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Background: Myeloid-derived suppressor cells (MDSCs) contribute to tumour escape from host immune surveillance and to tumour progression by producing tumour-promoting factors. We focused on clinical and analytical MDSCs-related issues as potential biomarkers and immune regulators involved in tumour progression. Patients and Methods: We analyzed 10 patients with advanced colorectal carcinoma (CRC) with (M1 subgroup) or without (M0 subgroup) distant metastases at diagnosis. Peripheral blood was collected at diagnosis prior to treatment and subsequently 12 months after therapy initiation. Using multicolour flow cytometry MDSC subsets were evaluated. Monocytic MDSCs (M-MDSCs) were detected as CD45 + CD11b + CD33 + HLA-DR low / − CD14 + CD15 - , granulocytic MDSCs (CD33 hi PMN-MDSC) were detected as CD45 + CD11b + CD33 hi HLA-DR low/− CD14 - CD15 + . For analytical and preanalytical studies random fresh blood specimens predominantly from cancer patients were analyzed. Results: Levels of circulating M-MDSCs were not associated with metastatic disease within advanced CRC patients. Levels of circulating CD33 hi PMN-MDSCs were elevated in patients with distant metastases compared to T3 M0 subgroup. Circulating M-MDSCs increased upon treatment initiation in 9 out of 10 patients. CD33 hi PMN-MDSCs substantially dropped upon treatment initiation in 5 out of 10 patients and substantially increased in 2 out of 10 patients. Analytical part showed that absolute and relative counts within each MDSC subset are correlated. Coefficient of variation (CV) for repeatability was 6-11 % for M-MDSCs and 25-44 % for CD33 hi PMN-MDSCs. CV for reproducibility was higher with 8-22 % for M-MDSCs and 35-79 % for CD33 hi PMN-MDSCs demonstrating that delay in measurement of MDSCs in whole blood specimen may distort quantification of circulating MDSC subsets. Conclusion: The quantification of MDSC subsets is substantially dependent on the type of specimen examined and its preanalytical processing. Exploratory analysis of M-MDSCs and CD33 hi PMN-MDSCs in colorectal carcinoma patients revealed different dynamics of M-MDSC and CD33 hi PMN-MDSC subsets in the context anti-cancer treatment.

We manufacture fully personalized monocytederived dendritic cell-based vaccines (DC-based vaccines) that are evaluated in investigator- initiated clinical trial “Combined antitumor therapy with ex vivo manipulated dendritic cells producing interleukin-12 in children, adolescents and young adults with progressive, recurrent or primarily metastatic high-risk tumors”. Detailed immunomonitoring of peripheral blood leucocytes subsets at the baseline and during vaccination is performed and includes routine examined peripheral blood parameters, monocyte subsets, MDSC, specific lymphocyte subsets including Tregs, effector Tcells, NKT-like cells.

We manufacture fully personalized monocyte-derived dendritic cell-based vaccines that are evaluated in investigator-initiated clinical trial „Combined antitumor therapy with ex vivo manipulated dendritic cells producing interleukin-12 in children, adolescents and young adults with progressive, recurrent of primarily metastatic high-risk tumours”. Immunobiological part of quality control (QC) of DC batches revealed variability in immunostimulatory properties. We analyzed DC parametres within the context of patients medical history and explore the effect of anti-cancer compound on monocyte functional properties.

γδ T cells present a minor population of the T cell family which basically differs in construction of their T cell receptor (TCR). Thanks to the features of γδ TCR, these cells can acquire unique effector functions and play a specific role (not only) in antitumor immune response. In this article, we describe the basic characteristics of this cell population and their connection to cancer. In the experimental part we performed exploratory analysis of circulating γδ T cells in reference population and comparison with melanoma and breast carcinoma patients. The median percentage of γδ T cells from all lymphocytes was 2.9% (interquartile range-IQR 1.7-4%). The median absolute numbers of γδ cells per liter of blood was 5.05×10(7) (IQR 2.9-7.84×10(7)). The median percentage of γδ cells between all CD3 T cells was 3.9% (IQR 2.3-5.6%). No correlation between γδ T cells levels and gender or age was observed in reference population. Detailed immunophenotyping was also conducted describing representation of memory subsets (using CD45RO and CD27 markers) and presence of surface markers HLADr, CD69, CD25, CD28, CCR7, CTLA 4, ICOS, PD 1L and PD 1 between γδ T cells of the controls and breast carcinoma patients. From this analysis, it is evident that γδ T cells do not represent a uniform population but they differ in surface markers as well as in their effector functions.