Evaluation of a universal template for single‐platform absolute T‐lymphocyte subset enumeration

Bergeron, Marcelle; Faucher, Sylvie; Ding, Tao; Phaneuf, S.; Mandy, Francis

doi:10.1002/cyto.10089

Cited by 17 publications

(10 citation statements)

References 12 publications

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“…ϩ LCs from a single tube (4,6,12,34,35). Botswana, where the prevalence of HIV infection is one of the highest in the world, began a national antiretroviral treatment program in the public sector in January 2002.…”

Section: Cd4mentioning

confidence: 99%

Low CD4 ⁺ T-Lymphocyte Values in Human Immunodeficiency Virus-Negative Adults in Botswana

Bussmann

Wester

Masupu

et al. 2004

Clin Vaccine Immunol

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CD4+-lymphocyte counts (LCs) play a crucial role in the management and monitoring of HIV infection. Variability in CD4+ LCs has been reported to occur as a result of measurement techniques and/or biological variations. We report on the CD4+ LCs of healthy human immunodeficiency virus (HIV)-seronegative adults in Botswana. Samples were obtained from HIV-seronegative blood donors. The median CD4+ LC was 726 cells/mm3 (for females, 782 cells/mm3; for males, 698 cells/mm3). The median CD8+ LC was 488 cells/mm3 (for females, 494 cells/mm3; for males, 485 cells/mm3). The median CD4+-to-CD8+ ratio was 1.57 (for females, 1.66; for males, 1.51). Our findings of low CD4+ LCs among HIV-negative adults in Botswana are significant and have important implications for the management of HIV disease in the population of this sub-Saharan African country

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Section: Cd4mentioning

confidence: 99%

Low CD4 ⁺ T-Lymphocyte Values in Human Immunodeficiency Virus-Negative Adults in Botswana

Bussmann

Wester

Masupu

et al. 2004

Clin Vaccine Immunol

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“…Further manipulation of the specimens to obtain absolute white cell count and differential count by a separated automatic hematology cell counter are avoided by the incorporation of a known quantity of microfluorospheres in the flow cytometry step (2). The whole procedure is more robust, convenient, and reproducible than the traditional dual-platform flow cytometry.…”

mentioning

confidence: 99%

Reference Ranges for Lymphocyte Subsets among Healthy Hong Kong Chinese Adults by Single-Platform Flow Cytometry

Wong

Siu

et al. 2013

Clin Vaccine Immunol

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Lymphocyte subset enumeration is useful in the evaluation of hereditary and acquired immunodeficiency, assessment of immune reconstitution post-allogeneic stem cell transplantation, and also for monitoring CD4 count in HIV and AIDS patients. In particular, the CD4 count serves as a guide for initiation of antiretroviral therapy and prophylactic treatment of opportunistic infections (1-3) because it can provide valuable information for treatment response and disease progression. As a result, immunophenotyping of lymphocyte subsets by flow cytometry has been an indispensable tool in the management of HIV and AIDS patients. Various studies have shown that the reference ranges of lymphocyte subsets are subject to influences by age, sex, ethnicity, and environment (4-17). Flow cytometry is the "gold standard" for the enumeration of lymphocyte subsets, and it is important to establish the local reference ranges for interpretation of laboratory results and clinical decision making (13). Most of the published reference ranges were established by dual-platform flow cytometry (3,5,6,(9)(10)(11)(12)(14)(15)(16)(17).Healthy blood and apheresis donors from the local blood transfusion service were recruited for the establishment of the reference ranges of lymphocyte subsets by the use of single-platform flow cytometry. Single-platform flow cytometry allows simultaneous identification and enumeration of total CD3ϩ B cells, and CD56 ϩ natural killer cell population percentages and absolute cell counts. Further manipulation of the specimens to obtain absolute white cell count and differential count by a separated automatic hematology cell counter are avoided by the incorporation of a known quantity of microfluorospheres in the flow cytometry step (2). The whole procedure is more robust, convenient, and reproducible than the traditional dual-platform flow cytometry. Gender-and age-related differences in the lymphocyte subsets were evaluated. The results were also compared with those from different ethnic groups. The influence of sex and age on the lymphocyte composition and comparison with data from both Asian and non-Asian populations will be discussed.

