CD20 Antigen Research Articles

Allogeneic Off-the-Shelf CAR T-Cell Therapy for Relapsed or Refractory B-Cell Malignancies

Despite clinical benefit with the use of CAR T-cells, the need to manufacture patient-specific products severely limits the clinical utility of this therapy. To overcome this barrier, we developed an allogeneic, "off-the-shelf" CAR T-cell product using Epstein-Barr virus (EBV)-specific T cells (EBV-VSTs) genetically modified with a CD19-specific CAR (19-28z). Patients with relapsed/refractory (R/R) B-cell malignancies (n=16) were stratified into 3 treatment cohorts: cohort 1 (n=8; disease recurrence after allogeneic or autologous hematopoietic cell transplant (HCT)), cohort 2 (n=6; consolidative therapy after autologous HCT), or cohort 3 (n=2; consolidative therapy after allogeneic HCT) (NCT01430390). The primary objective of this trial was to determine safety of multiple CAR EBV-VST infusions. Most patients (n=12/16; 75%) received multiple doses (overall median 2.5; range 1-3) with 3x106 T-cells/kg determined to be the optimal dose enabling multiple treatments per manufactured cell line. Severe cytokine release syndrome or neurotoxicity did not occur post-infusion, and no dose limiting toxicity was observed in the trial. Median follow-up was 48 months (range 4-135 months) with 4 deaths due to disease progression. Overall survival of all patients was 81% (95% confidence interval: 64-100%) at 12 months and 75% (57-100%) at 36 months. Post-infusion expansion and persistence were limited, and CAR EBV-VSTs demonstrated a unique T-cell phenotype as compared to autologous 19-28z CAR T-cells. Our study demonstrates the feasibility and safety of an allogeneic, "off-the-shelf" CAR EBV-VST product with favorable outcomes for patients with R/R B-cell malignancies when targeting the CD19 antigen.

Recurrent bacterial pneumonia in Irish Wolfhounds - histopathology and characterization of bronchial cartilage changes.

An increased incidence of bacterial pneumonia (BP) has been reported in Irish Wolfhounds (IWHs) and the disease typically recurs repeatedly. The aetiology is unclear but the development of bronchiectasis in the absence of clinical findings indicative of interstitial lung disease is common in affected IWHs. This observation led us to hypothesize that the primary cause could be at bronchial level. We evaluated bronchial cartilage structure and composition in IWHs (n=10) with previous episodes of pneumonia compared with IWH control dogs (n=5) and other large-breed control dogs (n=5) without a history of respiratory disease. Histological evaluation of lung sections was performed with haematoxylin and eosin, Masson's trichrome and Prussian blue staining as well as with immunohistochemistry (IHC) for CD3 and CD79a antigens and pancytokeratin. Bronchial cartilage proteoglycan content was evaluated in sections stained with safranin 'O' (SO) and chondrocyte apoptosis by IHC for cleaved caspase-3. Image analysis was used to measure the bronchial cartilage to lumen area ratio as well as the SO staining intensity and caspase-3-positive cell count in digital whole slide images. The most consistent histological finding in the lungs of IWHs with recurrent BP was chronic lymphoplasmacytic inflammation in the peribronchial and perivascular pulmonary interstitium, along with remodelling of the bronchial walls. The bronchial cartilage to lumen area ratio was significantly lower in all IWHs than in control dogs of other breeds (P=0.011), suggesting less cartilage support in the bronchi of the IWH breed. The SO staining intensity of the bronchial cartilage was significantly higher (P=0.008) in affected IWHs than in control dogs of other breeds. Analysis of caspase-3 immunolabelling of large bronchial cartilage revealed significantly fewer apoptotic chondrocytes in affected IWHs than in all control dogs (P=0.045). Based on these findings, altered cartilage metabolism, as a breed-related feature or secondary to chronic inflammation, may play a role in the pathogenesis of bronchiectasis development and recurrent lung infections in IWHs.

