CN116183933A - 生物标志物组合物及其在制备评价机体免疫功能的产品中的应用 - Google Patents
生物标志物组合物及其在制备评价机体免疫功能的产品中的应用 Download PDFInfo
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Abstract
本发明公开了生物标志物组合物及其在制备评价机体免疫功能的产品中的应用,生物标志物组合物包含膜联蛋白Annexin A1、膜结合的II型C‑凝集素受体分化抗原簇CD69和白介素2受体α链CD25、细胞因子IFNγ中的任意两种或者任意两种以上的组合。通过检测Annexin A1、CD69、CD25的蛋白表达水平,IFNγ的分泌水平,对检测数据构建评分体系,根据评分体系对免疫功能进行综合性评价,以Annexin A1的蛋白表达水平与年龄进行线性拟合,当检测的Annexin A1蛋白表达水平大于等于线性拟合计算所得数值的60%时,判定免疫细胞功能存在异常;当CD69与CD25表达峰值小于103时,判定细胞活化功能受损失调;当分泌IFNγ的细胞亚群小于10%时,判定细胞在体外无法被有效激活,可用于综合评价机体免疫功能。
Description
技术领域
本发明属于医学诊断领域,具体涉及生物标志物组合物及其在制备评价机体免疫功能的产品中的应用。
背景技术
目前,对于评价机体免疫功能的应用方案为检测各项免疫细胞亚群在外周血中的数量比例,比如CD3+T细胞、CD4+T细胞、NK细胞、巨噬细胞等的比例,来判断免疫系统是否活跃。特殊的应用是针对特定肿瘤或者HIV感染等,通过检测抗原特异性的CD8+T细胞(靶向特定病毒或肿瘤的)数量来进行检查。
但是,这些检测并不是直接针对CD8+T细胞在发挥效应过程中直接相关的各项指标,因此,检测结果存在不精确的缺陷,无法综合评价机体的免疫功能。
发明内容
本发明的目的是克服现有技术中生物标志物无法综合评价机体免疫功能的缺陷。
为了达到上述目的,本发明提供了一种生物标志物组合物,所述生物标志物组合物包含膜联蛋白Annexin A1、膜结合的II型C-凝集素受体分化抗原簇CD69和白介素2受体α链CD25、细胞因子IFNγ中的任意两种或者任意两种以上的组合。
较佳地,所述生物标志物组合物包含膜联蛋白Annexin A1、膜结合的II型C-凝集素受体分化簇CD69和白介素2受体α链CD25、细胞因子IFNγ。
本发明还提供了一种如上所述的生物标志物组合物用于评价机体免疫功能的产品中的应用。
较佳地,所述产品包含评价机体免疫功能的生物标志物组合物检测试剂盒,该试剂盒包含检测所述生物标志物组合物的试剂。
较佳地,所述试剂包含用于检测所述Annexin A1蛋白表达水平的第一试剂,用于检测所述CD69和CD25蛋白表达水平的第二试剂,用于检测所述IFNγ的分泌水平的第三试剂。
较佳地,所述评价机体免疫功能的方法包含以下步骤:S1,提取外周血样本中单核细胞,分离其中的CD8+T细胞;S2,体外激活并培养所述CD8+T细胞;S3,分别使用所述第一试剂检测Annexin A1的蛋白表达水平、使用所述第二试剂检测CD69、CD25的蛋白表达水平以及使用所述第三试剂检测IFNγ的分泌水平,对得到的检测数据构建评分体系,根据评分体系对免疫功能进行综合性评价。
较佳地,S3中,检测之前还包含使用染色抗体混合物A对Annexin A1进行染色、使用染色抗体混合物B对CD69和CD25进行染色、使用染色抗体混合物C对IFNγ进行染色;其中,所述染色抗体混合物A包含DAPI、抗人CD3抗体、抗人CD8抗体、抗人Annexin A1抗体;所述染色抗体混合物B包含DAPI、抗人CD3抗体、抗人CD8抗体、抗人CD69抗体、抗人CD25抗体;所述染色抗体混合物C包含live/dead blue、抗人CD3抗体、抗人CD8抗体。
较佳地,S3中,所述综合性评价包含:以Annexin A1的蛋白表达水平与年龄进行线性拟合,当实际检测的Annexin A1的蛋白表达水平大于等于线性拟合计算所得数值的60%时,判定免疫细胞功能存在异常;当CD69与CD25的表达峰值小于103时,判定细胞活化功能受损失调;当分泌IFNγ的细胞亚群小于10%时,判定细胞在体外无法被有效激活。
较佳地,所述机体包括但不限于健康人、感染病人、癌症病人。
本发明的有益效果:
