CN111549126A - 一种用于预测或诊断gvhd的试剂盒、gvhd的风险计算方法和应用 - Google Patents

一种用于预测或诊断gvhd的试剂盒、gvhd的风险计算方法和应用 Download PDF

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CN111549126A
CN111549126A CN202010558859.XA CN202010558859A CN111549126A CN 111549126 A CN111549126 A CN 111549126A CN 202010558859 A CN202010558859 A CN 202010558859A CN 111549126 A CN111549126 A CN 111549126A
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elafin
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CN111549126B (zh
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王昭
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Beijing Friendship Hospital
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Abstract

本发明公开了一种用于预测或诊断GVHD的试剂盒、GVHD的风险计算方法和应用，涉及GVHD相关领域。具体地，该应用包括将用于检测TIM‑3，MMP‑3，IL‑2R，IL‑6，IL‑8，CD30，ST2，TNFRI，HGF，REG3a，MIG，Elafin、骨调素和颗粒酶A等细胞因子组合的试剂应用于制备诊断GVHD的试剂中，通过该细胞因子组合可高通量、快速有效地测定GVHD，通过动态监测细胞因子组合的变化，可直接或间接地分析和判断GVHD的程度，为诊断治疗GVHD提供一种新的途径。

Description

一种用于预测或诊断GVHD的试剂盒、GVHD的风险计算方法和 应用
技术领域
本发明涉及GVHD相关领域，具体而言，涉及一种用于预测或诊断GVHD的试剂盒、GVHD的风险计算方法和应用。
背景技术
GVHD(graft-versus-host disease，移植物抗宿主病)是异基因造血干细胞移植术后最常见、最重要的一种并发症，也是造血干细胞移植术后的主要死亡原因。
GVHD可分为急性GVHD和慢性GVHD，急性GVHD一般发生在移植后3个月内，主要表现为皮疹、肠炎伴腹泻、肝功能损伤等；慢性GVHD通常发生在移植3个月后，临床主要表现为硬皮肤样皮肤疾病，皮肤黏膜干燥综合症，慢性肝病等。
目前，GVHD的诊断仍然建立在病理活检基础上，如急性GVHD病理特点为皮肤、肝脏和消化道细胞坏死，慢性GVHD的病理特点则为受累器官的纤维化和萎缩。国际上很多研究集中在能否避免有创操作检查而仅通过外周血检查即有效诊断GVHD。GVHD本身就是一种炎症过程，而且其严重程度往往与参与炎症过程的T调节细胞的数量和比例等相关。
目前，研究中与GVHD最可能关系密切的就是IL-2和TNF-α，其中IL-2的升高可以预测急性GVHD的产生，并且sIL-2R与GVHD的严重程度相关。而可溶性TNF受体的升高程度甚至与预后相关。IL-12、IL-18、IFN-γ等Th1相关因子也在部分研究中被证实可能与急性GVHD相关，其中IL-18在GVHD有效治疗后会出现显著下降，可以提示治疗效果。
