Journal: Cell Reports Methods

Article Title: Generation, expansion, gene delivery, and single-cell profiling in rhesus macaque plasma B cells

doi: 10.1016/j.crmeth.2024.100878

Figure Lengend Snippet:

Article Snippet: anti-CD38 FITC clone: OKT10 , Caprico Biotechnologies , 100815.

Techniques: Enzyme-linked Immunospot, Isolation, Enzyme-linked Immunosorbent Assay, Recombinant, Software

Journal: iScience

Article Title: CD38 is a key mediator of NAD + depletion in the brain of ZIKV-infected mice

doi: 10.1016/j.isci.2025.114018

Figure Lengend Snippet: CD38 expression and activity increase at late stages of ZIKV infection and correlate with NAD + decline in the brain (A) Total NADase activity measured in the brains of ZIKV-infected and mock-injected mice over the course of infection, showing significant increases from 18 dpi onward ( n ≥ 7 mice per group). (B) Overlay of NADase activity (red line) and NAD + levels (gray line), both relative to mock controls (dashed line), showing an inverse temporal association. (C–H) Relative mRNA expression of Sarm1 , Cd157 , and Cd38 , respectively, in the brains of ZIKV-infected and control mice ( n ≥ 4 mice per group). (D, F, H) Overlays of mRNA expression profiles of Sarm1 (D), Cd157 (F), and Cd38 (H) with NAD + levels (gray line) and ZIKV genomic RNA (yellow line), indicating that Cd38 induction temporally coincides with NAD + decline, while Sarm1 and Cd157 do not. (I) Linear regression shows a positive correlation between total NADase activity and Cd38 mRNA expression ( n = 50). (J) CD38-dependent NADase activity, calculated as the fraction inhibited by the specific CD38 inhibitor 78c ( n ≥ 6 mice per group). (K) CD38-independent NADase activity, which remains low and unchanged during infection ( n ≥ 6 mice per group). (L) On the right, representative Western blot of CD38 protein expression in brain extracts at 24 dpi, with α-tubulin as loading control (representative bands from the same experiment shown in the full blot in ). On the left, quantification of the Western blot bands’ intensities (CD38/α-tubulin) relative to mock ( n ≥ 4 mice per group). Data in panels A, C, E, G, J, K, and L are presented as mean ± SD; panels B, D, F, and H as mean ± SEM (shaded area). Statistical significance was determined by unpaired Student’s t test or Mann–Whitney test, as appropriate. ∗ p ≤ 0.05; ∗∗ p ≤ 0.01; ∗∗∗ p ≤ 0.001.

Article Snippet: Proteins were separated by electrophoresis on 15% SDS–polyacrylamide gels (SDS-PAGE), transferred to nitrocellulose membranes (Bio-Rad, California, USA), and probed with primary antibodies against CD38 (AF4947, R&D Systems, Minnesota, USA) or NAMPT (AG-20A-0034-C050, AdipoGen Life Sciences, California, USA), using concentrations recommended by the manufacturers and previously validated in-house.

Techniques: Expressing, Activity Assay, Infection, Injection, Control, Western Blot, MANN-WHITNEY

Journal: iScience

Article Title: CD38 is a key mediator of NAD + depletion in the brain of ZIKV-infected mice

doi: 10.1016/j.isci.2025.114018

Figure Lengend Snippet: CD38 inhibition prevents NAD + depletion in the brains of ZIKV-infected mice (A) Schematic representation of the experimental design. Neonatal mice were subcutaneously infected with ZIKV at postnatal day 3 (P3). At 21 days post-infection (dpi), animals received a unilateral intracerebroventricular (i.c.v.) injection of the CD38-blocking antibody Ab68 (5.76 μg) or vehicle (Veh), and brains were collected at 24 dpi for analysis. (B) NAD + hydrolase activity in brain tissue. (C) Quantification of total NAD + levels in brain tissue. Data are presented as mean ± SD. Statistical analyses were performed using an unpaired Student’s t test ( n ≥ 7 mice per group). ∗p ≤ 0.05; ∗∗∗∗p ≤ 0.0001.