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“…Seven SWBPs (Table 1) with high and low level of CD4 T-cells were tested using thirteen reagent combinations, including a standardised reference method [13], on seven enumeration platforms (Table 2). This assessment took place between 2009 and 2011.…”

Section: Methodsmentioning

confidence: 99%

“…The reference test method is a universal template for single platform T-cell enumeration previously evaluated for a wide array of instruments and immunophenotyping settings within the Canadian Clinical Trial Network laboratories [13]. This method uses a double anchor gating strategy based on two cell lineage specific markers (CD45 and CD3).…”

Section: Methodsmentioning

confidence: 99%

“…The reference test method has been used since 2002, by the National Laboratory for HIV Immunology of the Public Health Agency of Canada which is certified by United States National Institute of Allergy and Infectious Diseases (NIAID) CD4 Immunology Quality Assessment Program (IQAP, https://iqa.center.duke.edu). Additionally, the reference method has been used by the National Laboratory for HIV Immunology during their participation in the external quality assurance programs; UK-NEQAS for Leucocyte Immunophenotyping Program (www.ukneqasli.co.uk) and Flow Cytometry: CD34+ Stem Cell Enumeration Program (www.wiv-isp.be) [8], [13].…”

Section: Methodsmentioning

confidence: 99%

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Compatibility of Stabilized Whole Blood Products with CD4 Technologies and Their Suitability for Quality Assessment Programs

et al. 2014

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BackgroundCD4 T cell enumeration is the most widely used prognostic marker for management of HIV disease. Internal quality control and external quality assessment (EQA) programs are critical to ensure reliability of clinical measurements. The utility of stabilized whole blood products (SWBP) as a test reagent for EQA programs such as Quality Assessment and Standardization for Immunological measures relevant to HIV/AIDS (QASI) program have been demonstrated previously. Since then, several new commercial SWBPs and alternative CD4 enumeration technologies have become available. Seven SWBPs were evaluated on seven different enumeration platforms to determine which product(s) are most suitable for EQA programs that support multiple analytical technologies.MethodAssessment of SWBPs was based on two criteria: (1) accuracy of CD4 T cell measurements and; (2) stability under sub optimal storage conditions.ResultsThree SWBPs (Multi-Check, StatusFlow and CD4 Count) showed accurate CD4 T-cell absolute count and percentage values across six of the enumeration platforms. All products retain stability up to 18 days at 21–23°C with the exception of Multi-Check-high on FacsCount and Multi-Check-Low and StatusFlow-Low on Pima. One of the products (CD4 Count) retained stability for three days on all platforms tested when stored at 37°C.ConclusionThis study demonstrated that the characteristics of commercially available SWBPs vary across multiple CD4 platforms. The compatibility of testing panels for EQA programs with multiple analytical platforms needs to be carefully considered, especially in large multiplatform CD4 EQA programs. The selection of a suitable cross-platform SWBP is an increasing challenge as more reagents and platforms are introduced for CD4 T-cell enumeration.

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Evaluation of a universal template for single‐platform absolute T‐lymphocyte subset enumeration

Cited by 17 publications

References 12 publications

Low CD4 ⁺ T-Lymphocyte Values in Human Immunodeficiency Virus-Negative Adults in Botswana

Low CD4 ⁺ T-Lymphocyte Values in Human Immunodeficiency Virus-Negative Adults in Botswana

Reference Ranges for Lymphocyte Subsets among Healthy Hong Kong Chinese Adults by Single-Platform Flow Cytometry

Compatibility of Stabilized Whole Blood Products with CD4 Technologies and Their Suitability for Quality Assessment Programs

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Evaluation of a universal template for single‐platform absolute T‐lymphocyte subset enumeration

Cited by 17 publications

References 12 publications

Low CD4 + T-Lymphocyte Values in Human Immunodeficiency Virus-Negative Adults in Botswana

Low CD4 + T-Lymphocyte Values in Human Immunodeficiency Virus-Negative Adults in Botswana

Reference Ranges for Lymphocyte Subsets among Healthy Hong Kong Chinese Adults by Single-Platform Flow Cytometry

Compatibility of Stabilized Whole Blood Products with CD4 Technologies and Their Suitability for Quality Assessment Programs

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Low CD4 ⁺ T-Lymphocyte Values in Human Immunodeficiency Virus-Negative Adults in Botswana

Low CD4 ⁺ T-Lymphocyte Values in Human Immunodeficiency Virus-Negative Adults in Botswana