Abstract
The monitoring of immune cells gained great significance in prognosis and prediction of therapy responses. For analyzing blood samples, the multicolor flow cytometry has become the method of choice as it combines high specificity on single cell level with multiple parameters and high throughput. Here, we present a modular assay for the detailed immunophenotyping of blood (DIoB) that was optimized for an easy and direct application in whole blood samples. The DIoB assay characterizes 34 immune cell subsets that circulate the peripheral blood including all major immune cells such as T cells, B cells, natural killer (NK) cells, monocytes, dendritic cells (DCs), neutrophils, eosinophils, and basophils. In addition, it evaluates their functional state and a few non-leukocytes that also have been associated with the outcome of cancer therapy. This DIoB assay allows a longitudinal and close-meshed monitoring of a detailed immune status in patients requiring only 2.0 mL of peripheral blood and it is not restricted to peripheral blood mononuclear cells. It is currently applied for the immune monitoring of patients with glioblastoma multiforme (IMMO-GLIO-01 trial, NCT02022384), pancreatic cancer (CONKO-007 trial, NCT01827553), and head and neck cancer (DIREKHT trial, NCT02528955) and might pave the way for immune biomarker identification for prediction and prognosis of therapy outcome.
Highlights
In the last decades, immunotherapy (IT) has become a prominent part in multimodal cancer therapy complementing the classical treatments of surgery, chemotherapy (CT) and radiotherapy (RT)
We present here a multicolor flow cytometry-based assay that examines the detailed immune status covering 34 different immune cell subsets and three non-immune cell subsets in only 2 mL of human peripheral blood
The detailed immunophenotyping of blood (DIoB) assay was designed in a modular system comprising 12 different panels (Table 1)
Summary
Immunotherapy (IT) has become a prominent part in multimodal cancer therapy complementing the classical treatments of surgery, chemotherapy (CT) and radiotherapy (RT). We present here a multicolor flow cytometry-based assay that examines the detailed immune status covering 34 different immune cell subsets and three non-immune cell subsets in only 2 mL of human peripheral blood It was optimized for a direct staining of whole blood samples which on the one hand allows the detection of all circulating immune cells and on the other hand reduces the required preparation steps. The DIoB assay was designed in a modular principle comprising 12 panels which each is dedicated to determine one specific type of cell and its subsets It is suited for the monitoring of the detailed immune status of patients in short intervals paving the way for optimization or even individualization of multimodal cancer therapies
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