Kayamuro Hiroyuki, Abe Yasuhiro, Yoshioka Yasuo, Katayama Kazufumi, Nomura Tetsuya, Yoshida Tokuyuki, Yamashita Kohei, Yoshikawa Tomoaki, Kawai Yuichi, Mayumi Tadanori, Hiroi Takachika, Itoh Norio, Nagano Kazuya, Kamada Haruhiko, Tsunoda Shin-ichi, Tsutsumi Yasuo
Laboratory of Pharmaceutical Proteomics, National Institute of Biomedical Innovation, 7-6-8 Saito-Asagi, Ibaraki, Osaka 567-0085, Japan.
Biomaterials. 2009 Oct;30(29):5869-76. doi: 10.1016/j.biomaterials.2009.07.009. Epub 2009 Jul 30.
Safe and potent adjuvants are required in order to establish effective mucosal vaccines. Cytokines are promising adjuvants because they are human-derived safe biomaterial and display immune-modulating functions. We have created a mutant tumor necrosis factor-alpha (TNF-alpha), mTNF-K90R, that exhibits high bioactivity and resistance to proteases. Here, we examined the potential of mTNF-K90R as a mucosal adjuvant. Initially, we showed that intranasal co-administration of mTNF-K90R with ovalbumin (OVA) potently produced OVA-specific Immunoglobulin (Ig) G antibodies (Abs) in serum and IgA Abs both at local and distal mucosal sites compared to co-administration with wild-type TNF-alpha. The OVA-specific immune response was characterized by high levels of serum IgG1 and increased production of interleukin-4 (IL-4), IL-5 and IL-10 from splenocytes of immunized mice, suggesting a Th2 response. Furthermore, intranasal immunization with an antigen from influenza virus plus mTNF-K90R exhibited mucosal adjuvant activity for induction of both systemic and mucosal immune responses. Importantly, histopathological examination of the nasal tissue of mTNF-K90R treated mice detected no signs of toxicity. These findings suggest that mTNF-K90R is safe and effective mucosal adjuvant and this system may have potential application as a universal mucosal adjuvant system for mucosal vaccines improving the immune response to a variety of viral antigens.
为了建立有效的黏膜疫苗，需要安全且有效的佐剂。细胞因子是很有前景的佐剂，因为它们是源自人类的安全生物材料并具有免疫调节功能。我们创造了一种突变型肿瘤坏死因子-α（TNF-α），即mTNF-K90R，它具有高生物活性且对蛋白酶具有抗性。在此，我们研究了mTNF-K90R作为黏膜佐剂的潜力。首先，我们发现与野生型TNF-α共同给药相比，mTNF-K90R与卵清蛋白（OVA）经鼻共同给药能在血清中高效产生OVA特异性免疫球蛋白（Ig）G抗体（Abs），并在局部和远端黏膜部位产生IgA Abs。OVA特异性免疫反应的特征是血清IgG1水平高，且免疫小鼠脾细胞产生的白细胞介素-4（IL-4）、IL-5和IL-10增加，提示为Th2反应。此外，用流感病毒抗原加mTNF-K90R进行鼻内免疫表现出黏膜佐剂活性，可诱导全身和黏膜免疫反应。重要的是，对mTNF-K90R处理小鼠的鼻组织进行组织病理学检查未发现毒性迹象。这些发现表明mTNF-K90R是一种安全有效的黏膜佐剂，该系统可能具有作为通用黏膜佐剂系统的潜在应用，用于黏膜疫苗，改善对多种病毒抗原的免疫反应。
