Immune monitoring

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Inter- and Intra-laboratory Variability of Polyfunctional Responses Assessed by Intracellular Staining Assay as Part of the External Quality Assurance Program Oversight Laboratory Flow Cytometry Proficiency Testing Program

Friday, June 16
6:15 PM - 7:30 PM

Polyfunctional ICS responses play a significant role in the context of immune monitoring to evaluate vaccines and immunotherapies. Studies suggest that polyfunctional ICS responses are associated with immunologic control of HIV and represent correlates of protection post vaccination. Additionally, many laboratories are currently utilizing ICS assays to profile anti-tumor responses in the context of cancer vaccine trials and novel immunotherapies.


While polyfunctional cells are routinely measured in the context of clinical trials, there is a paucity of data describing the variability of these measures between and within laboratories. A limited number of proficiency programs assess the performance of ICS assays across laboratories, albeit these programs have focused primarily on assessing total functional ICS responses, rather than polyfunctional responses.


The EQAPOL Flow Cytometry Program assesses the proficiency of NIH/NIAID/DAIDS-supported laboratories in performing ICS assays. The EQAPOL Program currently assesses site performance of ICS assays based on total functional responses. Introducing an evaluation of polyfunctional ICS responses represents the next logical step for advancing the EQAPOL Program. Therefore, we devised a pilot study to determine the feasibility of assessing polyfunctional ICS responses as part of the EQAPOL Flow Cytometry Proficiency Testing Program. EQAPOL compared total functional and polyfunctional ICS data, collected across three proficiency testing surveys with an average of 16 participating laboratories. Respectively, inter- and intra-laboratory variance estimate ranges were 0.000-0.032 and 0.000-0.005 for polyfunctional and 0.008-0.035 and 0.000-0.009 for total cytokine ICS responses. Overall, the inter- and intra-laboratory variability of polyfunctional ICS responses were similar to total functional ICS responses.

Janet S. Staats

Scientific/Research Laboratory Manager
Duke University

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    Jennifer H. Enzor

    Laboratory Research Analyst, Sr
    Duke University

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      Robert Rountree

      Biostatistician
      Duke University

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        John Bainbridge

        Biostatistician III
        Duke University

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          Thomas N. Denny

          Professor, Dept of Medicine
          Duke University

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            Kent J. Weinhold

            Professor, Depts of Surgery and Immunology
            Duke University

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