Hap2, a novel gene in Babesia bigemina is expressed in tick stages, and specific antibodies block zygote formation
Abstract Background Bovine babesiosis is a tick-borne disease caused by the protozoan parasites of the genus Babesia. In their host vector, Babesia spp. undergo sexual reproduction. Therefore, the development of sexual stages and the subsequent formation of the zygote are essential for the parasite...
| Main Authors: | , , , , , , , , |
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| Format: | Article |
| Language: | English |
| Published: |
BMC
2017-11-01
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| Series: | Parasites & Vectors |
| Subjects: | |
| Online Access: | http://link.springer.com/article/10.1186/s13071-017-2510-0 |
| _version_ | 1818257381408112640 |
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| author | Minerva Camacho-Nuez Diego Josimar Hernández-Silva Elizabeth Jacqueline Castañeda-Ortiz María Elena Paredes-Martínez Marisol Karina Rocha-Martínez María Elizbeth Alvarez-Sánchez Ricardo Francisco Mercado-Curiel Gabriela Aguilar-Tipacamu Juan Mosqueda |
| author_facet | Minerva Camacho-Nuez Diego Josimar Hernández-Silva Elizabeth Jacqueline Castañeda-Ortiz María Elena Paredes-Martínez Marisol Karina Rocha-Martínez María Elizbeth Alvarez-Sánchez Ricardo Francisco Mercado-Curiel Gabriela Aguilar-Tipacamu Juan Mosqueda |
| author_sort | Minerva Camacho-Nuez |
| collection | DOAJ |
| description | Abstract Background Bovine babesiosis is a tick-borne disease caused by the protozoan parasites of the genus Babesia. In their host vector, Babesia spp. undergo sexual reproduction. Therefore, the development of sexual stages and the subsequent formation of the zygote are essential for the parasite to invade the intestinal cells of the vector tick and continue its life-cycle. HAP2/GCS1 is a protein identified in plants, protozoan parasites and other organisms that has an important role during membrane fusion in fertilization processes. The identification and characterization of HAP-2 protein in Babesia would be very significant to understand the biology of the parasite and to develop a transmission-blocking vaccine in the future. Results To isolate and sequence the hap2 gene DNA from an infected bovine with Babesia bigemina was purified. The hap2 gene was amplified, cloned and sequenced. The sequences of hap2 from four geographically different strains showed high conservation at the amino acid level, including the typical structure with a signal peptide and the HAP2/GSC domain. Antisera anti-HAP2 against the conserved extracellular region of the HAP2 amino acid sequence were obtained from rabbits. The expression of hap2 in the host and vector tissues was analyzed by using semi-quantitative RT-PCR, and the protein was examined by western blot and immunofluorescence. Based on the RT-PCR and WB results, HAP2 is expressed in both, sexual stages induced in vitro, and in infected ticks as well. We did not detect any expression in asexual erythrocytic stages of B. bigemina, relevantly anti-HAP2 specific antibodies were able to block zygotes formation in vitro. Conclusion Babesia bigemina HAP2 is expressed only in tick-infecting stages, and specific antibodies block zygote formation. Further studies regarding the function of HAP2 during tick infection may provide new insights into the molecular mechanisms of sexual reproduction of the parasite. |
| first_indexed | 2024-12-12T17:42:45Z |
| format | Article |
| id | doaj.art-352de4fd7ab34aa7a156a84a8c37c833 |
| institution | Directory Open Access Journal |
| issn | 1756-3305 |
| language | English |
| last_indexed | 2024-12-12T17:42:45Z |
| publishDate | 2017-11-01 |
| publisher | BMC |
| record_format | Article |
| series | Parasites & Vectors |
| spelling | doaj.art-352de4fd7ab34aa7a156a84a8c37c8332022-12-22T00:17:01ZengBMCParasites & Vectors1756-33052017-11-011011910.1186/s13071-017-2510-0Hap2, a novel gene in Babesia bigemina is expressed in tick stages, and specific antibodies block zygote formationMinerva Camacho-Nuez0Diego Josimar Hernández-Silva1Elizabeth Jacqueline Castañeda-Ortiz2María Elena Paredes-Martínez3Marisol Karina Rocha-Martínez4María Elizbeth Alvarez-Sánchez5Ricardo Francisco Mercado-Curiel6Gabriela Aguilar-Tipacamu7Juan Mosqueda8Posgrado en Ciencias Genómicas, Universidad Autónoma de la Ciudad de MéxicoFacultad de Ciencias Naturales, Universidad Autónoma de QuerétaroPosgrado en Ciencias Genómicas, Universidad Autónoma de la Ciudad de MéxicoPosgrado en Ciencias Genómicas, Universidad Autónoma de la Ciudad de MéxicoPosgrado en Ciencias Genómicas, Universidad Autónoma de la Ciudad de MéxicoPosgrado en Ciencias Genómicas, Universidad Autónoma de la Ciudad de MéxicoFacultad de Medicina, Universidad Autónoma de QuerétaroFacultad de Ciencias Naturales, Universidad Autónoma de QuerétaroFacultad de Ciencias Naturales, Universidad Autónoma de QuerétaroAbstract Background Bovine babesiosis is a tick-borne disease caused by the protozoan parasites of the genus Babesia. In their host vector, Babesia spp. undergo sexual reproduction. Therefore, the development of sexual stages and the subsequent formation of the zygote are essential for the parasite to invade the intestinal cells of the vector tick and continue its life-cycle. HAP2/GCS1 is a protein identified in plants, protozoan parasites and other organisms that has an important role during membrane fusion in fertilization processes. The identification and characterization of HAP-2 protein in Babesia would be very significant to understand the biology of the parasite and to develop a transmission-blocking vaccine in the future. Results To isolate and sequence the hap2 gene DNA from an infected bovine with Babesia bigemina was purified. The hap2 gene was amplified, cloned and sequenced. The sequences of hap2 from four geographically different strains showed high conservation at the amino acid level, including the typical structure with a signal peptide and the HAP2/GSC domain. Antisera anti-HAP2 against the conserved extracellular region of the HAP2 amino acid sequence were obtained from rabbits. The expression of hap2 in the host and vector tissues was analyzed by using semi-quantitative RT-PCR, and the protein was examined by western blot and immunofluorescence. Based on the RT-PCR and WB results, HAP2 is expressed in both, sexual stages induced in vitro, and in infected ticks as well. We did not detect any expression in asexual erythrocytic stages of B. bigemina, relevantly anti-HAP2 specific antibodies were able to block zygotes formation in vitro. Conclusion Babesia bigemina HAP2 is expressed only in tick-infecting stages, and specific antibodies block zygote formation. Further studies regarding the function of HAP2 during tick infection may provide new insights into the molecular mechanisms of sexual reproduction of the parasite.http://link.springer.com/article/10.1186/s13071-017-2510-0Bovine babesiosisBabesia bigeminaHAP2Gamete fusion |
| spellingShingle | Minerva Camacho-Nuez Diego Josimar Hernández-Silva Elizabeth Jacqueline Castañeda-Ortiz María Elena Paredes-Martínez Marisol Karina Rocha-Martínez María Elizbeth Alvarez-Sánchez Ricardo Francisco Mercado-Curiel Gabriela Aguilar-Tipacamu Juan Mosqueda Hap2, a novel gene in Babesia bigemina is expressed in tick stages, and specific antibodies block zygote formation Parasites & Vectors Bovine babesiosis Babesia bigemina HAP2 Gamete fusion |
| title | Hap2, a novel gene in Babesia bigemina is expressed in tick stages, and specific antibodies block zygote formation |
| title_full | Hap2, a novel gene in Babesia bigemina is expressed in tick stages, and specific antibodies block zygote formation |
| title_fullStr | Hap2, a novel gene in Babesia bigemina is expressed in tick stages, and specific antibodies block zygote formation |
| title_full_unstemmed | Hap2, a novel gene in Babesia bigemina is expressed in tick stages, and specific antibodies block zygote formation |
| title_short | Hap2, a novel gene in Babesia bigemina is expressed in tick stages, and specific antibodies block zygote formation |
| title_sort | hap2 a novel gene in babesia bigemina is expressed in tick stages and specific antibodies block zygote formation |
| topic | Bovine babesiosis Babesia bigemina HAP2 Gamete fusion |
| url | http://link.springer.com/article/10.1186/s13071-017-2510-0 |
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