目前，缺乏能有效精确诊断GVHD的方法，鉴于此，提出本申请。
发明内容
本发明的目的在于提供一种用于预测或诊断GVHD的试剂盒、GVHD的风险计算方法和应用。
本发明提供一种技术方案：
第一方面，本发明实施例提供了一种用于检测细胞因子表达水平的试剂在制备诊断GVHD的试剂盒中的应用试剂盒，所述细胞因子包括：TIM-3，MMP-3，IL-2R，IL-6，IL-8，CD30，ST2，TNFRI，HGF，REG3a，MIG，Elafin、骨调素和颗粒酶A。
第二方面，本发明实施例提供了一种用于预测或诊断GVHD的试剂盒，其包括用于检测GVHD相关细胞因子表达的试剂，所述细胞因子包括：TIM-3，MMP-3，IL-2R，IL-6，IL-8，CD30，ST2，TNFRI，HGF，REG3a，MIG，Elafin、骨调素和颗粒酶A。
第三方面，本发明实施例提供了一种GVHD风险分数的计算方法，应用于电子设备，所述计算方法包括：
根据获取待测样品的细胞因子的表达量，计算所述待测样品对应的患GVHD的风险分数；
所述细胞因子包括：TIM-3，MMP-3，IL-2R，IL-6，IL-8，CD30，ST2，TNFRI，HGF，REG3a，MIG，Elafin、骨调素和颗粒酶A。
有益效果：
本发明实施例提供了用于检测细胞因子表达水平的试剂在制备诊断GVHD的试剂盒中的应用试剂试剂盒，该应用包括将用于检测TIM-3，MMP-3，IL-2R，IL-6，IL-8，CD30，ST2，TNFRI，HGF，REG3a，MIG，Elafin、骨调素和颗粒酶A等细胞因子组合的试剂应用于制备诊断GVHD的试剂中，通过该细胞因子组合可高通量、快速有效地测定GVHD，通过动态监测细胞因子组合的变化，可直接或间接地分析和判断GVHD的程度，为诊断治疗GVHD提供一种新的途径。
附图说明
为了更清楚地说明本发明实施例的技术方案，下面将对实施例中所需要使用的附图作简单地介绍，应当理解，以下附图仅示出了本发明的某些实施例，因此不应被看作是对范围的限定，对于本领域普通技术人员来讲，在不付出创造性劳动的前提下，还可以根据这些附图获得其他相关的附图。
图1为本发明验证例1中的TIM-3、MMP-3、IL-2R和IL-6的水平检测结果图；
图2为本发明验证例1中的IL-8、CD30、ST2和TNFRI的水平检测结果图；
图3为本发明验证例1中的HGF、REG3a、MIG和Elafin的水平检测结果图；
图4为本发明验证例1中的Osteopontin和Granzyme A的水平检测结果图。
具体实施方式
为使本发明实施例的目的、技术方案和优点更加清楚，下面将对本发明实施例中的技术方案进行清楚、完整地描述。实施例中未注明具体条件者，按照常规条件或制造商建议的条件进行。所用试剂或仪器未注明生产厂商者，均为可以通过市售购买获得的常规试剂。
以下结合实施例对本发明的特征和性能作进一步的详细描述。
本发明实施例提供了用于检测细胞因子表达水平的试剂在制备诊断GVHD的试剂盒中的应用试剂试剂盒，所述细胞因子包括：TIM-3，MMP-3，IL-2R，IL-6，IL-8，CD30，ST2，TNFRI，HGF，REG3a，MIG，Elafin、骨调素和颗粒酶A。
通过上述细胞因子的组合，可有效达到早期、快速诊断GVHD的目的。IL-6、CD30、IL-2a、TNFRI(TNFR-1)和Elafin在移植后早期的升高可以预测急性GVHD的发生；出现急性GVHD后，IL-6和HGF的升高提示急性GVHD有更严重的程度，更差的预后；REG3a及颗粒酶A在急性GVHD中的升高，则提示GVHD的主要累及部位为肠道；IL-8、ST2(IL-33R)，MIG(CXCL9)，MMP-3和骨调素(Osteopontin)在移植后晚期的升高则可以预测慢性GVHD的发生。同时，其动态变化亦可表示GVHD的严重程度和治疗反应等疾病状态。
