Takasuka Naomi, Fujii Hideki, Takahashi Yoshimasa, Kasai Masataka, Morikawa Shigeru, Itamura Shigeyuki, Ishii Koji, Sakaguchi Masahiro, Ohnishi Kazuo, Ohshima Masamichi, Hashimoto Shu-ichi, Odagiri Takato, Tashiro Masato, Yoshikura Hiroshi, Takemori Toshitada, Tsunetsugu-Yokota Yasuko
Department of Immunology, National Institute of Infectious Diseases, Toyama 1-23-1, Shinjuku-ku, Tokyo 162-8640, Japan.
Int Immunol. 2004 Oct;16(10):1423-30. doi: 10.1093/intimm/dxh143. Epub 2004 Aug 16.
The recent emergence of severe acute respiratory syndrome (SARS) was caused by a novel coronavirus, SARS-CoV. It spread rapidly to many countries and developing a SARS vaccine is now urgently required. In order to study the immunogenicity of UV-inactivated purified SARS-CoV virion as a vaccine candidate, we subcutaneously immunized mice with UV-inactivated SARS-CoV with or without an adjuvant. We chose aluminum hydroxide gel (alum) as an adjuvant, because of its long safety history for human use. We observed that the UV-inactivated SARS-CoV virion elicited a high level of humoral immunity, resulting in the generation of long-term antibody secreting and memory B cells. With the addition of alum to the vaccine formula, serum IgG production was augmented and reached a level similar to that found in hyper-immunized mice, though it was still insufficient to elicit serum IgA antibodies. Notably, the SARS-CoV virion itself was able to induce long-term antibody production even without an adjuvant. Anti-SARS-CoV antibodies elicited in mice recognized both the spike and nucleocapsid proteins of the virus and were able to neutralize the virus. Furthermore, the UV-inactivated virion induced regional lymph node T-cell proliferation and significant levels of cytokine production (IL-2, IL-4, IL-5, IFN-gamma and TNF-alpha) upon restimulation with inactivated SARS-CoV virion in vitro. Thus, a whole killed virion could serve as a candidate antigen for a SARS vaccine to elicit both humoral and cellular immunity.
严重急性呼吸综合征（SARS）近期的出现是由一种新型冠状病毒SARS-CoV引起的。它迅速传播到许多国家，因此迫切需要研发一种SARS疫苗。为了研究经紫外线灭活的纯化SARS-CoV病毒粒子作为候选疫苗的免疫原性，我们用经紫外线灭活的SARS-CoV对小鼠进行皮下免疫，免疫时添加或不添加佐剂。我们选择氢氧化铝凝胶（明矾）作为佐剂，因为其在人类使用方面有着悠久的安全历史。我们观察到，经紫外线灭活的SARS-CoV病毒粒子引发了高水平的体液免疫，导致产生长期分泌抗体的记忆B细胞。在疫苗配方中添加明矾后，血清IgG产量增加，并达到与高度免疫小鼠相似的水平，不过仍不足以引发血清IgA抗体。值得注意的是，即使没有佐剂，SARS-CoV病毒粒子本身也能够诱导长期的抗体产生。在小鼠体内引发的抗SARS-CoV抗体能够识别该病毒的刺突蛋白和核衣壳蛋白，并能够中和病毒。此外，经紫外线灭活的病毒粒子在体外经灭活的SARS-CoV病毒粒子再次刺激后，可诱导局部淋巴结T细胞增殖和显著水平的细胞因子产生（IL-2、IL-4、IL-5、IFN-γ和TNF-α）。因此，完整的灭活病毒粒子可以作为SARS疫苗的候选抗原，以引发体液免疫和细胞免疫。
