The Validation of Antibodies Suitable for Flow Cytometric Analysis and Immunopeptidomics of Peptide–MHC Complexes in the Outbred Swiss Albino Mouse Strain
Antigen presentation on major histocompatibility complex (MHC) molecules is central to the initiation of immune responses, and a lot of our understanding about the antigen processing and presentation pathway has been gained through studies in mice. MHC molecules are the most genetically diverse gene...
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2025-04-01
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| author | Shanzou Chung Isambard G. Knox-Johnson Sarah E. Gazzard Runqiu Song Ngoc H. Le Luise A. Cullen-McEwen John F. Bertram Anthony W. Purcell Asolina Braun |
| author_facet | Shanzou Chung Isambard G. Knox-Johnson Sarah E. Gazzard Runqiu Song Ngoc H. Le Luise A. Cullen-McEwen John F. Bertram Anthony W. Purcell Asolina Braun |
| author_sort | Shanzou Chung |
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| description | Antigen presentation on major histocompatibility complex (MHC) molecules is central to the initiation of immune responses, and a lot of our understanding about the antigen processing and presentation pathway has been gained through studies in mice. MHC molecules are the most genetically diverse genes; consequently, mouse strains differ substantially in their MHC make up and resulting antigen presentation. Swiss mice are commonly used in pharmacological research, yet our understanding of antigen presentation in this strain is surprisingly limited. Here, we have tested a range of anti-MHC antibodies and present a range of clones suitable to analyse MHC class I and class II molecules in Swiss mice who have the H2-q MHC haplotype. Moreover, we demonstrate using immunopeptidomics that clones 28-12-8, 34-1-2, MKD6, and N22 are also suited to isolate MHC class I and class II ligands in this mouse strain. Thus, this work also establishes a first experimental account of the H2-q-derived thymus and spleen immunopeptidome in Swiss mice which bears strong resemblance with ligands isolated from the H2-d MHC haplotype of Balb/C mice. The analysis of source proteins shows common but also organ- and function-specific antigen presentation in line with the involvement of the thymus in tolerance induction and the function of the spleen as a site of immune responses. |
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| language | English |
| publishDate | 2025-04-01 |
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| spelling | doaj-art-1de3f45db0f548ffb24f4289e9a0036f2025-08-20T02:21:14ZengMDPI AGMethods and Protocols2409-92792025-04-01834310.3390/mps8030043The Validation of Antibodies Suitable for Flow Cytometric Analysis and Immunopeptidomics of Peptide–MHC Complexes in the Outbred Swiss Albino Mouse StrainShanzou Chung0Isambard G. Knox-Johnson1Sarah E. Gazzard2Runqiu Song3Ngoc H. Le4Luise A. Cullen-McEwen5John F. Bertram6Anthony W. Purcell7Asolina Braun8Department of Biochemistry and Molecular Biology, Infection and Immunity Program, Biomedicine Discovery Institute, Monash University, Clayton, VIC 3800, AustraliaDepartment of Biochemistry and Molecular Biology, Infection and Immunity Program, Biomedicine Discovery Institute, Monash University, Clayton, VIC 3800, AustraliaDepartment of Anatomy and Developmental Biology, Monash Biomedicine Discovery Institute, Clayton, VIC 3800, AustraliaDepartment of Biochemistry and Molecular Biology, Infection and Immunity Program, Biomedicine Discovery Institute, Monash University, Clayton, VIC 3800, AustraliaDepartment of Biochemistry and Molecular Biology, Infection and Immunity Program, Biomedicine Discovery Institute, Monash University, Clayton, VIC 3800, AustraliaDepartment of Anatomy and Developmental Biology, Monash Biomedicine Discovery Institute, Clayton, VIC 3800, AustraliaDepartment of Anatomy and Developmental Biology, Monash Biomedicine Discovery Institute, Clayton, VIC 3800, AustraliaDepartment of Biochemistry and Molecular Biology, Infection and Immunity Program, Biomedicine Discovery Institute, Monash University, Clayton, VIC 3800, AustraliaDepartment of Biochemistry and Molecular Biology, Infection and Immunity Program, Biomedicine Discovery Institute, Monash University, Clayton, VIC 3800, AustraliaAntigen presentation on major histocompatibility complex (MHC) molecules is central to the initiation of immune responses, and a lot of our understanding about the antigen processing and presentation pathway has been gained through studies in mice. MHC molecules are the most genetically diverse genes; consequently, mouse strains differ substantially in their MHC make up and resulting antigen presentation. Swiss mice are commonly used in pharmacological research, yet our understanding of antigen presentation in this strain is surprisingly limited. Here, we have tested a range of anti-MHC antibodies and present a range of clones suitable to analyse MHC class I and class II molecules in Swiss mice who have the H2-q MHC haplotype. Moreover, we demonstrate using immunopeptidomics that clones 28-12-8, 34-1-2, MKD6, and N22 are also suited to isolate MHC class I and class II ligands in this mouse strain. Thus, this work also establishes a first experimental account of the H2-q-derived thymus and spleen immunopeptidome in Swiss mice which bears strong resemblance with ligands isolated from the H2-d MHC haplotype of Balb/C mice. The analysis of source proteins shows common but also organ- and function-specific antigen presentation in line with the involvement of the thymus in tolerance induction and the function of the spleen as a site of immune responses.https://www.mdpi.com/2409-9279/8/3/43Swiss miceantigen presentationimmunopeptidomicsmajor histocompatibility complexMHC ligandsflow cytometry |
| spellingShingle | Shanzou Chung Isambard G. Knox-Johnson Sarah E. Gazzard Runqiu Song Ngoc H. Le Luise A. Cullen-McEwen John F. Bertram Anthony W. Purcell Asolina Braun The Validation of Antibodies Suitable for Flow Cytometric Analysis and Immunopeptidomics of Peptide–MHC Complexes in the Outbred Swiss Albino Mouse Strain Methods and Protocols Swiss mice antigen presentation immunopeptidomics major histocompatibility complex MHC ligands flow cytometry |
| title | The Validation of Antibodies Suitable for Flow Cytometric Analysis and Immunopeptidomics of Peptide–MHC Complexes in the Outbred Swiss Albino Mouse Strain |
| title_full | The Validation of Antibodies Suitable for Flow Cytometric Analysis and Immunopeptidomics of Peptide–MHC Complexes in the Outbred Swiss Albino Mouse Strain |
| title_fullStr | The Validation of Antibodies Suitable for Flow Cytometric Analysis and Immunopeptidomics of Peptide–MHC Complexes in the Outbred Swiss Albino Mouse Strain |
| title_full_unstemmed | The Validation of Antibodies Suitable for Flow Cytometric Analysis and Immunopeptidomics of Peptide–MHC Complexes in the Outbred Swiss Albino Mouse Strain |
| title_short | The Validation of Antibodies Suitable for Flow Cytometric Analysis and Immunopeptidomics of Peptide–MHC Complexes in the Outbred Swiss Albino Mouse Strain |
| title_sort | validation of antibodies suitable for flow cytometric analysis and immunopeptidomics of peptide mhc complexes in the outbred swiss albino mouse strain |
| topic | Swiss mice antigen presentation immunopeptidomics major histocompatibility complex MHC ligands flow cytometry |
| url | https://www.mdpi.com/2409-9279/8/3/43 |
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