Article Snippet: Proteins were separated by electrophoresis on 15% SDS–polyacrylamide gels (SDS-PAGE), transferred to nitrocellulose membranes (Bio-Rad, California, USA), and probed with primary antibodies against CD38 (AF4947, R&D Systems, Minnesota, USA) or NAMPT (AG-20A-0034-C050, AdipoGen Life Sciences, California, USA), using concentrations recommended by the manufacturers and previously validated in-house.

Techniques: Inhibition, Infection, Injection, Blocking Assay, Activity Assay

Journal: iScience

Article Title: CD38 is a key mediator of NAD + depletion in the brain of ZIKV-infected mice

doi: 10.1016/j.isci.2025.114018

Figure Lengend Snippet: NAMPT is induced in the brain of ZIKV-infected mice (A) Relative mRNA expression of Nampt in the brains of ZIKV-infected and mock-injected mice across time points post-infection ( n ≥ 4 mice per group). (B) Overlay of Nampt mRNA expression (blue line), total NADase activity (red line), NAD + levels (gray line), and ZIKV genomic RNA (yellow line), all relative to mock controls (dashed line). (C–F) Correlation analyses between Nampt mRNA expression and (C) ZIKV genomic RNA, (D) Parp12 , (E) Parp10 , and (F) Cd38 mRNA expression. Nampt shows a strong correlation with viral load and early-induced Parps , but a weak correlation with Cd38 . (G) Representative Western blot of NAMPT protein expression in the brains of ZIKV-infected and mock-injected mice at 24 days post-infection (dpi; representative bands from the same experiment shown in the full blot in ).. HPRT was used as a loading control. The right panel shows quantification of NAMPT protein levels (NAMPT/HPRT ratio), normalized to mock controls ( n ≥ 4 mice per group). Data in panels A and G are presented as mean ± SD; panel B as mean ± SEM (shaded area). Correlations were assessed by linear regression analysis. Statistical analyses were performed using an unpaired Student’s t test or Mann–Whitney test, as appropriate. ∗ p ≤ 0.05; ∗∗ p ≤ 0.01; ∗∗∗ p ≤ 0.001.

Article Snippet: Proteins were separated by electrophoresis on 15% SDS–polyacrylamide gels (SDS-PAGE), transferred to nitrocellulose membranes (Bio-Rad, California, USA), and probed with primary antibodies against CD38 (AF4947, R&D Systems, Minnesota, USA) or NAMPT (AG-20A-0034-C050, AdipoGen Life Sciences, California, USA), using concentrations recommended by the manufacturers and previously validated in-house.

Techniques: Infection, Expressing, Injection, Activity Assay, Western Blot, Control, MANN-WHITNEY

Journal: iScience

Article Title: CD38 is a key mediator of NAD + depletion in the brain of ZIKV-infected mice

doi: 10.1016/j.isci.2025.114018

Figure Lengend Snippet: Proinflammatory cytokine expression precedes CD38 induction in the brains of ZIKV-infected mice (A, C, E) Relative mRNA expression of Il6 and Tnf (linear scale), and Ccl5/Rantes (Log 10 -transformed) in the brains of ZIKV-infected and mock-injected mice across time points post-infection ( n ≥ 3 mice per group). All three inflammatory mediators were significantly upregulated during the early and mid-stages of infection. (B, D, F) Overlay of Il6 (B), Tnf (D), and Rantes – log 10 scale (F) mRNA expression profiles (purple line) with Cd38 mRNA (red line), NAD + levels (gray line), and ZIKV genomic RNA (yellow line), all relative to mock controls (dashed line). (G) Brain IL-6 protein levels determined by ELISA ( n ≥ 5 mice per group). The temporal pattern shows that cytokine and chemokine induction precede Cd38 expression, suggesting that neuroinflammation may contribute to the upregulation of CD38. Data in panels A, C, and E are presented as mean ± SD; panels B, D, and F as mean ± SEM (shaded area). Statistical analyses were performed using an unpaired Student’s t test or Mann–Whitney test, as appropriate. ∗ p ≤ 0.05; ∗∗ p ≤ 0.01; ∗∗∗ p ≤ 0.001.

Article Snippet: Proteins were separated by electrophoresis on 15% SDS–polyacrylamide gels (SDS-PAGE), transferred to nitrocellulose membranes (Bio-Rad, California, USA), and probed with primary antibodies against CD38 (AF4947, R&D Systems, Minnesota, USA) or NAMPT (AG-20A-0034-C050, AdipoGen Life Sciences, California, USA), using concentrations recommended by the manufacturers and previously validated in-house.

