Validation of Cytomegalovirus Immune Competence Assays for the Characterization of CD8+ T Cell Responses Posttransplant. (16th December 2012)

• Record Type: Journal Article
• Authors: Ravkov, Eugene V.; Pavlov, Igor Y.; Hanson, Kimberly E.; Delgado, Julio C.
• Other Names: Cavallo Rossana Academic Editor.
• Is Part Of: Clinical & developmental immunology. Volume 2012(2012)
• Publication Date: 2012-12-16
• Subjects: Developmental immunology -- Periodicals; Clinical immunology -- Periodicals; Immune System -- immunology -- Periodicals; Immune System -- growth & development -- Periodicals; Immune System Diseases -- immunology -- Periodicals; 571.9638
• DOI: 10.1155/2012/451059
• Languages: English
• ISSNs: 1740-2522

Abstract

Abstract : 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/ μ L of blood for tetramer staining, 0−0.23 cell/ μ L for IFN- γ, and 0.11−0.98 cells/ μ L for CD107a/b. The assays were linear and specific. The reference interval with 95% confidence level was 0−18 cells/ μ L for tetramer staining, 0−2 cells/ μ L for IFN- γ, and 0–3 cells/ μ L 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 μ L of blood.