Amblyostatin-1, the first salivary cystatin with host immunomodulatory and anti-inflammatory properties from the Neotropical tick Amblyomma sculptum, vector of Brazilian spotted fever
IntroductionThe Neotropical tick Amblyomma sculptum is the primary vector of Rickettsia rickettsii, the causative agent of Brazilian spotted fever, a disease associated with high fatality rates. Tick saliva, a complex mixture of bioactive molecules essential for successful blood feeding, facilitates...
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2025-07-01
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| author | Wilson Santos Molari Mohamed Amine Jmel Mohamed Amine Jmel Josiane Betim Assis Alan Frazão-Silva Júlia Moura Bernardi Gretta Huamanrayme José María Medina Eliane Esteves Solange Cristina Antão Gabriel Cerqueira Alves Costa Aparecida Sadae Tanaka Aparecida Sadae Tanaka Andréa Cristina Fogaça Andréa Cristina Fogaça Zdenek Franta Lucas Tirloni Michalis Kotsyfakis Michalis Kotsyfakis Anderson Sá-Nunes Anderson Sá-Nunes |
| author_facet | Wilson Santos Molari Mohamed Amine Jmel Mohamed Amine Jmel Josiane Betim Assis Alan Frazão-Silva Júlia Moura Bernardi Gretta Huamanrayme José María Medina Eliane Esteves Solange Cristina Antão Gabriel Cerqueira Alves Costa Aparecida Sadae Tanaka Aparecida Sadae Tanaka Andréa Cristina Fogaça Andréa Cristina Fogaça Zdenek Franta Lucas Tirloni Michalis Kotsyfakis Michalis Kotsyfakis Anderson Sá-Nunes Anderson Sá-Nunes |
| author_sort | Wilson Santos Molari |
| collection | DOAJ |
| description | IntroductionThe Neotropical tick Amblyomma sculptum is the primary vector of Rickettsia rickettsii, the causative agent of Brazilian spotted fever, a disease associated with high fatality rates. Tick saliva, a complex mixture of bioactive molecules essential for successful blood feeding, facilitates pathogen transmission and modulates host immune responses. A comprehensive evaluation of the salivary gland transcriptome database reveals that protease inhibitors are abundantly expressed molecules in tick saliva during feeding. Thus, this study aims to describe and characterize the most expressed member of the cystatin family identified in Amblyomma sculptum salivary transcriptome, named Amblyostatin-1. MethodsBioinformatic tools were employed for in silico analysis of the Amblyostatin-1 sequence and structure. A recombinant version of Amblyostatin-1 was expressed in an Escherichia coli system, evaluated against a panel of cysteine proteases in biochemical assays, and used to generate antibodies in immunized mice. The biological activities of Amblyostatin-1 were assessed by its effects on dendritic cell maturation in vitro and in a carrageenan-induced inflammation model in vivo.ResultsBased on its sequence and predicted three-dimensional structure, Amblyostatin-1 is classified as an I25B cystatin, and its recombinant form selectively inhibits cathepsins L, C, and S at different rates, with a low nanomolar Ki value of 0.697 ± 0.22 nM against cathepsin L. Regarding its biological activities, recombinant Amblyostatin-1 partially affects LPS-induced dendritic cell maturation by downmodulating the costimulatory molecules CD80 and CD86 at higher micromolar concentrations (3 µM) while promoting IL-10 production at nanomolar concentrations (100 nM). The apparent lack of Amblyostatin-1-specific antibody responses in immunized mice suggests an impairment of antigen processing and presentation in vivo. Furthermore, in a carrageenan-induced inflammation model, Amblyostatin-1 decreased edema formation and neutrophil infiltration into the skin without affecting other myeloid cells.DiscussionThese findings establish Amblyostatin-1 as a novel salivary cystatin with immunomodulatory and anti-inflammatory properties, highlighting its potential as an immunobiological agent. |
| format | Article |
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| institution | Kabale University |
