Validation of Cytomegalovirus Immune Competence Assays for the Characterization of CD8+ T Cell Responses Posttransplant
Cytomegalovirus (CMV) infection is one of the most important infectious complications of transplantation. Monitoring CMV-specific CD8 T cell immunity is useful for predicting active CMV infection and for directing targeted antiviral therapy. In this study, we examined four basic parameters for valid...
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Wiley
2012-01-01
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| Series: | Clinical and Developmental Immunology |
| Online Access: | http://dx.doi.org/10.1155/2012/451059 |
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| author | Eugene V. Ravkov Igor Y. Pavlov Kimberly E. Hanson Julio C. Delgado |
| author_facet | Eugene V. Ravkov Igor Y. Pavlov Kimberly E. Hanson Julio C. Delgado |
| author_sort | Eugene V. Ravkov |
| collection | DOAJ |
| description | Cytomegalovirus (CMV) infection is one of the most important infectious complications of transplantation. Monitoring CMV-specific CD8 T cell immunity is useful for predicting active CMV infection and for directing targeted antiviral therapy. In this study, we examined four basic parameters for validation of CMV-specific tetramer staining and peptide stimulation assays that cover five most frequent HLA class I alleles. We also examined the potential use of CMV-specific CD8+ T cell numbers and functional and cytolytic responses in two autologous HSCT recipients treated for multiple myeloma. The coefficient of variation (CV %) of the precision within assays was 3.1−24% for HLA-tetramer staining, 2.5−47% for IFN-γ, and 3.4−59.7% for CD107a/b production upon peptide stimulation. The precision between assays was 5−26% for tetramer staining, 4−24% for IFN-γ, and 5−48% for CD107a/b. The limit of detection was 0.1−0.23 cells/μL of blood for tetramer staining, 0−0.23 cell/μL for IFN-γ, and 0.11−0.98 cells/μL for CD107a/b. The assays were linear and specific. The reference interval with 95% confidence level was 0−18 cells/μL for tetramer staining, 0−2 cells/μL for IFN-γ, and 0–3 cells/μL for CD107a/b. Our results provide acceptable measures of test performance for CMV immune competence assays for the characterization of CD8+ T cell responses posttransplant measured in the absolute cell count per μL of blood. |
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| institution | OA Journals |
| issn | 1740-2522 1740-2530 |
| language | English |
| publishDate | 2012-01-01 |
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| series | Clinical and Developmental Immunology |
| spelling | doaj-art-caaeea07aa5149228acb512793d5ee9c2025-08-20T02:19:34ZengWileyClinical and Developmental Immunology1740-25221740-25302012-01-01201210.1155/2012/451059451059Validation of Cytomegalovirus Immune Competence Assays for the Characterization of CD8+ T Cell Responses PosttransplantEugene V. Ravkov0Igor Y. Pavlov1Kimberly E. Hanson2Julio C. Delgado3Department of Pathology, ARUP Institute for Clinical and Experimental Pathology, School of Medicine, University of Utah, 500 Chipeta Way, Salt Lake City, UT 84108, USADepartment of Pathology, ARUP Institute for Clinical and Experimental Pathology, School of Medicine, University of Utah, 500 Chipeta Way, Salt Lake City, UT 84108, USADepartment of Pathology, ARUP Institute for Clinical and Experimental Pathology, School of Medicine, University of Utah, 500 Chipeta Way, Salt Lake City, UT 84108, USADepartment of Pathology, ARUP Institute for Clinical and Experimental Pathology, School of Medicine, University of Utah, 500 Chipeta Way, Salt Lake City, UT 84108, USACytomegalovirus (CMV) infection is one of the most important infectious complications of transplantation. Monitoring CMV-specific CD8 T cell immunity is useful for predicting active CMV infection and for directing targeted antiviral therapy. In this study, we examined four basic parameters for validation of CMV-specific tetramer staining and peptide stimulation assays that cover five most frequent HLA class I alleles. We also examined the potential use of CMV-specific CD8+ T cell numbers and functional and cytolytic responses in two autologous HSCT recipients treated for multiple myeloma. The coefficient of variation (CV %) of the precision within assays was 3.1−24% for HLA-tetramer staining, 2.5−47% for IFN-γ, and 3.4−59.7% for CD107a/b production upon peptide stimulation. The precision between assays was 5−26% for tetramer staining, 4−24% for IFN-γ, and 5−48% for CD107a/b. The limit of detection was 0.1−0.23 cells/μL of blood for tetramer staining, 0−0.23 cell/μL for IFN-γ, and 0.11−0.98 cells/μL for CD107a/b. The assays were linear and specific. The reference interval with 95% confidence level was 0−18 cells/μL for tetramer staining, 0−2 cells/μL for IFN-γ, and 0–3 cells/μL for CD107a/b. Our results provide acceptable measures of test performance for CMV immune competence assays for the characterization of CD8+ T cell responses posttransplant measured in the absolute cell count per μL of blood.http://dx.doi.org/10.1155/2012/451059 |
| spellingShingle | Eugene V. Ravkov Igor Y. Pavlov Kimberly E. Hanson Julio C. Delgado Validation of Cytomegalovirus Immune Competence Assays for the Characterization of CD8+ T Cell Responses Posttransplant Clinical and Developmental Immunology |
| title | Validation of Cytomegalovirus Immune Competence Assays for the Characterization of CD8+ T Cell Responses Posttransplant |
| title_full | Validation of Cytomegalovirus Immune Competence Assays for the Characterization of CD8+ T Cell Responses Posttransplant |
| title_fullStr | Validation of Cytomegalovirus Immune Competence Assays for the Characterization of CD8+ T Cell Responses Posttransplant |
| title_full_unstemmed | Validation of Cytomegalovirus Immune Competence Assays for the Characterization of CD8+ T Cell Responses Posttransplant |
| title_short | Validation of Cytomegalovirus Immune Competence Assays for the Characterization of CD8+ T Cell Responses Posttransplant |
| title_sort | validation of cytomegalovirus immune competence assays for the characterization of cd8 t cell responses posttransplant |
| url | http://dx.doi.org/10.1155/2012/451059 |
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