CN114859060A - 一种细胞因子组及其应用 - Google Patents
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Abstract
本发明公开了一种细胞因子组及其应用，细胞因子组包括IFN‑γ、IL‑17和IFN‑α细胞因子；细胞因子组的应用为将细胞因子组用于检测中，例如将细胞因子的浓度数据用于非疾病诊断的检测中，获得每一项细胞因子在待测样本经干涉后和经干涉前的浓度比值；每一项细胞因子的浓度比值＞各自的界值，为干涉无效，至少一项细胞因子的浓度比值≤各自的界值，为干涉有效；该细胞因子组在人体发生变化，可用于样本状态的判断。
Description
技术领域
本发明涉及一种细胞因子组及其应用，属于生物技术辅助的检测领域。
背景技术
结核病(Tuberculosis，TB)是一种慢性消耗性传染病，已成为威胁人类健康的全球性问题。WHO发布的《2021年全球结核病报告》显示，2020年，全球结核潜伏感染人群接近20亿，新发结核病患者987万，发病率为127/10万，估算发病数和发病率持续呈现下降趋势，但下降幅度较往年有所减缓。排除新冠肺炎疫情的影响，结核病曾是单一传染源的头号死亡原因，也是全球第13大死因。现今结核病治疗效果的评估主要依赖于症状好转、痰菌转阴、病灶吸收。对于症状不明显或痰菌阴性的患者，治疗效果的评估主要依赖于病灶吸收，然而病灶吸收过程漫长，且不易观察，如果不能准确判断患者情况，则难以做到及时指导临床医生调整治疗方案。
发明内容
为了克服现有技术的不足，本发明的第一个目的在于提供一种细胞因子组，该细胞因子组在人体发生变化，可用于样本状态的判断。
本发明的第二个目的在于提供一种上述细胞因子组的量，通过细胞因子组对样本进行状态判断。
本发明的第三个目的在于提供一种上述细胞因子组的应用，将细胞因子组用于检测中。
实现本发明的第一个目的可以通过采取如下技术方案达到：一种细胞因子组，包括IFN-γ、IL-17和IFN-α细胞因子。
实现本发明的第二个目的可以通过采取如下技术方案达到：一种细胞因子组的量，细胞因子组的量为IFN-γ、IL-17和IFN-α细胞因子分别在血清中的浓度。
实现本发明的第三个目的可以通过采取如下技术方案达到：一种细胞因子组的应用，为将细胞因子组用于检测中；细胞因子组包括IFN-γ、IL-17和IFN-α细胞因子。
进一步地，获得每一项细胞因子在血清中的浓度，将浓度数据用于非疾病诊断的检测中。
进一步地，获得每一项细胞因子在待测样本经干涉后和经干涉前的浓度比值；每一项细胞因子的浓度比值＞各自的界值，为干涉无效，至少一项细胞因子的浓度比值≤各自的界值，为干涉有效。
进一步地，干涉为非治疗的干涉。
进一步地，获得每一项细胞因子在血清中的浓度，将浓度数据用于肺结核治疗效果的检测中。
进一步地，获得每一项细胞因子在待测样本经治疗后和经治疗前的浓度比值；每一项细胞因子的浓度比值＞各自的界值，为治疗无效，至少一项细胞因子的浓度比值≤各自的界值，为治疗有效。
进一步地，IFN-γ的界值为0.64、IL-17的界值为0.71；IFN-α的界值为0.65。
相比现有技术，本发明的有益效果在于：
1、本发明细胞因子组在人体发生变化，可用于样本状态的判断；
2、利用本发明细胞因子组进行检测，准确度高；
3、本发明细胞因子组的应用揭示了IFN-γ、IL-17和IFN-α细胞因子的变化，为检测提供新的指导方向。
具体实施方式
下面，结合具体实施方式，对本发明做进一步描述：
一种细胞因子组的应用，为将细胞因子组用于检测中；细胞因子组包括IFN-γ、IL-17和IFN-α细胞因子，具体如下：
获得每一项细胞因子在血清中的浓度，将浓度数据用于肺结核治疗效果的检测中，获得每一项细胞因子在待测样本经治疗后和经治疗前的浓度比值；每一项细胞因子的浓度比值＞各自的界值，为治疗无效，至少一项细胞因子的浓度比值≤各自的界值，为治疗有效；IFN-γ的界值为0.64、IL-17的界值为0.71；IFN-α的界值为0.65。
本技术的工作原理：
从机体抗结核免疫反应的微观层面来评估结核病情是否得到控制及好转，可缩短抗结核治疗效果评估的时间，提高抗结核治疗效果评估的灵敏度及准确度。结核发病过程中的免疫反应主要由巨噬细胞、T细胞等免疫细胞及其表达分泌的细胞因子介导。血清中细胞因子可由多种细胞共同产生，同时细胞因子对免疫细胞的免疫活性具有一定的调节作用。肺结核治疗过程中伴随着结核的控制及病情好转，患者血清中IFN-γ、IL-17和IFN-α细胞因子表达水平也会出现相应变化，这种变化可以一定程度上反应其抗痨治疗的效果。
实施例：
一、样本的选择：
患者纳入标准：临床确诊为肺结核的患者。排除标准：(1)既往结核病史或耐药结核病；(2)肺部其它疾病；(3)其它感染性疾病；(4)自身免疫性疾病(如HIV，类风湿关节炎、强直性脊柱炎、哮喘、干燥综合征、系统性红斑狼疮、硬皮病、多发性肌炎以及皮肌炎等；(5)近三个月内服用免疫抑制剂，激素类药物；(6)肝肾等脏器存在器质性病变、血液病、恶性肿瘤等；(7)妊娠期或哺乳期妇女。
二、样本收集和保存：
所有受试者在治疗前及强化期治疗2月后，均拍肺部X线胸片，并抽取静脉血3mL。静脉血置于抗凝管中，静置1h后，2000rpm离心20min，分装血清于冻存管中，-80℃保存。共入组病例56例。
