Novel Insights Into Leishmania (Viannia) braziliensis In Vitro Fitness Guided by Temperature Changes Along With Its Subtilisins and Oligopeptidase B
Proteases are virulence factors with a recognized impact on the Leishmania spp. life cycle. This study considers a set of analyses measuring phenotypic factors of L. (V.) braziliensis clinical isolates as promastigotes growth curves, murine peritoneal macrophages infection, inflammatory mediators pr...
| Main Authors: | , , , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
Frontiers Media S.A.
2022-04-01
|
| Series: | Frontiers in Cellular and Infection Microbiology |
| Subjects: | |
| Online Access: | https://www.frontiersin.org/articles/10.3389/fcimb.2022.805106/full |
| _version_ | 1818049425243635712 |
|---|---|
| author | Anabel Zabala-Peñafiel Lea Cysne-Finkelstein Fatima Conceição-Silva Aline Fagundes Luciana de Freitas Campos Miranda Franklin Souza-Silva Franklin Souza-Silva Artur A. M. L. Brandt Artur A. M. L. Brandt Geovane Dias-Lopes Carlos Roberto Alves |
| author_facet | Anabel Zabala-Peñafiel Lea Cysne-Finkelstein Fatima Conceição-Silva Aline Fagundes Luciana de Freitas Campos Miranda Franklin Souza-Silva Franklin Souza-Silva Artur A. M. L. Brandt Artur A. M. L. Brandt Geovane Dias-Lopes Carlos Roberto Alves |
| author_sort | Anabel Zabala-Peñafiel |
| collection | DOAJ |
| description | Proteases are virulence factors with a recognized impact on the Leishmania spp. life cycle. This study considers a set of analyses measuring phenotypic factors of L. (V.) braziliensis clinical isolates as promastigotes growth curves, murine peritoneal macrophages infection, inflammatory mediators production, and serine proteases gene expression (subtilisin 13: S13, subtilisin 28: S28, oligopeptidase B: OPB) assessing these isolates’ fitness on in vitro conditions. Parasites had different behavior during the early growth phase from day zero to day three, and all isolates reached the stationary growth phase between days four and seven. Macrophages infection showed two tendencies, one of decreased infection rate and number of parasites per macrophage (Infection Index <1000) and another with a constant infection index (≥1400). TNF-α (≥10 pg/mL) detected in infections by 75% of isolates, IL-6 (≥80 pg/mL) by 30% of isolates and low levels of NO (≥0.01µM) in almost all infections. Gene expression showed higher values of S13 (≥2RQ) in the intracellular amastigotes of all the isolates evaluated. On the contrary, S28 expression was low (≤1RQ) in all isolates. OPB expression was different between promastigotes and intracellular amastigotes, being significantly higher (≥2RQ) in the latter form of 58% of the isolates. Predictive structural assays of S13 and OPB were performed to explore temperature influence on gene expression and the encoded proteases. Gene expression data is discussed based on in silico predictions of regulatory regions that show plasticity in the linearity index of secondary structures of S13 and OPB 3’-untranslated regions of mRNA, dependent on temperature changes. While hairpin structures suggest an active region of mRNA for both genes above 26°C, pseudoknot structure found in S13 is an indication of a particular profile of this gene at mammalian host temperatures (37°C). Furthermore, the predicted 3D structures are in accordance with the influence of these temperatures on the catalytic site stability of both enzymes, favoring their action over peptide substrates. Data gathered here suggest that L. (V.) braziliensis serine proteases can be influenced by the temperature conditions affecting parasite fitness throughout its life cycle. |
| first_indexed | 2024-12-10T10:37:22Z |
| format | Article |
| id | doaj.art-a0234b75763f4641b5b2e480b1fc40c3 |
| institution | Directory Open Access Journal |
| issn | 2235-2988 |
| language | English |
| last_indexed | 2024-12-10T10:37:22Z |
| publishDate | 2022-04-01 |
