Juillard V, Villefroy P, Godfrin D, Pavirani A, Venet A, Guillet J G
Laboratoire d'Immunologie des Interactions Cellulaires et Moléculaires, INSERM Unité 152, Paris, France.
Eur J Immunol. 1995 Dec;25(12):3467-73. doi: 10.1002/eji.1830251239.
This study examines the suitability of replication-defective adenovirus vectors for engineering recombinant vaccines. The immunological abilities and limitations of E1-deleted adenoviruses containing the lacZ gene (Ad-beta-gal) were investigated by examining the humoral and cellular immune responses to the beta-galactosidase protein. BALB/c mice (H-2d) were given in a single injection of recombinant adenovirus. The cytotoxic T lymphocyte (CTL) response of spleen cells was evaluated. Recognized target cells were H-2d-derived tumor cells transfected by the lac Z gene, or incubated with the 876-884 beta-galactosidase peptide known to be restricted by the Ld molecule of the major histocompatibility complex. A long-lasting beta-galactosidase-specific cytotoxic T cell response was obtained. By contrast, CTL from mice immunized with the Ld-restricted peptide were less specific for the endogenous epitope presented by the transfectants expressing beta-galactosidase. Ad-beta-gal-immunized mice were also protected against an intra-cerebral challenge with a recombinant vaccinia virus expressing the lac-Z gene. These results suggest that Ad-beta-gal-induced CTL have protective abilities in vivo. The induction of beta-galactosidase-specific T helper lymphocytes and humoral IgG responses were also examined. A proliferative response occurred only late after immunization and the primed T lymphocytes produced interleukin-2, but no interleukin-4. A humoral IgG response to the beta-galactosidase protein was detected 15-30 days after a single immunization and remained stable for 6 months without boosting. Lastly, we followed the evolution of the immune response over the course of successive immunizations. The magnitude and kinetics of the cellular and humoral responses were similar to those obtained after a single immunization. Consistent with these observations, an adenovirus-specific neutralizing antibody response was detected as early as the second immunization. Thus, a single immunization with a replication-defective adenovirus recombinant vector induces long-lasting humoral and cellular immune responses specific to the transgene product.
本研究考察了复制缺陷型腺病毒载体用于构建重组疫苗的适用性。通过检测对β-半乳糖苷酶蛋白的体液免疫和细胞免疫反应，研究了含lacZ基因的E1缺失腺病毒（Ad-β-gal）的免疫能力及局限性。给BALB/c小鼠（H-2d）单次注射重组腺病毒，评估脾细胞的细胞毒性T淋巴细胞（CTL）反应。识别的靶细胞是经lacZ基因转染的H-2d来源的肿瘤细胞，或与已知受主要组织相容性复合体Ld分子限制的876 - 884β-半乳糖苷酶肽一起孵育。获得了持久的β-半乳糖苷酶特异性细胞毒性T细胞反应。相比之下，用Ld限制肽免疫的小鼠的CTL对表达β-半乳糖苷酶的转染细胞所呈递的内源性表位特异性较低。用Ad-β-gal免疫的小鼠也受到保护，可抵御表达lac-Z基因的重组痘苗病毒的脑内攻击。这些结果表明，Ad-β-gal诱导的CTL在体内具有保护能力。还检测了β-半乳糖苷酶特异性T辅助淋巴细胞的诱导及体液IgG反应。仅在免疫后晚期出现增殖反应，致敏的T淋巴细胞产生白细胞介素-2，但不产生白细胞介素-4。单次免疫后15 - 30天检测到对β-半乳糖苷酶蛋白的体液IgG反应，且在未加强免疫的情况下持续6个月保持稳定。最后，我们跟踪了连续免疫过程中免疫反应的演变。细胞免疫和体液免疫反应的强度及动力学与单次免疫后相似。与这些观察结果一致，早在第二次免疫时就检测到腺病毒特异性中和抗体反应。因此，用复制缺陷型腺病毒重组载体进行单次免疫可诱导针对转基因产物的持久体液免疫和细胞免疫反应。