Techniques: Expressing, Infection, Transformation Assay, Injection, Enzyme-linked Immunosorbent Assay, MANN-WHITNEY

Journal: iScience

Article Title: CD38 is a key mediator of NAD + depletion in the brain of ZIKV-infected mice

doi: 10.1016/j.isci.2025.114018

Figure Lengend Snippet: Infiltrating immune cells contribute to increased CD38 expression in the brains of ZIKV-infected mice (A) On the left, representative dot plots show the gating strategy used to identify CD45 lo CD11b + (putative resting microglia), CD45 hi CD11b + (infiltrating myeloid cells), and CD45 + CD11b − (lymphoid cells) populations. On the right, graphs showing the frequency of each population in ZIKV-infected and mock-injected mice ( n ≥ 5 mice per group). (B) Representative dot plots and quantification of CD11b + TMEM119 + , CD11b + TMEM119 + , and CD11b + TMEM119 - populations in the brains of infected and control mice ( n ≥ 6 mice per group), with the respective graphs showing the frequency of each population. (C–E) Representative histograms and quantification of CD38 expression (median fluorescence intensity, MFI) in CD11b + TMEM119 + (C) CD11b − TMEM119 + (D), and CD11b + TMEM119 - (E) populations. (F and G) Gating and quantification of CD3 + T cells (F) and corresponding CD38 expression (G). (H and I) Gating and quantification of CD19 + B cells (H) and corresponding CD38 expression (I). All graphs represent mean ± SD. Gating was based on negative controls; histogram quantification was performed using MFI, and curves were normalized to unit area. Statistical analyses were performed using an unpaired Student’s t test or Mann–Whitney test, as appropriate. ∗ p ≤ 0.05; ∗∗ p ≤ 0.01; ∗∗∗∗ p ≤ 0.0001.

Article Snippet: Proteins were separated by electrophoresis on 15% SDS–polyacrylamide gels (SDS-PAGE), transferred to nitrocellulose membranes (Bio-Rad, California, USA), and probed with primary antibodies against CD38 (AF4947, R&D Systems, Minnesota, USA) or NAMPT (AG-20A-0034-C050, AdipoGen Life Sciences, California, USA), using concentrations recommended by the manufacturers and previously validated in-house.

Techniques: Expressing, Infection, Injection, Control, Fluorescence, MANN-WHITNEY

Journal: bioRxiv

Article Title: Germ-free piglets display variable neuroinflammatory-like perturbations in prefrontal cortical microglia

doi: 10.64898/2026.03.22.713463

Figure Lengend Snippet: (A) Volcano plot of significant upregulated and downregulated genes in GF tissue, using p < 0.05 and a threshold of 1.2-fold change in expression as a cutoff for analysis. CD38 (p < 0.001, FC = 1.55), RSAD2 (p < 0.001, FC = 0.08), GABRR1 (p < 0.001, FC = 0.08), FN1 (p < 0.001, FC = 0.54), IFI6 (p < 0.001, FC = 0.37), IRAK1BP1 (p < 0.001, FC = 1.74), RGS1 (p < 0.001, FC = 3.22) are labeled in bold. (B) Graphical reconstruction of immune activation Ingenuity Pathway Analysis result, showing differentially expressed genes in our tissue related to this pathway. Panel made with BioRender. (C) Representative immunohistochemical images of the PFCII-III in control and GF animals. CD38 + (red), DAPI + (blue), and Iba1 + (white) staining (left, scale bar 200µm), a middle inset of the leftmost image (middle, scale bar 50µm), and a single channel (CD38) of this inset (right, scale bar 50µm) for each group n=3/animals per group. Data supported by Extended Data Table 6-1.

Article Snippet: Sections were blocked with 5% donkey serum, 1% BSA in PBST for 1 hour, and then stained with Iba1 (Wako, catalog #NC9288364), Ki67 (Abcam, catalog #ab16667), and CD38 (Proteintech, catalog #60006-1-Ig) overnight at 4°C.

