Tags

Type your tag names separated by a space and hit enter

Detection of human cytomegalovirus-specific T lymphocytes in human blood: comparison of two methods.
Cytotherapy. 2008; 10(8):834-41.C

Abstract

BACKGROUND

Human cytomegalovirus (HCMV) infection remains a major cause of morbidity and mortality in immunocompromised patients undergoing allogeneic stem cell transplantation (SCT). In the case of HCMV reactivation, the well-defined detection of virus-specific effector cells in patients might positively impact antiviral treatment.

METHODS

We examined blood samples from healthy volunteers serologically typed for HCMV IgG. Based on multicolor flow cytometry analysis, we addressed HCMV-specific CD8(+) effector T lymphocytes using HCMV-specific tetramers for the respective major histocompatibility complex (MHC) class I type. As a second approach, we employed the cytokine secretion assay (CSA), which allows the indirect detection of target-specific CD4(+) and CD8(+) T cells via their interferon (IFN)-gamma secretion upon HCMV pp65 in vitro stimulation.

RESULTS

We hypothesized the detection of HCMV-specific lymphocytes in >50% of healthy Caucasians that were IgG-seropositive for HCMV. In terms of specificity, both assays showed comparably good results (specificity 100%, confidence interval >95%). Regarding sensitivity, both assays met the zero hypothesis. However, with 45/52 (86.5%) the tetramer technology was superior to the CSA, which detected 34/52 (65.4%) based on CD8(+) T cells and 41/52 (78.8%) based on both CD4(+) and CD8(+) T cells.

DISCUSSION

A good correlation was observed between both assays, although the tetramers addressed only CD8(+) HCMV-specific T cells, whereas IFN-gamma secretion was detected on all T-cell types. Disadvantages of the CSA are the time-consuming stimulation, the extensive cell washing steps and the fact that the target cells are detected indirectly. The analysis with tetramers is rapid and reliable but their general use is hampered because of the restriction to a few HLA types.

Authors+Show Affiliations

Clinical Immunology Laboratory, Intercell AG, Vienna, Austria.No affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info available

Pub Type(s)

Comparative Study
Journal Article

Language

eng

PubMed ID

19016371

Citation

Schermann, C M., et al. "Detection of Human Cytomegalovirus-specific T Lymphocytes in Human Blood: Comparison of Two Methods." Cytotherapy, vol. 10, no. 8, 2008, pp. 834-41.
Schermann CM, Fischer G, Witt V, et al. Detection of human cytomegalovirus-specific T lymphocytes in human blood: comparison of two methods. Cytotherapy. 2008;10(8):834-41.
Schermann, C. M., Fischer, G., Witt, V., Kurz, M., Potschger, U., & Fritsch, G. (2008). Detection of human cytomegalovirus-specific T lymphocytes in human blood: comparison of two methods. Cytotherapy, 10(8), 834-41. https://doi.org/10.1080/14653240802474315
Schermann CM, et al. Detection of Human Cytomegalovirus-specific T Lymphocytes in Human Blood: Comparison of Two Methods. Cytotherapy. 2008;10(8):834-41. PubMed PMID: 19016371.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Detection of human cytomegalovirus-specific T lymphocytes in human blood: comparison of two methods. AU - Schermann,C M, AU - Fischer,G, AU - Witt,V, AU - Kurz,M, AU - Potschger,U, AU - Fritsch,G, PY - 2008/11/20/entrez PY - 2008/11/20/pubmed PY - 2009/7/18/medline SP - 834 EP - 41 JF - Cytotherapy JO - Cytotherapy VL - 10 IS - 8 N2 - BACKGROUND: Human cytomegalovirus (HCMV) infection remains a major cause of morbidity and mortality in immunocompromised patients undergoing allogeneic stem cell transplantation (SCT). In the case of HCMV reactivation, the well-defined detection of virus-specific effector cells in patients might positively impact antiviral treatment. METHODS: We examined blood samples from healthy volunteers serologically typed for HCMV IgG. Based on multicolor flow cytometry analysis, we addressed HCMV-specific CD8(+) effector T lymphocytes using HCMV-specific tetramers for the respective major histocompatibility complex (MHC) class I type. As a second approach, we employed the cytokine secretion assay (CSA), which allows the indirect detection of target-specific CD4(+) and CD8(+) T cells via their interferon (IFN)-gamma secretion upon HCMV pp65 in vitro stimulation. RESULTS: We hypothesized the detection of HCMV-specific lymphocytes in >50% of healthy Caucasians that were IgG-seropositive for HCMV. In terms of specificity, both assays showed comparably good results (specificity 100%, confidence interval >95%). Regarding sensitivity, both assays met the zero hypothesis. However, with 45/52 (86.5%) the tetramer technology was superior to the CSA, which detected 34/52 (65.4%) based on CD8(+) T cells and 41/52 (78.8%) based on both CD4(+) and CD8(+) T cells. DISCUSSION: A good correlation was observed between both assays, although the tetramers addressed only CD8(+) HCMV-specific T cells, whereas IFN-gamma secretion was detected on all T-cell types. Disadvantages of the CSA are the time-consuming stimulation, the extensive cell washing steps and the fact that the target cells are detected indirectly. The analysis with tetramers is rapid and reliable but their general use is hampered because of the restriction to a few HLA types. SN - 1477-2566 UR - https://www.unboundmedicine.com/medline/citation/19016371/Detection_of_human_cytomegalovirus_specific_T_lymphocytes_in_human_blood:_comparison_of_two_methods_ L2 - https://linkinghub.elsevier.com/retrieve/pii/905663391 DB - PRIME DP - Unbound Medicine ER -