The tissue microenvironment in chronic lymphocytic leukaemia (CLL) plays a key role in the pathogenesis of CLL, but the complex blood microenvironment in CLL has not yet been fully characterised. Therefore, immunophenotyping of circulating immune cells in 244 CLL patients and 52 healthy controls was performed using flow cytometry and analysed by multivariate Patient Similarity Networks (PSNs). Our study revealed high inter-individual heterogeneity in the distribution and activation of bystander immune cells in CLL, depending on the bulk of the CLL cells. High CLL counts were associated with low activation on circulating monocytes and T cells and vice versa. The highest activation of immune cells, particularly of intermediate and non-classical monocytes, was evident in patients treated with novel agents. PSNs revealed a low activation of immune cells in CLL progression, irrespective of IgHV status, Binet stage and TP53 disruption. Patients with high intermediate monocytes (> 5.4%) with low activation were 2.5 times more likely (95% confidence interval 1.421-4.403, P = 0.002) to had shorter time-to-treatment than those with low monocyte counts. Our study demonstrated the association between the activation of circulating immune cells and the bulk of CLL cells. The highest activation of bystander immune cells was detected in patients with slow disease course and in those treated with novel agents. The subset of intermediate monocytes showed predictive value for time-to-treatment in CLL.

1st Department of Medicine Department of Hematology 1st Faculty of Medicine Charles University and General University Hospital Prague Praha Czech Republic

Department of Clinical Hematology Hospital in Havirov Havirov Czech Republic

Department of Computer Science Faculty of Electrical Engineering and Computer Science VSB Technical University of Ostrava Ostrava Czech Republic

Department of Hematology and Oncology University Hospital Plzen Plzen Czech Republic

Department of Hematology Oncology Faculty of Medicine and Dentistry Palacký University and University Hospital Olomouc Olomouc Czech Republic

Department of Immunology Faculty of Medicine and Dentistry Palacký University and University Hospital Olomouc Hnevotinska 3 775 15 Olomouc Czech Republic

Department of Internal Medicine Hematology 3rd Faculty of Medicine Charles University and Faculty Hospital Kralovske Vinohrady Praha Czech Republic

Department of Internal Medicine Hematology and Oncology University Hospital Brno and Faculty of Medicine Masaryk University Brno Czech Republic

Department of Internal Medicine Hematology University Hospital and Medical School Hradec Kralove Hradec Kralove Czech Republic

Laboratory of Molecular and Cellular Immunology Institute of Molecular Biology NAS RA Yerevan Armenia

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