Identification and characterization of intestinal antigen-presenting cells involved in uptake and processing of a nontoxic recombinant chimeric mucosal immunogen based on cholera toxin using imaging flow cytometry.
Identification and characterization of intestinal antigen-presenting cells involved in uptake and processing of a nontoxic recombinant chimeric mucosal immunogen based on cholera toxin using imaging flow cytometry.
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使用成像流式细胞术对参与摄取和加工基于霍乱毒素的无毒重组嵌合粘膜免疫原的肠道抗原呈递细胞进行鉴定和表征。
DOI：
10.1128/cvi.00452-13
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发表时间：
2014
期刊：
影响因子：
--
通讯作者：
Russell,MichaelW
中科院分区：
文献类型：
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作者：
Zhao,Weiwei;Minderman,Hans;Russell,MichaelW
将变形链球菌抗原AgI/II的唾液结合区（SBR）与霍乱毒素（CT）的A2/B亚单位构建成嵌合免疫原SBR-CTA 2/B，经胃内免疫小鼠，诱导产生抗变形链球菌的唾液抗体和循环抗体。防止龋齿的变异蛋白。我们先前发现SBR-CTA 2/B激活派伊尔集合淋巴结（PP）和肠系膜淋巴结（MLN）中的树突状细胞（DC）。为了鉴定参与SBR-CTA 2/B的肠摄取和免疫应答的起始的细胞，用荧光素标记的SBR-CTA 2/B或SBR灌胃免疫小鼠，并在荧光染色细胞表面标记物后通过成像流式细胞术检查肠细胞。SBR-CTA 2/B在PP中优先被CD 103 +DC吸收，在肠固有层（LP）中优先被CD 103+和CD 11 c +DC吸收，而SBR在2 - 16 h内被PP CD 11 c +DC吸收的程度较低。到16小时，在MLN中发现含有荧光素标记的SBR-CTA 2/B的CD 103+和CD 11 c +DC，并且显示趋化因子受体CCR 7的上调。大量含SBR-CTA 2/B的DC与CD 4+（辅助性T）细胞相互作用，这些细胞核转录因子T-bet或RORγt共染色，鉴定为Th 1或Th 17细胞。相反，含有SBR的CD 11 c +DC优先与GATA 3+（Th 2）细胞相互作用。未发现含有SBR或SBR-CTA 2/B的DC与Foxp 3+（T调节）细胞相互作用。我们的结论是，SBR的耦合CTA 2/B增强其免疫原性，通过促进DC在PP和LP的摄取，这些含抗原的DC迁移到MLN和相互作用优先与Th 1和Th 17细胞诱导主动免疫反应。
Intragastric immunization with recombinant chimeric immunogen, SBR-CTA2/B, constructed from the saliva-binding region (SBR) of Streptococcus mutans antigen AgI/II and the A2/B subunits of cholera toxin (CT) induces salivary and circulating antibodies against S. mutans that protect against dental caries. We previously found that SBR-CTA2/B activated dendritic cells (DC) in the Peyer's patches (PP) and mesenteric lymph nodes (MLN). To identify the cells involved in the intestinal uptake of SBR-CTA2/B and the initiation of immune responses, mice were immunized intragastrically with fluorescein-labeled SBR-CTA2/B or SBR, and intestinal cells were examined by imaging flow cytometry after fluorescent staining for cell surface markers. SBR-CTA2/B was preferentially taken up by CD103+DC in the PP and by both CD103+and CD11c+DC in intestinal lamina propria (LP), whereas SBR was taken up to a lesser extent by PP CD11c+DC, within 2 to 16 h. By 16 h, CD103+and CD11c+DC containing fluorescein-labeled SBR-CTA2/B were found in MLN and showed upregulation of the chemokine receptor CCR7. Large numbers of SBR-CTA2/B-containing DC were found interacting with CD4+(T helper) cells, which costained for nuclear transcription factors T-bet or RORγt, identifying them as Th1 or Th17 cells. In contrast, SBR-containing CD11c+DC interacted preferentially with GATA3+(Th2) cells. No SBR- or SBR-CTA2/B-containing DC were found interacting with Foxp3+(T regulatory) cells. We conclude that the coupling of SBR to CTA2/B enhances its immunogenicity by promoting uptake by DC in both PP and LP and that these antigen-containing DC migrated to MLN and interacted preferentially with Th1 and Th17 cells to induce active immune responses.