Techniques: Expressing, Labeling, Activation Assay, Immunohistochemical staining, Control, Staining

Journal: The Journal of Biological Chemistry

Article Title: CD38 mediates nicotinamide mononucleotide base exchange to yield nicotinic acid mononucleotide

doi: 10.1016/j.jbc.2025.108248

Figure Lengend Snippet: CD38 mediates NMN base exchange with nicotinic acid . A , experimental scheme to identify NMN base exchange. Unlabeled NMN (M + 0) and 13 C 6 -labeled–nicotinic acid (M + 6) are coincubated, and the formation of M + 6–labeled nicotinic acid mononucleotide (NaMN) is indicative of base exchange of the unlabeled nicotinamide group from NMN for an M + 6–labeled nicotinic acid. B , targeted mass spectrometry for these metabolites in the presence or absence of recombinant CD38 protein, with or without cotreatment with the small molecule CD38 inhibitor 78c , showing CD38-dependent formation of M + 6–labeled NaMN. This base exchange is preferential for nicotinic acid over nicotinamide, as ( C ) coincubation of unlabeled NMN with both 13 C 6 -nicotinic acid (M + 6) and D 4 -nicotinamide (M + 4) leads to the greater formation of M + 6 NaMN over M + 4 NMN. D , technical control to show the absence of labelled NaMN in the absence of 13 C 6 -nicotinic acid (M + 6). E , scheme to test whether CD38-mediated base exchange is selective to nicotinic acid. Double-labeled NMN containing D 2 -ribose and D 4 -nicotinamide, for an overall mass shift of M + 6 was coincubated with nicotinic acid or its structural orthologs picolinic acid or isonicotinic acid, which are identical in molecular weight. CD38-mediated base exchange would result in exchange of the D 4 -nicotinamide label for an unlabeled group, resulting in the formation of a product with a molecular weight identical to NaMN plus an M + 2 mass shift, due to retention of the ribose label. Base exchange was not observed for ( F ) picolinic acid, but was observed for ( G ) nicotinic acid and ( H ) isonicotinic acid. I , chromatograms showing the detection of M + 2 ribose-labeled NaMN in each group. Data points represent technical replicates within a single experiment, and experiments were performed three times. Error bars are SDs. NMN, nicotinamide mononucleotide.

Article Snippet: CD38 base-exchange activity was assessed in vitro using recombinant human CD38 (UniProt P28907, Val54–Ile300) corresponding to its extracellular domain, with a C-terminal 6xHis tag produced in NS0-derived mouse myeloma cell line from R&D Systems (catalog number RDS2404-AC), with activity of >2500 pmol/min/μg, as measured by its ability to convert nicotinamide guanine dinucleotide to cyclic GDP ribose.

Techniques: Labeling, Mass Spectrometry, Recombinant, Control, Molecular Weight

Journal: The Journal of Biological Chemistry

Article Title: CD38 mediates nicotinamide mononucleotide base exchange to yield nicotinic acid mononucleotide

doi: 10.1016/j.jbc.2025.108248

Figure Lengend Snippet: CD38 base exchange is selective to NMN, and not NaMN, NR, or NaR . CD38-mediated base exchange was confirmed through three different labelling combinations. A , 5,5-D 2 -ribose labelled NMN (M + 2) was coincubated with 13 C 6 -nicotinic acid, leading to the formation of D 2 -ribose, 13 C 6 -nicotinic acid labeled NaMN (M + 8). B , 5,5-D 2 -ribose, D 4 -nicotinamide–labeled NMN (M + 6) was coincubated with unlabeled nicotinic acid, leading to the formation of D 2 -ribose–labeled NaMN (M + 2). C , unlabeled NMN was coincubated with 13 C 6 -nicotinic acid, leading to the formation of 13 C 6 -nicotinic acid–labeled NaMN (M + 6). D , to test whether CD38 could mediate base exchange on NaMN rather than NMN, unlabeled NaMN was coincubated with D 4 -labeled nicotinamide (M + 4), however M + 4–labeled NMN was not identified. E , to test whether there is direct base exchange between NMN and NaMN, D 4 -nicotinic acid–labeled NaMN (M + 4) was coincubated with D 2 -ribose–labeled NMN (M + 2), however the hypothetical base exchange products were not detected. F and G , to test whether nicotinamide riboside (NR) or nicotinic acid riboside (NaR) could act as a substrate for base exchange, ( F ) unlabeled NR was coincubated with 13 C 6 -nicotinic acid (M + 6), and ( G ) unlabeled NaR was coincubated with D 4 -nicotinamide (M + 4), with no base-exchange products observed in either case. Experiments were performed three separate times; data are shown from a single experiment that included all reaction combinations in parallel, using one technical observation per reaction condition. NaMN, nicotinic acid mononucleotide; NMN, nicotinamide mononucleotide.

