A nontoxic mutant of cholera toxin elicits Th2-type responses for enhanced mucosal immunity
A nontoxic mutant of cholera toxin elicits Th2-type responses for enhanced mucosal immunity
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DOI：
10.1073/pnas.94.10.5267
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发表时间：
1997-05-13
影响因子：
11.1
通讯作者：
McGhee, JR
中科院分区：
文献类型：
--
作者：
Yamamoto, S;Kiyono, H;McGhee, JR
我们的特点是无毒突变霍乱毒素（CT）作为粘膜佐剂在小鼠中。突变体CT是通过在A亚基（S61 F）的61位丝氨酸被苯丙氨酸取代而产生的，这导致ADP核糖基转移酶活性和毒性的丧失。用卵清蛋白、破伤风类毒素或流感病毒单独或与突变CT S61 F、天然CT或重组CT-B一起鼻内免疫小鼠。用这些蛋白加上S61 F免疫的小鼠显示出与天然CT诱导的那些相当的蛋白特异性IgG和伊加抗体的高血清滴度。此外，在鼻和阴道洗液、唾液和粪便提取物中观察到高蛋白特异性伊加抗体应答，以及用这些蛋白和S61 F或天然CT鼻内免疫的小鼠的颈部淋巴结和肺组织中IgG和伊加抗体形成细胞的数量增加，但单独用重组CT-B或蛋白则没有。S61 F和天然CT均增强了肺和脾组织中卵清蛋白特异性CD 4（+）T细胞的诱导，这些T细胞产生Th 2型细胞因子模式的白细胞介素4（IL-4）、IL-5、IL-6和IL-10，通过分泌蛋白分析和精氨酸特异性mRNA定量测定。这些结果表明，突变CT S61 F在鼻内给药时是一种有效的粘膜佐剂，并诱导由CD 4（+）Th 2型细胞介导的粘膜和全身抗体反应。
We have characterized a nontoxic mutant of cholera toxin (CT) as a mucosal adjuvant in mice. The mutant CT was made by substitution of serine with phenylalanine at position 61 of the A subunit (S61F), which resulted in loss of ADP ribosyltransferase activity and toxicity. Mice were intranasally immunized with ovalbumin, tetanus toxoid, or influenza virus either alone or together with mutant CT S61F, native CT, or recombinant CT-B. Mice immunized with these proteins plus S61F showed high serum titers of protein-specific IgG and IgA antibodies that were comparable to those induced by native CT. Further, high protein-specific IgA antibody responses were observed in nasal and vaginal washes, saliva, and fecal extracts as well as increased numbers of IgG and IgA antibody forming cells in cervical lymph nodes and lung tissues of mice intranasally immunized with these proteins and S61F or native CT, but not with recombinant CT-B or protein alone. Both S61F and native CT enhanced the induction of ovalbumin-specific CD4(+) T cells in lung and splenic tissues, and these T cells produced a Th2-type cytokine pattern of interleukin 4 (IL-4), IL-5, IL-6, and IL-10 as determined by analysis of secreted proteins and by quantitation of cytokine-specific mRNA. These results have shown that mutant CT S61F is an effective mucosal adjuvant when administrated intranasally and induces mucosal and systemic antibody responses which are mediated by CD4(+) Th2-type cells.