(1)生物标志物Annexin A1在正常人群内其表达水平随着年龄增长而升高,其表达水平与T细胞状态与免疫功能密切相关,能够更加独特、深入地评价受检者免疫系统状态;生物标志物CD69、CD25为T细胞活化的特异性标志物,其表达水平在特定时间点随着细胞受到激活后的活化强度的增强而增加;生物标志物IFNγ是由CD8+杀伤性T细胞分泌的效应性细胞因子,在维持组织稳态、介导炎症反应以及肿瘤免疫监视中具有重要作用。当受检者实际检测的Annexin A1的蛋白表达水平大于等于60%,提示其免疫细胞功能存在异常;当受检者的CD69、CD25的表达峰值均小于103,提示CD8+T细胞在体外刺激条件下不能被有效激活,细胞活化功能受损失调;当受检者分泌IFNγ的细胞亚群小于10%时,提示细胞在体外无法被有效激活。通过检测受检者外周血中上述三类生物标志物的表达水平,可以构建综合评价人体免疫功能的实验方法及应用体系,为衡量人体抗感染、抗肿瘤能力提供体外评价模型。
(2)相较于现有技术中检测的其他免疫指标,本发明的三种生物标志物的检测均是与CD8+T细胞发挥功能直接相关的各项指标,根据上述三类生物标志物制备的试剂盒可以同时检测三种生物标志物的表达水平,进而有助于免疫功能的早期筛查与干预。
(3)本发明提供的检测方案完善,检测指标相关性显著,且受检者的外周血样本易得、易处理,能够随体检进行实时检测。
附图说明
图1为本发明的Annexin A1蛋白在总CD8+T细胞内的表达水平与年龄呈正相关示意图。
图1的A为代表性样本的外周血CD8+T细胞检测图。
图1的B为Annexin A1蛋白在代表性样本的外周血CD8+T细胞内的表达峰值图。
图1的C为Annexin A1蛋白在237例不同年龄样本内的表达水平与其年龄的回归线图。
图1的D为不同年龄段Annexin A1蛋白表达水平的统计图,对应C图数据。
图1的E为样本生物学年龄与Annexin A1蛋白表达强度(MFI)的相关性统计图。
图2为T细胞活化的表面标志物CD69与CD25的表达随年龄增长而下降的示意图。
图2的A为CD69在10例代表性样本的CD8+T细胞内的表达峰值图。
图2的B为CD69在两个年龄段内的表达统计图,对应A图数据。
图2的C为CD25在10例代表性样本的CD8+T细胞内的表达峰值图。
图2的D为CD25在两个年龄段内的表达统计图,对应C图数据。
图3为T细胞激活后分泌效应分子IFNγ的水平随年龄增长而下降的示意图。
图3的A为IFNγ在代表性样本的外周血CD8+T细胞内的分泌水平图。
图3的B为IFNγ在6例不同年龄的代表性样本内分泌水平的峰值图。
图3的C为分泌IFNγ的效应性CD8+T细胞在6例不同年龄的代表性样本内的比例统计图,对应A、B图数据。
图3的D为6例不同年龄代表性样本的CD8+T细胞分泌IFNγ的强度统计图,对应B图数据。
具体实施方式
以下结合附图和实施例对本发明的技术方案做进一步的说明。
本发明未提交的实验材料均为市售取得、本发明未提及的实验仪器均为本领域常规仪器、本发明未提及的实验方法均为本领域常规操作。
T细胞是适应性免疫系统的关键组成部分,在机体抵抗细菌、病毒感染及抗肿瘤过程中发挥重要作用。静息状态的T细胞在活化后通过增殖及向效应性细胞分化以发挥免疫应答功能,其中CD8+T细胞接受MHC I类分子的信号刺激而活化,进而克隆扩增以形成表型和功能异质性效应细胞,发挥抗病毒、抗肿瘤功能。细胞毒性CD8+T淋巴细胞(CTL)对靶细胞的效应过程十分关键,当其有效被激活活化后,可促使储存在胞质溶胶中的穿孔素和颗粒酶合成,特异性靶向邻近的细胞并发挥杀伤功能。其中,衡量效应表型的特征是细胞毒性强并可生成细胞因子如IFNγ。
膜联蛋白A1(Annexin A1)是膜联蛋白超家族的成员,在CD8+T细胞等多种免疫细胞中均表达,参与糖皮质激素诱导的免疫调节过程。当CD8+T细胞受到双信号刺激活化后,Annexin A1的表达水平影响细胞增殖与效应细胞分化方向,干预炎症反应,如小鼠关节炎模型发病时期出现症状加剧,以及类风湿性关节炎患者血液中CD4+T细胞出现Annexin A1表达水平的显著升高等。这些结果共同表明,Annexin A1是TCR信号传导的分子“调谐器”,其表达水平与T细胞状态与免疫功能密切相关,是调节适应性免疫反应的重要介质。Annexin A1影响T细胞免疫反应能力与体内炎症水平,其表达水平与年龄正相关,能够更加独特、深入地评价受检者免疫系统状态,因此Annexin A1是免疫系统功能检测的特定生物标志物。
