Performance of new pp65-IGRA for the quantification of HCMV-specific CD4+ T-cell response in healthy subjects and in solid organ transplant recipients
Immune control of human cytomegalovirus (HCMV) replication is critical in bone marrow and solid organ transplant recipients, where uncontrolled replication can lead to high mortality. Current commercial immune monitoring tools have several limitations, such as a lack of appropriate test cutoff value...
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Frontiers Media S.A.
2025-05-01
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| Series: | Frontiers in Immunology |
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| Online Access: | https://www.frontiersin.org/articles/10.3389/fimmu.2025.1553305/full |
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| author | Dalila Mele Federica Zavaglio Federica Bergami Marilena Gregorini Marilena Gregorini Domenica Federica Briganti Domenica Federica Briganti Carlo Pellegrini Carlo Pellegrini Giuditta Comolli Irene Cassaniti Irene Cassaniti Daniele Lilleri Fausto Baldanti Fausto Baldanti |
| author_facet | Dalila Mele Federica Zavaglio Federica Bergami Marilena Gregorini Marilena Gregorini Domenica Federica Briganti Domenica Federica Briganti Carlo Pellegrini Carlo Pellegrini Giuditta Comolli Irene Cassaniti Irene Cassaniti Daniele Lilleri Fausto Baldanti Fausto Baldanti |
| author_sort | Dalila Mele |
| collection | DOAJ |
| description | Immune control of human cytomegalovirus (HCMV) replication is critical in bone marrow and solid organ transplant recipients, where uncontrolled replication can lead to high mortality. Current commercial immune monitoring tools have several limitations, such as a lack of appropriate test cutoff values and the inability to characterise antigen-specific T cells. The main aim of our study was to develop a new interferon-γ (IFN-γ) release assay (IGRA), easy to use, to quantify and characterise the HCMV-specific T-cell response (pp65-IGRA). Secondary analyses included an evaluation of the performance of pp65-IGRA to assess whether its specificity and sensitivity were equal to or greater than those of the intracellular cytokine staining (ICS) and enzyme-linked immunospot (ELISpot) assays. In the study, 76 immunocompetent donors and nine solid organ transplant recipients were enrolled. Blood samples or peripheral blood mononuclear cells were stimulated with HCMV pp65-recombinant protein or with a complete pool of overlapping pp65 peptides. IFN-γ production was analysed by enzyme-linked immunoassay, ELISpot assays, and flow cytometry. For each assay, appropriate cutoff values were calculated. Our data demonstrate the suitability of pp65-IGRA for the quantification of HCMV-specific CD4+ T-cell responses and may support its use in routine clinical practice to improve the management of immunocompromised patients. |
| format | Article |
| id | doaj-art-7d01f6635626465d8fffd0378c2ba0ab |
| institution | OA Journals |
| issn | 1664-3224 |
| language | English |
| publishDate | 2025-05-01 |
| publisher | Frontiers Media S.A. |
| record_format | Article |
| series | Frontiers in Immunology |
| spelling | doaj-art-7d01f6635626465d8fffd0378c2ba0ab2025-08-20T01:49:58ZengFrontiers Media S.A.Frontiers in Immunology1664-32242025-05-011610.3389/fimmu.2025.15533051553305Performance of new pp65-IGRA for the quantification of HCMV-specific CD4+ T-cell response in healthy subjects and in solid organ transplant recipientsDalila Mele0Federica Zavaglio1Federica Bergami2Marilena Gregorini3Marilena Gregorini4Domenica Federica Briganti5Domenica Federica Briganti6Carlo Pellegrini7Carlo Pellegrini8Giuditta Comolli9Irene Cassaniti10Irene Cassaniti11Daniele Lilleri12Fausto Baldanti13Fausto Baldanti14Molecular Virology Unit, Department of Microbiology and Virology, Fondazione IRCCS Policlinico San Matteo, Pavia, ItalyMolecular Virology Unit, Department of Microbiology and Virology, Fondazione IRCCS Policlinico San Matteo, Pavia, ItalyMolecular Virology Unit, Department of Microbiology and Virology, Fondazione IRCCS Policlinico San Matteo, Pavia, ItalyUnit of Nephrology, Dialysis and Transplantation, Fondazione IRCCS Policlinico San Matteo, Pavia, ItalyUOS Transplant Center, Fondazione IRCCS Policlinico San Matteo, Pavia, ItalyUOS