Journal: Molecular Therapy Oncology

Article Title: Point-of-care manufacturing of anti-CD19 CAR-T cells using a closed production platform: Experiences of an academic in Thailand

doi: 10.1016/j.omton.2024.200889

Figure Lengend Snippet: Functionality of the final cell product (A) Cytotoxicity of the manufactured SiCF-019 (effector) cells against CD19 + and CD19 − tumor (target) cells. (Top left) Flow cytometric gating strategy for specifically detecting the cell death of PKH67-labeled tumor cells after incubation with SiCF-019 cells by annexin V/7-AAD assay. Percentages of total cell death of K562, Z-138, and REH cells, comprising annexin V + and/or 7-AAD + cells, at various E:T ratios at 4 h are plotted. ∗ p < 0.05; ∗∗ p < 0.01; ∗∗∗ p < 0.001 versus basal control (without SiCF-019); # p < 0.05; ## p < 0.01; ### p < 0.001 versus indicated groups; one-way ANOVA with Tukey’s multiple comparison test. The culture conditions are also labeled. (B) Quantitative measurement of TNF-⍺ and IFN-γ by ELISA in cell-free supernatant collected from the coexposure of untransduced T cells or SiCF-019 cells with tumor cells at various E:T ratios at 4 h. ∗ p < 0.05; ∗∗ p < 0.01; ∗∗∗ p < 0.001 versus indicated groups; one-way ANOVA with Tukey’s multiple comparison test. NS, not significant.

Article Snippet: Cytotoxicity assay of the manufactured SiCF-019 cells against various target tumor cells, including CD19 + human ALL-derived REH cells (American Type Culture Collection [ATCC], Manassas, VA), CD19 + human mantle cell lymphoma-derived Z-138 cells (ATCC), and CD19 − human chronic myeloid leukemia-derived K562 cells (Japanese Collection of Research Bioresources Cell Bank, Osaka, Japan), was performed.

Techniques: Labeling, Incubation, Control, Comparison, Enzyme-linked Immunosorbent Assay

Journal: Cell Host & Microbe

Article Title: Discovery of ultrapotent broadly neutralizing antibodies from SARS-CoV-2 elite neutralizers

doi: 10.1016/j.chom.2021.12.010

Figure Lengend Snippet:

Article Snippet: CD19 MicroBeads, human , Miltenyi Biotec , Catalog# 130-050-301.

Techniques: Purification, Bioprocessing, Virus, Variant Assay, Clinical Proteomics, Recombinant, Transfection, Expressing, Random Hexamer, Blocking Assay, Lysis, Staining, Enzyme-linked Immunosorbent Assay, Mutagenesis, Antibody Labeling, Microscopy, Software, Electron Microscopy

Journal: bioRxiv

Article Title: Elucidating genes sufficient for viral entry into cells through sequential genome-wide CRISPR activation screens

doi: 10.64898/2026.03.06.710083

Figure Lengend Snippet: (A) Summary of gene deletion vs. gene overexpression approaches. (B) Rationale for the present study. (C) Experimental outline of the present study. (D) HEK293 and Jurkat cells were inoculated with different volumes of Ebola or rabies pseudovirus encoding a cell-surface marker (mCD19t; a truncated mutant of mouse CD19), followed by flow cytometry to determine the percentage of infected cells. This revealed that Jurkat cells are largely refractory to Ebola or rabies pseudovirus entry, relative to HEK293 cells. (E) Construction of a clonal Jurkat cell line, known as “Jurkat C6”, stably expressing a degron-tagged CRISPRa construct ( left ). Jurkat C6 cells were transduced with sgRNA targeting the endogenous human CD19 gene, in the presence or absence of TMP (1 μM) for 3 days, followed by flow cytometry to detect hCD19 expression ( right ). (F) At each of the indicated points of the genome-wide CRISPRa screen, the cell population was challenged with Ebola or rabies pseudovirus, and cell infectivity was evaluated by flow cytometry. Upon successive rounds of the screen, the cell population became progressively more susceptible to infection to either Ebola or rabies pseudovirus, respectively. (G) sgRNA distribution upon successive rounds of the genome-wide CRISPRa screen for Ebola pseudovirus entry. (H) sgRNA distribution upon successive rounds of the genome-wide CRISPRa screen for rabies pseudovirus entry.

Article Snippet: Cells were infected with pseudotyped lentiviruses, and 72 hours later, were harvested and stained with antibodies against the relevant surface marker: mouse CD19 (Miltenyi Biotec, 130-111-884), mouse H2Kk (Miltenyi Biotec, 130-117-235), or human EGFR (R&D Systems, FAB9577R-100), and viability staining was performed using DAPI (Thermo Fisher Scientific, D1306).

Techniques: Over Expression, Marker, Mutagenesis, Flow Cytometry, Infection, Stable Transfection, Expressing, Construct, Transduction, Genome Wide

Journal: bioRxiv

Article Title: Elucidating genes sufficient for viral entry into cells through sequential genome-wide CRISPR activation screens

doi: 10.64898/2026.03.06.710083

Figure Lengend Snippet: A) Plasmids used to pseudotype non-replicating lentiviruses with either Ebola or rabies envelope proteins. EBOV-GP: glycoprotein of Ebola virus, Makona variant. RABV-GP N2C: glycoprotein of rabies virus, N2C variant. B) HEK293 and Jurkat cells were inoculated with different volumes of VSV envelope protein-pseudotyped lentivirus encoding a cell-surface marker (mCD19t; a truncated mutant of mouse CD19), followed by flow cytometry to determine the percentage of infected cells. This served as a positive control to confirm that Jurkat cells and HEK293 cells are both susceptible to VSV pseudovirus entry. C) Jurkat C6 cells were transduced with sgRNA targeting the endogenous human CD19 gene, in the presence of different TMP concentrations (0-4 μM) for 2-3 days, followed by flow cytometry to detect human CD19 expression.

