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>技术资源 > 前沿速递 >【技术干货】T细胞激活最佳搭档 —— CD3/CD28抗体偶联磁珠

【技术干货】T细胞激活最佳搭档 —— CD3/CD28抗体偶联磁珠

2022-04-22 09:49    浏览量:1169


T细胞激活

T淋巴细胞如何被激活?

在机体内,T淋巴细胞的活化,在免疫应答中扮演着相当重要的角色,研究表明,诱导T细胞的活化与增殖需要两种信号:第一信号是TCR/CD3与抗原提呈细胞(APCs)表面特异的MHC分子抗原肽复合物结合产生的特异性抗原刺激信号;第二信号是非特异性的共刺激信号,由APCs表面多对共刺激分子和T细胞的相应受体相互作用后产生(如:CD28CTLA-4CD80CD86, 4-1BB4-1BBLCD40CD40LPD-1PD-L1等),其中CD28是最为重要的共刺激分子,第二信号可使T细胞完全活化,分泌细胞因子和表达细胞因子受体[1], 如果缺乏共刺激信号,第一信号非但不能有效激活特异性T细胞,反而导致T细胞失能[2]而在体外,联合使用CD3CD28的抗体刺激T细胞,模拟体内T细胞活化的双信号作用,是目前体外进行T细胞激活与扩增应用最广泛的方法[3]

T细胞激活



Tips:T细胞在经过第一、第二信号刺激完全活化后,还有赖于多种细胞因子(IL-1IL-2IL-4IL-6IL-10IL-12IL-15 IFN-γ等)的作用才能进一步增殖活化,如果没有细胞因子,活化T细胞不能增殖和分化,导致T细胞的活化后凋亡[2]

>>>点击了解GMP级别细胞因子

细胞治疗领域中T细胞的激活

CAR-T和TCR-T疗法是目前过继性免疫细胞疗法的研究热点,无论是CAR-T还是TCR-T细胞的体外培养,T细胞都需要被激活后才能进行后续操作步骤,因此合适的T细胞激活试剂是开发细胞治疗药物的必需原料,目前市场上体外激活T细胞的方法主要有:①使用可溶性抗体, 如CD3/CD28抗体联合细胞因子如IL-2等进行刺激,但有研究显示,IL-2浓度过高的情况下,可能会导致T细胞产物耗尽,进入功能障碍阶段,显示不良的效应功能,并迅速凋亡[4]; ②使用结合在固相载体上的抗体, 如CD3/CD28抗体偶联磁珠,研究显示,使用抗CD3/CD28抗体包被的磁珠作为人工抗原递呈颗粒,比用OKT3(抗CD3抗体)/IL-2激活可保留更多的T细胞记忆表型[5],且由于磁珠可持续刺激T细胞,细胞因子的产生比其他方法(如用OKT3和IL-2激活)高10-100倍,这表明使用磁珠的激活作用更强[6],另外,也有研究表明,与OKT3和IL-2相比,用抗CD3/CD28磁珠激活可以使得T细胞耗竭更少,疗效更加持久[7]
抗CD3/CD28抗体
为支持细胞治疗药物的开发,ACROBiosystems 自主开发了高质量的 CD3/CD28抗体偶联磁珠产品(货号:MBS-C001,经细胞水平验证,可高效刺激扩增T细胞,更好的助力细胞治疗药物的开发进程。

产品列表

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产品特色

★ 5.5 μm大小,可更好的模拟APC,刺激T细胞

★ 磁性强,轻松分离,磁珠不易残留

★ 超低内毒素(< 2EU/mg),对T细胞无伤害

★ 经细胞水平验证,可高效激活扩增T细胞

产品数据

 Anti-CD3/CD28 Antibody-coupled Magnetic Beads (Cat. No. MBS-C001) 可高效激活T细胞

The purified human T cells were activated using Anti-CD3/CD28 Antibody-coupled Magnetic Beads (Cat. No. MBS-C001) at a ratio of 1:1 beads-to-cells for 24 hours with RPMI1640 supplemented with 10% of FBS. The negative control experiment was performed with adding the Negative Control Beads coupled HSA. Cells were fluorescently stained using PE labeled anti-human CD25 antibody and labeled FITC anti-human CD69 antibody and analyzed by flow cytometry.