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| language | English |
| publishDate | 2025-07-01 |
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| spelling | doaj-art-665ee3ef89584e33a4b68709692afc252025-08-20T03:50:53ZengFrontiers Media S.A.Frontiers in Immunology1664-32242025-07-011610.3389/fimmu.2025.15857031585703Amblyostatin-1, the first salivary cystatin with host immunomodulatory and anti-inflammatory properties from the Neotropical tick Amblyomma sculptum, vector of Brazilian spotted feverWilson Santos Molari0Mohamed Amine Jmel1Mohamed Amine Jmel2Josiane Betim Assis3Alan Frazão-Silva4Júlia Moura Bernardi5Gretta Huamanrayme6José María Medina7Eliane Esteves8Solange Cristina Antão9Gabriel Cerqueira Alves Costa10Aparecida Sadae Tanaka11Aparecida Sadae Tanaka12Andréa Cristina Fogaça13Andréa Cristina Fogaça14Zdenek Franta15Lucas Tirloni16Michalis Kotsyfakis17Michalis Kotsyfakis18Anderson Sá-Nunes19Anderson Sá-Nunes20Department of Immunology, Institute of Biomedical Sciences, University of São Paulo, São Paulo, BrazilInstitute of Parasitology, Biology Centre, Czech Academy of Sciences, České Budějovice, CzechiaUniversité de Lorraine, CNRS, IMoPA, Nancy, FranceDepartment of Immunology, Institute of Biomedical Sciences, University of São Paulo, São Paulo, BrazilDepartment of Immunology, Institute of Biomedical Sciences, University of São Paulo, São Paulo, BrazilDepartment of Immunology, Institute of Biomedical Sciences, University of São Paulo, São Paulo, BrazilDepartment of Immunology, Institute of Biomedical Sciences, University of São Paulo, São Paulo, BrazilDepartamento de Genética, Facultad de Ciencias, Universidad de Granada, Granada, SpainDepartment of Microbiology and Immunology, Frederick P. Whiddon College of Medicine, University of South Alabama, Mobile, AL, United StatesDepartment of Parasitology, Institute of Biomedical Sciences, University of São Paulo, São Paulo, SP, BrazilDepartment of Biochemistry, Federal University of São Paulo (UNIFESP), São Paulo, BrazilDepartment of Biochemistry, Federal University of São Paulo (UNIFESP), São Paulo, BrazilNational Institute of Science and Technology in Molecular Entomology, National Council for Scientific and Technological Development, Rio de Janeiro, BrazilDepartment of Parasitology, Institute of Biomedical Sciences, University of São Paulo, São Paulo, SP, BrazilNational Institute of Science and Technology in Molecular Entomology, National Council for Scientific and Technological Development, Rio de Janeiro, BrazilDepartment of Chemistry, Faculty of Science, University of South Bohemia, České Budějovice, Czechia0Tick-Pathogen Transmission Unit, Laboratory of Bacteriology, Division of Intramural Research, National Institute of Allergy and Infectious Diseases, Hamilton, MT, United StatesInstitute of Parasitology, Biology Centre, Czech Academy of Sciences, České Budějovice, Czechia1Institute of Molecular Biology and Biotechnology, Foundation for Research and Technology-Hellas, Heraklion, GreeceDepartment of Immunology, Institute of Biomedical Sciences, University of São Paulo, São Paulo, BrazilNational Institute of Science and Technology in Molecular Entomology, National Council for Scientific and Technological Development, Rio de Janeiro, BrazilIntroductionThe Neotropical tick Amblyomma sculptum is the primary vector of Rickettsia rickettsii, the causative agent of Brazilian spotted fever, a disease associated with high fatality rates. Tick saliva, a complex mixture of bioactive molecules essential for successful blood feeding, facilitates pathogen transmission and modulates host immune responses. A comprehensive evaluation of the salivary gland transcriptome database reveals that protease inhibitors are abundantly expressed molecules in tick saliva during feeding. Thus, this study aims to describe and characterize the most expressed member of the cystatin family identified in Amblyomma sculptum salivary transcriptome, named