T细胞的活化是机体适应性免疫应答的核心部分,T细胞的活化、增殖及分化需要接受双信号刺激:第一信号由T细胞受体转导并由粘附分子增强;协同刺激信号由抗原递呈细胞表面协同刺激分子和T细胞相应受体相互作用产生。在体外实验中,往往使用抗CD3抗体和抗CD28抗体作为双信号刺激以激活T细胞。
T淋巴细胞上表达的最重要的激活分子可分为早期激活标志物,如CD69和CD25,以及晚期激活标志物,如CD62L和HLA-DR。其中,分化抗原簇69(CD69)是一种膜结合的II型C-凝集素受体,在淋巴细胞受到刺激后迅速出现在质膜表面,是淋巴细胞活化早期的经典标志物。CD69参与调节性T(Treg)细胞的分化以及IFNγ、IL-17和IL-22的分泌过程,从而调节免疫反应。体外实验中,当T细胞接受抗CD3/CD28联合抗体刺激后,CD69的表达最早可在刺激后2-3小时检测到,是T细胞激活最早期的细胞表面标志物。
白介素2受体α链(IL2RA,CD25)由IL2RA基因编码,是在活化淋巴细胞细胞表面发现的65kDa跨膜糖蛋白,可与β链、γ链一起构成高亲和力的IL2受体,介导T细胞参与的细胞免疫反应。作为T细胞激活的第二标志物,CD25在静息状态下的淋巴细胞内表达很低,在细胞接受活化信号刺激后其表达在3到12小时之间以时间依赖性方式增加,与CD69一起是指征细胞活化程度的理想指标。
干扰素γ(Interferon gamma,IFNγ)是主要由CD8+杀伤性T细胞分泌的效应性细胞因子,在维持组织稳态、介导炎症反应以及肿瘤免疫监视中具有重要作用。IFNγ在T细胞内的表达水平是衡量CD8+T细胞发挥其溶解靶细胞潜力的重要标志物。当CD8+T细胞接受到抗原刺激后产生IFNγ作用于其受体激活Janus激酶(JAK)信号转导和转录激活因子(STAT)信号通路,以诱导具有关键免疫效应子功能的经典干扰素刺激基因(ISGs)的表达。随着时间的推移,IFNγ引起的细胞反应通过影响各种酶的表达、功能以及代谢、染色质和转录的调节来诱导靶细胞状态的改变。试验发现,IFNγ能活化固有免疫细胞,进而达到抑制病毒数量的效果。在治疗A型流感小鼠试验中,当流感感染小鼠缺失干扰素后,其死亡率大幅提升且体内病毒数量维持高浓度水平;相较之下,体内有干扰素存在的小鼠,流感病毒数量随着干扰素的生成而逐渐下降。对此,评估CD8+T细胞在接受刺激信号后分泌IFNγ的水平是检测细胞能否正常发挥免疫功能的关键指标。
Annexin A1、CD69及CD25、IFNγ是与CD8+T细胞发挥功能直接相关的各项指标,因此,检测上述生物标志物能够更加具体、精确、深入地检测人体免疫功能。
1.实验材料及仪器:
1)抗体:live/dead blue(Thermo Scientific,L23105),DAPI(Biosciences,#564907,BD),抗人CD3抗体(anti-human CD3 PE,BioLegend,#300441),抗人CD8抗体(anti-human CD8 FITC,BD Biosciences,#561947),抗人Annexin A1(anti-human Annexin A1APC,BioLegend,#831604),抗人CD69抗体(anti-human CD69 PE-Cy7,BioLegend,#310911),抗人CD25抗体(anti-human CD25 BV421,eBioscience,#562442),抗人IFNγ抗体(anti-human IFNγPE-Cy7,BioLegend,#502527)。
2)阴性分选的试剂盒:DynabeadsTMUntouchedTMHuman CD8 T Cells Kit,(Thermo Scientific,11348D)。
3)激活CD8+T细胞所用的抗体磁珠:DynabeadsTMHuman T-Activator CD3/CD28for T Cell Expansion and Activation(Thermo Scientific,11161D)。
4)仪器:流式细胞仪(LSR Fortessa,BD)。
2.实验方法:
2.1受检者外周血样本中单核细胞提取及CD8+T细胞的分离
采集符合预设标准的人群外周血液样本2mL于抗凝管内,加入等体积PBS稀释后,通过ficoll溶液进行梯度离心,获得外周血内单核细胞总群,使用分离液重悬、计数,按照108个/mL重悬于分离液。