三、血清中细胞因子浓度测定：
血清解冻后离心取上清，流式细胞管中加入细胞因子联合检测试剂盒(免疫荧光法，赛基生物)中的捕获微球混合液25μL，标品管中加入梯度稀释的标品25μL，样本管中加入血清25μL；所有管中加入25μL荧光检测试剂，涡旋混匀，室温避光孵育2.5h；反应结束加入1mL PBS，200g离心5min后，弃掉上清，加入200μL PBS。样品按照标准品管、样品管的顺序上流式细胞仪(BD，FACSAriaT MⅡ)检测，利用试剂盒自带软件做标曲，并分析血清中的白细胞介素IL-1β、IL-2、IL-4、IL-5、IL-6、IL-8、IL-10、IL-12P70、IL-17、肿瘤坏死因子(TNF)-α、干扰素(IFN)-γ和IFN-α的浓度。
四、数据处理：
将病例治疗2月时的胸片病灶情况与治疗前相比，病灶无变化或变大的患者归为无效组(N＝25)，病灶吸收的患者归为有效组(N＝31)。Ratio值：治疗2月时血清中各细胞因子浓度与治疗前浓度的比值。
采用SPSS 20.0统计软件进行统计学分析，依据受试者工作特征曲线(receiveroperating characteristic curve，ROC)探索Ratio值的界值及检验效能，结果如表格1所示：
表格1 12种细胞因子Ratio值对肺结核治疗有效性评估的检验效能分析
组合：对于单个细胞因子来说，以该细胞因子的界值为界，Rati o值大于该界值认为治疗无效，即对于该细胞因子来说为阳性。将12种细胞因子进行串联组合，具有较好检验效能的8种组合如下：
A：IFN-γ、IL-17、IFN-α；
B：IL-6、TNF、IFN-γ、IL-17、IL-1β；
C：IL-6、IFN-α；
D：IL-6、IL-8；
E：IL-6、IFN-γ、IL-17；
F：IFN-γ、IL-17、IL-8；
G：IFN-α、IL-8；
H：IL-12P70、IL-8；
若组合内所有细胞因子的Ratio值均大于其界值则为阳性，即治疗无效；否则为阴性，即治疗有效；组合的检验效能分析如表格2所示：
表格2细胞因子串联组合对肺结核治疗有效性评估的检验效能分析
其中，A组可在治疗2月时，判断肺结核的治疗效果不依赖于痰菌阴转(患者必须为痰菌阳性的患者，所占比例仅约30％-50％)、症状好转(患者必须症状明显，具有一定主观性)及影像学结果(好转较慢、灵敏度低)，该方法具有适用性广、客观准确的优点。同时，揭示了肺结核在治疗过程中IFN-γ、IL-17和IFN-α细胞因子的变化，为肺外结核、耐多药结核等难治性结核治疗效果的评估及更快评估治疗效果提供了新的指导方向。
对于本领域的技术人员来说，可根据以上描述的技术方案以及构思，做出其它各种相应的改变以及变形，而所有的这些改变以及变形都应该属于本发明权利要求的保护范围之内。
Claims (9)
1.一种细胞因子组，其特征在于，包括IFN-γ、IL-17和IFN-α细胞因子。
2.一种细胞因子组的量，其特征在于，所述细胞因子组的量为IFN-γ、IL-17和IFN-α细胞因子分别在血清中的浓度。
3.一种细胞因子组的应用，其特征在于，为将细胞因子组用于检测中；所述细胞因子组包括IFN-γ、IL-17和IFN-α细胞因子。
4.如权利要求3所述的细胞因子组的应用，其特征在于，获得每一项细胞因子在血清中的浓度，将浓度数据用于非疾病诊断的检测中。
5.如权利要求4所述的细胞因子组的应用，其特征在于，获得每一项细胞因子在待测样本经干涉后和经干涉前的浓度比值；每一项细胞因子的浓度比值＞各自的界值，为干涉无效，至少一项细胞因子的浓度比值≤各自的界值，为干涉有效。
6.如权利要求5所述的细胞因子组的应用，其特征在于，所述干涉为非治疗的干涉。
7.如权利要求3所述的细胞因子组的应用，其特征在于，获得每一项细胞因子在血清中的浓度，将浓度数据用于肺结核治疗效果的检测中。
8.如权利要求7所述的细胞因子组的应用，其特征在于，获得每一项细胞因子在待测样本经治疗后和经治疗前的浓度比值；每一项细胞因子的浓度比值＞各自的界值，为治疗无效，至少一项细胞因子的浓度比值≤各自的界值，为治疗有效。
9.如权利要求3或8所述的细胞因子组的应用，其特征在于，IFN-γ的界值为0.64、IL-17的界值为0.71；IFN-α的界值为0.65。
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| M. LICHTNER 等: "Circulating dendritic cells and interferon-a production in patients with tuberculosis: correlation with clinical outcome and treatment response", 《CLINICAL AND EXPERIMENTAL IMMUNOLOGY》, vol. 143, 31 December 2005 (2005-12-31), pages 329 - 337, XP071080088, DOI: 10.1111/j.1365-2249.2005.02994.x * |
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