en) * | 2022-07-11 | 2023-06-09 | 复旦大学附属儿科医院 | Malat 1-mediated polymorphonuclear myeloid-derived suppressor cell production and its use in sepsis |
| CN116380755A (en) * | 2023-03-20 | 2023-07-04 | 广州市第一人民医院（广州消化疾病中心、广州医科大学附属市一人民医院、华南理工大学附属第二医院） | Application and diagnostic kit of CD127+PMN-MDSCs in the diagnosis of bronchopulmonary dysplasia |
Non-Patent Citations (19)
| Title |
|---|
| CHIARA CAMISASCHI等: "Targeting Immune Regulatory Networks to Counteract Immune Suppression in Cancer", VACCINES (BASEL), vol. 4, no. 4, 4 November 2006 (2006-11-04), pages 38 * |
| ECE TAVUKCUOGLU等: "Human splenic polymorphonuclear myeloid-derived suppressor cells (PMN-MDSC) are strategically located immune regulatory cells in cancer", EUR J IMMUNOL, vol. 50, no. 12, 31 December 2020 (2020-12-31), pages 2067 - 2074 * |
| LINDA WANG等: "Increased myeloid-derived suppressor cells in gastric cancer correlate with cancer stage and plasma S100A8/A9 proinflammatory proteins", J IMMUNOL, vol. 190, no. 2, 15 January 2023 (2023-01-15), pages 794 - 804, XP055570349, DOI: 10.4049/jimmunol.1202088 * |
| SHUIPING TU等: "Overexpression of interleukin-1beta induces gastric inflammation and cancer and mobilizes myeloid-derived suppressor cells in mice", CANCER CELL, vol. 14, no. 5, 30 November 2008 (2008-11-30), pages 408 - 419 * |
| ZHOU XINGYU等: "PMN-MDSCs accumulation induced by CXCL1 promotes CD8+ T cells exhaustion in gastric cancer", CANCER LETTERS, vol. 532, 28 April 2022 (2022-04-28), pages 215598 * |
| 刘鹏等: "肿瘤免疫细胞因子与慢性萎缩性胃炎"炎-癌转化"研究进展", 中华中医药杂志, vol. 37, no. 6, 9 August 2022 (2022-08-09), pages 3346 - 3351 * |
| 叶妮: "免疫抑制性细胞Treg/Th17、MDSCs在胃癌发生发展中作用的临床研究", 万方学位论文, 16 September 2013 (2013-09-16) * |
| 周佳杰: "髓源性抑制细胞在胃癌患者外周血和肿瘤灶中的表达水平及临床意义", 万方学位论文, 31 October 2012 (2012-10-31) * |
| 屠妍霞等: "髓源性抑制细胞在胃癌发生发展中的作用与应用", 哈尔滨医药, vol. 40, no. 05, 25 October 2020 (2020-10-25), pages 490 - 493 * |
| 张君等: "慢性炎症介导胃癌发生、发展的研究进展", 世界最新医学信息文摘, vol. 17, no. 68, 31 December 2017 (2017-12-31) * |
| 朱立宁: "幽门螺杆菌感染诱导髓源抑制细胞致胃癌肿瘤免疫逃逸的临床研究", 万方学位论文, 3 September 2012 (2012-09-03), pages 4 - 1 * |
| 朱立宁等: "髓源抑制细胞与慢性胃炎至胃癌演变的相关性", 中华消化杂志, vol. 32, no. 9, 31 December 2012 (2012-12-31) * |
| 朱立宁等: "髓源抑制细胞与慢性胃炎至胃癌演变的相关性", 中华消化杂志, vol. 32, no. 9, 5 December 2012 (2012-12-05) * |
| 李晓宁: "胃癌患者外周血MDSC特点及其与中医证型关系的研究", 万方学位论文, 27 March 2019 (2019-03-27) * |
| 杨梅: "髓源性抑制细胞在胃癌发展机制中的研究", 中国优秀硕士学位论文全文数据库 医药卫生科技辑, no. 1, 15 January 2020 (2020-01-15), pages 1 - 2 * |
| 王文娟: "LTBP4在胃腺癌中的表达与相应的机制研究", 中国优秀硕士学位论文全文数据库 医药卫生科技辑, no. 05, 15 May 2023 (2023-05-15) * |
| 郑志华: "基于MDSCs探讨慢性萎缩性胃炎表型特征及健脾清热活血方的干预研究", 中国优秀硕士学位论文全文数据库 医药卫生科技辑, no. 01, 15 January 2023 (2023-01-15) * |
| 郭维华等: "《口腔细胞实验操作技术》", 30 June 2021, 四川大学出版社, pages: 145 * |
| 鲁翔等: "《癌症的免疫治疗》", 31 October 2019, 东南大学出版社, pages: 467 - 468 * |
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