将500μL溶于分离液的外周血单核细胞转移至5mL流式分离管内,加入100μL未经过热激活的胎牛血清(FBS),另加入100μL阴性分选CD8+T细胞所需的抗体混合物混匀,4度孵育20分钟。
加入2mL分离液进行离心,4度条件下以350xg离心8分钟,弃上清。
使用500μL分离液重悬细胞,加入500μL分离CD8+T细胞所需的磁珠与管内细胞结合,室温条件下孵育15分钟。
加入2mL分离液重悬细胞,将流式管插入磁铁内进行细胞分离,收集上清液,即得到目标CD8+T细胞群。
其中,所述的阴性分选CD8+T细胞所需的抗体混合物为抗人CD4、CD14、CD16a、CD16b、CD19、CD36、CD56、CD123和CD235a(血型糖蛋白A)的单克隆抗体,用于结合外周血内的非CD8+T细胞。
2.2体外激活并培养分离后的CD8+T细胞
计数2.1中分离得的CD8+T细胞后,取一半细胞按照4:1的比例配制含anti-humanCD3及anti-human CD28抗体磁珠的溶液,溶剂为细胞培养基,使用该溶液重悬细胞,加入溶液体积千分之一的IL-2细胞因子,将细胞置于培养箱培养24小时,用于后续检测CD69与CD25指标。
另取一半细胞直接使用培养基重悬,加入佛波醇12-豆蔻酸13-乙酸酯(PMA)、离子霉素、Brefeldin A、莫能菌素和蛋白转运抑制剂的混合物,将细胞置于培养箱培养4-6小时,用于后续检测IFNγ的分泌。
2.3检测Annexin A1、CD69与CD25蛋白表达水平及细胞因子IFNγ的分泌强度
将染色抗体混合物A加入2.1中的剩余细胞,室温避光孵育15分钟进行Annexin A1染色。
混合物A包括DAPI、anti-human CD3 PE、anti-human CD8 FITC、anti-humanAnnexin A1 APC。
将染色抗体混合物B加入2.2第一部分培养24小时后的细胞,4度避光孵育30分钟进行CD69与CD25的染色。
混合物B包括DAPI、anti-human CD3 PE、anti-human CD8 FITC、anti-human CD69PE-Cy7、anti-human CD25 APC。
将染色抗体混合物C加入2.2第二部分培养4-6小时后的细胞,4度避光孵育30分钟进行细胞因子IFNγ的第一步染色。
混合物C包括live/dead blue、anti-human CD3 PE、anti-human CD8 FITC。
完成第一步染色的细胞离心后使用细胞破膜固定液重选,4度避光孵育20分钟,使用对应的清洗液清洗后加入anti-human IFNγPE-Cy7进行染色,4度避光孵育30分钟。
所有细胞即可用于流式细胞仪上机检测,所得结果如图1~3内各部分所示。
2.4根据评分体系对受检者免疫功能进行综合性评价
如图1的C所示,为Annexin A1蛋白在237例不同年龄样本内的表达水平与其年龄的回归线图,将237例预设正常人外周血样本的Annexin A1表达水平Y与其年龄X进行线性拟合,得到回归公式Y=6.888X+271.908,当受检者实际检测的Y值小于等于其计算所得的30%时,此项评价为优;30%~60%评价为良;大于等于60%评价为预警,提示其免疫细胞功能存在异常。
如图2所示为T细胞活化的表面标志物CD69与CD25的表达随年龄增长而下降的示意图,根据图2的数据判断CD8+T细胞能否被有效激活,当细胞CD69与CD25的表达峰值图均超过104评价为优,表明CD8+T细胞能够接受体外信号刺激而活化增殖;当CD69的表达峰值图均超过104而CD25的表达峰值图仅部分超过104评价为良,表明CD8+T细胞接受在体外信号刺激后能够激活但活化增殖能力中等;当CD69与CD25的表达峰值图均小于103评价为预警,表明CD8+T细胞在体外刺激条件下不能被有效激活,细胞活化功能受损失调。
如图3所示为T细胞激活后分泌效应分子IFNγ的水平随年龄增长而下降的示意图,根据图3的数据评价CD8+T细胞能否发挥其杀伤性效应功能。当分泌IFNγ的细胞亚群比例大于25%时评价为优,提示细胞可被强烈激活发挥效应功能;当比例为10%~25%时评价为良;小于10%时评价为预警,表明细胞在体外无法被有效激活。