| publisher | Frontiers Media S.A. |
| record_format | Article |
| series | Frontiers in Cellular and Infection Microbiology |
| spelling | doaj.art-a0234b75763f4641b5b2e480b1fc40c32022-12-22T01:52:24ZengFrontiers Media S.A.Frontiers in Cellular and Infection Microbiology2235-29882022-04-011210.3389/fcimb.2022.805106805106Novel Insights Into Leishmania (Viannia) braziliensis In Vitro Fitness Guided by Temperature Changes Along With Its Subtilisins and Oligopeptidase BAnabel Zabala-Peñafiel0Lea Cysne-Finkelstein1Fatima Conceição-Silva2Aline Fagundes3Luciana de Freitas Campos Miranda4Franklin Souza-Silva5Franklin Souza-Silva6Artur A. M. L. Brandt7Artur A. M. L. Brandt8Geovane Dias-Lopes9Carlos Roberto Alves10Laboratório de Biologia Molecular e Doenças Endêmicas, Instituto Oswaldo Cruz, Fundação Oswaldo Cruz, Rio de Janeiro, BrazilLaboratório de Imunoparasitologia, Instituto Oswaldo Cruz, Fundação Oswaldo Cruz, Rio de Janeiro, BrazilLaboratório de Imunoparasitologia, Instituto Oswaldo Cruz, Fundação Oswaldo Cruz, Rio de Janeiro, BrazilLaboratório de Pesquisa Clínica e Vigilância em Leishmanioses, Instituto Nacional de Infectologia Evandro Chagas, Fundação Oswaldo Cruz, Rio de Janeiro, BrazilLaboratório de Pesquisa Clínica e Vigilância em Leishmanioses, Instituto Nacional de Infectologia Evandro Chagas, Fundação Oswaldo Cruz, Rio de Janeiro, BrazilCentro de Desenvolvimento Tecnológico em Saúde, Fundação Oswaldo Cruz, Rio de Janeiro, BrazilUniversidade Iguaçu, Dom Rodrigo, Nova Iguaçu, Rio de Janeiro, BrazilDepartamento de Computação e Sistemas, Faculdade de Educação Tecnológica do Estado do Rio de Janeiro, Rio de Janeiro, BrazilDepartamento de Ciência da Computação, Univeritas-Rio, Rio de Janeiro, BrazilLaboratório de Biologia Molecular e Doenças Endêmicas, Instituto Oswaldo Cruz, Fundação Oswaldo Cruz, Rio de Janeiro, BrazilLaboratório de Biologia Molecular e Doenças Endêmicas, Instituto Oswaldo Cruz, Fundação Oswaldo Cruz, Rio de Janeiro, BrazilProteases are virulence factors with a recognized impact on the Leishmania spp. life cycle. This study considers a set of analyses measuring phenotypic factors of L. (V.) braziliensis clinical isolates as promastigotes growth curves, murine peritoneal macrophages infection, inflammatory mediators production, and serine proteases gene expression (subtilisin 13: S13, subtilisin 28: S28, oligopeptidase B: OPB) assessing these isolates’ fitness on in vitro conditions. Parasites had different behavior during the early growth phase from day zero to day three, and all isolates reached the stationary growth phase between days four and seven. Macrophages infection showed two tendencies, one of decreased infection rate and number of parasites per macrophage (Infection Index <1000) and another with a constant infection index (≥1400). TNF-α (≥10 pg/mL) detected in infections by 75% of isolates, IL-6 (≥80 pg/mL) by 30% of isolates and low levels of NO (≥0.01µM) in almost all infections. Gene expression showed higher values of S13 (≥2RQ) in the intracellular amastigotes of all the isolates evaluated. On the contrary, S28 expression was low (≤1RQ) in all isolates. OPB expression was different between promastigotes and intracellular amastigotes, being significantly higher (≥2RQ) in the latter form of 58% of the isolates. Predictive structural assays of S13 and OPB were performed to explore temperature influence on gene expression and the encoded proteases. Gene expression data is discussed based on in silico predictions of regulatory regions that show plasticity in the linearity index of secondary structures of S13 and OPB 3’-untranslated regions of mRNA, dependent on temperature changes. While hairpin structures suggest an active region of mRNA for both genes above 26°C, pseudoknot structure found in S13 is an indication of a particular profile of this gene at mammalian host temperatures (37°C). Furthermore, the predicted 3D structures are in accordance with the influence of these temperatures on the catalytic site stability of both enzymes, favoring their action over peptide substrates. Data gathered here suggest that L. (V.) braziliensis serine proteases can be influenced by the temperature conditions affecting parasite fitness throughout its life cycle.https://www.frontiersin.org/articles/10.3389/fcimb.2022.805106/fullLeishmania (Viannia) braziliensisphenotypic characteristicsgene expressionserine proteasesmolecular dynamics |