Left atrial function parameters in patients with a first episode of myocardial infarction: relationship with infarct size and serological inflammatory biomarkers

Abstract Aim Left atrial (LA) function is linked with clinical events in different cardiovascular scenarios. Our aim was to evaluate the associated between LA function and different imaging and serological biomarkers in a cohort of patients with a first episode of acute myocardial infarction (AMI). Methods Prospective cohort of patients with a first episode of AMI treated with percutaneous revascularization in a tertiary care hospital in 2016. All patients underwent echocardiography and cardiac magnetic resonance and blood samples were taken for serological biomarkers analysis. LA function was assessed by means of LA stiffness (E/e’ divided by LA reservoir strain) and LA volumetric-mechanical coupling index [LACI] (LA volume index divided by tissue Doppler a’). Results Mean age of patients in our cohort (n=41) was 57.5±10 years, and 38 (93%) were male. 64% of patients had single vessel coronary artery disease, and median LVEF eas 58% (55-62). Regarding LA function, median LACI was 2.47 (1.99-3.14) and median LA stiffness was 0.27 (0.21-0.44), with 11 (26.8%) patients fulfilling criteria for high LA stiffness. The correlation between these parameters and different imaging and serological biomarkers is shown in Figure 1. There was a significant correlation between LA stiffness and ventricular-arterial coupling, infarct size, and T1 native values. LACI was associated with pulmonary artery systolic pressure, interleukin-6 and soluble CD-14 antigen. Both LA parameters were also significantly associated with left ventricular (LV) ejection fraction and LV global longitudinal strain (Figure 2). Conclusions In our cohort of patients with a first episode of AMI, echocardiographic parameters of LA function were significantly associated with LV function, infarct size and serological biomarkers of inflammation. These parameters might serve as an additional tool for risk stratification in patients with ischemic cardiomyopathy. Figure 1 Figure 2

Chronic Lymphocytic Leukemia: 2025 Update on the Epidemiology, Pathogenesis, Diagnosis, and Therapy.

Chronic lymphocytic leukemia (CLL) is the most frequent type of leukemia. It typically occurs in older patients and has a highly variable clinical course. Leukemic transformation is initiated by specific genomic alterations that interfere with the regulation of proliferation and apoptosis in clonal B-cells. The diagnosis is established by blood counts, blood smears, and immunophenotyping of circulating B-lymphocytes, which identify a clonal B-cell population carrying the CD5 antigen as well as typical B-cell markers. Two clinical staging systems, Rai and Binet, provide prognostic information by using the results of physical examination and blood counts. Various biological and genetic markers provide additional prognostic information. Deletions of the short arm of chromosome 17 (del(17p)) and/or mutations of the TP53 gene predict a shorter time to progression with most targeted therapies. The CLL international prognostic index (CLL-IPI) integrates genetic, biological, and clinical variables to identify distinct risk groups of patients with CLL. The CLL-IPI retains its significance in the era of targeted agents, but the overall prognosis of CLL patients with high-risk stages has improved. Only patients with active or symptomatic disease or with advanced Binet or Rai stages require therapy. When treatment is indicated, several therapeutic options exist: combinations of the BCL2 inhibitor venetoclax with obinutuzumab, or venetoclax with ibrutinib, or monotherapy with one of the inhibitors of Bruton tyrosine kinase (BTK). At relapse, the initial treatment may be repeated if the treatment-free interval exceeds 3 years. If the leukemia relapses earlier, therapy should be changed using an alternative regimen. Combinations of targeted agents now provide efficient therapies with a fixed duration that generate deep and durable remissions. These fixed-duration therapies have gained territory in the management of CLL, as they are cost-effective, avoid the emergence of resistance, and offer treatment free time to the patient. The cure rate of these novel combination regimens is unknown. Moreover, the optimal sequencing of targeted therapies remains to be determined. A medical challenge is to treat patients who are double-refractory to both BTK and BCL2 inhibitors. These patients need to be treated within experimental protocols using novel drugs.

Case report: Tongue metastasis as an initial sign of clear cell renal cell carcinoma and its prognosis

Clear cell renal cell carcinoma (ccRCC) is the most prevalent and lethal subtype of renal cell carcinoma (RCC), characterized by a poor prognosis and a high likelihood of distant metastasis. Nonetheless, metastasis of ccRCC to the tongue remains rare. Diagnosing and planning treatment for patients who initially present with tongue metastasis can be particularly challenging, as few cases have been reported in the literature. We present a case of a 62-year-old man who presented with a painful lump on the right anterior border of his tongue. Histological examination revealed lobulated and nested epithelial cell clusters with moderate dysplasia and frequent mitotic figures within the lamina propria. Immunohistochemistry showed positivity for vimentin, CD10, PAX-8, and epithelial membrane antigen (EMA), but negativity for PAX-2, calponin, S-100 protein, periodic acid-Schiff with diastase (PAS-D), P63, P40, and CK7, confirming the diagnosis of ccRCC metastasis to the tongue. After comprehensive evaluation and multidisciplinary team consultation, the patient underwent cytoreductive nephrectomy (CN), metastasectomy, and targeted therapy. According to the Response Evaluation Criteria in Solid Tumors (RECIST) Version 1.1, the patient maintained stable disease (SD) during systemic treatment. Unfortunately, treatment was discontinued due to adverse drug reactions, and the patient was transitioned to palliative care. His disease progressed to progressive disease (PD), and he ultimately succumbed to systemic infection, with a progression-free survival (PFS) of approximately 15 months. This case highlights the urgent need for improved therapeutic strategies to manage symptoms and prolong survival in patients with this rare metastatic presentation.