Article Snippet: CD38 base-exchange activity was assessed in vitro using recombinant human CD38 (UniProt P28907, Val54–Ile300) corresponding to its extracellular domain, with a C-terminal 6xHis tag produced in NS0-derived mouse myeloma cell line from R&D Systems (catalog number RDS2404-AC), with activity of >2500 pmol/min/μg, as measured by its ability to convert nicotinamide guanine dinucleotide to cyclic GDP ribose.

Techniques: Labeling

Journal: The Journal of Biological Chemistry

Article Title: CD38 mediates nicotinamide mononucleotide base exchange to yield nicotinic acid mononucleotide

doi: 10.1016/j.jbc.2025.108248

Figure Lengend Snippet: CD38 inhibition blocks NMN-induced increases in NaMN and NaAD in vivo . Mice received a single oral dose of the amidated NAD + precursor NMN (500 mg/kg) in the presence or absence of the small molecule CD38 inhibitor 78c (“CD38i”) (10 mg/kg). Two hours later, animals were euthanized and liver, muscle (quadriceps), and kidneys were collected for metabolomics to measure levels of the Preiss–Handler/ de novo pathway deamidated intermediates ( A ) nicotinic acid mononucleotide (NaMN) and ( B ) nicotinic acid adenine dinucleotide (NaAD). Each point represents data from a separate animal, data analyzed by two-way ANOVA followed by Tukey’s test for post hoc comparisons, n = 5/group, error bars are SD. NMN, nicotinamide mononucleotide.

Article Snippet: CD38 base-exchange activity was assessed in vitro using recombinant human CD38 (UniProt P28907, Val54–Ile300) corresponding to its extracellular domain, with a C-terminal 6xHis tag produced in NS0-derived mouse myeloma cell line from R&D Systems (catalog number RDS2404-AC), with activity of >2500 pmol/min/μg, as measured by its ability to convert nicotinamide guanine dinucleotide to cyclic GDP ribose.

Techniques: Inhibition, In Vivo

Journal: The Journal of Biological Chemistry

Article Title: CD38 mediates nicotinamide mononucleotide base exchange to yield nicotinic acid mononucleotide

doi: 10.1016/j.jbc.2025.108248

Figure Lengend Snippet: Mammalian NAD + biosynthesis and proposed role for CD38 . A , NAD + can be synthesized via the salvage pathway, which recycles the amidated precursor nicotinamide via an amidated intermediate, nicotinamide mononucleotide (NMN). In the Preiss–Handler pathway, nicotinic acid is incorporated into NAD + via the deamidated intermediates nicotinic acid mononucleotide (NaMN) and nicotinic acid adenine dinucleotide (NaAD). Treatment with the amidated precursor NMN can increase levels of the deamidated intermediates NaMN and NaAD. B , the enzyme CD38 is well-studied for its NAD + glycohydrolase activity, cleaving NAD + into free nicotinamide and ADP ribose (ADPR). C , CD38 also has base-exchange activity toward NADP + , yielding the Ca 2+ signaling intermediate NaADP + . D , this investigation identified NMN and nicotinic acid as new base-exchange substrates for CD38, yielding NaMN and explaining the crossover between the amidated salvage/recycling pathways and the deamidated Preiss–Handler/ de novo pathways of NAD + biosynthesis.

Article Snippet: CD38 base-exchange activity was assessed in vitro using recombinant human CD38 (UniProt P28907, Val54–Ile300) corresponding to its extracellular domain, with a C-terminal 6xHis tag produced in NS0-derived mouse myeloma cell line from R&D Systems (catalog number RDS2404-AC), with activity of >2500 pmol/min/μg, as measured by its ability to convert nicotinamide guanine dinucleotide to cyclic GDP ribose.

