Purpose

We recently identified disorganized muscle protein-1 of Brugia malayi (DIM-1bm) as a vaccine candidate for human lymphatic filariasis. The present study was aimed at investigating the localization of DIM-1bm in the life-stages of B. malayi to identify the tissue target of vaccine action.

Methods

Recombinant DIM-1bm (rDIM-1bm) was prepared and antibodies were raised in BALB/c mice. Immunoblots of SDS-PAGE resolved B. malayi infective 3rd stage larvae (L₃) and adult worm antigens and rDIM-1bm were prepared and reacted with anti-rDIM-1bm sera. Sections of adult female worms and whole-mount preparations of L₃ and microfilariae (mf) were stained by immunofluorescence using rDIM-1bm antibodies and Alexa Fluor 488 labeled secondary antibodies, and examined under a confocal microscope.

Results

Immunofluorescence staining showed that DIM-1bm is localized mainly in the subcuticular muscle layer in the L₃ and the adult worms; no fluorescent signal could be detected in mf.

Conclusion

The localization of DIM-1bm in the parasites’ muscle layer suggests that the immunoprophylactic efficacy of DIM-1 is evidently due to immobilization of the parasite and its subsequent immune elimination.

中文翻译：

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人类淋巴丝虫不同生命阶段无组织的肌肉蛋白 1 的免疫定位，马来亚。

目的

我们最近确定了马来丝虫的无序肌肉蛋白-1 (DIM-1bm) 作为人类淋巴丝虫病的候选疫苗。本研究旨在调查DIM-1BM的本地化的生命阶段马来丝虫，以确定疫苗行动组织的目标。

方法

制备重组 DIM-1bm (rDIM-1bm) 并在 BALB/c 小鼠中产生抗体。SDS-PAGE 的免疫印迹可分辨马来芽孢杆菌感染性第 3 阶段幼虫 (L ₃ ) 和成虫抗原和 rDIM-1bm，并与抗 rDIM-1bm 血清反应。使用 rDIM-1bm 抗体和 Alexa Fluor 488 标记的二抗，通过免疫荧光染色成体雌性蠕虫的切片和 L ₃和微丝蚴 (mf) 的整体制剂，并在共聚焦显微镜下进行检查。

结果

免疫荧光染色显示DIM-1bm主要定位于L ₃和成虫的皮下肌层；在 mf 中未检测到荧光信号。

结论

DIM-1bm 在寄生虫肌肉层的定位表明 DIM-1 的免疫预防功效显然是由于寄生虫的固定及其随后的免疫消除。