The Detection of Lung Cancer Cell Profiles in Mediastinal Lymph Nodes Using a Hematological Analyzer and Flow Cytometry Method

The presence of metastases in mediastinal lymph nodes (LNs) is essential for planning lung cancer treatment and assessing anticancer immune responses. The aim of the study was to assess LNs for the presence of neoplastic cells and evaluate lung cancer-selected antigen expression. LN aspirates were obtained during an EBUS/TBNA procedure. The cells were analyzed using a hematological analyzer and flow cytometry. It was possible to indicate the presence of cells characterized by high fluorescence connected with high metabolic activity using a hematological analyzer and to determine their non-hematopoietic origin using flow cytometry. Using these methods together, we detected very quickly a high proportion of cancer cells in LNs. We noticed that it was possible to determine a high expression of EpCAM, TTF-1, Ki67, cytokeratin, HER, and differences between non-small-cell (NSCLC) and small-cell lung cancer (SCLC) for the antigens MUC-1, CD56, HLA-DR, CD39, CD184, PD-L1, PD-L2 and CTLA-4 on tumor cells. We report, for the first time, that the detection of tumor cells in LNs with the expression of specific antigens is easy to evaluate using a hematological analyzer and flow cytometry in EBUS/TBNA samples. Such precise characteristics of non-hematopoietic cells in LNs may be of great diagnostic importance in the detection of micrometastases.

Prostaglandin D2 delays CD8+ T-cell responses and respiratory syncytial virus clearance in geriatric cotton rats.

Respiratory syncytial virus (RSV) infection is associated with increased rates of severe disease, hospitalization, and death in elderly individuals. Clearance of RSV is frequently delayed within this demographic, contributing to the more severe disease course. Geriatric cotton rats mimic this prolonged clearance kinetic and serve as a useful animal model for studying age-associated immunological deficits during RSV infection. Treatment with the cyclooxygenase (COX) inhibitor ibuprofen restores RSV clearance, indicating that inflammation contributes to impaired clearance in geriatric cotton rats. Here, we further characterize a compromised immune response in geriatric cotton rats and identify an inflammatory pathway that contributes to this deficiency. Dendritic cell (DC) activation and migration to mediastinal lymph nodes are decreased during early infection in geriatric cotton rats, resulting in delayed generation of cytotoxic T cells and virus clearance. Prostaglandin D2 (PGD2), which reduces DC migration through the elevation of D-type prostanoid 1 receptor (DP1 receptor), is elevated in the airways of infected geriatric cotton rats. Reducing PGD2 production by inhibiting COX-2 or PGD2 synthase improves RSV clearance kinetics through DC activation and RSV-specific CD8+ T-cell responses in geriatric cotton rats, whereas activation of DP1 receptor through an agonist resulted in delayed viral clearance in adult cotton rats. These results indicate that PGD2 contributes to delayed antigen presentation and CD8+ T-cell responses to RSV in geriatric cotton rats. Inhibiting PGD2 generation or signaling may be a useful mechanism of therapeutic intervention in elderly individuals.IMPORTANCEElderly adults are at increased risk of severe disease resulting from infection with respiratory syncytial virus (RSV), characterized in part by delayed clearance (removal of the virus from airways). Understanding the immunological factors that lead to this delayed clearance may allow for the development of therapies to improve disease outcomes in elderly individuals infected with RSV and other respiratory viruses. Here, we describe an inflammatory pathway in geriatric cotton rats, the preferred small animal laboratory model for RSV, that impairs the generation of an effective immune response. We show that inhibiting this inflammatory pathway in geriatric cotton rats improves immune parameters and speeds clearance of RSV. These results contribute to our understanding of delayed RSV clearance in elderly individuals with possible applications for improving immune responses to RSV in clinical settings.

A Review of CAR T Cells and Adoptive T-Cell Therapies in Lymphoid and Solid Organ Malignancies.

Chimeric antigen receptor (CAR) T cells are genetically engineered T lymphocytes that express a synthetic receptor that recognizes a tumor cell surface antigen, which causes the T lymphocyte to kill the tumor cell. As of December 2024, the US Food and Drug Administration (FDA) approved six CAR T-cell therapies, with ten CAR T-cell therapies commercially available globally, which target the CD19 and B-cell maturation antigen (BCMA) molecules and with approved indications that include B-cell acute lymphoblastic leukemia (ALL), large B-cell lymphoma (LBCL), follicular lymphoma, mantle cell lymphoma, chronic lymphocytic leukemia (CLL), and multiple myeloma. Pharmaceutical and economic forecasts have shown that the global CAR T-cell therapy market was worth USD 4.6 billion in 2024, with a projected USD 25 billion by 2035. However, there are several challenges in treating hematologic malignancies with CAR T-cell therapy, which include reduced treatment efficacy and durability in some patients, acute and long-term adverse effects, lack of effective salvage treatments, limited access to CAR T-cell therapies due to cost and availability, and the rare association with developing myeloid malignancies. A tumor-infiltrating lymphocyte (TIL) therapy, lifileucel, is FDA-approved for advanced melanoma. The T-cell receptor (TCR) therapy, afamitresgene autoleucel, is FDA-approved for advanced synovial sarcoma. The results from ongoing studies and clinical trials are awaited in solid tumors (melanoma, sarcomas, and carcinomas). This article reviews recent developments and ongoing challenges in adoptive T-cell therapies, including CAR T-cell therapies, in lymphoid and solid organ malignancies.

Clinical characteristics and prognosis of acute erythroleukemia in children

To investigate the clinical characteristics and prognosis of acute erythroleukemia (AEL) in children. A retrospective analysis was conducted on the clinical data, treatment, and prognosis of 8 children with AEL treated at the First Affiliated Hospital of Zhengzhou University from January 2013 to December 2023. Among the 7 patients with complete bone marrow morphological analysis, 4 exhibited trilineage dysplasia, with a 100% incidence of erythroid dysplasia (7/7), a 71% incidence of myeloid dysplasia (5/7), and a 57% incidence of megakaryocytic dysplasia (4/7). Immunophenotyping revealed that myeloid antigens were primarily expressed as CD13, CD33, CD117, CD38, and CD123, with 4 cases expressing erythroid antigens CD71 and 2 cases expressing CD235a. Chromosomal analysis indicated that 2 cases presented with abnormal karyotypes, including +8 in one case and +4 accompanied by +6 in another; no complex karyotypes were observed. Genetic abnormalities were detected in 4 cases, with fusion genes including one case each of dup MLL positive and EVI1 positive, as well as mutations involving KRAS, NRAS, WT1, and UBTF. Seven patients received chemotherapy, with 6 achieving remission after one course of treatment; 2 underwent hematopoietic stem cell transplantation, and all had disease-free survival. Follow-up (median follow-up time of 6 months) showed that only 3 patients survived (2 cases after hematopoietic stem cell transplantation and 1 case during treatment). Children with AEL have unique clinical and biological characteristics, exhibit poor treatment response, and have a poor prognosis; however, hematopoietic stem cell transplantation may improve overall survival rates.

Imaging Flow Cytometric Identification of Chromosomal Defects in Paediatric Acute Lymphoblastic Leukaemia.

Acute lymphoblastic leukaemia is the most common childhood malignancy that remains a leading cause of death in childhood. It may be characterised by multiple known recurrent genetic aberrations that inform prognosis, the most common being hyperdiploidy and t(12;21) ETV6::RUNX1. We aimed to assess the applicability of a new imaging flow cytometry methodology that incorporates cell morphology, immunophenotype, and fluorescence in situ hybridisation (FISH) to identify aneuploidy of chromosomes 4 and 21 and the translocation ETV6::RUNX1. We evaluated this new "immuno-flowFISH" platform on 39 cases of paediatric ALL of B-lineage known to have aneuploidy of chromosomes 4 and 21 and the translocation ETV6::RUNX1. After identifying the leukaemic population based on immunophenotype (i.e., expression of CD34, CD10, and CD19 antigens), we assessed for copy numbers of loci for the centromeres of chromosomes 4 and 21 and the ETV6 and RUNX1 regions using fluorophore-labelled DNA probes in more than 1000 cells per sample. Trisomy 4 and 21, tetrasomy 21, and translocations of ETV6::RUNX1, as well as gains and losses of ETV6 and RUNX1, could all be identified based on FISH spot counts and digital imagery. There was variability in clonal makeup in individual cases, suggesting the presence of sub-clones. Copy number alterations and translocations could be detected even when the cell population comprised less than 1% of cells and included cells with a mature B-cell phenotype, i.e., CD19-positive, lacking CD34 and CD10. In this proof-of-principle study of 39 cases, this sensitive and specific semi-automated high-throughput imaging flow cytometric immuno-flowFISH method has been able to show that alterations in ploidy and ETV6::RUNX1 could be detected in the 39 cases of paediatric ALL. This imaging flow cytometric FISH method has potential applications for diagnosis and monitoring disease and marrow regeneration (i.e., distinguishing residual ALL from regenerating haematogones) following chemotherapy.

