Yamada Takaya, Kataoka Shingo, Ogasawara Keiko, Ishimitsu Ryotaro, Hashigucci Kazuhiro, Suzuki Tatsuo, Kawauchi Hideyuki
Department of Otolaryngology, Shimane University, Izumo City, Japan.
Rhinology. 2005 Sep;43(3):190-8.
The ideal vaccine therapy has been warranted for activation of the mucosal immune response in the upper respiratory tract against various types of microbial infection. However, the precise study in regard to the mucosal route of vaccine administration and its mechanism of action remains to be further investigated. Therefore, to better understand the exact mechanism of nasopharyngeal mucosal immunology, from T-cell aspects, the antigen-specific antibody response was investigated in T cell receptor transgenic (OVA23-3) mice (Tg-mice) and wild type BALB/c mice, in comparison, which were stimulated with repeated nasal antigen challenges of ovalbumin (OVA) together with cholera toxin (CT) or OVA alone. OVA-specific IgA and IgG antibodies were not detected in nasal washings of BALB/c mice when these mice were intranasally stimulated with OVA alone. But they were detected in those of BALB/c mice stimulated with OVA and CT, as we have already reported. Interestingly, OVA-specific IgA and IgG antibodies were significantly higher in nasal washings of Tg-mice stimulated with OVA and CT or OVA alone rather than those of BALB/c mice stimulated with OVA and CT. In line with data of the antibody response, OVA-specific IgA and IgG antibody-producing cells significantly increased in number in nasal passage (NP), nasopharyngeal-associated lymphoreticular tissue (NALT), cervical lymph node (CLN), and spleen (SP) of these mice. In nasal washings of Tg-mice, interferon (IFN)-gamma and interleukin (IL)-4 was detected even with a small amount of antigen. To see the cytokine profile of NALT, NP, CLN, and SP of these mice, various cytokine concentrations were measured in supernatants of these cells cultured in vitro with OVA. As a result, IFN-gamma was detected at significantly higher levels in culture supernatants of lymphocytes sampled from NP, CLN, SP as well as NALT of mice having increased antibody titers in nasal washings. On the other hand, Th2 type cytokines such as IL-4, IL-6 and IL-13 were efficiently detected in culture supernatants of NP, CLN, and SP cells from Tg-mice mice, but not in those from NALT cells of those mice. All these data taken together indicate that helper T cells recruited into nasal mucosa and locally activated in an antigen-specific fashion, as well as NALT T cells, are essential for mounting local antigen-specific antibody responses.
理想的疫苗疗法已被证实可激活上呼吸道的黏膜免疫反应，以抵御各种类型的微生物感染。然而，关于疫苗黏膜给药途径及其作用机制的精确研究仍有待进一步探究。因此，为了更好地从T细胞方面理解鼻咽黏膜免疫学的确切机制，我们在T细胞受体转基因（OVA23 - 3）小鼠（Tg小鼠）和野生型BALB/c小鼠中研究了抗原特异性抗体反应，并进行比较，这两种小鼠均用卵清蛋白（OVA）联合霍乱毒素（CT）或仅用OVA进行反复鼻腔抗原刺激。当单独用OVA对BALB/c小鼠进行鼻内刺激时，在其鼻腔灌洗液中未检测到OVA特异性IgA和IgG抗体。但正如我们之前所报道的，在OVA和CT刺激的BALB/c小鼠的鼻腔灌洗液中检测到了这些抗体。有趣的是，在用OVA和CT或仅用OVA刺激的Tg小鼠的鼻腔灌洗液中，OVA特异性IgA和IgG抗体显著高于用OVA和CT刺激的BALB/c小鼠。与抗体反应数据一致，这些小鼠的鼻腔（NP）、鼻咽相关淋巴组织（NALT）、颈淋巴结（CLN）和脾脏（SP）中产生OVA特异性IgA和IgG抗体的细胞数量显著增加。在Tg小鼠的鼻腔灌洗液中，即使使用少量抗原也能检测到干扰素（IFN）-γ和白细胞介素（IL）-4。为了观察这些小鼠的NALT、NP、CLN和SP的细胞因子谱，在用OVA体外培养这些细胞的上清液中测量了各种细胞因子的浓度。结果，在鼻腔灌洗液中抗体滴度升高的小鼠的NP、CLN、SP以及NALT的淋巴细胞培养上清液中，检测到IFN-γ的水平显著更高。另一方面，在Tg小鼠的NP、CLN和SP细胞的培养上清液中能有效检测到Th2型细胞因子，如IL-4、IL-6和IL-13，但在这些小鼠的NALT细胞的培养上清液中未检测到。所有这些数据表明，募集到鼻黏膜并以抗原特异性方式局部激活的辅助性T细胞以及NALT T细胞，对于产生局部抗原特异性抗体反应至关重要。
