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sPLA2 behaves like a prophylactic agent and mediates cellular and humoral immune responses in Plutella xylostella.
Archives of Insect Biochemistry and Physiology ( IF 1.5 ) Pub Date : 2020-03-20 , DOI: 10.1002/arch.21670 Miltan Chandra Roy 1 , Yonggyun Kim 1
Archives of Insect Biochemistry and Physiology ( IF 1.5 ) Pub Date : 2020-03-20 , DOI: 10.1002/arch.21670 Miltan Chandra Roy 1 , Yonggyun Kim 1
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Most immune effectors are inducible to microbial pathogen infection while some are already present to act as prophylactic immunity against as yet unseen infection. This study identified secretory phospholipase A2 (sPLA2) as a prophylactic factor in diamondback moth, Plutella xylostella. Western blotting using a polyclonal antibody raised against other lepidopteran sPLA2 reacted specifically with ∼25 kDa protein, which was present at approximately 0.4 mM in the plasma of naïve larvae. Interrogation of P. xylostella transcriptomes revealed an open‐reading frame for sPLA2 (Px‐sPLA2), exhibiting high homology with other Group III sPLA2s. Px‐sPLA2 was expressed in all developmental stages. In the larval stage, bacterial challenge induced its expression in hemocytes and fat body but not in gut or epidermis. RNA interference (RNAi) suppressed Px‐sPLA2 messenger RNA level and sPLA2 activity in plasma. An inhibition zone assay showed that Px‐sPLA2 exhibited antibacterial activities against different species, because specific RNAi knockdown impaired the activity. The RNAi treatment also suppressed the cellular immune response assessed by hemocyte nodule formation and humoral immune response assessed by antimicrobial peptide gene expression. Finally, benzylideneacetone (BZA, a specific sPLA2 inhibitor) treatment inhibited plasma sPLA2 activity of naive larvae in a dose‐dependent manner. An addition of BZA significantly increased the bacterial virulence of an entomopathogen, Bacillus thuringiensis. These results suggest that Px‐sPLA2 is an immune‐associated factor of P. xylostella and its relatively high level of concentration in the plasma of naive larvae strongly suggests its role as a prophylactic factor in defending against pathogens at early infection stages.
中文翻译：
sPLA2的行为类似于预防剂，并在小菜蛾中介导细胞和体液免疫反应。
大多数免疫效应物可诱导微生物病原体感染，而一些已经存在，可作为针对尚未见到的感染的预防性免疫。这项研究确定了分泌性磷脂酶A 2（sPLA 2）是小菜蛾小菜蛾的预防因子。使用针对其他鳞翅目sPLA 2的多克隆抗体进行的蛋白质印迹与约25 kDa的蛋白质发生了特异性反应，该蛋白质在幼稚的幼体血浆中的含量约为0.4 mM。对P. xylostella转录组的询问揭示了sPLA 2（Px-sPLA 2）的开放阅读框，与其他III类sPLA 2具有高度同源性s。Px-sPLA 2在所有发育阶段都有表达。在幼虫阶段，细菌攻击诱导其在血细胞和脂肪体内表达，但未在肠道或表皮中表达。RNA干扰（RNAi）抑制了血浆中Px-sPLA 2信使RNA水平和sPLA 2活性。抑制区分析表明，Px-sPLA 2表现出对不同物种的抗菌活性，因为特定的RNAi抑制作用削弱了该活性。RNAi处理还抑制了通过血细胞结节形成评估的细胞免疫反应和通过抗菌肽基因表达评估的体液免疫反应。最后，亚苄基丙酮（BZA，一种特定的sPLA 2抑制剂）处理以剂量依赖性方式抑制幼稚幼虫的血浆sPLA 2活性。添加BZA可显着增加昆虫病原体苏云金芽孢杆菌的细菌毒性。这些结果表明，Px-sPLA 2是小菜蛾的免疫相关因子，其在幼稚血浆中的浓度相对较高，强烈表明了其作为预防因子在早期感染阶段防御病原体的作用。
更新日期：2020-03-20
中文翻译：
sPLA2的行为类似于预防剂，并在小菜蛾中介导细胞和体液免疫反应。
大多数免疫效应物可诱导微生物病原体感染，而一些已经存在，可作为针对尚未见到的感染的预防性免疫。这项研究确定了分泌性磷脂酶A 2（sPLA 2）是小菜蛾小菜蛾的预防因子。使用针对其他鳞翅目sPLA 2的多克隆抗体进行的蛋白质印迹与约25 kDa的蛋白质发生了特异性反应，该蛋白质在幼稚的幼体血浆中的含量约为0.4 mM。对P. xylostella转录组的询问揭示了sPLA 2（Px-sPLA 2）的开放阅读框，与其他III类sPLA 2具有高度同源性s。Px-sPLA 2在所有发育阶段都有表达。在幼虫阶段，细菌攻击诱导其在血细胞和脂肪体内表达，但未在肠道或表皮中表达。RNA干扰（RNAi）抑制了血浆中Px-sPLA 2信使RNA水平和sPLA 2活性。抑制区分析表明，Px-sPLA 2表现出对不同物种的抗菌活性，因为特定的RNAi抑制作用削弱了该活性。RNAi处理还抑制了通过血细胞结节形成评估的细胞免疫反应和通过抗菌肽基因表达评估的体液免疫反应。最后，亚苄基丙酮（BZA，一种特定的sPLA 2抑制剂）处理以剂量依赖性方式抑制幼稚幼虫的血浆sPLA 2活性。添加BZA可显着增加昆虫病原体苏云金芽孢杆菌的细菌毒性。这些结果表明，Px-sPLA 2是小菜蛾的免疫相关因子，其在幼稚血浆中的浓度相对较高，强烈表明了其作为预防因子在早期感染阶段防御病原体的作用。
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