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  Anti-CD3/CD28 Antibody-coupled Magnetic Beads (Cat. No. MBS-C001) 刺激后,可对T细胞进行有效扩增


The purified human T cells were stimulated using Anti-CD3/CD28 Antibody-coupled Magnetic Beads (Cat. No. MBS-C001) at a ratio of 1:1 beads-to-cells. Cells were expanded in T cell culture medium supplemented with 4ng/mL of rhIL-2 Protein (Acrobiosystems, Cat. No. IL2-H4113). Activated Cells were expanded for up to 13 days (A) with high cell viability (B).

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竞品对比数据
 ACRO Anti-CD3/CD28 Antibody-coupled Magnetic Beads (Cat. No. MBS-C001) 与竞品分别激活T细胞,激活能力水平基本一致

The purified human T cells were activated using ACRO Anti-CD3/CD28 Antibody-coupled Magnetic Beads (Cat. No. MBS-C001) and competitor’s beads respectively at a ratio of 1:1 beads-to-cells for 24 hours with RPMI1640 supplemented with 10% of FBS. Cells were fluorescently stained using PE labeled anti-human CD25 antibody and labeled FITC anti-human CD69 antibody and analyzed by flow cytometry.

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与竞品相比,ACRO Anti-CD3/CD28 Antibody-coupled Magnetic Beads (Cat. No. MBS-C001),可对T细胞进行有效扩增,且有较高水平的扩增能力

The purified human T cells were stimulated using Anti-CD3/CD28 Antibody-coupled Magnetic Beads (Cat. No. MBS-C001) and competitor’s beads respectively. Cells were expanded in T cell culture medium supplemented with 4ng/mL of rhIL-2 Protein (Acrobiosystems, Cat. No. IL2-H4113). Activated Cells were expanded for up to 13 days (A) with high cell viability (B).

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更多T细胞激活试剂

Anti-CD3 antibody(clone: OKT3)
Anti-CD28 antibody

GMP级别细胞因子

CGT-solution


参考文献

[1] Kay, J.E., 1991. Mechanisms of T lymphocyteactivation. Immunology Letters 29, 51 – 54

[2] 医学免疫学-第7版

[3] Trickett A, Kwan YL. T cellstimulation and expansion using anti-CD3/CD28 beads. J Immunol Methods. 2003Apr 1;275(1-2):251-5.

[4] Kalos M, Levine BL, Porter DL, KatzS, Grupp SA, Bagg A, June CH: T cells with chimeric antigen receptors havepotent antitumor effects and can establish memory in patients with advancedleukemia. Sci Transl Med 2011, 3:95ra73.

[5] Hollyman D, Stefanski J, PrzybylowskiM, Bartido S, Borquez- Ojeada O, Taylor C, Yeh R, Capacio V, Olszewska M, HoseyJ et al.: Manufacturing validation of biologically functional T cells targetedto CD19 antigen for autologous adoptive cell therapy. J Immunother 2009, 32:169-180.

[6]Casati A, Varghaei-Nahvi A, FeldmanSA, Assenmacher M, Rosenberg SA, Dudley ME, Scheffold A: Clinical-scaleselection and viral transduction of human naı¨ve and central memory CD8+ T cells for adoptive cell therapy ofcancer patients. Cancer Immunol Immunother 2013, 62:1563-1573.

[7] Barrett DM, Singh N, Liu X, JiangS, June CH, Grupp SA, Zhao Y: Relation of clinical culture method to T-cellmemory status and efficacy in xenograft models of adoptive immunotherapy. Cytotherapy2014, 16:619-630.



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