Localization and RNA Interference-Driven Inhibition of a Brugia malayi-Encoded Interleukin-5 Receptor Binding Protein

• Published in: Infection and immunity 2022-05, Vol.90 (5), p.e0031721-e0031721
• Main Authors: Mejia, Rojelio, Bennuru, Sasisekhar, Oksov, Yelena, Lustigman, Sara, Munirathinam, Gnanasekar, Kalyanasundaram, Ramaswamy, Nutman, Thomas B
• Format: Article
• Language: English
• Subjects: Animals; Brugia malayi - genetics; Fungal and Parasitic Infections; Immunology; Interleukin-5 - genetics; Rabbits; Receptors, Interleukin-5 - genetics; Receptors, Interleukin-5 - metabolism; RNA Interference; RNA, Messenger - metabolism
• ISSN: 0019-9567; 1098-5522
• DOI: 10.1128/iai.00317-21

A molecule we termed Brugia malayi IL-5 receptor (IL-5R) binding protein (BmIL5Rbp; also known as Bm8757) was identified from B. malayi filarial worms and found to inhibit human interleukin-5 (IL-5) binding to its human receptor competitively. After the expression and purification of a recombinant BmIL5Rbp and generation of BmIL5Rbp-specific rabbit antibody, we localized the molecule on B. malayi worms through immunohistochemistry and immunoelectron microscopy. RNA interference (RNAi) was used to inhibit BmIL5Rbp mRNA and protein production. BmIL5Rbp was shown to localize to the cuticle of Brugia malayi and to be released in its excretory/secretory products. RNAi inhibited BmIL5Rbp mRNA production by 33%, reduced the surface protein expression by ~50%, and suppressed the release of BmIL5Rbp in the excretory/secretory products. RNAi has been used successfully to knock down the mRNA and protein expression of BmIL5Rbp in the early larval stages of B. malayi and provided a proof of principle for the local inhibition of the human IL-5R. These findings provide evidence that a parasite-encoded IL-5R antagonist may locally inhibit a vital host innate immune activation of IL-5 on eosinophils.