Flow Cytometric Assessment of the Influence of Cryopreservation on Vitality, Recovery and Immunophenotype of Camel Peripheral Blood Mononuclear Cells

Background: Immunophenotyping has been proven a valuable tool in diagnostic and research immunology laboratories to evaluate the immune status in health and disease. However, in some cases, cells cannot be analyzed immediately after separation and must be stored for later analysis. The present study was undertaken to investigate the impact of cryopreservation of camel PBMC on their vitality, recovery, shape, staining patterns with monoclonal antibodies and lymphocyte composition. Methods: Flow cytometry was employed to analyze cell apoptosis, necrosis, shape change, staining patterns with mAbs and cellular composition of freshly separated and cryopreserved camel PBMC. Result: The results showed similar lymphocytes and monocytes absolute numbers with comparable percentages of apoptotic and necrotic cells between fresh and preserved PBMC indicating no significant impact of cryopreservation on the recovery of camel PBMC. Although a cryopreservation-induced shape change was identified for camel lymphocytes, this change did not interfere with the identification of cryopreserved lymphocytes based on their SSC and FSC characteristics. In addition, the reactivity of camel PBMC with mAbs to the cell markers antigens CD45, CD44, CD11a, MHC-I, CD14, CD163 and CD172a remained unchanged for cryopreserved cells. However, mAbs to MHC-II and BAQ44A displayed modified binding to cryopreserved PBMC leading to significant changes in the positively stained B cells and increased MHC-II expression on monocytes. In conclusion, the results of the current study show that storing camel PBMC in cryopreservation media at -80 is an appropriate procedure for preserving the epitopes for all the used mAbs with modified binding to B cell markers.

Incidence of Acute Kidney Injury in Relapsed and Refractory Multiple Myeloma treated with Teclistamab versus CAR T-cells.

Teclistamab, a novel bispecific monoclonal antibody targeting CD3 and B-cell maturation antigen (BCMA), and chimeric antigen receptor T-cell (CAR-T) therapy are promising options for treating relapsed/refractory multiple myeloma (MM). However, the rates of acute kidney injury (AKI) associated with teclistamab remain inadequately characterized. This study aims to compare the incidence, severity, and outcomes of AKI between patients receiving teclistamab and CAR-T therapy. This was a retrospective study involving 64 patients with relapsed/refractory MM treated with either teclistamab or CAR-T therapy. All patients had previously received at least four lines of chemotherapy before being treated with either teclistamab or CAR-T. The primary outcome was the incidence of AKI, and secondary outcomes included AKI severity, kidney recovery rates, and mortality. Kaplan-Meier estimates for AKI-free survival were calculated, and hazard ratios (HRs) for AKI risk were determined using Cox proportional hazards models. Sixty-four patients met inclusion criteria for this study (30 received CAR-T and 34 received teclistamab therapy). Among these patients, 14 AKI events occurred in total (22%), with 10 events (29%) in the teclistamab group and 4 events (13%) in the CAR-T group. AKI-free survival estimates at 180 days after treatment initiation were 68% (95% confidence interval [CI]: 53%-87%) for teclistamab patients and 90% (95% CI: 79%-100%) for CAR-T patients. While patients receiving teclistamab were found to have an increased risk of an AKI event compared to those receiving CAR-T therapy, the results were not statistically significant (HR [95% CI]: 3.38 [0.93-12.31], P=0.065). This study suggests that patients treated with teclistamab may experience a higher incidence of AKI compared to those receiving CAR-T therapy. However, further research is required to determine whether this increased risk is attributable to disease progression or teclistamab itself. These results highlight the need for close kidney function monitoring in patients receiving teclistamab.

MicroRNA-21 as a Regulator of Cancer Stem Cell Properties in Oral Cancer.

