Journal: Nature Communications

Article Title: Simultaneous orthogonal cell engineering by a single CRISPR-Cas9 polyfunctional editor

doi: 10.1038/s41467-026-72846-2

Figure Lengend Snippet: A Polyfunctional editing strategy for the epi-silencing of B2M and TET2 and insertion of the CD19–28ζ CAR:ΔLNGFR construct into TRAC . B Left: percentages of the indicated T cell populations at day 14 after poly-editing (mean ± SD of 7 blood donors). TCR + : TCR-positive cells (green bar). TCR - : TCR-negative cells (light blue bar). TCR - /ΔLNGFR + : cells negative for endogenous TCRs and positive for ΔLNGFR (lilac bar). B2M - : B2M-negative cells (gray bar). B2M + : B2M-positive cells (white bar). Right: representative flow cytometry dot plots of poly-edited T cells showing expression of the TCRs and ΔLNGFR (left plot) and, within the TCR - /ΔLNGFR + cells, of B2M (right plot). C Fold-change in TET2 (light blue bars) and B2M (gray bars) expression in poly-edited vs . mock-treated cells (mean ± SD of 7 blood donors). D Schematic of the in vivo experiment. NSG mice were injected with GLuc.NALM-6 cells and, 7 days later, with the indicated T cell populations. Peripheral blood was collected at the indicated time points to measure tumor growth, phenotype human T cells, and quantify cytokine release. Mock: mock-transfected T cells. CAR-T Δ LNGFR : T cells expressing CD19–28ζ CAR:ΔLNGFR from TRAC . Poly-edited cells: T cells expressing CD19–28ζ CAR:ΔLNGFR from TRAC with epi-silencing of B2M and TET2 . E Tumor growth (left; mean ± SEM) and survival (right) curves over 20 days post-transplantation of the indicated T cell populations. n = 6 mice for Mock; n = 12 mice for each other group. ** p = 0.0075, *** p = 0.0008 by Mantel-Cox (log-rank) test. F Circos plots from CAST-Seq analyses of triple KO (left) and poly-edited (right) T cells ( n = 2 experimental replicates). Triple-KO cells were transfected with mRNA encoding Cas9 and the TRAC gRNA, together with gRNA B#6 and gRNA TE#19 . For poly-editing, the TRAC gRNA and the selected guide combinations for B2M and TET2 were used. Aberrations at the TRAC locus are in violet; translocations between TRAC and either B2M or TET2 are in blue; translocations between TRAC and OT sites of all gRNAs are in gray. G Number of unique CAST-Seq reads corresponding to either aberrations (violet bars) or translocations (gray bars) at the TRAC locus in the indicated treatments. Graphs were generated using GraphPad Prism (GraphPad Software). Source data are provided as a file.

Article Snippet: LNGFR; 130-113-422, Miltenyi), CD3 (345763, BD Biosciences), HLA-ABC (565332, BD Biosciences), HLA-E (130-117-402, Miltenyi), and CD19 CAR FMC63 Idiotype (130-127-342, Miltenyi).

Techniques: Construct, Flow Cytometry, Expressing, In Vivo, Injection, Transfection, Transplantation Assay, Generated, Software

Journal: Nature Communications

Article Title: Simultaneous orthogonal cell engineering by a single CRISPR-Cas9 polyfunctional editor

doi: 10.1038/s41467-026-72846-2

Figure Lengend Snippet: A Scatter plot comparing whole-transcriptome analyses of T cells treated with the tripartite ETR and either full-length or truncated gRNAs targeting CD3D (left) or TGFBR2 (right) ( n = 2 experimental replicates). Data are expressed as log 2 TPM of mapped reads. B Polyfunctional editing strategy used to epi-silence CD3D and TGFBR2 and insert the scHLA-E:CD19–28ζ CAR cassette into exon 1 of B2M . C Left: percentages of the indicated T cell populations as measured by flow cytometry 14 days after polyfunctional editing (mean ± SD of 7 blood donors). HLA-ABC + : HLA-ABC-positive cells (light yellow bars); HLA-ABC - : HLA-ABC-negative cells (blue bars); CD19-CAR - : cells negative for the expression of the CD19–28ζ CAR (gray bars); CD19-CAR + : cells positive for the expression of the CD19–28ζ CAR (lilac bars); CD3 + : CD3-positive cells; CD3 - : CD3-negative cells; HLA-E - : HLA-E-negative cells; HLA-E + : HLA-E-positive cells. Right: representative flow cytometry dot plots of poly-edited T cells showing the expression levels of the HLA-ABC and CD3 (left plot) and, within the double-negative cells, of HLA-E and CD19-CAR (right plot). D Fold-change in the expression levels of CD3D (green bars) and TGFBR2 (orange bars) in poly-edited cells relative to mock-treated cells, 14 days post-editing (mean ± SD of 7 blood donors). Graphs were generated using GraphPad Prism (GraphPad Software). Source data are provided as a file.

