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From research tool to routine test: CD38 monitoring in HIV patients.
Cytometry B Clin Cytom. 2009 Nov; 76(6):375-84.CB

Abstract

BACKGROUND

CD38 expression on CD8+ T lymphocytes in HIV-infected patients is monitored by flow cytometry (FCM). There is however no consensus re CD38 protocols, analyses or result reporting within/between laboratories. Internal quality control measures (QC) were established for a standardized CD38 protocol and a system proposed for reporting CD38 fluctuation in longitudinal HIV+ patient monitoring.

METHODS

A single-platform (SP) CD38/CD8 protocol was "piggy-backed" onto the standardized "panleucogating" CD45/CD4+ protocol. A weekly QC was established to monitor instrument stability (FlowSET) and absolute cell count accuracy and reproducibility (stabilized blood product, Immuno-Trol). The Mean Fluorescence Intensity (MFI) of CD38 expression on CD8(+)-lymphocytes was monitored on both stabilized blood and HIV-control samples. Linearized MFI values were determined from biological controls, i.e. healthy donor monocytes and granulocytes, and tested as a method of reporting CD38 expression on selected HIV+ patients on ART.

RESULTS

The CD45/CD4/CD8/CD3 method for lymphocyte enumeration compared well with the CD38 protocol (CD45/CD4/CD8/CD38) with excellent similarity (+/-100%) and precision for absolute CD4 and CD8 counts (CVs < 5%). Fluorosphere MFI- (FlowSet, FlowCount) and color compensation values were exceptionally stable over time. CD38 MFI values established on monocytes as biological control was 4.0 and <2.0 for HIV-control lymphocytes.

CONCLUSIONS

Monitoring FCM with fluorosphere MFI values, color compensation, and biological controls, can ensure that CD38 analyses are technologically stable. Flow cytometry is thus the preferred method to monitor fluctuations in CD38 MFI (CD38 molecules/cell) associated with HIV-disease progression and/or response to ART and has potential for application across instruments and centers.

Authors+Show Affiliations

National Health Laboratory Service and Department of Molecular Medicine and Haematology, Faculty of Health Sciences, University of the Witwatersrand, Johannesburg, South Africa.No affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info available

Pub Type(s)

Journal Article
Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

19422053

Citation

Coetzee, Lindi M., et al. "From Research Tool to Routine Test: CD38 Monitoring in HIV Patients." Cytometry. Part B, Clinical Cytometry, vol. 76, no. 6, 2009, pp. 375-84.
Coetzee LM, Tay SS, Lawrie D, et al. From research tool to routine test: CD38 monitoring in HIV patients. Cytometry B Clin Cytom. 2009;76(6):375-84.
Coetzee, L. M., Tay, S. S., Lawrie, D., Janossy, G., & Glencross, D. K. (2009). From research tool to routine test: CD38 monitoring in HIV patients. Cytometry. Part B, Clinical Cytometry, 76(6), 375-84. https://doi.org/10.1002/cyto.b.20478
Coetzee LM, et al. From Research Tool to Routine Test: CD38 Monitoring in HIV Patients. Cytometry B Clin Cytom. 2009;76(6):375-84. PubMed PMID: 19422053.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - From research tool to routine test: CD38 monitoring in HIV patients. AU - Coetzee,Lindi M, AU - Tay,Szun Szun, AU - Lawrie,Denise, AU - Janossy,George, AU - Glencross,Deborah K, PY - 2009/5/8/entrez PY - 2009/5/8/pubmed PY - 2009/12/16/medline SP - 375 EP - 84 JF - Cytometry. Part B, Clinical cytometry JO - Cytometry B Clin Cytom VL - 76 IS - 6 N2 - BACKGROUND: CD38 expression on CD8+ T lymphocytes in HIV-infected patients is monitored by flow cytometry (FCM). There is however no consensus re CD38 protocols, analyses or result reporting within/between laboratories. Internal quality control measures (QC) were established for a standardized CD38 protocol and a system proposed for reporting CD38 fluctuation in longitudinal HIV+ patient monitoring. METHODS: A single-platform (SP) CD38/CD8 protocol was "piggy-backed" onto the standardized "panleucogating" CD45/CD4+ protocol. A weekly QC was established to monitor instrument stability (FlowSET) and absolute cell count accuracy and reproducibility (stabilized blood product, Immuno-Trol). The Mean Fluorescence Intensity (MFI) of CD38 expression on CD8(+)-lymphocytes was monitored on both stabilized blood and HIV-control samples. Linearized MFI values were determined from biological controls, i.e. healthy donor monocytes and granulocytes, and tested as a method of reporting CD38 expression on selected HIV+ patients on ART. RESULTS: The CD45/CD4/CD8/CD3 method for lymphocyte enumeration compared well with the CD38 protocol (CD45/CD4/CD8/CD38) with excellent similarity (+/-100%) and precision for absolute CD4 and CD8 counts (CVs < 5%). Fluorosphere MFI- (FlowSet, FlowCount) and color compensation values were exceptionally stable over time. CD38 MFI values established on monocytes as biological control was 4.0 and <2.0 for HIV-control lymphocytes. CONCLUSIONS: Monitoring FCM with fluorosphere MFI values, color compensation, and biological controls, can ensure that CD38 analyses are technologically stable. Flow cytometry is thus the preferred method to monitor fluctuations in CD38 MFI (CD38 molecules/cell) associated with HIV-disease progression and/or response to ART and has potential for application across instruments and centers. SN - 1552-4957 UR - https://www.unboundmedicine.com/medline/citation/19422053/From_research_tool_to_routine_test:_CD38_monitoring_in_HIV_patients_ L2 - https://doi.org/10.1002/cyto.b.20478 DB - PRIME DP - Unbound Medicine ER -