具体地，TIM-3为免疫检查点分子，游离型的TIM-3与肠道重型GVHD相关；CD30、IL-2a、TNFRI(TNFR-1)和Elafin均对aGVHD(急性GVHD)的诊断作用；IL-8、ST2(IL-33R)，MIG(CXCL9)，MMP-3和骨调素(Osteopontin)：对慢性GVHD的诊断作用；HGF为肝细胞生长因子，在aGVHD中升高，且与aGVHD严重程度程正相关；REG3a作为肠道急性移植物抗宿主病(GI-aGVHD)特异性标志物。颗粒酶A在急性GVHD中升高，尤其是在肠道急性GVHD中。IL-6水平升高能够预测aGVHD的发生，并且其水平的高低与预后相关。
在可选实施方式中，所述试剂包括引物对1～14中的任意一对或多对的组合。具体地，引物对1～14分别用于检测TIM-3，MMP-3，IL-2R，IL-6，IL-8，CD30，IL-33R，TNFRI，HGF，REG3a，MIG，Elafin、骨调素和颗粒酶A的基因表达水平。
在可选实施方式中，所述试剂包括探针1～14中的任意一个或多个的组合，探针1～14分别用于检测TIM-3，MMP-3，IL-2R，IL-6，IL-8，CD30，IL-33R，TNFRI，HGF，REG3a，MIG，Elafin、骨调素和颗粒酶A的基因表达水平。
在可选实施方式中，所述试剂包括抗体1～14中的任意一个或多个的组合，抗体1～14分别用于检测TIM-3，MMP-3，IL-2R，IL-6，IL-8，CD30，IL-33R，TNFRI，HGF，REG3a，MIG，Elafin、骨调素和颗粒酶A的蛋白表达水平。
需要说明的是，将上述引物对1～14、上述抗体1～14或上述探针1～14进行随机组合，以达到能检测上述细胞因子表达水平的技术方案均属于本申请的保护范围。
在可选实施方式中，所述诊断GVHD为诊断急性GVHD。
本发明实施例还提供了一种用于预测或诊断GVHD的试剂盒，其包括用于检测细胞因子表达水平的试剂，所述细胞因子包括：TIM-3，MMP-3，IL-2R，IL-6，IL-8，CD30，IL-33R，TNFRI，HGF，REG3a，MIG，Elafin、骨调素和颗粒酶A。
在可选实施方式中，所述试剂盒包括引物对1～14中的任意一对或多对的组合，引物对1～14分别用于检测TIM-3，MMP-3，IL-2R，IL-6，IL-8，CD30，IL-33R，TNFRI，HGF，REG3a，MIG，Elafin、骨调素和颗粒酶A的基因表达水平。
在可选实施方式中，所述试剂盒包括探针1～14中的任意一个或多个的组合，探针1～14分别用于检测TIM-3，MMP-3，IL-2R，IL-6，IL-8，CD30，IL-33R，TNFRI，HGF，REG3a，MIG，Elafin、骨调素和颗粒酶A的基因表达水平。
在可选实施方式中，所述试剂盒包括抗体1～14中的任意一个或多个的组合，抗体1～14分别用于检测TIM-3，MMP-3，IL-2R，IL-6，IL-8，CD30，IL-33R，TNFRI，HGF，REG3a，MIG，Elafin、骨调素和颗粒酶A的蛋白表达水平。
此外，本发明实施例还提供了一种GVHD风险分数的计算方法，应用于电子设备，所述计算方法包括：
根据获取待测样品的细胞因子的表达量，计算所述待测样品对应的患GVHD的风险分数；
