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The role of extracellular vesicles (EVs) proteome in diffuse large B-cell lymphoma (DLBCL) pathology, subclassification, and patient screening is unexplored. We analyzed by state-of-the-art mass spectrometry the whole cell and secreted extracellular vesicles (EVs) proteomes of different molecular subtypes of DLBCL, germinal center B cell (GCB subtype), and activated B cell (ABC subtype). After quality control assessment, we compared whole-cell and secreted EVs proteomes of the two cell-of-origin (COO) categories, GCB and ABC subtypes, resulting in 288/1115 significantly differential expressed proteins from the whole-cell proteome and 228/608 proteins from EVs (adjust p-value < 0.05/p-value < 0.05). In our preclinical model system, we demonstrated that the EV proteome and the whole-cell proteome possess the capacity to separate cell lines into ABC and GCB subtypes. KEGG functional analysis and GO enrichment analysis for cellular component, molecular function, and biological process of differential expressed proteins (DEP) between ABC and GCB EVs showed a significant enrichment of pathways involved in immune response function. Other enriched functional categories for DEPs constitute cellular signaling and intracellular trafficking such as B-cell receptor (BCR), Fc_gamma R-mediated phagocytosis, ErbB signaling, and endocytosis. Our results suggest EVs can be explored as a tool for patient diagnosis, follow-up, and disease monitoring. Finally, this study proposes novel drug targets based on highly expressed proteins, for which antitumor drugs are available suggesting potential combinatorial therapies for aggressive forms of DLBCL. Data are available via ProteomeXchange with identifier PXD028267.

Introduction: 3-(4,5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide (MTT) assay is a safe, convenient, and low-cost technique with high applications for the measurement of cell proliferation rate in researches and clinical laboratories. Our aim was to evaluate the proliferation rate of human peripheral blood mononuclear cells (PBMCs) and production rate of Tumor necrosis factor alpha (TNF-α) by these cells after various mitogens stimulation in different situations. Methods: The MTT test was performed with various concentrations of mitogens including concanavalin A (ConA), lipopolysaccharide (LPS) and phytohemagglutinin (PHA) on the PBMCs. The cells were incubated for 24, 48, 72, and 96 hours in the culture medium and TNF-α cytokine assay was performed on the supernatant of the cultured splenocytes using the enzyme-linked immunosorbent assay (ELISA) method. Results: The optimal time and incubation of the PBMCs with the mixture of PHA-ConA were 5 μg/mL and 72 hours, respectively. The TNF-α level increased significantly after PHA-ConA and PHA stimulation. Conclusion:The results showed that the mixture of PHA-ConA (at the concentration of 5 μg/ mL) can give rise to the optimal results on stimulation of the PBMcs using the MTT assay after 72 hours incubation.

Infectious diseases are common life-threatening problems mediated by pathogen micro-organisms that cause morbidity and mortality worldwide. Currently, there is an increasing rate of the bacterial infections and emergence of the new antibiotic resistance in human societies. On the other hand, early detection of the bacterial infection present in biological samples suffers from extended time, high cost, and laborious methods. Therefore, there is a permanent need for robust diagnostic and therapeutic tools against bacterial agents. Recently, specific targeting bio-molecules, such as aptamer and nanobody have been appeared as specific and effective tools for biomedical application. They have excellent physicochemical parameters that make them superior to diagnosis and treatment of infectious agents achievable from diverse large libraries through systematic evolution of ligands by exponential enrichment (SELEX) or phage display process, respectively. The present study provides an overview of nanobody and aptamer and their method description. Main contexts of article focus on the application of nanobody and aptamer as an inhibiting moiety for some bacterial toxins.

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