Oral squamous cell carcinoma (OSCC) is a highly aggressive malignancy with poor prognosis, mainly due to the presence of cancer stem cells (CSCs), a small subpopulation of cells that contribute to therapy resistance and tumor progression. The principal objective of this study was to investigate the role of miRNA-21 in the maintenance of cancer cell stemness and the possibility of altering it. The CD44 antigen was used as a marker for CSC isolation from oral cancer cell cultures. CD44+ and CD44- populations were sorted via magnetic separation. miRNA-21 inhibition was performed in CD44+ cells via transfection. CD44+ cells possessed a significantly higher migration and invasion potential compared to CD44- cells, higher levels of miRNA-21 (p = 0.004) and β-catenin (p = 0.005), and lower levels of BAX (p = 0.015). miRNA-21 inhibition in CD44+ cells reduced migration, invasion, and colony formation while increasing apoptosis. Stemness markers were significantly downregulated following miRNA-21 inhibition: OCT4 (p = 0.013), SOX2 (p = 0.008), and NANOG (p = 0.0001), as well as β-catenin gene (CTNNB1) (p < 0.05), an important member of WNT signaling pathway. Apoptotic activity was enhanced, with a significant downregulation of the antiapoptotic Bcl-2 (p = 0.008) gene. In conclusion, miRNA-21 plays a critical role in the regulation of oral cancer CD44+ cells properties. Targeting and inhibiting miRNA-21 in CD44+ cells could represent a promising novel strategy in OSCC treatment.

Efficacy and Safety of CART Cell Therapy in Aggressive B-Cell Lymphomas Involving the Gastrointestinal Tract.

Currently, chimeric antigen receptor T-cell (CART) therapy represents a highly effective approach for relapsed/refractory B-cell lymphomas. However, it also carries treatment-related risks. Limited data are available on the risks associated with CART therapy in patients with gastrointestinal involvement in B-cell lymphomas. Therefore, we conducted a retrospective cohort study to address this gap in knowledge. During the period from May 2019 to August 2022, a total of 26 patients recurrent/refractory with recurrent/refractory B-cell lymphoma involving the gastrointestinal tract enrolled. Pathology confirmed CD19 antigen expression in tumor tissues. The disease status of patients who failed multiple lines of therapy was progressive disease (PD). Before CART cell infusion, patients received an FC regimen (fludarabine and cyclophosphamide) lymphodepletion. Quantitative PCR and flow cytometry were adopted for monitoring CART cell kinetics and function, with a focus on gastrointestinal AEs during treatment. The overall response rate (ORR) of the 26 patients was 61.5% (16/26), while the complete response rate (CR) was 23.1% (6/26). Their median follow-up time was 22.49 months, while the medians of overall survival (OS) and progression-free survival (PFS) were 10.88 and 5.47 months, respectively. The 1-year OS and PFS rates were 45% and 42.3%, respectively. The prevalence of gastrointestinal complications was 21/26 (80.7%), including gastrointestinal hemorrhage in 11/26 (42.3%), emesis and diarrhea in 9/26 (34.6%), as well as intestinal obstruction in 2/26 (7.7%). A total of three patients (3/26, 11.5%) died of gastrointestinal hemorrhage. The gastrointestinal hemorrhage group exhibited markedly lower ORR and inferior OS compared to the non-hemorrhage group. Generally, the CART cell therapy is valid in relapsed/refractory B-cell lymphoma with gastrointestinal involvement, but gastrointestinal bleeding is a unique risk factor that requires special attention, particularly in patients with high gastrointestinal tumor burden, as it is associated with poor efficacy and survival.

Overcoming CD226-related immune evasion in acute myeloid leukemia with CD38 CAR-engineered NK cells.

CD226 plays a vital role in natural killer (NK) cell cytotoxicity, interacting with its ligands CD112 and CD155 to initiate immune synapse formation, primarily through leukocyte function-associated-1 (LFA-1). Our study examined the role of CD226 in NK cell surveillance of acute myeloid leukemia (AML). NK cells in patients with AML had lower expression of CD226. CRISPR-Cas9 deletion of CD226 led to reduced LFA-1 recruitment, poor synapse formation, and decreased NK cell anti-leukemic activity. Engineering NK cells to express a chimeric antigen receptor targeting the AML antigen CD38 (CAR38) could overcome the need for CD226 to establish strong immune synapses. LFA-1 blockade reduced CAR38 NK cell activity, and this depended on the CD38 expression levels of AML cells. This suggests parallel but potentially cooperative roles for LFA-1 and CAR38 in synapse formation. Our findings suggest that CAR38 NK cells could be an effective therapeutic strategy to overcome CD226-mediated immune evasion in AML.

Integration of CD200, CD43 and ROR1 in Multiparameter Flow Cytometry (MFC) Routine Panels for the Differential Diagnosis of B-cell lymphoproliferative Disorders (B-LPDs).