Techniques: Synthesized, Activity Assay

Journal: Blood Neoplasia

Article Title: CD38-CAR human NK cells in combination with ATRA enhance cytotoxicity against CD38-expressing hematologic malignancies ∗

doi: 10.1016/j.bneo.2024.100032

Figure Lengend Snippet: Generation of CRISPR-engineered CD38 KO /CD38-CAR human primary NK cells using Cas9/RNP and AAV. (A) Schemata of steps for CRISPR/RNP knockout of the CD38 gene and directed insertion of a CD38-CAR encoding DNA delivered by AAV6 vector with homology arms for CD38 targeting site. (B) CD38 (PE) and CAR (APC) expression levels measured by flow cytometry for binding of CD38 antigen, 7 days after stimulation. Constructs contain a 41BB signaling domain, a CD8α transmembrane domain/hinge, a CD3ζ stimulatory domain, and reversed orderings of light and heavy chain orientations. (C) Relative percentage and intensity of CD38-CAR expression (n = 10; mean ± standard deviation [SD]). (D) Fold expansion of WT and CD38-CAR NK cells over 12 days after activation with irradiated, modified mbIL21-K562 cells and IL-2 show no significant change from WT human NK cells (n = 10; mean ± SD). P values were calculated using a 2-way analysis of variance (ANOVA); ∗ P = .0332; ∗∗ P = .0021; ∗∗∗ P = .0002; ∗∗∗∗ P < .0001. (E) Cytotoxicity observed for V3 and V4 CD38 KO /CD38-CAR NK cells against high CD38-expressing MM (H929), BL (Raji), and AML (MV-11) (n = 5; mean ± SD). P values were calculated using a 2-way ANOVA; ∗ P < .05; ∗∗ P < .01; ∗∗∗ P = .001; ∗∗∗∗ P < .0001.

Article Snippet: Cells were stained in phosphate-buffered saline containing 2% FBS using anti-CD38/PE (Miltenyi Biotec; clone REA572) and CD38 Fc chimera protein (R&D Systems; rhCD38), followed by secondary staining with Alexa Fluor 647 AffiniPure goat anti-human immunoglobulin G (Jackson ImmunoResearch) and Tonbo Ghost Dye 450 (Tonbo Biosciences, San Diego, CA).

Techniques: CRISPR, Knock-Out, Plasmid Preparation, Expressing, Flow Cytometry, Binding Assay, Construct, Standard Deviation, Activation Assay, Irradiation, Modification

Journal: Blood Neoplasia

Article Title: CD38-CAR human NK cells in combination with ATRA enhance cytotoxicity against CD38-expressing hematologic malignancies ∗

doi: 10.1016/j.bneo.2024.100032

Figure Lengend Snippet: CD38-CAR NK cells exhibit enhanced cytotoxic function and cytokine secretion. CD38-CAR NK cells were tested against CD38-expressing AML, MM, and T-cell malignancies collected from patients at baseline. (A) NK and CD38-CAR NK killing of AML-1 (n = 3; mean ± SD). (B) NK and CD38-CAR NK killing of samples from patients with MM (MM-1 and MM-2; n = 1; mean ± SD). (C) NK and CD38-CAR NK killing of T-cell malignancies (hepatosplenic T-cell lymphoma and T-PLL [T-PLL-1 and T-PLL-2]; n = 1; mean ± SD). All cytotoxicity P values were calculated using a 2-way ANOVA; ∗ P < .05; ∗∗ P < .01; ∗∗∗ P = .001; ∗∗∗∗ P < .0001. (D) Bio-Plex Pro Human Cytokine assay was performed on the supernatant of WT and CD38-CAR NK cells cocultured with CD38 + malignancies (n = 9; mean ± SD). P values were calculated using a paired Student t test; ∗ P = .05; ∗∗ P = .01; ∗∗∗ P = .001. GM-CSF, IFN-γ, MCP-1, MIP-1α, TNF-α.

Article Snippet: Cells were stained in phosphate-buffered saline containing 2% FBS using anti-CD38/PE (Miltenyi Biotec; clone REA572) and CD38 Fc chimera protein (R&D Systems; rhCD38), followed by secondary staining with Alexa Fluor 647 AffiniPure goat anti-human immunoglobulin G (Jackson ImmunoResearch) and Tonbo Ghost Dye 450 (Tonbo Biosciences, San Diego, CA).

