A monoclonal antibody targeting spore wall protein 1 inhibits the proliferation of Nosema bombycis in Bombyx mori
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 s...
| Main Authors: | , , , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
American Society for Microbiology
2023-12-01
|
| Series: | Microbiology Spectrum |
| Subjects: | |
| Online Access: | https://journals.asm.org/doi/10.1128/spectrum.00681-23 |
| _version_ | 1797393607642054656 |
|---|---|
| author | Bin Yu Rong Zheng Maofei Bian Ting Liu Kun Lu Jialing Bao Guoqing Pan Zeyang Zhou Chunfeng Li |
| author_facet | Bin Yu Rong Zheng Maofei Bian Ting Liu Kun Lu Jialing Bao Guoqing Pan Zeyang Zhou Chunfeng Li |
| author_sort | Bin Yu |
| collection | DOAJ |
| description | 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. |
| first_indexed | 2024-03-09T00:06:17Z |
| format | Article |
| id | doaj.art-75baf097509f49729bd5afa63787f5f6 |
| institution | Directory Open Access Journal |
| issn | 2165-0497 |
| language | English |
| last_indexed | 2024-03-09T00:06:17Z |
| publishDate | 2023-12-01 |
| publisher | American Society for Microbiology |
| record_format | Article |
| series | Microbiology Spectrum |
| spelling | doaj.art-75baf097509f49729bd5afa63787f5f62023-12-12T13:17:20ZengAmerican Society for MicrobiologyMicrobiology Spectrum2165-04972023-12-0111610.1128/spectrum.00681-23A monoclonal antibody targeting spore wall protein 1 inhibits the proliferation of Nosema bombycis in Bombyx moriBin Yu0Rong Zheng1Maofei Bian2Ting Liu3Kun Lu4Jialing Bao5Guoqing Pan6Zeyang Zhou7Chunfeng Li8State Key Laboratory of Silkworm Genome Biology, Southwest University , Chongqing, ChinaState Key Laboratory of Silkworm Genome Biology, Southwest University , Chongqing, ChinaState Key Laboratory of Silkworm Genome Biology, Southwest University , Chongqing, ChinaState Key Laboratory of Silkworm Genome Biology, Southwest University , Chongqing, ChinaState Key Laboratory of Silkworm Genome Biology, Southwest University , Chongqing, ChinaState Key Laboratory of Silkworm Genome Biology, Southwest University , Chongqing, ChinaState Key Laboratory of Silkworm Genome Biology, Southwest University , Chongqing, ChinaState Key Laboratory of Silkworm Genome Biology, Southwest University , Chongqing, ChinaState Key Laboratory of Silkworm Genome Biology, Southwest University , Chongqing, ChinaABSTRACT 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.https://journals.asm.org/doi/10.1128/spectrum.00681-23microsporidiaNosema bombycisBombyx morimonoclonal antibodiestransgene |
| spellingShingle | Bin Yu Rong Zheng Maofei Bian Ting Liu Kun Lu Jialing Bao Guoqing Pan Zeyang Zhou Chunfeng Li A monoclonal antibody targeting spore wall protein 1 inhibits the proliferation of Nosema bombycis in Bombyx mori Microbiology Spectrum microsporidia Nosema bombycis Bombyx mori monoclonal antibodies transgene |
| title | A monoclonal antibody targeting spore wall protein 1 inhibits the proliferation of Nosema bombycis in Bombyx mori |
| title_full | A monoclonal antibody targeting spore wall protein 1 inhibits the proliferation of Nosema bombycis in Bombyx mori |
| title_fullStr | A monoclonal antibody targeting spore wall protein 1 inhibits the proliferation of Nosema bombycis in Bombyx mori |
| title_full_unstemmed | A monoclonal antibody targeting spore wall protein 1 inhibits the proliferation of Nosema bombycis in Bombyx mori |
| title_short | A monoclonal antibody targeting spore wall protein 1 inhibits the proliferation of Nosema bombycis in Bombyx mori |
| title_sort | monoclonal antibody targeting spore wall protein 1 inhibits the proliferation of nosema bombycis in bombyx mori |
| topic | microsporidia Nosema bombycis Bombyx mori monoclonal antibodies transgene |
| url | https://journals.asm.org/doi/10.1128/spectrum.00681-23 |
| work_keys_str_mv | AT binyu amonoclonalantibodytargetingsporewallprotein1inhibitstheproliferationofnosemabombycisinbombyxmori AT rongzheng amonoclonalantibodytargetingsporewallprotein1inhibitstheproliferationofnosemabombycisinbombyxmori AT maofeibian amonoclonalantibodytargetingsporewallprotein1inhibitstheproliferationofnosemabombycisinbombyxmori AT tingliu amonoclonalantibodytargetingsporewallprotein1inhibitstheproliferationofnosemabombycisinbombyxmori AT kunlu amonoclonalantibodytargetingsporewallprotein1inhibitstheproliferationofnosemabombycisinbombyxmori AT jialingbao amonoclonalantibodytargetingsporewallprotein1inhibitstheproliferationofnosemabombycisinbombyxmori AT guoqingpan amonoclonalantibodytargetingsporewallprotein1inhibitstheproliferationofnosemabombycisinbombyxmori AT zeyangzhou amonoclonalantibodytargetingsporewallprotein1inhibitstheproliferationofnosemabombycisinbombyxmori AT chunfengli amonoclonalantibodytargetingsporewallprotein1inhibitstheproliferationofnosemabombycisinbombyxmori AT binyu monoclonalantibodytargetingsporewallprotein1inhibitstheproliferationofnosemabombycisinbombyxmori AT rongzheng monoclonalantibodytargetingsporewallprotein1inhibitstheproliferationofnosemabombycisinbombyxmori AT maofeibian monoclonalantibodytargetingsporewallprotein1inhibitstheproliferationofnosemabombycisinbombyxmori AT tingliu monoclonalantibodytargetingsporewallprotein1inhibitstheproliferationofnosemabombycisinbombyxmori AT kunlu monoclonalantibodytargetingsporewallprotein1inhibitstheproliferationofnosemabombycisinbombyxmori AT jialingbao monoclonalantibodytargetingsporewallprotein1inhibitstheproliferationofnosemabombycisinbombyxmori AT guoqingpan monoclonalantibodytargetingsporewallprotein1inhibitstheproliferationofnosemabombycisinbombyxmori AT zeyangzhou monoclonalantibodytargetingsporewallprotein1inhibitstheproliferationofnosemabombycisinbombyxmori AT chunfengli monoclonalantibodytargetingsporewallprotein1inhibitstheproliferationofnosemabombycisinbombyxmori |