Epidermal powder immunization induces both cytotoxic T-lymphocyte and antibody responses to protein antigens of influenza and hepatitis B viruses
Epidermal powder immunization induces both cytotoxic T-lymphocyte and antibody responses to protein antigens of influenza and hepatitis B viruses
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DOI：
10.1128/jvi.75.23.11630-11640.2001
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发表时间：
2001-12-01
影响因子：
5.4
通讯作者：
Payne, LG
中科院分区：
文献类型：
--
作者：
Chen, DX;Weis, KF;Payne, LG
细胞毒性T淋巴细胞(CTL)在宿主抵抗病毒和细胞内细菌感染中起着至关重要的作用。然而，使用注射器和针头肌肉注射的非复制型疫苗主要引起体液反应，而不是CTL反应。在这里，我们报道了表皮粉末免疫(EPI)，这是一种使用无针粉末递送系统将1.5-2.5微米金粒上的抗原递送到表皮的技术，它能诱导CTL对非复制性抗原的反应。EPI后，在靶皮肤的组织学切片中，可在存活的表皮中发现大多数包被抗原的金粒子。进一步的透射电子显微镜研究揭示了金粒子在细胞内的定位。双色免疫荧光显微镜下可见接种部位的朗格汉斯细胞(LCs)内有大量包被抗原的颗粒，20h后引流淋巴结内可见LCs。在小鼠中研究了EPI后对几种病毒蛋白抗原的免疫应答。乙肝表面抗原(HBs)和流感病毒核蛋白(NP)合成肽的EPI可诱导抗原特异性CTL反应和抗体反应。在体外细胞耗竭实验中，我们证明了EPI诱导的抗HBs的CTL活性来自CD8(+)T细胞，而不是CD4(+)T细胞。作为对照，将乙肝表面抗原或NP多肽注射到更深的组织中只引起抗体反应，而不是CTL反应。我们进一步证明，用灭活的A/Aichi/68(H3N 2)或A/悉尼/97(H3N 2)流感病毒接种的计划免疫可对小鼠适应的A/Aichi/68病毒产生完全保护作用。综上所述，EPI将蛋白质抗原直接输送到皮肤中LC的胞浆中，并引发细胞和抗体反应。
Cytotoxic T lymphocytes (CTL) play a vital role in host defense against viral and intracellular bacterial infections. However, nonreplicating vaccines administered by intramuscular injection using a syringe and needle elicit predominantly humoral responses and not CTL responses. Here we report that epidermal powder immunization (EPI), a technology that delivers antigens on 1.5- to 2.5-mum gold particles to the epidermis using a needle-free powder delivery system, elicits CTL responses to nonreplicating antigens. Following EPI, a majority of the antigen-coated gold particles were found in the viable epidermis in the histological sections of the target skin. Further studies using transmission electron microscopy revealed the intracellular localization of the gold particles. Many Langerhans cells (LCs) at the vaccination site contained antigen-coated particles, as revealed by two-color immunofluorescence microscopy, and these cells were found in the draining lymph nodes 20 h later. Immune responses to several viral protein antigens after EPI were studied in mice. EPI with hepatitis B surface antigen (HBsAg) and a synthetic peptide of influenza virus nucleoprotein (NP peptide) elicited antigen-specific CTL responses as well as antibody responses. In an in vitro cell depletion experiment, we demonstrated that the CTL activity against HBsAg elicited by EPI was attributed to CD8(+), not CD4(+), T cells. As controls, needle injections of HBsAg or the NP peptide into deeper tissues elicited solely antibody, not CTL, responses. We further demonstrated that EPI with inactivated A/Aichi/68 (H3N2) or A/Sydney/97 (H3N2) influenza virus elicited complete protection against a mouse-adapted A/Aichi/68 virus. In summary, EPI directly delivers protein antigens to the cytosol of the LCs in the skin and elicits both cellular and antibody responses.