分别检测三部分即Annexin A1的表达、CD69与CD25的表达、IFNγ的分泌水平,对三部分数据分别进行评价后,最后从细胞效应潜力、细胞活化能力、细胞效应能力三部分综合评价,可以针对性提出免疫功能的缺陷点,提供精准的个体化医疗指导建议。
利用本发明的三种生物标志物进行制备评价机体免疫功能的产品,可以分别检测机体中Annexin A1的表达、CD69与CD25的表达、IFNγ的分泌水平,进而综合评价机体的免疫水平。所述产品至少包含有用于检测Annexin A1蛋白表达水平的第一试剂、用于检测CD69和CD25蛋白表达水平的第二试剂、用于检测IFNγ的分泌水平的第三试剂。
本发明提出的生物标志物组合物包含Annexin A1、CD69与CD25、IFNγ,上述生物标志物均是与CD8+T细胞发挥功能直接相关的各项指标,Annexin A1的表达水平与年龄正相关,影响T细胞免疫反应能力与体内炎症水平,CD69与CD25可以表征细胞活化程度,IFNγ可以表征细胞在体外是否被有效激活;通过分别检测上述生物标志物的表达水平,当受检者实际检测的Annexin A1的表达水平大于等于线性拟合计算所得的60%时评价预警,提示其免疫细胞功能存在异常,当CD69与CD25的表达峰图均小于103评价预警,提示CD8+T细胞在体外刺激条件下不能被有效激活,细胞活化功能受损失调,当检测到的分泌IFNγ的细胞亚群比例小于10%时评价为预警,提示细胞在体外无法被有效激活。从上述三方面综合评价,可以针对性提出免疫功能的优缺点,提供精准的个体化医疗指导建议。
尽管本发明的内容已经通过上述优选实施例作了详细介绍,但应当认识到上述的描述不应被认为是对本发明的限制。在本领域技术人员阅读了上述内容后,对于本发明的多种修改和替代都将是显而易见的。因此,本发明的保护范围应由所附的权利要求来限定。
Claims (9)
1.生物标志物组合物,其特征在于,所述生物标志物组合物包含膜联蛋白Annexin A1、膜结合的II型C-凝集素受体分化抗原簇CD69和白介素2受体α链CD25、细胞因子IFNγ中的任意两种或者任意两种以上的组合。
2.如权利要求1所述的生物标志物组合物,其特征在于,所述生物标志物组合物包含膜联蛋白Annexin A1、膜结合的II型C-凝集素受体分化簇CD69和白介素2受体α链CD25、细胞因子IFNγ。
3.一种根据权利要求1或2所述的生物标志物组合物用于制备评价机体免疫功能的产品中的应用。
4.如权利要求3所述的应用,其特征在于,所述产品包含评价机体免疫功能的生物标志物组合物检测试剂盒,该试剂盒包含检测所述生物标志物组合物的试剂。
5.如权利要求3所述的应用,其特征在于,所述试剂包含用于检测所述Annexin A1蛋白表达水平的第一试剂,用于检测所述CD69和CD25蛋白表达水平的第二试剂,用于检测所述IFNγ的分泌水平的第三试剂。
6.如权利要求5所述的应用,其特征在于,所述评价机体免疫功能的方法包含以下步骤:
S1,提取外周血样本中单核细胞,分离其中的CD8+T细胞;
S2,体外激活并培养所述CD8+T细胞;
S3,分别使用所述第一试剂检测Annexin A1的蛋白表达水平、使用所述第二试剂检测CD69、CD25的蛋白表达水平以及使用所述第三试剂检测IFNγ的分泌水平,对得到的检测数据构建评分体系,根据评分体系对免疫功能进行综合性评价。
7.如权利要求6所述的应用,其特征在于,S3中,检测之前还包含使用染色抗体混合物A对Annexin A1进行染色、使用染色抗体混合物B对CD69和CD25进行染色、使用染色抗体混合物C对IFNγ进行染色;其中,所述染色抗体混合物A包含DAPI、抗人CD3抗体、抗人CD8抗体、抗人Annexin A1抗体;所述染色抗体混合物B包含DAPI、抗人CD3抗体、抗人CD8抗体、抗人CD69抗体、抗人CD25抗体;所述染色抗体混合物C包含live/dead blue、抗人CD3抗体、抗人CD8抗体。
8.如权利要求6所述的应用,其特征在于,S3中,所述综合性评价包含:以Annexin A1的蛋白表达水平与年龄进行线性拟合,当实际检测的Annexin A1的蛋白表达水平大于等于线性拟合计算所得数值的60%时,判定免疫细胞功能存在异常;当CD69与CD25的表达峰值小于103时,判定细胞活化功能受损失调;当分泌IFNγ的细胞亚群小于10%时,判定细胞在体外无法被有效激活。
9.如权利要求6所述的应用,特征在于,所述机体包括但不限于健康人、感染病人、癌症病人。
