Natural killer/T-cell lymphoma (NKTCL) is a rare subtype of non-Hodgkin lymphoma that is associated with a poor outcome. Currently, the treatment needs of NKTCL remain unmet, and efforts to further improve treatment are urgently needed. Herein, seven patients with NKTCL who failed to respond to various types of chemotherapies were treated with the anti-programmed death 1 (anti-PD-1) antibody pembrolizumab at 100 mg every 3 weeks. After a median of four cycles of treatment (range 2–18), four out of seven patients responded (two complete response, two partial response, overall response rate 57%). Expression of PD1-ligand available was 50, 20, 30, 70, and 30% of five patients respectively. It is negative in one patient and not tested in one patient. Adverse events, which mostly ranged from grade I to grade III, were tolerable and could be safely handled, although immune-related pneumonitis was notable. Overall, PD-1 blockade with pembrolizumab represents a favorable strategy for the treatment of refractory/relapsed NKTCL.

Neutrophil extracellular traps (NETs) are web‑like structures made of chromatin and have been identified to have a role in the host's immune defense. Differentiated human promyelocytic leukemia HL‑60 cells (dHL‑60) have been used to study the mechanisms of NETs formation, as neutrophils have a short lifespan that limits their use. However, dHL‑60 cells are inefficient at generating NETs and therefore are not ideal replacements for neutrophils in studying of NET formation. In the present study, the optimal cell culture conditions and differentiation time that result in the most effective release of NETs from dHL‑60 cells upon stimulation were determined. HL‑60 cells were cultured in serum (FBS) or serum‑free (X‑VIVO) medium and differentiated using all‑trans retinoic acid (ATRA) or dimethyl sulfoxide (DMSO). dHL‑60 cells were stimulated with phorbol 12‑myristate 13‑acetate (PMA) or Ca2+ ionophore (CI). Cell differentiation and apoptosis, as well as the formation of reactive oxygen species (ROS) and citrullinated histone H3 (citH3) were analyzed using flow cytometry. NETs were visualized using fluorescence microscopy and NET quantification was performed using PicoGreen. Induction of HL‑60 cells for five days produced the best results in terms of differentiation markers and cell viability. Both ATRA‑ and DMSO‑induced dHL‑60 cells were able to release NETs upon PMA and CI stimulation; dHL‑60 cells in serum‑free medium produced more NETs than those in serum‑containing medium. DMSO‑dHL‑60 (X‑VIVO) cells were most efficient at producing NETs and ROS upon stimulation with PMA, while ATRA‑dHL‑60 (X‑VIVO) cells were most efficient at producing NETs and citH3 upon stimulation with CI. It was concluded that DMSO‑dHL‑60 (X‑VIVO) may be a model for the study of ROS‑high NETosis and ATRA‑dHL‑60 (X‑VIVO) may be suitable for ROS‑low NETosis.

Prognostic impact of c-Rel nuclear expression and REL amplification and crosstalk between c-Rel and the p53 pathway in diffuse large B-cell lymphoma Prognostic impact of c-Rel nuclear expression and REL amplification and crosstalk between c-Rel and the p53 pathway in diffuse large B-cell lymphoma

Background: Although chimeric antigen receptor T (CAR-T) cells have been proven to be an effective way of treating B cell malignancies, a lot of patients could not benefit from it because of failure in CAR-T cell manufacturing, disease progression, and unaffordable price. The study aimed to explore universal CAR-T cell products to extend the clinical accessibility. Methods: The antitumor activity of CRISPR/Cas9-edited allogeneic anti-CD19 CAR-T (CAR-T19) cells was assessed in vitro, in animal models, and in patients with relapsed/refractory (R/R) acute B cell lymphoblastic leukemia (B-ALL) or diffuse large B cell lymphoma. Results: B2M⁻/TRAC⁻ universal CAR-T19 (U-CAR-T19) cells exhibited powerful anti-leukemia abilities both in vitro and in animal models, as did primary CD19⁺ leukemia cells from leukemia patients. However, expansion, antitumor efficacy, or graft-versus-host-disease (GvHD) was not observed in six patients with R/R B cell malignancies after U-CAR-T19 cell infusion. Accordingly, significant activation of natural killer (NK) cells by U-CAR-T19 cells was proven both clinically and in vitro. HLA-A⁻/B⁻/TRAC⁻ novel CAR-T19 (nU-CAR-T19) cells were constructed with similar tumoricidal capacity but resistance to NK cells in vitro. Surprisingly, robust expansion of nU-CAR-T19 cells, along with rapid eradication of CD19⁺ abnormal B cells, was observed in the peripheral blood and bone marrow of another three patients with R/R B-ALL. The patients achieved complete remission with no detectable minimal residual disease 14 days after the infusion of nU-CAR-T19 cells. Two of the three patients had grade 2 cytokine release syndrome, which were managed using an IL-6 receptor blocker. Most importantly, GvHD was not observed in any patient, suggesting the safety of TRAC-disrupted CAR-T cells generated using the CRISPR/Cas9 method for clinical application. Conclusions: The nU-CAR-T19 cells showed a strong response in R/R B-ALL. nU-CAR-T19 cells have the potential to be a promising new approach for treating R/R B cell malignancies.

Summary: Purpose Apatinib, a new tyrosine kinase inhibitor targeting vascular endothelial growth factor receptor-2, has shown promising efficacy against several solid cancers, but evidence of its efficacy against relapsed and refractory nasopharyngeal carcinoma is limited. We investigated the efficacy and safety of apatinib for relapsed and refractory nasopharyngeal carcinoma in an open-label, single-arm, phase II clinical trial. Fifty-one patients with relapsed and refractory nasopharyngeal carcinoma in the First Affiliated Hospital, Zhengzhou University, who met the inclusion criteria were enrolled in the study. All patients received apatinib at an initial dose of 500 mg daily (1 cycle = 28 days). The primary and secondary endpoints were overall response rate, progression-free survival, and overall survival. We evaluated treatment effects and recorded apatinib-related adverse events by performing regular follow-ups and workup. The overall response rate (complete and partial responses) was 31.37% (16/51). The median overall survival and progression-free survival were 16 (95% CI, 9.32–22.68) and 9 months (95% CI, 5.24–12.76), respectively. Most patients tolerated treatment-related adverse events of grades 1 and 2; hypertension (29, 56.86%), proteinuria (25, 49.02%), and hand–foot syndrome (27, 52.94%) were the most common adverse events. There were no treatment-related deaths. Apatinib showed good efficacy and safety in patients with relapsed and refractory NPC.