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：CD28、CTLA-4和CD80、CD86, 4-1BB和4-1BBL，CD40和CD40L，PD-1和PD-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CD3、CD28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]。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]。
★ 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
☛ 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.
☛ 经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). 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.
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).
GMP级别细胞因子
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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