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Development of a Modular Assay for Detailed Immunophenotyping of Peripheral Human Whole Blood Samples by Multicolor

Paul F Rühle1, Rainer Fietkau2, Udo S Gaipl3

  • 1Department of Radiation Oncology, Universitätsklinikum Erlangen, Friedrich-Alexander-Universität Erlangen-Nürnberg, Erlangen 91054, Germany. paul.ruehle@uk-erlangen.de.

International Journal of Molecular Sciences
|August 17, 2016
PubMed

Insights

A new assay precisely monitors 34 immune cell subsets in peripheral blood, aiding cancer therapy response prediction. This detailed immune profiling uses minimal blood volume for longitudinal patient studies.

Area of Science:

  • Immunology
  • Biotechnology
  • Medical Diagnostics

Background:

  • Immune cell monitoring is crucial for cancer therapy prognosis and predicting treatment response.
  • Multicolor flow cytometry is the preferred method for analyzing blood samples due to its specificity, multi-parameter capability, and high throughput.
  • Detailed immunophenotyping of blood (DIoB) is needed for comprehensive immune status assessment.

Purpose of the Study:

  • To present a modular assay for detailed immunophenotyping of blood (DIoB) optimized for direct use in whole blood samples.
  • To characterize a broad range of immune cell subsets and their functional states.
  • To enable longitudinal and close-meshed immune monitoring in patients.

Main Methods:

  • Development of a modular assay for detailed immunophenotyping of blood (DIoB).
  • Application of the assay to whole blood samples for direct analysis.
  • Characterization of 34 distinct peripheral blood immune cell subsets, including T cells, B cells, NK cells, monocytes, dendritic cells (DCs), neutrophils, eosinophils, and basophils.
  • Evaluation of immune cell functional states and select non-leukocytes.

Main Results:

  • The DIoB assay successfully characterizes 34 immune cell subsets in peripheral blood.
  • The assay requires only 2.0 mL of peripheral blood for analysis.
  • It is applicable to longitudinal and close-meshed immune monitoring, not limited to peripheral blood mononuclear cells.
  • The assay is currently used in clinical trials for glioblastoma, pancreatic cancer, and head and neck cancer.

Conclusions:

  • The DIoB assay provides a detailed immune status overview for patients.
  • It facilitates easy and direct application in whole blood samples.
  • This assay may significantly contribute to identifying immune biomarkers for predicting and prognosing therapy outcomes in cancer patients.

Keywords:
adaptive immune systemimmune monitoringimmunophenotypinginnate immune systemliquid biopsymulticolor flow cytometrywhole blood

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