Maleimide conjugation markedly enhances the immunogenicity of both human and murine idiotype-KLH vaccines.
Maleimide conjugation markedly enhances the immunogenicity of both human and murine idiotype-KLH vaccines.
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马来酰亚胺缀合显着增强了人和鼠独特型-KLH 疫苗的免疫原性。
DOI：
10.1016/j.molimm.2008.10.020
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发表时间：
2009-01
影响因子：
3.6
通讯作者：
Timmerman JM
中科院分区：
文献类型：
--
作者：
Kafi K;Betting DJ;Yamada RE;Bacica M;Steward KK;Timmerman JM
B细胞淋巴瘤（独特型，Id）表达的肿瘤特异性克隆免疫球蛋白的可变区内存在的表位集合可以作为主动免疫治疗的靶标。传统上，使用戊二醛将肿瘤来源的 Id 蛋白与免疫原性外源载体蛋白匙孔血蓝蛋白 (KLH) 化学缀合，作为治疗性疫苗。虽然这种方法为一些早期临床试验中治疗的患者提供了有希望的结果，但戊二醛 Id-KLH 疫苗未能在许多接种疫苗的受试者中诱导免疫和临床反应。我们最近描述了一种采用马来酰亚胺-硫氢基化学的替代缀合方法，该方法显着提高了 Id-KLH 疫苗在三种不同的小鼠 B 细胞淋巴瘤模型中的治疗效果，并由 CD8+ T 细胞或抗体介导保护。我们现在详细定义了对于增强人和鼠 Id-KLH 结合疫苗的体内免疫原性至关重要的方法和参数。确定了 Id 巯基预还原的最佳条件，马来酰亚胺 Id-KLH 缀合物即使在长期储存后也能保持稳定性和效力。 Id-KLH 粒径的场流分级分析表明，马来酰亚胺缀合物的尺寸比戊二醛缀合物均匀得多。在日益严格的条件下，马来酰亚胺 Id-KLH 疫苗在治疗已形成的播散性鼠淋巴瘤方面保持了优于戊二醛 Id-KLH 的功效。更重要的是，人马来酰亚胺 Id-KLH 缀合物在诱导 Id 特异性抗体和 T 细胞反应方面始终优于戊二醛 Id-KLH 缀合物。所描述的方法应该很容易适应用于 B 细胞恶性肿瘤人体试验的临床级疫苗的生产。
The collection of epitopes present within the variable regions of the tumor-specific clonal immunoglobulin expressed by B cell lymphomas (idiotype, Id) can serve as a target for active immunotherapy. Traditionally, tumor-derived Id protein is chemically-conjugated to the immunogenic foreign carrier protein keyhole limpet hemocyanin (KLH) using glutaraldehyde to serve as a therapeutic vaccine. While this approach offered promising results for some patients treated in early clinical trials, glutaraldehyde Id-KLH vaccines have failed to induce immune and clinical responses in many vaccinated subjects. We recently described an alternative conjugation method employing maleimide-sulfhydryl chemistry that significantly increased the therapeutic efficacy of Id-KLH vaccines in three different murine B cell lymphoma models, with protection mediated by either CD8+ T cells or antibodies. We now define in detail the methods and parameters critical for enhancing the in vivo immunogenicity of human as well as murine Id-KLH conjugate vaccines. Optimal conditions for Id sulfhydryl pre-reduction were determined, and maleimide Id-KLH conjugates maintained stability and potency even after prolonged storage. Field flow fractionation analysis of Id-KLH particle size revealed that maleimide conjugates were far more uniform in size than glutaraldehyde conjugates. Under increasingly stringent conditions, maleimide Id-KLH vaccines maintained superior efficacy over glutaraldehyde Id-KLH in treating established, disseminated murine lymphoma. More importantly, human maleimide Id-KLH conjugates were consistently superior to glutaraldehyde Id-KLH conjugates in inducing Id-specific antibody and T cell responses. The described methods should be easily adaptable to the production of clinical grade vaccines for human trials in B cell malignancies.
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DOI：
10.1073/pnas.75.5.2411
发表时间：
1978-01-01
影响因子：
11.1
作者：
LEVY, R;DILLEY, J
通讯作者：
DILLEY, J
影响因子：
4.4
作者：
Betting, David J.;Kafi, Kamran;Timmerman, John M.
通讯作者：
Timmerman, John M.
影响因子：
168.9
作者：
KWAK, LW;TAUB, DD;LONGO, DL
通讯作者：
LONGO, DL
影响因子：
4.6
作者：
Hurvitz, SA;Timmerman, JM
通讯作者：
Timmerman, JM
影响因子：
3.9
作者：
ELLMAN, GL
通讯作者：
ELLMAN, GL