Hammer Markus H, Meyer Sonja, Brestrich Gordon, Moosmann Andreas, Kern Florian, Tesfa Lydia, Babel Nina, Mittenzweig Alexa, Rooney Cliona M, Hammerschmidt Wolfgang, Volk Hans-Dieter, Reinke Petra
Department of Nephrology and Internal Intensive Care, Universitätsmedizin Berlin, Berlin, Germany.
Eur J Immunol. 2005 Jul;35(7):2250-8. doi: 10.1002/eji.200526230.
Adoptive immunotherapy with antigen-specific T cells has been successfully used to treat certain infectious diseases and cancers. Although more patients may profit from T cell therapy, its more frequent use is restricted by limitations in current T cell generation strategies. The most commonly applied peptide-based approaches rely on the knowledge of relevant epitopes. Therefore, T cells cannot be generated for diseases with unknown epitopes or for patients with unfavorable HLA types. We developed a peptide-based approach for HLA type-independent generation of specific T cells against various proteins. It is based on short-time stimulation with peptide libraries that cover most CD4(+) and CD8(+) T cell epitopes of given proteins. The procedure requires no prior knowledge of epitopes because libraries are synthesized solely on the basis of the protein's amino acid sequence. Stimulation is followed by immunomagnetic selection of activated IFN-gamma-secreting cells and nonspecific expansion. To evaluate the protocol, we generated autologous T cells specific for a well-characterized antigen, the human cytomegalovirus phosphoprotein 65 (pp65). Generated T cell lines consisted of pp65-specific CD4(+) and CD8(+) lymphocytes that displayed antigen-specific killing and proliferation. The protocol combines the biosafety of peptide-based approaches with HLA type independence and may help to advance adoptive immunotherapy in the future.
采用抗原特异性T细胞进行过继性免疫治疗已成功用于治疗某些传染病和癌症。尽管更多患者可能从T细胞疗法中获益，但其更广泛的应用受到当前T细胞生成策略局限性的限制。最常用的基于肽的方法依赖于相关表位的知识。因此，对于表位未知的疾病或HLA类型不利的患者，无法生成T细胞。我们开发了一种基于肽的方法，用于独立于HLA类型生成针对各种蛋白质的特异性T细胞。它基于用覆盖给定蛋白质的大多数CD4(+)和CD8(+)T细胞表位的肽库进行短时间刺激。该过程不需要事先了解表位，因为肽库仅根据蛋白质的氨基酸序列合成。刺激后通过免疫磁珠分选活化的分泌IFN-γ的细胞并进行非特异性扩增。为了评估该方案，我们生成了针对一种特征明确的抗原——人巨细胞病毒磷蛋白65（pp65）的自体T细胞。生成的T细胞系由具有pp65特异性的CD4(+)和CD8(+)淋巴细胞组成，这些细胞表现出抗原特异性杀伤和增殖能力。该方案将基于肽的方法的生物安全性与HLA类型独立性相结合，可能有助于在未来推动过继性免疫治疗的发展。