Amblyostatin-1. MethodsBioinformatic tools were employed for in silico analysis of the Amblyostatin-1 sequence and structure. A recombinant version of Amblyostatin-1 was expressed in an Escherichia coli system, evaluated against a panel of cysteine proteases in biochemical assays, and used to generate antibodies in immunized mice. The biological activities of Amblyostatin-1 were assessed by its effects on dendritic cell maturation in vitro and in a carrageenan-induced inflammation model in vivo.ResultsBased on its sequence and predicted three-dimensional structure, Amblyostatin-1 is classified as an I25B cystatin, and its recombinant form selectively inhibits cathepsins L, C, and S at different rates, with a low nanomolar Ki value of 0.697 ± 0.22 nM against cathepsin L. Regarding its biological activities, recombinant Amblyostatin-1 partially affects LPS-induced dendritic cell maturation by downmodulating the costimulatory molecules CD80 and CD86 at higher micromolar concentrations (3 µM) while promoting IL-10 production at nanomolar concentrations (100 nM). The apparent lack of Amblyostatin-1-specific antibody responses in immunized mice suggests an impairment of antigen processing and presentation in vivo. Furthermore, in a carrageenan-induced inflammation model, Amblyostatin-1 decreased edema formation and neutrophil infiltration into the skin without affecting other myeloid cells.DiscussionThese findings establish Amblyostatin-1 as a novel salivary cystatin with immunomodulatory and anti-inflammatory properties, highlighting its potential as an immunobiological agent.https://www.frontiersin.org/articles/10.3389/fimmu.2025.1585703/fullAmblyostatin-1Amblyomma sculptumimmunomodulationinflammationtick-host interactiontick saliva |
| spellingShingle | Wilson Santos Molari Mohamed Amine Jmel Mohamed Amine Jmel Josiane Betim Assis Alan Frazão-Silva Júlia Moura Bernardi Gretta Huamanrayme José María Medina Eliane Esteves Solange Cristina Antão Gabriel Cerqueira Alves Costa Aparecida Sadae Tanaka Aparecida Sadae Tanaka Andréa Cristina Fogaça Andréa Cristina Fogaça Zdenek Franta Lucas Tirloni Michalis Kotsyfakis Michalis Kotsyfakis Anderson Sá-Nunes Anderson Sá-Nunes Amblyostatin-1, the first salivary cystatin with host immunomodulatory and anti-inflammatory properties from the Neotropical tick Amblyomma sculptum, vector of Brazilian spotted fever Frontiers in Immunology Amblyostatin-1 Amblyomma sculptum immunomodulation inflammation tick-host interaction tick saliva |
| title | Amblyostatin-1, the first salivary cystatin with host immunomodulatory and anti-inflammatory properties from the Neotropical tick Amblyomma sculptum, vector of Brazilian spotted fever |
| title_full | Amblyostatin-1, the first salivary cystatin with host immunomodulatory and anti-inflammatory properties from the Neotropical tick Amblyomma sculptum, vector of Brazilian spotted fever |
| title_fullStr | Amblyostatin-1, the first salivary cystatin with host immunomodulatory and anti-inflammatory properties from the Neotropical tick Amblyomma sculptum, vector of Brazilian spotted fever |
| title_full_unstemmed | Amblyostatin-1, the first salivary cystatin with host immunomodulatory and anti-inflammatory properties from the Neotropical tick Amblyomma sculptum, vector of Brazilian spotted fever |
| title_short | Amblyostatin-1, the first salivary cystatin with host immunomodulatory and anti-inflammatory properties from the Neotropical tick Amblyomma sculptum, vector of Brazilian spotted fever |
| title_sort | amblyostatin 1 the first salivary cystatin with host immunomodulatory and anti inflammatory properties from the neotropical tick amblyomma sculptum vector of brazilian spotted fever |
| topic | Amblyostatin-1 Amblyomma sculptum immunomodulation inflammation tick-host interaction tick saliva |
| url | https://www.frontiersin.org/articles/10.3389/fimmu.2025.1585703/full |
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