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Citations (8)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN1444043A (zh) * | 2002-12-25 | 2003-09-24 | 帕弗瑞生物技术(北京)有限公司 | 一种个体化特异性免疫细胞功能测定方法 |
US20060051358A1 (en) * | 2004-05-24 | 2006-03-09 | Baylor Research Institute | Immune response assessment method |
US20060275752A1 (en) * | 2005-06-03 | 2006-12-07 | University Of Pittsburgh Of The Commonwealth System Of Higher Education | Multiparameteric method for assessing immune system status |
US20080038283A1 (en) * | 2004-02-17 | 2008-02-14 | Topham David J | Methods of Evaluating Efficacy of an Immune Response by Assesing Alpha-1 Integrin Expression |
US20110207134A1 (en) * | 2008-11-07 | 2011-08-25 | Sequenta, Inc. | Monitoring health and disease status using clonotype profiles |
CN102272156A (zh) * | 2008-12-02 | 2011-12-07 | 玛丽皇后和威斯特-弗尔德学院 | 通过调控膜联蛋白-1对自身免疫疾病的治疗 |
US20150152474A1 (en) * | 2012-03-09 | 2015-06-04 | Caris Life Sciences Switzerland Holdings Gmbh | Biomarker compositions and methods |
CN114791411A (zh) * | 2022-04-21 | 2022-07-26 | 广州先康达生物科技有限公司 | 评估人体免疫功能的指标组合、试剂盒和方法 |
-
2023
- 2023-02-21 CN CN202310143454.3A patent/CN116183933A/zh active Pending
Patent Citations (8)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN1444043A (zh) * | 2002-12-25 | 2003-09-24 | 帕弗瑞生物技术(北京)有限公司 | 一种个体化特异性免疫细胞功能测定方法 |
US20080038283A1 (en) * | 2004-02-17 | 2008-02-14 | Topham David J | Methods of Evaluating Efficacy of an Immune Response by Assesing Alpha-1 Integrin Expression |
US20060051358A1 (en) * | 2004-05-24 | 2006-03-09 | Baylor Research Institute | Immune response assessment method |
US20060275752A1 (en) * | 2005-06-03 | 2006-12-07 | University Of Pittsburgh Of The Commonwealth System Of Higher Education | Multiparameteric method for assessing immune system status |
