Ibrutinib, a first‐generation Bruton's tyrosine kinase (BTK) inhibitor, could improve immunity of relapsed or refractory (R/R) chronic lymphocytic leukemia/small lymphocytic lymphoma (CLL/SLL) patients. Whether zanubrutinib, a second‐generation selective BTK inhibitor, has similar effects as ibrutinib remains to be determined. Dynamics of number and immunophenotype of immune cells during zanubrutinib treatment in 25 R/R CLL/SLL patients were examined by flow cytometry and blood routine tests. The expression intensity of programmed death‐1 (PD‐1) on total CD4+ (P < .01), total CD8+ (P < .01), and T helper cells (P < .05) and cytotoxic T lymphocyte‐associated antigen‐4 (CTLA‐4) on total CD4+ (P = .010) and regulatory T cells (P < .05) reduced after treatment. There were significant differences in expression intensity of CD19 (P < .01), C‐X‐C chemokine receptor type 5 (CXCR5) (P < .01), and CD49d (P < .05) on B cells before and after treatment. Downregulation of PD‐1 on T cells and CXCR5 and CD19 on B cells were observed in nearly all patients after zanubrutinib treatment. Programmed death‐ligand 1 expression downregulated, especially in the female, CLL, normal spleen, normal β2‐macroglobulin (β2‐MG) and abnormal lactate dehydrogenase (LDH) subgroups, and CTLA‐4 expression on CD4+ T cells tended to decrease in the male, old, CLL, splenomegaly, abnormal β2‐MG, normal LDH, IGHV‐mutated and wild‐type tumor protein 53 subgroups after zanubrutinib treatment. These findings suggest that zanubrutinib can regulate immunity primarily by improving T cell exhaustion, inhibiting suppressor cells and disrupting CLL cells migration through downregulation of adhesion/homing receptors. Furthermore, favorable changes in cell number and immunophenotype were preferably observed in patients without adverse prognostic factors.

Background Chronic lymphocytic leukemia (CLL) results in increased susceptibility to infections. T cell dysfunction is not associated with CLL in all patients; therefore, it is important to identify CLL patients with T cell defects. The role of B-cell lymphoma-2 (BCL-2) in CLL has been explored; however, few studies have examined its role in T cells in CLL patients. Herein, we have investigated the regulatory role of BCL-2 in T cells in the CLL tumor microenvironment. Methods The expression of BCL-2 in T cells was evaluated using flow cytometry. The regulatory roles of BCL-2 were investigated using single-cell RNA sequencing (scRNA-seq) and verified using multi-parameter flow cytometry on CD4 and CD8 T cells. The clinical features of BCL-2 expression in T cells in CLL were also explored. Results We found a significant increase in BCL-2 expression in the T cells of CLL patients (n = 266). Single cell RNA sequencing (scRNA-seq) indicated that BCL-2+CD4+ T cells had the gene signature of increased regulatory T cells (Treg); BCL-2+CD8+ T cells showed the gene signature of exhausted cytotoxic T lymphocytes (CTL); and increased expression of BCL-2 was associated with T cell activation and cellular adhesion. The results from scRNA-seq were verified in peripheral T cells from 70 patients with CLL, wherein BCL-2+CD4+ T cells were enriched with Tregs and had higher expression of interleukin-10 and transforming growth factor-β than BCL-2−CD4+ T cells. BCL-2 expression in CD8+T cells was associated with exhausted cells (PD-1+Tim-3+) and weak expression of granzyme B and perforin. T cell–associated cytokine profiling revealed a negative association between BCL-2+ T cells and T cell activation. Decreased frequencies and recovery functions of BCL-2+T cells were observed in CLL patients in complete remission after treatment with venetoclax. Conclusion BCL-2 expression in the T cells of CLL patients is associated with immunosuppression via promotion of Treg abundance and CTL exhaustion.

Objectives: Multiple myeloma (MM) involves a clinically and biologically heterogeneous malignancy of plasma cells. It is difficult to predict the prognosis of MM. The presence of circulating clonal plasma cells (CPC) has been associated with a worse prognosis in patients with MM. Methods: This study retrospectively analysed CPC in 108 newly diagnosed MM patients by 8colour flow cytometry to investigate their value for predicting the outcome and combined the level of CPC with the revised International Staging System (R-ISS) to stratify the MM patients according to risk. Results: CPC were detected in 58/108 patients (53.7%). The optimum cut-off for the prediction of overall survival was determined to be 0.105%. Patients with higher R-ISS stages seemed to harbour more CPC. A level of CPC≥0.105% was an independent risk factor for adverse outcomes (P<0.001). The combination of the R-ISS staging system and level of CPC was used to stratify MM patients according to risk, and the combination of R-ISS stage III and a level of CPC≥0.105% defined the ultra-high-risk group. Conclusion:This study suggests that a high proportion of CPC is associated with aggressive disease and that the use of the current R-ISS system in conjunction with assessment of the level of CPC may facilitate the stratification of newly diagnosed MM patients into clinically relevant prognostic subgroups.