| spellingShingle | Anabel Zabala-Peñafiel Lea Cysne-Finkelstein Fatima Conceição-Silva Aline Fagundes Luciana de Freitas Campos Miranda Franklin Souza-Silva Franklin Souza-Silva Artur A. M. L. Brandt Artur A. M. L. Brandt Geovane Dias-Lopes Carlos Roberto Alves Novel Insights Into Leishmania (Viannia) braziliensis In Vitro Fitness Guided by Temperature Changes Along With Its Subtilisins and Oligopeptidase B Frontiers in Cellular and Infection Microbiology Leishmania (Viannia) braziliensis phenotypic characteristics gene expression serine proteases molecular dynamics |
| title | Novel Insights Into Leishmania (Viannia) braziliensis In Vitro Fitness Guided by Temperature Changes Along With Its Subtilisins and Oligopeptidase B |
| title_full | Novel Insights Into Leishmania (Viannia) braziliensis In Vitro Fitness Guided by Temperature Changes Along With Its Subtilisins and Oligopeptidase B |
| title_fullStr | Novel Insights Into Leishmania (Viannia) braziliensis In Vitro Fitness Guided by Temperature Changes Along With Its Subtilisins and Oligopeptidase B |
| title_full_unstemmed | Novel Insights Into Leishmania (Viannia) braziliensis In Vitro Fitness Guided by Temperature Changes Along With Its Subtilisins and Oligopeptidase B |
| title_short | Novel Insights Into Leishmania (Viannia) braziliensis In Vitro Fitness Guided by Temperature Changes Along With Its Subtilisins and Oligopeptidase B |
| title_sort | novel insights into leishmania viannia braziliensis in vitro fitness guided by temperature changes along with its subtilisins and oligopeptidase b |
| topic | Leishmania (Viannia) braziliensis phenotypic characteristics gene expression serine proteases molecular dynamics |
| url | https://www.frontiersin.org/articles/10.3389/fcimb.2022.805106/full |
| work_keys_str_mv | AT anabelzabalapenafiel novelinsightsintoleishmaniavianniabraziliensisinvitrofitnessguidedbytemperaturechangesalongwithitssubtilisinsandoligopeptidaseb AT leacysnefinkelstein novelinsightsintoleishmaniavianniabraziliensisinvitrofitnessguidedbytemperaturechangesalongwithitssubtilisinsandoligopeptidaseb AT fatimaconceicaosilva novelinsightsintoleishmaniavianniabraziliensisinvitrofitnessguidedbytemperaturechangesalongwithitssubtilisinsandoligopeptidaseb AT alinefagundes novelinsightsintoleishmaniavianniabraziliensisinvitrofitnessguidedbytemperaturechangesalongwithitssubtilisinsandoligopeptidaseb AT lucianadefreitascamposmiranda novelinsightsintoleishmaniavianniabraziliensisinvitrofitnessguidedbytemperaturechangesalongwithitssubtilisinsandoligopeptidaseb AT franklinsouzasilva novelinsightsintoleishmaniavianniabraziliensisinvitrofitnessguidedbytemperaturechangesalongwithitssubtilisinsandoligopeptidaseb AT franklinsouzasilva novelinsightsintoleishmaniavianniabraziliensisinvitrofitnessguidedbytemperaturechangesalongwithitssubtilisinsandoligopeptidaseb AT arturamlbrandt novelinsightsintoleishmaniavianniabraziliensisinvitrofitnessguidedbytemperaturechangesalongwithitssubtilisinsandoligopeptidaseb AT arturamlbrandt novelinsightsintoleishmaniavianniabraziliensisinvitrofitnessguidedbytemperaturechangesalongwithitssubtilisinsandoligopeptidaseb AT geovanediaslopes novelinsightsintoleishmaniavianniabraziliensisinvitrofitnessguidedbytemperaturechangesalongwithitssubtilisinsandoligopeptidaseb AT carlosrobertoalves novelinsightsintoleishmaniavianniabraziliensisinvitrofitnessguidedbytemperaturechangesalongwithitssubtilisinsandoligopeptidaseb |