US20110207134A1 (en) * | 2008-11-07 | 2011-08-25 | Sequenta, Inc. | Monitoring health and disease status using clonotype profiles |
CN102272156A (zh) * | 2008-12-02 | 2011-12-07 | 玛丽皇后和威斯特-弗尔德学院 | 通过调控膜联蛋白-1对自身免疫疾病的治疗 |
US20150152474A1 (en) * | 2012-03-09 | 2015-06-04 | Caris Life Sciences Switzerland Holdings Gmbh | Biomarker compositions and methods |
CN114791411A (zh) * | 2022-04-21 | 2022-07-26 | 广州先康达生物科技有限公司 | 评估人体免疫功能的指标组合、试剂盒和方法 |
Non-Patent Citations (9)
Title |
---|
AFONSO BEZERRA RIBEIRO等: "Expression of annexin-A1 in blood and tissue leukocytes of leprosy patients", REV SOC BRAS MED TROP, vol. 53, 25 November 2020 (2020-11-25), pages 20200277 * |
ANGELA A SENA等: "Lack of TNFRI signaling enhances annexin A1 biological activity in intestinal inflammation", BIOCHEM PHARMACOL, vol. 98, no. 3, 16 September 2015 (2015-09-16), pages 422 - 431 * |
NIKOLAOS PASCHALIDIS等: "Role of endogenous annexin-A1 in the regulation of thymocyte positive and negative selection", CELL CYCLE, vol. 9, no. 4, 17 February 2010 (2010-02-17), pages 784 - 793, XP055054841, DOI: 10.4161/cc.9.4.10673 * |
YICHENG TAO等: "Functional modulation of CD8+ T cell by approved novel immune enhancer: Nocardia rubra Cell-Wall Skeletons (Nr-CWS)", INT IMMUNOPHARMACOL, vol. 78, 24 December 2019 (2019-12-24), pages 106023, XP085973894, DOI: 10.1016/j.intimp.2019.106023 * |
YUAN H YANG等: "Deficiency of annexin A1 in CD4+ T cells exacerbates T cell-dependent inflammation", J IMMUNOL, vol. 190, no. 3, 24 December 2012 (2012-12-24), pages 997 - 1007 * |
熊言骏: "构建肌层浸润性膀胱癌预后相关蛋白风险评分模型", 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 2023 (2023-01-15) * |
白永恒等: "《肿瘤微环境与免疫耐受》", 30 June 2020, 天津科学技术出版社, pages: 95 * |
辛欢欢等: "白花蛇舌草黄酮注射液对小鼠脾淋巴细胞的免疫增强作用", 中国兽医科学, vol. 40, no. 07, 31 December 2010 (2010-12-31) * |
黄海华: "《药学细胞生物学》", 31 January 2006, 中国医药科技出版社, pages: 460 * |
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