Development and characterization of monoclonal antibodies against Besnoitia besnoiti tachyzoites
This is the first report on the development and characterization of eight monoclonal antibodies (MABs) generated against whole- and membrane-enriched tachyzoite extracts of the apicomplexan parasite Besnoitia besnoiti. Confocal laser scanning immunofluorescence microscopy was used to localize respec...
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| Veröffentlicht in: | Parasitology 2019-02, Vol.146 (2), p.187-196 |
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| creator | García-Lunar, P. Schares, G. Sanz-Fernández, A. Jiménez-Meléndez, A. García-Soto, I. Regidor-Cerrillo, J. Pastor-Fernández, I. Hemphill, A. Fernández-Álvarez, M. Ortega-Mora, L. M. Álvarez-García, G. |
| description | This is the first report on the development and characterization of eight monoclonal antibodies (MABs) generated against whole- and membrane-enriched tachyzoite extracts of the apicomplexan parasite Besnoitia besnoiti. Confocal laser scanning immunofluorescence microscopy was used to localize respective epitopes in B. besnoiti tachyzoites along the lytic cycle. A pattern compatible with dense granule staining was observed with MABs 2.A.12, 2.F.3 and 2.G.4, which could be confirmed by immunogold electron microscopy for MABs 2.A.12 and 2.F.3. In particular, MABs 2.F.3 and 2.G.4 were secreted during early invasion, proliferation and egress phases. MABs 3.10.8 and 5.5.11 labelled the tachyzoite surface, whilst MABs 1.17.8, 8.9.2 and 2.G.A recognized the apical tip, which is reminiscent for microneme localization. Besides, the epitopes recognized by the latter two (MABs 8.9.2 and 2.G.A) exhibited a redistribution from the anterior part across the parasite surface towards the posterior end during invasion. Most MABs developed were genus-specific. Indeed, the MABs cross-reacted neither with T. gondii nor with N. caninum tachyzoites. In summary, we have generated MABs that will be useful to study the key processes in the lytic cycle of the parasite and with additional promising diagnostic value. However, the molecular identity of the antigens recognized remains to be elucidated. |
| doi_str_mv | 10.1017/S0031182018001336 |
| format | Article |
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M. ; Álvarez-García, G.</creator><creatorcontrib>García-Lunar, P. ; Schares, G. ; Sanz-Fernández, A. ; Jiménez-Meléndez, A. ; García-Soto, I. ; Regidor-Cerrillo, J. ; Pastor-Fernández, I. ; Hemphill, A. ; Fernández-Álvarez, M. ; Ortega-Mora, L. M. ; Álvarez-García, G.</creatorcontrib><description>This is the first report on the development and characterization of eight monoclonal antibodies (MABs) generated against whole- and membrane-enriched tachyzoite extracts of the apicomplexan parasite Besnoitia besnoiti. Confocal laser scanning immunofluorescence microscopy was used to localize respective epitopes in B. besnoiti tachyzoites along the lytic cycle. A pattern compatible with dense granule staining was observed with MABs 2.A.12, 2.F.3 and 2.G.4, which could be confirmed by immunogold electron microscopy for MABs 2.A.12 and 2.F.3. In particular, MABs 2.F.3 and 2.G.4 were secreted during early invasion, proliferation and egress phases. MABs 3.10.8 and 5.5.11 labelled the tachyzoite surface, whilst MABs 1.17.8, 8.9.2 and 2.G.A recognized the apical tip, which is reminiscent for microneme localization. Besides, the epitopes recognized by the latter two (MABs 8.9.2 and 2.G.A) exhibited a redistribution from the anterior part across the parasite surface towards the posterior end during invasion. Most MABs developed were genus-specific. Indeed, the MABs cross-reacted neither with T. gondii nor with N. caninum tachyzoites. In summary, we have generated MABs that will be useful to study the key processes in the lytic cycle of the parasite and with additional promising diagnostic value. However, the molecular identity of the antigens recognized remains to be elucidated.