Protocol for identifying immune mediators during Leishmania infection in mice using metabolomic analysis
Summary: Here, we present a protocol for identifying immune mediators during Leishmania infection in mice using metabolomic analysis. We describe steps for Leishmania infection, harvesting infected tissues, and performing liquid chromatography-mass spectrometry (LC-MS) to identify enriched metabolic...
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| Format: | Article |
| Language: | English |
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Elsevier
2025-09-01
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| Series: | STAR Protocols |
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| Online Access: | http://www.sciencedirect.com/science/article/pii/S2666166725002230 |
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| author | Thalia Pacheco-Fernandez Laura Klenow Nazli Azodi Hannah Markle Matthew Bernier Greta Volpedo Timur Oljuskin Parna Bhattacharya Shinjiro Hamano Greg Matlashewski Hira L. Nakhasi Abhay R. Satoskar Sreenivas Gannavaram |
| author_facet | Thalia Pacheco-Fernandez Laura Klenow Nazli Azodi Hannah Markle Matthew Bernier Greta Volpedo Timur Oljuskin Parna Bhattacharya Shinjiro Hamano Greg Matlashewski Hira L. Nakhasi Abhay R. Satoskar Sreenivas Gannavaram |
| author_sort | Thalia Pacheco-Fernandez |
| collection | DOAJ |
| description | Summary: Here, we present a protocol for identifying immune mediators during Leishmania infection in mice using metabolomic analysis. We describe steps for Leishmania infection, harvesting infected tissues, and performing liquid chromatography-mass spectrometry (LC-MS) to identify enriched metabolic pathways and altered metabolites. We then detail procedures for verifying the metabolite role in immune response via PCR and ELISA. This protocol focuses on cutaneous leishmaniasis models, but the analyses here are applicable to other infection models.For complete details on the use and execution of this protocol, please refer to Volpedo et al.1 and Oljuskin et al.2 : Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics. |
| format | Article |
| id | doaj-art-c3d9e4e74ef94896acc5f4edaa7ac69f |
| institution | Kabale University |
| issn | 2666-1667 |
| language | English |
| publishDate | 2025-09-01 |
| publisher | Elsevier |
| record_format | Article |
| series | STAR Protocols |
| spelling | doaj-art-c3d9e4e74ef94896acc5f4edaa7ac69f2025-08-20T03:51:13ZengElsevierSTAR Protocols2666-16672025-09-016310381710.1016/j.xpro.2025.103817Protocol for identifying immune mediators during Leishmania infection in mice using metabolomic analysisThalia Pacheco-Fernandez0Laura Klenow1Nazli Azodi2Hannah Markle3Matthew Bernier4Greta Volpedo5Timur Oljuskin6Parna Bhattacharya7Shinjiro Hamano8Greg Matlashewski9Hira L. Nakhasi10Abhay R. Satoskar11Sreenivas Gannavaram12Division of Emerging and Transfusion Transmitted Diseases, Office of Blood Research and Review, Center of Biologics Research and Evaluation, Food and Drug Administration, Silver Spring, MD 20993, USADivision of Emerging and Transfusion Transmitted Diseases, Office of Blood Research and Review, Center of Biologics Research and Evaluation, Food and Drug Administration, Silver Spring, MD 20993, USADivision of Emerging and Transfusion Transmitted Diseases, Office of Blood Research and Review, Center of Biologics Research and Evaluation, Food and Drug Administration, Silver Spring, MD 20993, USADivision of Emerging and Transfusion Transmitted Diseases, Office of Blood Research and Review, Center of Biologics Research and Evaluation, Food and Drug Administration, Silver Spring, MD 20993, USAThe Ohio State University, Columbus, OH 43201, USADepartment of Neurosciences, Rehabilitation, Ophthalmology, Genetics, Maternal and Child Health, University of Genoa, 16126 Genoa, ItalyAnimal Parasitic Diseases Laboratory, Agricultural Research Service, USDA, Beltsville, MD 20705, USACenter for Devices and Radiological Health, Food and Drug Administration, Silver Spring, MD 20993, USADepartment of Parasitology, Institute of Tropical Medicine (NEKKEN), The Joint Usage/Research Center on Tropical Disease, Nagasaki University, Sakamoto, Nagasaki 852-8523, JapanDepartment of Microbiology and Immunology, McGill University, Montreal, QC H3A2B4, CanadaDivision of Emerging and Transfusion Transmitted Diseases, Office of Blood Research and Review, Center of Biologics Research and Evaluation, Food and Drug Administration, Silver Spring, MD 20993, USAThe Ohio State University, Columbus, OH 43201, USADivision of Emerging and Transfusion Transmitted Diseases, Office of Blood Research and Review, Center of Biologics Research and Evaluation, Food and Drug Administration, Silver Spring, MD 20993, USA; Corresponding authorSummary: Here, we present a protocol for identifying immune mediators during Leishmania infection in mice using metabolomic analysis. We describe steps for Leishmania infection, harvesting infected tissues, and performing liquid chromatography-mass spectrometry (LC-MS) to identify enriched metabolic pathways and altered metabolites. We then detail procedures for verifying the metabolite role in immune response via PCR and ELISA. This protocol focuses on cutaneous leishmaniasis models, but the analyses here are applicable to other infection models.For complete details on the use and execution of this protocol, please refer to Volpedo et al.1 and Oljuskin et al.2 : Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics.http://www.sciencedirect.com/science/article/pii/S2666166725002230metabolomicsmicrobiologymodel organismsmolecular biologymass spectrometry |
| spellingShingle | Thalia Pacheco-Fernandez Laura Klenow Nazli Azodi Hannah Markle Matthew Bernier Greta Volpedo Timur Oljuskin Parna Bhattacharya Shinjiro Hamano Greg Matlashewski Hira L. Nakhasi Abhay R. Satoskar Sreenivas Gannavaram Protocol for identifying immune mediators during Leishmania infection in mice using metabolomic analysis STAR Protocols metabolomics microbiology model organisms molecular biology mass spectrometry |
| title | Protocol for identifying immune mediators during Leishmania infection in mice using metabolomic analysis |
| title_full | Protocol for identifying immune mediators during Leishmania infection in mice using metabolomic analysis |
| title_fullStr | Protocol for identifying immune mediators during Leishmania infection in mice using metabolomic analysis |
| title_full_unstemmed | Protocol for identifying immune mediators during Leishmania infection in mice using metabolomic analysis |
| title_short | Protocol for identifying immune mediators during Leishmania infection in mice using metabolomic analysis |
| title_sort | protocol for identifying immune mediators during leishmania infection in mice using metabolomic analysis |
| topic | metabolomics microbiology model organisms molecular biology mass spectrometry |
| url | http://www.sciencedirect.com/science/article/pii/S2666166725002230 |
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