ImmunoPET Imaging of Murine CD4(+) T Cells Using Anti-CD4 Cys-Diabody: Effects of Protein Dose on T Cell Function and Imaging.
ImmunoPET Imaging of Murine CD4(+) T Cells Using Anti-CD4 Cys-Diabody: Effects of Protein Dose on T Cell Function and Imaging.
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DOI：
10.1007/s11307-016-1032-z
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发表时间：
2017-08
影响因子：
3.1
通讯作者：
Wu AM
中科院分区：
文献类型：
--
作者：
Freise AC;Zettlitz KA;Salazar FB;Lu X;Tavaré R;Wu AM
全身 CD4+ T 细胞的分子成像对于监测自身免疫性疾病和癌症免疫治疗具有重要意义。鉴于这些细胞在调节免疫方面的关键作用，开发一种生物惰性探针非常重要。 GK1.5 cys-双抗体 (cDb) 是一种先前开发的抗小鼠 CD4 抗体片段，在不同剂量下进行了测试，以评估其对正电子发射断层扫描 (PET) 成像和 CD4+ T 细胞活力、增殖、CD4 表达和功能的影响。通过给小鼠施用不同量的 89Zr 标记的 GK1.5 cDb，然后进行 PET 成像和离体生物分布分析，评估蛋白质剂量对图像对比度（淋巴组织与肌肉比率）的影响。为了评估 GK1.5 cDb 对 T 细胞生物学的影响，将 GK1.5 cDb 在体外与 T 细胞一起孵育，或以多种蛋白质剂量静脉内给予 C57BL/6 小鼠。使用流式细胞术分析 CD4 表达和 T 细胞增殖，并测定细胞因子。对于免疫 PET 成像，与 12 µg 蛋白质剂量相比，2 µg 89Zr 标记的 GK1.5 cDb 的最低蛋白质剂量在腹股沟淋巴结 (ILN) 和脾脏中产生显着更高的注射剂量/克百分比。体内给予 40 µg 高剂量的 GK1.5 cDb 导致脾脏、血液、淋巴结和胸腺中 CD4 表达短暂降低，注射后 3 天内恢复；当服用 2 µg 时，这种作用虽然没有消除，但有所减弱。注射 40 µg GK1.5 cDb 可抑制体内 ILN 的增殖，但不会抑制脾脏的增殖。 GK1.5 cDb 浓度超过 25 nM 会在体外显着抑制 CD4+ T 细胞增殖和干扰素-γ 产生。总体而言，使用低剂量 GK1.5 cDb 最大限度地减少了对 CD4+ T 细胞的生物效应。低剂量 GK1.5 cDb 产生高对比度免疫 PET 图像，对体外和体内 T 细胞生物学影响最小，并且可能是在临床前疾病模型背景下研究 CD4+ T 细胞的有用工具。未来最小化生物效应的方法可能包括创建单价片段或选择针对替代表位的抗 CD4 抗体。
Molecular imaging of CD4+ T cells throughout the body has implications for monitoring autoimmune disease and immunotherapy of cancer. Given the key role of these cells in regulating immunity, it is important to develop a biologically inert probe. GK1.5 cys-diabody (cDb), a previously developed anti-mouse CD4 antibody fragment, was tested at different doses to assess its effects on positron emission tomography (PET) imaging and CD4+ T cell viability, proliferation, CD4 expression, and function. The effect of protein dose on image contrast (lymphoid tissue-to-muscle ratio) was assessed by administering different amounts of 89Zr-labeled GK1.5 cDb to mice followed by PET imaging and ex vivo biodistribution analysis. To assess impact of GK1.5 cDb on T cell biology, GK1.5 cDb was incubated with T cells in vitro or administered intravenously to C57BL/6 mice at multiple protein doses. CD4 expression and T cell proliferation were analyzed with flow cytometry and cytokines were assayed. For immunoPET imaging, the lowest protein dose of 2 µg 89Zr-labeled GK1.5 cDb resulted in significantly higher % injected dose/gram in inguinal lymph nodes (ILN) and spleen compared to the 12 µg protein dose. In vivo administration of GK1.5 cDb at the high dose of 40 µg caused a transient decrease in CD4 expression in spleen, blood, lymph nodes, and thymus, which recovered within 3 days post-injection; this effect was reduced, although not abrogated, when 2 µg was administered. Proliferation was inhibited in vivo in ILN but not the spleen by injection of 40 µg GK1.5 cDb. Concentrations of GK1.5 cDb in excess of 25 nM significantly inhibited CD4+ T cell proliferation and interferon-γ production in vitro. Overall, using low dose GK1.5 cDb minimized biological effects on CD4+ T cells. Low dose GK1.5 cDb yields high-contrast immunoPET images with minimal effects on T cell biology in vitro and in vivo, and may be a useful tool for investigating CD4+ T cells in the context of preclinical disease models. Future approaches to minimizing biological effects may include the creation of monovalent fragments or selecting anti-CD4 antibodies which target alternative epitopes.
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影响因子：
3.6
作者：
Freise AC;Wu AM
通讯作者：
Wu AM
影响因子：
3.1
作者：
Steinhoff, K.;Pierer, M.;Sabri, O.
通讯作者：
Sabri, O.
DOI：
10.1111/j.2517-6161.1995.tb02031.x
发表时间：
1995-01-01
影响因子：
5.8
作者：
BENJAMINI, Y;HOCHBERG, Y
通讯作者：
HOCHBERG, Y
影响因子：
14.8
作者：
Quah, Ben J. C.;Warren, Hilary S.;Parish, Christopher R.
通讯作者：
Parish, Christopher R.
DOI：
10.1364/josaa.1.000612
发表时间：
1984-01-01
影响因子：
1.9
作者：
FELDKAMP, LA;DAVIS, LC;KRESS, JW
通讯作者：
KRESS, JW