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Performance of calibration standards for antigen quantitation with flow cytometry in chronic lymphocytic leukemia.
Cytometry B Clin Cytom. 2007 Nov; 72(6):450-7.CB

Abstract

BACKGROUND

The fluorescence intensities of CD3, CD4 on T cells and CD20, CD22 molecules on B cells were quantitatively measured on lymphocytes from chronic lymphocytic leukemia (CLL) patients and healthy donors.

METHODS

The performance of three different types of microbeads was compared, i.e. Quantum molecules of equivalent soluble fluorochrome (Q-MESF), Quantum simply cellular (QSC), and QuantiBRITE (QB). As all PE-conjugates had a F/P ratio of 1:1, the MESF units represented also the antibody binding capacity (ABC).

RESULTS

The ABCs of CD4 and CD20 antigens estimated with QSC (ABC(QSC)) were higher than those assigned with QB (ABC(QB)) with an average difference 49%. Higher numbers of antigenic sites were obtained with Q-MESF than with QSC for CD20 antigen. On the contrary, CD4 antigenic sites numbers estimated with QSC were higher than those estimated with Q-MESF. ABC values estimated with Quantum MESF PE (ABC(Q-MESF)) were approximately 15% higher than ABC(QSC), whereas ABC(Q-MESF) was approximately 49% higher than ABC(QB). Statistically significant correlations were found between the values obtained using various standards. The present study is the first to report down-regulation of CD3 antigen on T cells from patients with CLL.

CONCLUSIONS

This study emphasizes the relevance of quantitative measurement of fluorescence intensity by flow cytometry as a standardized approach to measure and interpret the expression of some CLL markers and reduce variability of results obtained at different sites in multi-center clinical studies.

Authors+Show Affiliations

Department of Oncology (Radiumhemmet), Karolinska University Hospital, Stockholm, Sweden. eva.rossmann@karolinska.seNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info available

Pub Type(s)

Comparative Study
Journal Article

Language

eng

PubMed ID

17565749

Citation

Rossmann, Eva D., et al. "Performance of Calibration Standards for Antigen Quantitation With Flow Cytometry in Chronic Lymphocytic Leukemia." Cytometry. Part B, Clinical Cytometry, vol. 72, no. 6, 2007, pp. 450-7.
Rossmann ED, Lenkei R, Lundin J, et al. Performance of calibration standards for antigen quantitation with flow cytometry in chronic lymphocytic leukemia. Cytometry B Clin Cytom. 2007;72(6):450-7.
Rossmann, E. D., Lenkei, R., Lundin, J., Mellstedt, H., & Osterborg, A. (2007). Performance of calibration standards for antigen quantitation with flow cytometry in chronic lymphocytic leukemia. Cytometry. Part B, Clinical Cytometry, 72(6), 450-7.
Rossmann ED, et al. Performance of Calibration Standards for Antigen Quantitation With Flow Cytometry in Chronic Lymphocytic Leukemia. Cytometry B Clin Cytom. 2007;72(6):450-7. PubMed PMID: 17565749.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Performance of calibration standards for antigen quantitation with flow cytometry in chronic lymphocytic leukemia. AU - Rossmann,Eva D, AU - Lenkei,Rodica, AU - Lundin,Jeanette, AU - Mellstedt,Håkan, AU - Osterborg,Anders, PY - 2007/6/15/pubmed PY - 2007/12/7/medline PY - 2007/6/15/entrez SP - 450 EP - 7 JF - Cytometry. Part B, Clinical cytometry JO - Cytometry B Clin Cytom VL - 72 IS - 6 N2 - BACKGROUND: The fluorescence intensities of CD3, CD4 on T cells and CD20, CD22 molecules on B cells were quantitatively measured on lymphocytes from chronic lymphocytic leukemia (CLL) patients and healthy donors. METHODS: The performance of three different types of microbeads was compared, i.e. Quantum molecules of equivalent soluble fluorochrome (Q-MESF), Quantum simply cellular (QSC), and QuantiBRITE (QB). As all PE-conjugates had a F/P ratio of 1:1, the MESF units represented also the antibody binding capacity (ABC). RESULTS: The ABCs of CD4 and CD20 antigens estimated with QSC (ABC(QSC)) were higher than those assigned with QB (ABC(QB)) with an average difference 49%. Higher numbers of antigenic sites were obtained with Q-MESF than with QSC for CD20 antigen. On the contrary, CD4 antigenic sites numbers estimated with QSC were higher than those estimated with Q-MESF. ABC values estimated with Quantum MESF PE (ABC(Q-MESF)) were approximately 15% higher than ABC(QSC), whereas ABC(Q-MESF) was approximately 49% higher than ABC(QB). Statistically significant correlations were found between the values obtained using various standards. The present study is the first to report down-regulation of CD3 antigen on T cells from patients with CLL. CONCLUSIONS: This study emphasizes the relevance of quantitative measurement of fluorescence intensity by flow cytometry as a standardized approach to measure and interpret the expression of some CLL markers and reduce variability of results obtained at different sites in multi-center clinical studies. SN - 1552-4949 UR - https://www.unboundmedicine.com/medline/citation/17565749/Performance_of_calibration_standards_for_antigen_quantitation_with_flow_cytometry_in_chronic_lymphocytic_leukemia_ L2 - https://doi.org/10.1002/cyto.b.20359 DB - PRIME DP - Unbound Medicine ER -