A monoclonal antibody targeting spore wall protein 1 inhibits the proliferation of Nosema bombycis in Bombyx mori

Abstract

ABSTRACT Microsporidia are obligate intracellular single-cell eukaryotic parasites that can infect almost all kinds of animals and cause microsporidiosis. However, there are a few achievements of microsporidiosis treatments. Nosema bombycis could cause a microsporidiosis condition, pébrine, in the silkworm. Treatment and prevention of pébrine are still the research hotspots in sericulture. In this study, N. bombycis was treated with K2CO3, the spore walls were removed by centrifugation, and then, the alkali-soluble germination proteins were used as antigens to develop monoclonal antibodies (mAbs). Three mAbs were successfully screened and one mAb named G9 showed the highest titer among these monoclonal antibodies after enzyme-linked-immunosorbent serologic assay (ELISA). Mass spectrometry analysis confirmed that the mAb G9 could recognize the N. bombycis spore wall protein 1. Furthermore, the heavy chain and light chain sequences of the G9 monoclonal antibody were cloned, respectively. The vectors that expressing the intact antibodies and the single-chain variable fragments (scFvs) of G9 were constructed, and then, these vectors were used to develop the transgenic silkworm cell lines or transgenic silkworms. The inhibitory effects against N. bombycis were evaluated by the count of microsporidia and qPCR. The scFvs showed better effect on blocking the proliferation of N. bombycis than the intact antibody, and the scFv without the secretory signal peptide was more effective than that with signal peptide. Our study has provided novel strategies for microsporidiosis control and the essential groundwork for the future development of N. bombycis-resistant transgenic silkworms. IMPORTANCE There are a few reports on the resistance of microsporidia, including Nosema bombycis. Here, the alkali-soluble germination proteins of N. bombycis were used as immunogens to prepare a monoclonal antibody, and its single-chain variable fragments effectively blocked microsporidia infection. Our study has provided novel strategies for microsporidiosis control and demonstrated a useful method for the potential treatment of other microsporidia diseases.

Keywords

• microsporidia
• Nosema bombycis
• Bombyx mori
• monoclonal antibodies
• transgene