Transplant Center, Fondazione IRCCS Policlinico San Matteo, Pavia, ItalyDepartment of Clinical, Surgical, Diagnostic and Pediatric Sciences, University of Pavia, Pavia, ItalyDepartment of Clinical, Surgical, Diagnostic and Pediatric Sciences, University of Pavia, Pavia, ItalyDepartment of Cardiothoracic Surgery, Fondazione IRCCS Policlinico San Matteo, Pavia, ItalyMolecular Virology Unit, Department of Microbiology and Virology, Fondazione IRCCS Policlinico San Matteo, Pavia, ItalyMolecular Virology Unit, Department of Microbiology and Virology, Fondazione IRCCS Policlinico San Matteo, Pavia, ItalyDepartment of Clinical, Surgical, Diagnostic and Pediatric Sciences, University of Pavia, Pavia, ItalyMolecular Virology Unit, Department of Microbiology and Virology, Fondazione IRCCS Policlinico San Matteo, Pavia, ItalyMolecular Virology Unit, Department of Microbiology and Virology, Fondazione IRCCS Policlinico San Matteo, Pavia, ItalyDepartment of Clinical, Surgical, Diagnostic and Pediatric Sciences, University of Pavia, Pavia, ItalyImmune control of human cytomegalovirus (HCMV) replication is critical in bone marrow and solid organ transplant recipients, where uncontrolled replication can lead to high mortality. Current commercial immune monitoring tools have several limitations, such as a lack of appropriate test cutoff values and the inability to characterise antigen-specific T cells. The main aim of our study was to develop a new interferon-γ (IFN-γ) release assay (IGRA), easy to use, to quantify and characterise the HCMV-specific T-cell response (pp65-IGRA). Secondary analyses included an evaluation of the performance of pp65-IGRA to assess whether its specificity and sensitivity were equal to or greater than those of the intracellular cytokine staining (ICS) and enzyme-linked immunospot (ELISpot) assays. In the study, 76 immunocompetent donors and nine solid organ transplant recipients were enrolled. Blood samples or peripheral blood mononuclear cells were stimulated with HCMV pp65-recombinant protein or with a complete pool of overlapping pp65 peptides. IFN-γ production was analysed by enzyme-linked immunoassay, ELISpot assays, and flow cytometry. For each assay, appropriate cutoff values were calculated. Our data demonstrate the suitability of pp65-IGRA for the quantification of HCMV-specific CD4+ T-cell responses and may support its use in routine clinical practice to improve the management of immunocompromised patients.https://www.frontiersin.org/articles/10.3389/fimmu.2025.1553305/fullHCMVIFN-γIGRApp65HCMV-specific T cell response |
| spellingShingle | Dalila Mele Federica Zavaglio Federica Bergami Marilena Gregorini Marilena Gregorini Domenica Federica Briganti Domenica Federica Briganti Carlo Pellegrini Carlo Pellegrini Giuditta Comolli Irene Cassaniti Irene Cassaniti Daniele Lilleri Fausto Baldanti Fausto Baldanti Performance of new pp65-IGRA for the quantification of HCMV-specific CD4+ T-cell response in healthy subjects and in solid organ transplant recipients Frontiers in Immunology HCMV IFN-γ IGRA pp65 HCMV-specific T cell response |
| title | Performance of new pp65-IGRA for the quantification of HCMV-specific CD4+ T-cell response in healthy subjects and in solid organ transplant recipients |
| title_full | Performance of new pp65-IGRA for the quantification of HCMV-specific CD4+ T-cell response in healthy subjects and in solid organ transplant recipients |
| title_fullStr | Performance of new pp65-IGRA for the quantification of HCMV-specific CD4+ T-cell response in healthy subjects and in solid organ transplant recipients |
| title_full_unstemmed | Performance of new pp65-IGRA for the quantification of HCMV-specific CD4+ T-cell response in healthy subjects and in solid organ transplant recipients |
| title_short | Performance of new pp65-IGRA for the quantification of HCMV-specific CD4+ T-cell response in healthy subjects and in solid organ transplant recipients |
| title_sort | performance of new pp65 igra for the quantification of hcmv specific cd4 t cell response in healthy subjects and in solid organ transplant recipients |
| topic | HCMV IFN-γ IGRA pp65 HCMV-specific T cell response |
| url | https://www.frontiersin.org/articles/10.3389/fimmu.2025.1553305/full |
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