所述细胞因子包括：TIM-3，MMP-3，IL-2R，IL-6，IL-8，CD30，IL-33R，TNFRI，HGF，REG3a，MIG，Elafin、骨调素和颗粒酶A。
在可选实施方式中，所述计算方法还包括采用如上述任一实施方式所述的试剂对待测样品的细胞因子的表达水平进行检测。
下面结合具体实施例对本发明的技术方案进行说明。
实施例1
本实施例提供一种用于预测或诊断GVHD的试剂盒，其包括：
用于分别检测TIM-3，MMP-3，IL-2R，IL-6，IL-8，CD30，IL-33R，TNFRI，HGF，REG3a，MIG，Elafin、骨调素和颗粒酶A的试剂。
实施例2
采用实施例1提供的试剂盒对待测样品进行GVHD进行检测，具体包括以下步骤。
方法：使用Luminex200检测受试者血清中上述细胞因子的浓度情况。细胞因子的正常参考值如表1所示。
表1 正常参考值
项目 正常参考值pg/ml
TIM-3 66～1031
MMP-3 0～455
IL-2R 0～7593
IL-6 0～216
IL-8 0～55
CD30 0～1457
ST2/IL-33R 0～2204
TNFRI 0～7286
HGF 0～521
REG3a 0～3645
MIG 0～54
Elafin 0～460
Osteopontin 0～18956
Granzyme A 0～270
。
验证例1
采用实施例1提供的用于预测或诊断GVHD的试剂盒，按照实施例2提供的检测方法，对5个样本(5个aGVHD患者样本和5个同期未患aGVHD患者)进行检测。
检测结果请参照表2和附图1～4。
表2 aGVHD患者急性期与同期未患aGVHD患者细胞因子比较结果
细胞因子 aGVHD急性期/正常值 同期未患GVHD/正常值
TIM-3 2.3 1.8
MMP-3 2.8 1.8
IL-2R 6.2 2.8
IL-6 1.2 1.0
IL-8 1.1 0.9
CD30 3.7 2.5
ST2/IL-33R 3.2 0.9
TNFRI 2.7 4.4
HGF 2.2 3.0
REG3a 1.7 1.7
MIG 5.1 2.7
Elafin 1.6 1.8
Osteopontin 1.4 2.4
Granzyme A 3.4 1.7
由表2和图1～4可知，aGVHD患者与同期未患aGVHD患者细胞因子水平变化比较(虚线：aGVHD患者，实线：同期非aGVHD患者)。可以看出，在出现aGVHD急性期(d31，横坐标31)前的1-2周，本发明实施例1中细胞因子组合中的细胞因子已经开始升高；在出现aGVHD后仍继续升高至高值(d43，横坐标43)；在aGVHD得到缓解(d54，横坐标54)后，细胞因子水平逐渐下降。说明本细胞因子可以早期诊断，甚至早期预测aGVHD的发生，并能够提示aGVHD的疾病状态。
综上，本发明实施例提供了一种用于检测细胞因子表达水平的试剂在制备诊断GVHD的试剂盒中的应用，该应用包括将用于检测TIM-3，MMP-3，IL-2R，IL-6，IL-8，CD30，ST2，TNFRI，HGF，REG3a，MIG，Elafin、骨调素和颗粒酶A等细胞因子组合的试剂应用于制备诊断GVHD的试剂中，通过该细胞因子组合可高通量、快速有效地测定GVHD，通过动态监测细胞因子组合的变化，可直接或间接地分析和判断GVHD的程度，为诊断治疗GVHD提供一种新的途径。
此外，本发明实施例还提供了用于检测GVHD相关细胞因子表达的试剂在制备诊断GVHD的试剂盒中的应用和GVHD风险分数的计算方法。
以上所述仅为本发明的优选实施例而已，并不用于限制本发明，对于本领域的技术人员来说，本发明可以有各种更改和变化。凡在本发明的精神和原则之内，所作的任何修改、等同替换、改进等，均应包含在本发明的保护范围之内。