Clonal mature B-cell lymphoproliferative disorders (B-LPDs) are a heterogeneous group of neoplasia characterized by the proliferation of mature B lymphocytes in the peripheral blood, bone marrow and/or lymphoid tissues. B-LPDs classification into different subtypes and their diagnosis is based on a multiparametric approach. However, accurate diagnosis may be challenging, especially in cases of ambiguous interpretation. Multiparameter flow cytometry (MFC) represents an extensively used technique to detect the presence of different cellular lines in immunology and hematology. MFC results provide an essential contribution to the B-LPDs diagnostic process, even more so considering that panels are constantly integrating novel markers to improve diagnostic accuracy. The aim was to evaluate the contributing role of MFC routinary studies by analyzing the expression and the mean fluorescence intensity (MFI) of CD200, ROR1, and CD43 in various B-LPDs to evaluate their usefulness in the differential diagnosis of these diseases. We retrospectively evaluated 2615 consecutive cases of newly collected samples (mostly from patients with lymphocytosis) analyzed by MFC carried out in the B-LPD diagnostic process referred to the Division of Hematology of the Sapienza University of Rome. We compared the results of CD200, ROR1, and CD43 expression percentage and their MFI between different subtypes of B-LPDs. In chronic lymphocytic leukemia (CLL), CD200, ROR1, and CD43 were always expressed with bright intensity. CLL samples presented high CD200 expression and MFI [CD200%, mean: 100 (range, 24-100); positivity rate: 100%; MFI, median = 125 (range, 10-1200)] statistically higher than mantle cell lymphoma (MCL) (p<0.001), which is usually negative for CD200, and variant hairy cell leukemia (vHCL, according to 2022 ICC) (p<0.001), but comparable with classic HCL (cHCL) (p>0.9). ROR1 resulted expressed in all CLL [ROR1%, mean: 100 (range, 52-100), positivity rate: 100%; MFI, median=50 (range, 10-202)] and MCL cases with comparable MFI (p>0.9). CD43 expression and MFI were significantly higher in CLL [CD43%, mean 99 (range, 59-100); positivity rate: 100%; MFI, median = 130 (range, 41-980)] than in MCL, vHCL, cHCL, and all the others mature B-cell neoplasia (p<0.001). CD200 and CD43 expression and MFI were significantly higher in cHCL compared to vHCL. Among the other mature B-cell neoplasia, CD200 was variably expressed in follicular lymphoma (FL), marginal zone lymphoma (MZL), diffuse large B-cell lymphoma (DLBCL), and lymphoplasmacytic lymphoma (LPL). ROR1 and CD43 presented a very low expression percentage in this latter group, being mostly negative. Persistent polyclonal B-cell lymphocytosis (PPBL) resulted in uniformly positive for CD200 and negative for ROR1 and CD43. Our data suggest that evaluating CD200, ROR1, and CD43 antigens and their intensity of expression, along with commonly used markers in MFC routine panels for B-LPDs, might be extremely useful for prompt diagnostic evaluation in the differential diagnosis of these diseases.

The Expected Medical Uses of Saudi Arabia Coffee Constituents Through Computer-Aided Drug Design

Background:-In this research work, we have investigated the molecular structure of 17 Arabian coffee flavonoids to identify their potential protein targets using Swiss Target Prediction platform. The obtained results indicated that aldose reductase, carbonic anhydrase II and lymphocyte differentiation antigen CD38 are the primary targets for the tested flavonoids. Methods:-Absorption, distribution, metabolism and excretion predictions were also performed to explore the pharmacokinetic profile of the Arabian coffee flavonoid content. Molecular docking investigations were also performed with aldose reductase, carbonic anhydrase II and lymphocyte differentiation antigen CD38. Results:-It has shown docking scores between -13.07 and -20.20 kcal/mol for aldose reductase, -11.76 to -18.27 kcal/mol for carbonic anhydrase II, and -10.65 to -19.44 kcal/mol for lymphocyte differentiation antigen CD38. Metabolic pathway analysis for Arabian coffee flavonoids predicted potential metabolites of epigallocatechin gallate, delphinidin 3-(6"-malonyl-glucoside), cyanidin-3-O-glucoside and isoquercitrin. Conclusion:-Arabian coffee flavonoids have the potential to be used in the development of medical treatments targeting proteins such as aldose reductase, carbonic anhydrase II and lymphocyte differentiation antigen CD38.