Article Snippet: Cells were infected with pseudotyped lentiviruses, and 72 hours later, were harvested and stained with antibodies against the relevant surface marker: mouse CD19 (Miltenyi Biotec, 130-111-884), mouse H2Kk (Miltenyi Biotec, 130-117-235), or human EGFR (R&D Systems, FAB9577R-100), and viability staining was performed using DAPI (Thermo Fisher Scientific, D1306).

Techniques: Virus, Variant Assay, Marker, Mutagenesis, Flow Cytometry, Infection, Positive Control, Transduction, Expressing

Journal: Cell reports

Article Title: Clonal Hematopoiesis Before, During, and After Human Spaceflight

doi: 10.1016/j.celrep.2020.108458

Figure Lengend Snippet: (A) Longitudinal trajectories of CH mutations discovered in the astronaut subjects. Allele frequency (%) is the VAF of each clone (vertical axis) visualized across time (horizontal axis). Subject TW (spaceflight) is compared to subject HR (ground). Flight start and end for the NASA Twins Study are denoted by the gray shaded region (left). Baseline (2015) time point corresponds to lymphocyte-depleted MNC fraction. Inflight time points and after flight 2016 time points were collected and processed by density gradient to obtain peripheral blood mononuclear cells, and the 2019 time point corresponds to buffy coat, spun-down blood cells. (B) VAF in distinct cell populations. The three significant mutations (rows) in DNMT3A and LPL for HR (top) and the two mutations found in subject TW (bottom). Shown mutations are present in bulk cell populations (bulk), as well as almost all subfractions of T cells (CD4, CD8) and B cells (CD19). VAF range is show in fraction of color (red to blue, up to 0.1), and asterisks (*) indicate detection of the variant but below 1%. Triangles indicate the beginning and end of the year-long mission.

Article Snippet: CD19 , Miltenyi Biotec , Cat# 130-097-055.

Techniques: Variant Assay

Journal: Cell reports

Article Title: Clonal Hematopoiesis Before, During, and After Human Spaceflight

doi: 10.1016/j.celrep.2020.108458

Figure Lengend Snippet:

Article Snippet: CD19 , Miltenyi Biotec , Cat# 130-097-055.

Techniques: Hybridization, Binding Assay, Purification, RNA Sequencing, Sequencing, Mutagenesis, Software, Variant Assay

Journal: Cancers

Article Title: Advancing CAR T-Cell Therapy for Solid Tumors: Lessons Learned from Lymphoma Treatment

doi: 10.3390/cancers12010125

Figure Lengend Snippet: Recent achievements in the field of CAR T-cell therapy against solid tumors. CNS—central nervous system; CR—complete response; PSCA—prostate stem cell antigen; MPM—malignant pleural mesothelioma; PR—partial remission; SD—stable disease; LD—lymphodepletion; PD—progressive disease; PD-1—programmed cell death protein 1; OS—overall survival; TCR—T-cell receptor; EGFRvIII—epidermal growth factor receptor variant III; HER2—human epidermal growth factor receptor 2; GD2—disialoganglioside.

Article Snippet: FDA-approved anti-CD19 Yescarta ® and Kymriah ® are produced from the whole population of peripheral blood mononuclear cells (PBMCs) without separation of specific T-cell subtypes [ , ].

Techniques: Variant Assay

Journal: Cancers

Article Title: Advancing CAR T-Cell Therapy for Solid Tumors: Lessons Learned from Lymphoma Treatment

doi: 10.3390/cancers12010125

Figure Lengend Snippet: CAR T-cell therapies for the treatment of hematological cancers. ALL—acute lymphoblastic leukemia; AML—acute myeloid leukemia; BCMA—B-cell maturation antigen; MM—multiple myeloma; NHL—В-cell non-Hodgkin’s lymphoma; PFS—progression-free survival.

Article Snippet: FDA-approved anti-CD19 Yescarta ® and Kymriah ® are produced from the whole population of peripheral blood mononuclear cells (PBMCs) without separation of specific T-cell subtypes [ , ].

Techniques:

Journal: Cancers

Article Title: Advancing CAR T-Cell Therapy for Solid Tumors: Lessons Learned from Lymphoma Treatment

doi: 10.3390/cancers12010125

Figure Lengend Snippet: The highlights of experience from treatment of lymphoma with CAR T-cells. Clinical administration of anti-CD19 CAR T-cells resulted in accumulation of vast experience specifying potential predictors of therapeutic response. Some of them are presented in this figure. ECM—extracellular matrix.

Article Snippet: FDA-approved anti-CD19 Yescarta ® and Kymriah ® are produced from the whole population of peripheral blood mononuclear cells (PBMCs) without separation of specific T-cell subtypes [ , ].

Techniques: