Neutrophils have limited utility as a fundamental research model and transfusion product due to their short lifespan. Here, we cultured CD34+ hematopoietic stem and progenitor cell (HSPC)-derived neutrophils and compared them to peripheral blood neutrophils in terms of morphology, phenotype, and function. Our culture system resulted in morphologically mature CD15+CD11b+CD16high neutrophils with effector functions almost indistinguishable from blood neutrophils, confirmed by a high similarity in transcription and protein abundance patterns. While exhibiting microbial killing capacity and antibody-dependent cellular cytotoxicity, these cells were deficient in myeloid-derived suppressor cell activity. This deficiency in immunosuppressive activity correlated with a distinct granular composition in comparison to blood neutrophils, instead of immunosuppressive characteristics that are currently held to define neutrophil-MDSC phenotype. Taken together, our cultured neutrophils closely resemble blood neutrophils, offering a repository for fundamental research and a step toward an effective transfusion product with limited immunosuppressive activity as a functional property.
© 2025 The Authors.

Monomorphic epitheliotropic intestinal T-cell lymphoma (MEITL) is a rare and aggressive primary intestinal lymphoma with a poor prognosis. MEITL can metastasize to the central nervous system, liver and spleen, but gallbladder involvement has not yet been reported. The present study describes the case of a 57-year-old woman who presented with abdominal distention, pain and vomiting. Contrast-enhanced computed tomography revealed thickening and perforation of the small intestinal wall, and a gallbladder mass. Histopathological analysis of the affected small intestine and gallbladder revealed a dense infiltrate of medium-sized monomorphic lymphocytes with a CD3+, CD4-, CD8+ and TIA-1+ phenotype. Based on the absence of celiac disease, aggressive clinical course, and characteristic histopathological and immunophenotypic features, a diagnosis of MEITL with gallbladder involvement was established. The patient underwent small intestinal resection and cholecystectomy, followed by chemotherapy, which was completed without gastrointestinal or gallbladder perforation. Diagnostic resection is currently the best approach for suspected malignant lymphoma of the gallbladder. This rare case of MEITL with gallbladder involvement highlights the importance of considering this diagnosis in similar clinical scenarios and the role of cholecystectomy, which can serve both diagnostic and therapeutic purposes.
Copyright: © 2025 Okuda et al.

Early development of broadly neutralizing antibodies (bNAbs) targeting the hepatitis C virus (HCV) envelope glycoprotein E2 is associated with spontaneous clearance of infection, so induction of bNAbs is a major goal of HCV vaccine development. However, the molecular antibody features important for broad neutralization are not known.
To identify B cell repertoire features associated with broad neutralization, we performed RNA sequencing of the B cell receptors (BCRs) of HCV E2-reactive B cells of HCV-infected individuals with either high or low plasma neutralizing breadth. We then produced a monoclonal antibody (mAb) expressed by pairing the most abundant heavy and light chains from public clonotypes identified among clearance, high neutralization subjects.
We found distinctive BCR features associated with broad neutralization of HCV, including long heavy chain complementarity determining region 3 (CDRH3) regions, specific VH gene usage, increased frequencies of somatic hypermutation, and particular VH gene mutations. Most intriguing, we identified many E2-reactive public BCR clonotypes (heavy and light chain clones with the same V and J-genes and identical CDR3 sequences) present only in subjects who produced highly neutralizing plasma. The majority of these public clonotypes were shared by two subjects who cleared infection. A mAb expressing the most abundant public heavy and light chains from these clearance, high neutralization subjects had features enriched in high neutralization clonotypes, such as increased somatic hypermutation frequency and usage of IGHV1-69, and was cross-neutralizing.
Together, these results demonstrate distinct BCR repertoires associated with high plasma neutralizing capacity. Further characterization of the molecular features and function of these antibodies can inform HCV vaccine development.
Copyright © 2023 Skinner, Ogega, Frumento, Clark, Yegnasubramanian, Schuebel, Meyers, Gupta, Wheelan, Cox, Crowe, Ray and Bailey.

Chimeric antigen receptor (CAR) T-cell therapy targeting CD19 has been a clinical breakthrough for pediatric B-cell acute lymphoblastic leukemia (B-ALL), and loss of the CD19 target antigen on leukemic cells represents a major mechanism of relapse. Previous studies have observed CD19 mutations specific to CD19- relapses, and we sought to clarify and strengthen this relationship using deep whole-exome sequencing in leukemic cells expanded in a patient-derived xenograft. By assessing pre-treatment and relapse cells from 13 patients treated with CAR T-cell therapy, 8 of whom developed CD19- relapse and 5 of whom developed CD19+ relapse, we demonstrate that relapse-specific single-nucleotide variants and small indels with high allele frequency combined with deletions in the CD19 gene in a manner specific to those patients with CD19- relapse. Before CAR T-cell infusion, one patient was found to harbor a pre-existing CD19 deletion in the context of genomic instability, which likely represented the first hit leading to the patient's subsequent CD19- relapse. Across patients, preexisting mutations and genomic instability were not significant predictors of subsequent CD19- relapse across patients, with sample size as a potential limiting factor. Together, our results clarify and strengthen the relationship between genomic events and CD19- relapse, demonstrating this intriguing mechanism of resistance to a targeted cancer immunotherapy.
©2022 The Authors; Published by the American Association for Cancer Research.

Research with animals and humans has demonstrated that chronic stress exposure can impact key biological aging pathways such as inflammation and DNA damage, suggesting a mechanism through which stress may increase risk for age-related disease. However, it is less clear whether these effects extend to other hallmarks of the aging process, such as cellular senescence. Male SCID mice were exposed to 14 days of restraint stress, with (n ​= ​6) or without (n ​= ​10) propranolol administration, or a non-stress control condition (n ​= ​10). Normal femoral bone marrow leukocytes were isolated from engrafted leukemia cells that had been injected prior to the stressor, as the mice were also under a cancer challenge. We performed whole genome transcriptional profiling to assess indicators of biological aging: cell stress, DNA damage repair, cellular senescence markers p16INK4a and p21, and the pro-inflammatory senescence-associated secretory phenotype (SASP). ANCOVAs that adjusted for tumor load and Fisher's pairwise comparisons revealed that stressed mice had enhanced p16INK4a (p ​= ​.02) and p21 (p ​= ​.004), lower DNA damage repair (p ​< ​.001), and higher SASP (p ​= ​.03) gene expression than control mice. Stressed mice also showed up-regulated beta-adrenergic (CREB) and inflammatory (NF-кB, AP-1) and down-regulated cell stress (Nrf2) transcription factor activity relative to control mice (ps ​< ​.01). Propranolol reversed CREB and Nrf2 activity (ps ​< ​.03). Findings suggest that chronic stress exposure can impact several key biological aging pathways within bone marrow leukocytes and these effects may be partially mediated by sympathetic beta-adrenergic receptor activation.
© 2022 The Authors.