Claims (10)

1.用于检测细胞因子表达水平的试剂在制备诊断GVHD的试剂盒中的应用，其特征在于，所述细胞因子包括：TIM-3，MMP-3，IL-2R，IL-6，IL-8，CD30，ST2，TNFRI，HGF，REG3a，MIG，Elafin、骨调素和颗粒酶A。
2.根据权利要求1所述的用于检测细胞因子表达水平的试剂在制备诊断GVHD的试剂盒中的应用，其特征在于，所述试剂包括引物对1～14中的任意一对或多对的组合，引物对1～14分别用于检测TIM-3，MMP-3，IL-2R，IL-6，IL-8，CD30，ST2，TNFRI，HGF，REG3a，MIG，Elafin、骨调素和颗粒酶A的基因表达水平。
3.根据权利要求1所述的用于检测细胞因子表达水平的试剂在制备诊断GVHD的试剂盒中的应用，其特征在于，所述试剂包括探针1～14中的任意一个或多个的组合，探针1～14分别用于检测TIM-3，MMP-3，IL-2R，IL-6，IL-8，CD30，ST2，TNFRI，HGF，REG3a，MIG，Elafin、骨调素和颗粒酶A的基因表达水平。
4.根据权利要求1所述的用于检测细胞因子表达水平的试剂在制备诊断GVHD的试剂盒中的应用，其特征在于，所述试剂包括抗体1～14中的任意一个或多个的组合，抗体1～14分别用于检测TIM-3，MMP-3，IL-2R，IL-6，IL-8，CD30，ST2，TNFRI，HGF，REG3a，MIG，Elafin、骨调素和颗粒酶A的蛋白表达水平。
5.根据权利要求1所述的用于检测细胞因子表达水平的试剂在制备诊断GVHD的试剂盒中的应用，其特征在于，所述诊断GVHD为诊断急性GVHD。
6.一种用于预测或诊断GVHD的试剂盒，其特征在于，其包括用于检测细胞因子表达水平的试剂，所述细胞因子包括：TIM-3，MMP-3，IL-2R，IL-6，IL-8，CD30，ST2，TNFRI，HGF，REG3a，MIG，Elafin、骨调素和颗粒酶A。
7.根据权利要求6所述的用于预测或诊断GVHD的试剂盒，其特征在于，所述试剂盒包括引物对1～14中的任意一对或多对的组合，引物对1～14分别用于检测TIM-3，MMP-3，IL-2R，IL-6，IL-8，CD30，ST2，TNFRI，HGF，REG3a，MIG，Elafin、骨调素和颗粒酶A的基因表达水平。
8.根据权利要求6所述的用于预测或诊断GVHD的试剂盒，其特征在于，所述试剂盒包括探针1～14中的任意一个或多个的组合，探针1～14分别用于检测TIM-3，MMP-3，IL-2R，IL-6，IL-8，CD30，ST2，TNFRI，HGF，REG3a，MIG，Elafin、骨调素和颗粒酶A的基因表达水平。
9.根据权利要求6所述的用于预测或诊断GVHD的试剂盒，其特征在于，所述试剂盒包括抗体1～14中的任意一个或多个的组合，抗体1～14分别用于检测TIM-3，MMP-3，IL-2R，IL-6，IL-8，CD30，ST2，TNFRI，HGF，REG3a，MIG，Elafin、骨调素和颗粒酶A的蛋白表达水平。
10.一种GVHD风险分数的计算方法，其特征在于，应用于电子设备，所述计算方法包括：
根据获取待测样品的细胞因子的表达量，计算所述待测样品对应的患GVHD的风险分数；
所述细胞因子包括：TIM-3，MMP-3，IL-2R，IL-6，IL-8，CD30，ST2，TNFRI，HGF，REG3a，MIG，Elafin、骨调素和颗粒酶A。
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Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN110988364A (zh) * 2019-12-31 2020-04-10 天津美瑞特医疗科技有限公司 一种应用流式细胞术检测移植后gvhd相关细胞因子的方法及检测试剂盒

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* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN110988364A (zh) * 2019-12-31 2020-04-10 天津美瑞特医疗科技有限公司 一种应用流式细胞术检测移植后gvhd相关细胞因子的方法及检测试剂盒

Non-Patent Citations (3)

* Cited by examiner, † Cited by third party
Title
CHEN， Y-B等: "Biomarkers for acute GVHD: can we predict the unpredictable?", 《BONE MARROW TRANSPLANTATION》 *
FIEMA, B.等: "High Throughput Sequential ELISA for Validation of Biomarkers of Acute Graft-Versus-Host Disease", 《JOURNAL OF VISUALIZED EXPERIMENTS》 *
ZAID, M.A.等: "Plasma biomarkers of risk for death in a multicenter phase 3 trial with uniform transplant characteristics post–allogeneic HCT", 《BLOOD》 *

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