Techniques: Expressing, Cytokine Assay

Journal: Blood Neoplasia

Article Title: CD38-CAR human NK cells in combination with ATRA enhance cytotoxicity against CD38-expressing hematologic malignancies ∗

doi: 10.1016/j.bneo.2024.100032

Figure Lengend Snippet: ATRA upregulates CD38 expression on tumor cells and can enhance antitumor activity. (A) CD38 cell surface expression as measured by flow cytometry across the hematologic malignancies MM, AML, BL, and T-ALL after treatment with 10 nM of ATRA for 48 hours. (B) Mean fluorescence intensity (MFI) of CD38 expression on cell lines with and without ATRA treatment. (C) Cytotoxicity assays performed by coculturing WT and CD38-CAR NK cells against AML, MM, BL, and T-cell malignancies with and without 48-hour, 10-nM ATRA pretreatments. MM1S (n = 4), H929 (n = 4), AML-10 (n = 4), MV4-11 (n = 4), Raji (n = 4), Daudi (n = 4), and primary cells from patients with T-ALL (n = 3; mean ± SD). P values were calculated using a 2-way ANOVA; ∗ P < .05; ∗∗ P <.01; ∗∗∗ P = .001; ∗∗∗∗ P < .0001.

Article Snippet: Cells were stained in phosphate-buffered saline containing 2% FBS using anti-CD38/PE (Miltenyi Biotec; clone REA572) and CD38 Fc chimera protein (R&D Systems; rhCD38), followed by secondary staining with Alexa Fluor 647 AffiniPure goat anti-human immunoglobulin G (Jackson ImmunoResearch) and Tonbo Ghost Dye 450 (Tonbo Biosciences, San Diego, CA).

Techniques: Expressing, Activity Assay, Flow Cytometry, Fluorescence

Journal: Blood Neoplasia

Article Title: CD38-CAR human NK cells in combination with ATRA enhance cytotoxicity against CD38-expressing hematologic malignancies ∗

doi: 10.1016/j.bneo.2024.100032

Figure Lengend Snippet: Mass cytometry analysis shows combination ATRA and CD38-CAR NK-cell treatment decreases live AML population. Mass cytometry analysis was performed 24 hours after the coculture of a primary AML cell line (AML-1) with WT or CD38-CAR NK cells with and without 48-hour, 10-nM ATRA pretreatment. Eight distinct conditions were analyzed: WT NK cells alone (A), CD38-CAR NK cells alone (B), AML cells alone (C), AML plus WT NK cells (D), AML plus CD38-CAR NK cells (E), AML cells with ATRA (F), AML plus ATRA plus WT NK cells (G), and AML plus ATRA plus CD38-CAR NK cells (H). (I) Heat map of surface marker appearing on the live AML cells after treatment analyzed by mass cytometry. Original values are ln(x)-transformed. Rows are centered; unit variance scaling is applied to rows. Both rows and columns are clustered using correlation distance and average linkage using ClustVis.

Article Snippet: Cells were stained in phosphate-buffered saline containing 2% FBS using anti-CD38/PE (Miltenyi Biotec; clone REA572) and CD38 Fc chimera protein (R&D Systems; rhCD38), followed by secondary staining with Alexa Fluor 647 AffiniPure goat anti-human immunoglobulin G (Jackson ImmunoResearch) and Tonbo Ghost Dye 450 (Tonbo Biosciences, San Diego, CA).

Techniques: Mass Cytometry, Marker, Transformation Assay

Journal: Blood Neoplasia

Article Title: CD38-CAR human NK cells in combination with ATRA enhance cytotoxicity against CD38-expressing hematologic malignancies ∗

doi: 10.1016/j.bneo.2024.100032

Figure Lengend Snippet: CD38 KO /CD38-CAR T cells generated by Cas9/RNP and AAV6 show antitumor activity. (A) CD38 (PE) and CAR (APC) expression levels measured by flow cytometry. (B) Relative percentage of CD38-CAR expression (n = 5; mean ± SD). (C) Ten-day fold expansion of generated CD38-CAR T cells with matched donors (n = 4; mean ± SD). P values were calculated using a 2-way ANOVA; ∗ P = .0332; ∗∗ P = .0021; ∗∗∗ P = .0002; ∗∗∗∗ P < .0001. (D) Cytotoxicity of CD38 KO /CD38-CAR T cells toward H929, Raji, and MV4-11 cell lines (n = 4; mean ± SD). P values were calculated using a 2-way ANOVA; ∗ P < .05; ∗∗ P < .01; ∗∗∗ P = .001; ∗∗∗∗ P < .0001.

Article Snippet: Cells were stained in phosphate-buffered saline containing 2% FBS using anti-CD38/PE (Miltenyi Biotec; clone REA572) and CD38 Fc chimera protein (R&D Systems; rhCD38), followed by secondary staining with Alexa Fluor 647 AffiniPure goat anti-human immunoglobulin G (Jackson ImmunoResearch) and Tonbo Ghost Dye 450 (Tonbo Biosciences, San Diego, CA).

Techniques: Generated, Activity Assay, Expressing, Flow Cytometry

Journal: Blood Neoplasia

Article Title: CD38-CAR human NK cells in combination with ATRA enhance cytotoxicity against CD38-expressing hematologic malignancies ∗

doi: 10.1016/j.bneo.2024.100032

Figure Lengend Snippet: AAVS1KO/CD38-CAR NK cells avoid fratricide. (A) CD38 (PE) and CAR (APC) expression levels measured by flow cytometry for the CD38-CAR inserted into different loci. All CAR NK cells were generated with matched donors. (B) Fold expansion of WT and CD38-CAR NK cells over 12 days (n = 6; mean ± SD). P values were calculated using a 2-way ANOVA; ∗ P < .05; ∗∗ P < .01; ∗∗∗ P = .001; ∗∗∗∗ P < .0001. (C) Relative percentage of CD38-CAR expression (n = 6; mean ± SD). (D) Reverse-transcription qPCR was performed using CD38 primer probes to detect transcription of the CD38 gene in the WT and AAVS1KO/CD38-CAR NK cells. (E) CD38 expression measured by flow cytometry using a polyclonal anti-CD38 antibody.

Article Snippet: Cells were stained in phosphate-buffered saline containing 2% FBS using anti-CD38/PE (Miltenyi Biotec; clone REA572) and CD38 Fc chimera protein (R&D Systems; rhCD38), followed by secondary staining with Alexa Fluor 647 AffiniPure goat anti-human immunoglobulin G (Jackson ImmunoResearch) and Tonbo Ghost Dye 450 (Tonbo Biosciences, San Diego, CA).

Techniques: Expressing, Flow Cytometry, Generated, Reverse Transcription

Journal: Blood Neoplasia

Article Title: CD38-CAR human NK cells in combination with ATRA enhance cytotoxicity against CD38-expressing hematologic malignancies ∗

doi: 10.1016/j.bneo.2024.100032

Figure Lengend Snippet: AAVS1KO/CD38-CAR NK cells display enhanced cytotoxicity and metabolism. CD38-CAR NK cells were tested against CD38-expressing AML, MM, and BL. (A) CD38-CAR NK-cell killing of CD38-expressing cell lines (n = 3; mean ± SD). P values were calculated using a 2-way ANOVA; ∗ P < .05; ∗∗ P < .01; ∗∗∗ P = .001; ∗∗∗∗ P < .0001. (B) Oxygen consumption rate (OCR) for CD38-CAR NK cells. (C) Glycolytic capacity, measured by the extracellular acidification rate (ECAR), observed in CD38-CAR NK cells. (D) Spare respiratory capacity, a measure of the cell's ability to produce adenosine triphosphate (ATP) in response to stress, measured in CAR NK cells. (E) CAR NK-cell maximal respiration rates compared with WT.

Article Snippet: Cells were stained in phosphate-buffered saline containing 2% FBS using anti-CD38/PE (Miltenyi Biotec; clone REA572) and CD38 Fc chimera protein (R&D Systems; rhCD38), followed by secondary staining with Alexa Fluor 647 AffiniPure goat anti-human immunoglobulin G (Jackson ImmunoResearch) and Tonbo Ghost Dye 450 (Tonbo Biosciences, San Diego, CA).

Techniques: Expressing