New line of mesenchymal stem cell isolated from warton’s jelly of the umbical cord of male human donor

A new non-immortalized fibroblast-like cell line, named MSCWJ-3, was generated and characterized. Characteristics during long-term cultivation (6–24 passages) confirm the status of MSCs. It is shown: 1) a gradual increase in the proportion of senescent cells during long-term cultivation; 2) a significant decrease in the proliferation index by the 24th passage; 3) preservation of the normal diploid karyotype of the man (46, XY) during the entire period of cultivation, trisomy for different autosomes in single cells, absence of structural chromosomal rearrangements; 4) a high proportion of cells carrying surface antigens characteristic of MSCs: CD44, CD73, CD90, CD105, HLA-ABC and a low proportion with antigens CD34, CD45 and HLA-DR over 24 passages. Cells of the MSCWJ-3 line are capable of differentiation in the osteogenic and adipogenic directions at early and late passages; differentiation in the chondrogenic direction is absent. In general, there are some differences with previously obtained lines isolated from the same source and are associated mainly with the degree of expression of a number of status characteristics.

The effect of rituximab therapy and the risk of hypogammaglobulinamea on immunoglobulin levels in patients with rheumatoid arthritis

Background. Rituximab (RTX) is a biological monoclonal antibody targeting the CD20 antigen on B cells, effectively decreasing disease activity and progression in patients with rheumatoid arthritis (RA). RTX treatment can lead to a decline in plasma cells that produce immunoglobulin, potentially resulting in hypogammaglobulinemia (HGG). This study aims to assess the effect of RTX on immunoglobulin levels and determine the prevalence of HGG in RA patients after receiving different doses and cycles of RTX. Additionally, immunoglobulin levels were compared between patients treated with RTX and those treated with traditional disease-modifying antirheumatic drugs (DMARDs). We also sought to identify patient, disease, and therapy factors associated with HGG to evaluate the importance of monitoring immunoglobulin levels. Method. A single-center observational cross-sectional study was conducted at Baghdad Teaching Hospital between November 2023 and May 2024. Patients with RA were recruited and divided into two groups. The first group consisted of 45 RA patients receiving RTX, and the second group included 45 patients receiving DMARDs. All patients met the American College of Rheumatology (ACR) 1987 or ACR/European League Against Rheumatism (EULAR) 2010 classification criteria for RA. Patients were excluded if they had received RTX for conditions other than RA or if they had been treated with RTX for less than six months. Quantitative IgG and IgA levels were measured via enzyme-linked immunosorbent assay (ELISA) to evaluate the effect of RTX on immunoglobulin levels and compare it with the effect of DMARDs. HGG was defined as an immunoglobulin level below two standard deviations from the mean of age-matched healthy controls. Results. Ninety patients were enrolled in the study, 45 of whom were treated with RTX. The mean number of RTX cycles was 3.67, ranging from 2 to 10 cycles. A significant decrease in immunoglobulin levels was observed post-RTX therapy, with 84.4% of patients exhibiting low IgG levels (&lt;5 μg/ml), leading to HGG. The IgG levels in RTX-treated patients ranged from 4.3 to 22.0 μg/ml, with a mean of 7.28 ± 3.95 μg/ml. In contrast, patients treated with DMARDs maintained normal immunoglobulin levels similar to healthy individuals, with IgG levels ranging from 19.7 to 43.7 μg/ml. RTX also affected IgA levels, but to a lesser extent than IgG. Conclusion. Lower immunoglobulin levels following RTX therapy frequently result in HGG, emphasizing the importance of immunologic surveillance before and after treatment. DMARD therapy did not affect immunoglobulin levels, and concurrent use of DMARDs with RTX offered no protection against HGG. Additionally, HGG was not associated with an increased incidence of infections.

Potassium/sodium cation carriers robustly up-regulate CD20 antigen by targeting MYC, and synergize with anti-CD20 immunotherapies to eliminate malignant B cells.

Our investigation uncovers that nanomolar concentrations of salinomycin, monensin, nigericin, and narasin (a group of potassium/sodium cation carriers) robustly enhance surface expression of CD20 antigen in B-cell-derived tumor cells, including primary malignant cells of chronic lymphocytic leukemia and diffuse large B-cell lymphoma. Experiments in vitro, ex vivo, and animal model reveal a novel approach of combining salinomycin or monensin with therapeutic anti-CD20 monoclonal antibodies or anti-CD20 CAR-T cells, significantly improving non- Hodgkin lymphoma (NHL) therapy. The results of RNA-seq, genetic editing, and chemical inhibition delineate the molecular mechanism of CD20 upregulation, at least partially, to the downregulation of MYC, the transcriptional repressor of the MS4A1 gene encoding CD20. Our findings propose the cation carriers as compounds targeting MYC oncogene, which can be combined with anti-CD20 antibodies or adoptive cellular therapies to treat NHL and mitigate resistance, which frequently depends on the CD20 antigen loss, offering new solutions to improve patient outcomes.