</description><identifier>ISSN: 0031-1820</identifier><identifier>EISSN: 1469-8161</identifier><identifier>DOI: 10.1017/S0031182018001336</identifier><identifier>PMID: 30113011</identifier><language>eng</language><publisher>Cambridge, UK: Cambridge University Press</publisher><subject>Animals ; Antibodies, Monoclonal - biosynthesis ; Antibodies, Monoclonal - immunology ; Antigens ; Biomarkers ; Blotting, Western ; Cloning ; Cysts ; Diagnostic systems ; Egress ; Electron microscopy ; Electrophoresis, Polyacrylamide Gel ; Epitopes ; Epitopes - biosynthesis ; Epitopes - immunology ; Hybridomas - immunology ; Immunofluorescence ; Immunoglobulins ; Immunohistochemistry ; Investigations ; Kinases ; Localization ; Mice ; Mice, Inbred BALB C ; Microscopy ; Microscopy, Confocal ; Microscopy, Electron, Transmission ; Monoclonal antibodies ; Multiple myeloma ; Parasites ; Proteins ; Sarcocystidae - immunology ; Sarcocystidae - ultrastructure ; Tachyzoites</subject><ispartof>Parasitology, 2019-02, Vol.146 (2), p.187-196</ispartof><rights>Copyright © Cambridge University Press 2018</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c373t-e0ccc4d728cd4f6ffa4049367b9a02f93b94c82e31697843abb4b4831d69a3f23</citedby><cites>FETCH-LOGICAL-c373t-e0ccc4d728cd4f6ffa4049367b9a02f93b94c82e31697843abb4b4831d69a3f23</cites><orcidid>0000-0002-6627-7577 ; 0000-0002-0622-2128 ; 0000-0001-5831-6970</orcidid></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktohtml>$$Uhttps://www.cambridge.org/core/product/identifier/S0031182018001336/type/journal_article$$EHTML$$P50$$Gcambridge$$H</linktohtml><link.rule.ids>164,314,776,780,27901,27902,55603</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/30113011$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>García-Lunar, P.</creatorcontrib><creatorcontrib>Schares, G.</creatorcontrib><creatorcontrib>Sanz-Fernández, A.</creatorcontrib><creatorcontrib>Jiménez-Meléndez, A.</creatorcontrib><creatorcontrib>García-Soto, I.</creatorcontrib><creatorcontrib>Regidor-Cerrillo, J.</creatorcontrib><creatorcontrib>Pastor-Fernández, I.</creatorcontrib><creatorcontrib>Hemphill, A.</creatorcontrib><creatorcontrib>Fernández-Álvarez, M.</creatorcontrib><creatorcontrib>Ortega-Mora, L. M.</creatorcontrib><creatorcontrib>Álvarez-García, G.</creatorcontrib><title>Development and characterization of monoclonal antibodies against Besnoitia besnoiti tachyzoites</title><title>Parasitology</title><addtitle>Parasitology</addtitle><description>This is the first report on the development and characterization of eight monoclonal antibodies (MABs) generated against whole- and membrane-enriched tachyzoite extracts of the apicomplexan parasite Besnoitia besnoiti. Confocal laser scanning immunofluorescence microscopy was used to localize respective epitopes in B. besnoiti tachyzoites along the lytic cycle. A pattern compatible with dense granule staining was observed with MABs 2.A.12, 2.F.3 and 2.G.4, which could be confirmed by immunogold electron microscopy for MABs 2.A.12 and 2.F.3. In particular, MABs 2.F.3 and 2.G.4 were secreted during early invasion, proliferation and egress phases. MABs 3.10.8 and 5.5.11 labelled the tachyzoite surface, whilst MABs 1.17.8, 8.9.2 and 2.G.A recognized the apical tip, which is reminiscent for microneme localization. Besides, the epitopes recognized by the latter two (MABs 8.9.2 and 2.G.A) exhibited a redistribution from the anterior part across the parasite surface towards the posterior end during invasion. Most MABs developed were genus-specific. Indeed, the MABs cross-reacted neither with T. gondii nor with N. caninum tachyzoites. In summary, we have generated MABs that will be useful to study the key processes in the lytic cycle of the parasite and with additional promising diagnostic value. However, the molecular identity of the antigens recognized remains to be elucidated.</description><subject>Animals</subject><subject>Antibodies, Monoclonal - biosynthesis</subject><subject>Antibodies, Monoclonal - immunology</subject><subject>Antigens</subject><subject>Biomarkers</subject><subject>Blotting, Western</subject><subject>Cloning</subject><subject>Cysts</subject><subject>Diagnostic systems</subject><subject>Egress</subject><subject>Electron microscopy</subject><subject>Electrophoresis, Polyacrylamide Gel</subject><subject>Epitopes</subject><subject>Epitopes - biosynthesis</subject><subject>Epitopes - immunology</subject><subject>Hybridomas - immunology</subject><subject>Immunofluorescence</subject><subject>Immunoglobulins</subject><subject>Immunohistochemistry</subject><subject>Investigations</subject><subject>Kinases</subject><subject>Localization</subject><subject>Mice</subject><subject>Mice, Inbred BALB C</subject><subject>Microscopy</subject><subject>Microscopy, Confocal</subject><subject>Microscopy, Electron, Transmission</subject><subject>Monoclonal antibodies</subject><subject>Multiple myeloma</subject><subject>Parasites</subject><subject>Proteins</subject><subject>Sarcocystidae - immunology</subject><subject>Sarcocystidae - ultrastructure</subject><subject>Tachyzoites</subject><issn>0031-1820</issn><issn>1469-8161</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2019</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><sourceid>BENPR</sourceid><recordid>eNp1kE9P3DAQxa2KqizQD8AFReLCJXQmNo59bPlbCYlDyzkdOw4YJfHW9laCT49XLCC16mE0I73fvNE8xvYRjhGw_fIDgCOqBlABIOfyA1ugkLpWKHGLLdZyvda32U5KDwAguWw-sW0OiOtasF9n7o8bw3Jyc65o7it7T5FsdtE_UfZhrsJQTWEOdgwzjQXJ3oTeu1TRHfk55eqbS3Pw2VNlNlOVyd4_PpXRpT32caAxuc-bvstuL85_nl7V1zeX30-_XteWtzzXDqy1om8bZXsxyGEgAUJz2RpN0AyaGy2sahxHqVslOBkjjFAce6mJDw3fZUcvvssYfq9cyt3kk3XjSLMLq9Q1oHSjJLZQ0MO_0IewiuW7QmFbMMlPdKHwhbIxpBTd0C2jnyg-dgjdOv7un_jLzsHGeWUm179tvOZdAL4xpclE39-599v_t30G68CPdw</recordid><startdate>201902</startdate><enddate>201902</enddate><creator>García-Lunar, P.</creator><creator>Schares, G.</creator><creator>Sanz-Fernández, A.</creator><creator>Jiménez-Meléndez, A.</creator><creator>García-Soto, I.</creator><creator>Regidor-Cerrillo, J.</creator><creator>Pastor-Fernández, I.</creator><creator>Hemphill, A.</creator><creator>Fernández-Álvarez, M.</creator><creator>Ortega-Mora, L. 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M.</au><au>Álvarez-García, G.</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Development and characterization of monoclonal antibodies against Besnoitia besnoiti tachyzoites</atitle><jtitle>Parasitology</jtitle><addtitle>Parasitology</addtitle><date>2019-02</date><risdate>2019</risdate><volume>146</volume><issue>2</issue><spage>187</spage><epage>196</epage><pages>187-196</pages><issn>0031-1820</issn><eissn>1469-8161</eissn><abstract>This is the first report on the development and characterization of eight monoclonal antibodies (MABs) generated against whole- and membrane-enriched tachyzoite extracts of the apicomplexan parasite Besnoitia besnoiti. Confocal laser scanning immunofluorescence microscopy was used to localize respective epitopes in B. besnoiti tachyzoites along the lytic cycle. A pattern compatible with dense granule staining was observed with MABs 2.A.12, 2.F.3 and 2.G.4, which could be confirmed by immunogold electron microscopy for MABs 2.A.12 and 2.F.3. In particular, MABs 2.F.3 and 2.G.4 were secreted during early invasion, proliferation and egress phases. MABs 3.10.8 and 5.5.11 labelled the tachyzoite surface, whilst MABs 1.17.8, 8.9.2 and 2.G.A recognized the apical tip, which is reminiscent for microneme localization. Besides, the epitopes recognized by the latter two (MABs 8.9.2 and 2.G.A) exhibited a redistribution from the anterior part across the parasite surface towards the posterior end during invasion. Most MABs developed were genus-specific. Indeed, the MABs cross-reacted neither with T. gondii nor with N. caninum tachyzoites. In summary, we have generated MABs that will be useful to study the key processes in the lytic cycle of the parasite and with additional promising diagnostic value. However, the molecular identity of the antigens recognized remains to be elucidated.</abstract><cop>Cambridge, UK</cop><pub>Cambridge University Press</pub><pmid>30113011</pmid><doi>10.1017/S0031182018001336</doi><tpages>10</tpages><orcidid>https://orcid.org/0000-0002-6627-7577</orcidid><orcidid>https://orcid.org/0000-0002-0622-2128</orcidid><orcidid>https://orcid.org/0000-0001-5831-6970</orcidid></addata></record> |
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| subjects | Animals Antibodies, Monoclonal - biosynthesis Antibodies, Monoclonal - immunology Antigens Biomarkers Blotting, Western Cloning Cysts Diagnostic systems Egress Electron microscopy Electrophoresis, Polyacrylamide Gel Epitopes Epitopes - biosynthesis Epitopes - immunology Hybridomas - immunology Immunofluorescence Immunoglobulins Immunohistochemistry Investigations Kinases Localization Mice Mice, Inbred BALB C Microscopy Microscopy, Confocal Microscopy, Electron, Transmission Monoclonal antibodies Multiple myeloma Parasites Proteins Sarcocystidae - immunology Sarcocystidae - ultrastructure Tachyzoites |
| title | Development and characterization of monoclonal antibodies against Besnoitia besnoiti tachyzoites |
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