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首页> 外文期刊>Infection and immunity >Adherence-Inhibitory Intestinal Immunoglobulin A Antibody Response in Baboons Elicited by Use of a Synthetic Intranasal Lectin-Based Amebiasis Subunit Vaccine
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Adherence-Inhibitory Intestinal Immunoglobulin A Antibody Response in Baboons Elicited by Use of a Synthetic Intranasal Lectin-Based Amebiasis Subunit Vaccine

机译：通过使用合成的鼻内凝集素为基础的阿米巴病亚单位疫苗诱发的狒狒中的黏附抑制性肠道免疫球蛋白A抗体反应

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We designed an amebiasis subunit vaccine that is constructed by using four peptide epitopes of the galactose-inhibitable lectin heavy subunit that were recognized by intestinal secretory immunoglobulin A (IgA) antibodies from immune human subjects. These epitopes are contained in the region encompassing amino acids 758 to 1134 of the lectin heavy subunit, designated LC3. Baboons (Papio anubis) are natural hosts for Entamoeba histolytica; naturally infected baboons raised in captivity possess serum IgA antibodies to the same four LC3 epitopes as humans. Uninfected, seronegative baboons received four intranasal immunizations at 7-day intervals with the synthetic peptide vaccine (400, 800, or 1,600 μg per nostril) with cholera toxin (20 μg) as the adjuvant. As determined by an enzyme-linked immunosorbent assay (ELISA), each dose of the peptide vaccine elicited antipeptide serum IgA and IgG and intestinal IgA antibody responses in all six immunized baboons by day 28, 7 days after the last immunization (P, <0.01 for each dose compared to the cholera toxin control). The peptide vaccine elicited serum IgG and intestinal IgA antibodies that recognized purified recombinant LC3 protein (P, <0.008 and 0.02, respectively) and native lectin protein (P < 0.01). In addition, an indirect immunofluorescence assay with whole trophozoites (P < 0.01) and Western blot analysis confirmed that serum IgG antibodies from vaccinated baboons recognized native lectin protein on the surfaces of axenic E. histolytica trophozoites or from solubilized amebae. All four synthetic peptides were immunogenic; the vaccine elicited dose- and time-dependent responses, as determined by ELISA optical density readings indicating the production of serum and intestinal antibodies (P, <0.02 for antipeptide and antilectin antibodies). As a positive control, intranasal immunization with purified recombinant LC3 protein with cholera toxin as the adjuvant elicited a serum anti-LC3 IgA and IgG antibody response (P, 0.05 and <0.0001, respectively); however, no intestinal anti-LC3 IgA antibody response was observed (P = 0.4). Of interest, serum IgA and IgG antibodies elicited by the recombinant LC3 vaccine did not recognize any of the four putatively protective LC3 peptide epitopes. Both serum and fecal antibodies elicited by the peptide vaccine exhibited neutralizing activity, as determined by their dose-dependent inhibition of the galactose-specific adherence of E. histolytica trophozoites to Chinese hamster ovary cells in vitro (P, <0.001 for each group of antibodies compared to the control). In summary, a lectin-based intranasal polylysine-linked synthetic peptide vaccine was effective in eliciting an adherence-inhibitory, intestinal antilectin IgA antibody response in baboons. Future studies with the baboon model will determine vaccine efficacy against asymptomatic E. histolytica intestinal infection.
机译：我们设计了一种氨虫病亚单位疫苗，该疫苗是通过使用半乳糖抑制性凝集素重亚基的四个肽表位构建的，这些表位被来自免疫人类受试者的肠道分泌性免疫球蛋白A（IgA）抗体识别。这些表位包含在包围被称为LC3的凝集素重亚基的氨基酸758-1134的区域中。狒狒（ Papio anubis ）是 Entomoeba histolytica 的天然宿主;人工饲养的自然感染的狒狒具有与人类相同的四个LC3表位的血清IgA抗体。未感染的血清阴性狒狒以霍乱毒素（20μg）为佐剂的合成肽疫苗（每个鼻孔400、800或1600μg）以7天间隔接受了四次鼻内免疫。如通过酶联免疫吸附测定（ELISA）所确定的，到最后一次免疫接种后第7天（第28天），每剂肽疫苗在第28天，第7天都在所有六个被免疫的狒狒中引发抗肽血清IgA和IgG以及肠道IgA抗体应答。 ，与霍乱毒素对照相比，每种剂量<0.01。该肽疫苗可引起血清IgG和肠IgA抗体，它们分别识别纯化的重组LC3蛋白（分别为 P ，<0.008和0.02）和天然凝集素蛋白（ P <0.01）。此外，用完整滋养体（ P <0.01）进行的间接免疫荧光分析和Western印迹分析证实，接种过狒狒的血清IgG抗体可识别轴突 E表面的天然凝集素蛋白。组织溶菌滋养体或溶解的变形虫。所有四种合成肽均具有免疫原性。根据ELISA光学密度读数确定，疫苗引起剂量和时间依赖性反应，表明血清和肠道抗体的产生（ P ，抗肽和抗凝集素抗体的产生<0.02）。作为阳性对照，以霍乱毒素为佐剂的纯化重组LC3蛋白进行鼻内免疫可引起血清抗LC3 IgA和IgG抗体应答（分别为 P ，0.05和<0.0001）;但是，未观察到肠道抗LC3 IgA抗体反应（ P = 0.4）。有趣的是，重组LC3疫苗引起的血清IgA和IgG抗体无法识别四个公认的保护性LC3肽表位中的任何一个。由肽疫苗引发的血清和粪便抗体均显示出中和活性，这取决于它们对 E的半乳糖特异性粘附的剂量依赖性抑制作用。组蛋白滋养体对中国仓鼠卵巢细胞的体外影响（ P ，与对照组相比，每组抗体<0.001）。总之，基于凝集素的鼻内多聚赖氨酸连接的合成肽疫苗可有效引起狒狒的粘附抑制性肠道抗凝集素IgA抗体反应。狒狒模型的进一步研究将确定针对无症状E疫苗的效力。组织溶菌肠道感染。

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