Automation for clinical CD4 T‐cell enumeration, a desirable tool in the hands of skilled operators

Diallo, Tamsir O.; Bergeron, Marcelle; Seely, Peggy; Yang, Xuefen; Ding, Tao; Plews, Margot; Sandstrom, Paul; Ball, T. Blake; Meyers, Adrienne F. A.

doi:10.1002/cyto.b.21370

Cited by 7 publications

(5 citation statements)

References 8 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…In QASI’s experience, the attractor algorithm associated with the software can have a limited capacity to adapt to some stabilised whole blood products provided as EQA quality control panels. 11 , 14 , 15 Fluorescence intensities are significantly different in these specimens as compared with fresh whole blood samples. This characteristic is not, however, exclusive to stabilised controls, as patient samples may behave similarly due to drug therapy or exposure to environmentally hostile conditions during transit.…”

Section: Resultsmentioning

confidence: 84%

QASI: A collaboration for implementation of an independent quality assessment programme in India

et al. 2016

View full text Add to dashboard Cite

ObjectiveThe HIV pandemic remains a significant global health concern. Accurate determination of CD4+ T-cells in patient samples relies on reliable CD4 enumeration. The Quality Assessment and Standardization programme for Immunological measures relevant to HIV/AIDS (QASI) programme of the Public Health Agency of Canada provides clinical laboratories from resource-limited countries with a mechanism to evaluate the quality of CD4 testing and develop the implementation of an independent national External Quality Assessment (EQA) programme. This study describes how QASI helped develop the capacity for managing a sustainable national CD4 EQA programme in India.DesignSupported by the Public Health Agency of Canada and Clinton Foundation HIV/AIDS Initiative, QASI engaged with the National AIDS Control Organization and the Indian National AIDS Research Institute to assist in technology transfer in preparation for the implementation/management of an independent CD4 EQA programme. Technology transfer training was provided to support corrective actions and to improve the quality of CD4 testing. Inter-laboratory variation of EQA surveys between pre- and post-skill development was compared.ResultsPrior to training, coefficient of variation values were 14.7% (mid-level CD4 count controls) and 39.0% (low-level). Following training, variation was reduced to 10.3% for mid-level controls and 20.0% for low-level controls.ConclusionThis training assisted the National AIDS Control Organization and the Indian National AIDS Research Institute in identifying the information necessary for management of an EQA programme, and developed the foundation for India to provide corrective actions for sites with challenges in achieving reliable results for CD4 enumeration. This led to a demonstrable improvement in CD4 testing quality and illustrates how country-specific training significantly improved CD4 enumeration performance for better clinical management of HIV care in India.

show abstract

Section: Resultsmentioning

confidence: 84%

QASI: A collaboration for implementation of an independent quality assessment programme in India

et al. 2016

View full text Add to dashboard Cite

show abstract

“…The loss of lymphocytes via automated light scatter gating in automated gating software was previously mentioned [33]. The automated program is optimized for sample analysis considering speci c conditions set by the manufacturer.…”

Section: Discussionmentioning

confidence: 99%

“…Furthermore, automated software excludes lymphocytes in certain circumstances, such as processing delays or environmental factors. This issue was resolved using a modi ed protocol that repaired all lost lymphocytes by increasing the LADJ area [33]; similar protocol was applied in the present study as well. Additionally, the percentage of total lymphocytes in the light scatter LADJ gate needs to be evaluated via automated gating program.…”

Section: Discussionmentioning

confidence: 99%

Comparison of Automated and Manual Gating of Lymphocyte Subsets in Hematopoietic Stem Cell Transplantation Recipients

et al. 2022

View full text Add to dashboard Cite

Background: Lymphocyte subset analysis is essential to evaluate the engraftment status in hematopoietic stem cell transplantation (HSCT). Automated gating tools are widely used for flow cytometry analysis. Unlike healthy individuals, different cell populations and aberrant expressions may occur in HSCT samples. In the present study, we evaluated the applicability of automated gating in HSCT recipients by comparing it to expertbased manual gating. Methods: Lymphocyte subset was performed using Beckman Coulter Navios (Beckman Coulter, USA) flow cytometry. Data files from 22 patients with hematologic malignancies were analyzed in parallel by manual gating and automated gating using Navios Tetra software. Quality control results and reproducibility were evaluated using IMMUNO-TROL controls. Results: Spearman rank correlation coefficients between the two gating methods were > 0.970 in all cell populations except CD8+ T cells. CD8+ T cell counts via automated gating were higher than those of manual gating in all cases due to the T cell populations with reduced CD8 expression. Automated gating program failed to identify CD4+CD8+ double-positive T cell population. Moreover, it excluded certain lymphocytes with low forward scatter (FSC) and high side scatter (SSC). Furthermore, two HSCT recipients revealed a high percentage of CD56−CD16+ NK cells, we found the need to add CD16 reagent to the Navios system. All coefficients of variation were < 10% except for CD56+ NK cells via automated gating. Conclusions: Manual gating confirmation via flow cytometry histogram is necessary to identify the aberrant phenotypes and unexpected cell populations in HSCT recipients.

show abstract

“…Guidelines for general method validation ( 114 , 118 – 120 ), as well as recommendations for specific applications are widely available ( 121 , 122 ). Furthermore, additional, unsupervised flow cytometry data evaluation methods have been developed in recent years ( 123 – 125 ) as the number of parameters that can be simultaneously interrogated by flow cytometry has increased ( 125 – 129 ). Therefore, there are few technical limitations preventing regular deployment of multiparametric flow cytometry for an in-depth and standardized characterization of cellular immunity in vaccine clinical trials.…”

Section: Measurements Of Cellular Immunitymentioning

confidence: 99%

Cellular Immunity Is Critical for Assessing COVID-19 Vaccine Effectiveness in Immunocompromised Individuals

Paramithiotis¹,

Sugden²,

Papp³

et al. 2022

Front. Immunol.

View full text Add to dashboard Cite

COVID-19 vaccine clinical development was conducted with unprecedented speed. Immunity measurements were concentrated on the antibody response which left significant gaps in our understanding how robust and long-lasting immune protection develops. Better understanding the cellular immune response will fill those gaps, especially in the elderly and immunocompromised populations which not only have the highest risk for severe infection, but also frequently have inadequate antibody responses. Although cellular immunity measurements are more logistically complex to conduct for clinical trials compared to antibody measurements, the feasibility and benefit of doing them in clinical trials has been demonstrated and so should be more widely adopted. Adding significant cellular response metrics will provide a deeper understanding of the overall immune response to COVID-19 vaccination, which will significantly inform vaccination strategies for the most vulnerable populations. Better monitoring of overall immunity will also substantially benefit other vaccine development efforts, and indeed any therapies that involve the immune system as part of the therapeutic strategy.

show abstract

Automation for clinical CD4 T‐cell enumeration, a desirable tool in the hands of skilled operators

Cited by 7 publications

References 8 publications

QASI: A collaboration for implementation of an independent quality assessment programme in India

QASI: A collaboration for implementation of an independent quality assessment programme in India

Comparison of Automated and Manual Gating of Lymphocyte Subsets in Hematopoietic Stem Cell Transplantation Recipients

Cellular Immunity Is Critical for Assessing COVID-19 Vaccine Effectiveness in Immunocompromised Individuals

Contact Info

Product

Resources

About