Article Snippet: LNGFR; 130-113-422, Miltenyi), CD3 (345763, BD Biosciences), HLA-ABC (565332, BD Biosciences), HLA-E (130-117-402, Miltenyi), and CD19 CAR FMC63 Idiotype (130-127-342, Miltenyi).

Techniques: Flow Cytometry, Expressing, Generated, Software

Journal: Frontiers in Oncology

Article Title: Clinical implementation of a one-step no-wash flow cytometry method allows for real-time monitoring of patients treated with autologous CAR-T cells

doi: 10.3389/fonc.2026.1774431

Figure Lengend Snippet: Determination of the limit of detection (LOD) and lower limit of quantification (LLOQ) for the two-step method. CAR-T cell absolute counts were measured in 31 negative control samples from patients not treated with CD19 CAR-T cells. The mean background signal is represented by the blue line. LOD was defined as mean + 3 standard deviations (SD), and LLOQ as mean + 10 SD.

Article Snippet: New CAR detection reagent (CDR) directly coupled to fluorochrome, either BCMA (BCMA CDR-PE, Miltenyi Biotec 130-133-888) or CD19 (CD19 CDR α-FMC63-PE, Miltenyi Biotec 130-127-342) was added extemporaneously in the analytical tube.

Techniques: Negative Control

Journal: Frontiers in Oncology

Article Title: Clinical implementation of a one-step no-wash flow cytometry method allows for real-time monitoring of patients treated with autologous CAR-T cells

doi: 10.3389/fonc.2026.1774431

Figure Lengend Snippet: Gating strategy for the single-step method. Absolute counting beads were excluded based on scatter and fluorescence properties. Dead cells were excluded using 7-aminoactinomycin D (7-AAD). CD45-positive leukocytes were selected, and lymphocytes were identified according to side scatter (SSC) characteristics. CD3-positive T cells were gated, and CAR-T cells were defined as viable CD45+/CD3+/CAR+ events using directly fluorochrome-conjugated CAR detection reagents (CD19 or BCMA). CD4 and CD8 subpopulations were subsequently identified within the CAR-positive T-cell compartment. Absolute quantification was calculated using TruCount beads according to the manufacturer’s formula.

Article Snippet: New CAR detection reagent (CDR) directly coupled to fluorochrome, either BCMA (BCMA CDR-PE, Miltenyi Biotec 130-133-888) or CD19 (CD19 CDR α-FMC63-PE, Miltenyi Biotec 130-127-342) was added extemporaneously in the analytical tube.

Techniques: Fluorescence, Quantitative Proteomics

Journal: Frontiers in Oncology

Article Title: Clinical implementation of a one-step no-wash flow cytometry method allows for real-time monitoring of patients treated with autologous CAR-T cells

doi: 10.3389/fonc.2026.1774431

Figure Lengend Snippet: Determination of LOD and LLOQ for the single-step method. CAR-T cell absolute counts were measured in 10 negative control samples from patients not treated with CD19 or BCMA CAR-T cells. The blue line represents the mean background signal. LOD was defined as mean + 3 SD and LLOQ (green dashed line) as mean + 10 SD.

Article Snippet: New CAR detection reagent (CDR) directly coupled to fluorochrome, either BCMA (BCMA CDR-PE, Miltenyi Biotec 130-133-888) or CD19 (CD19 CDR α-FMC63-PE, Miltenyi Biotec 130-127-342) was added extemporaneously in the analytical tube.

Techniques: Negative Control

Journal: Cancer Discovery

Article Title: Gut Microbiota Modulation through Akkermansia spp . Supplementation Increases CAR T-cell Potency

doi: 10.1158/2159-8290.CD-24-1230

Figure Lengend Snippet: Association between gut bacterial composition and response to CAR T-cell therapy. A, Schematic representation of the prospective longitudinal collection of biological samples (fecal material, blood, tumor tissues, and BM aspirations) from patients with B-cell malignancies treated with commercial anti-CD19 CAR T cells. Feces collection occurred at three time points: visit 1 (prior to lymphodepleting chemotherapy), visit 2 (7–15 days after CAR T-cell infusion), and visit 3 (3 months after CAR T-cell infusion). B, α-Diversity analysis using the Shannon index (left) and the species richness index (right) across visits 1, 2, and 3. Data are presented as means ± SEM, with statistical significance evaluated using Wilcoxon signed-rank tests. C, Comparison of bacterial alpha diversity between ORR + and ORR − at visits 1, 2, and 3, calculated using the Shannon index (left) and the species richness index (right). Results are shown as means ± SEM, with statistical analysis by Wilcoxon signed-rank tests. D, β-Diversity analysis using the Bray–Curtis metric to measure microbial community distances, with PERMANOVA tests performed at visits 1, 2, and 3. E, Comparison of bacterial β-diversity between ORR + and ORR − at visits 1, 2, and 3 using PERMANOVA tests. F, PFS curves comparing patients classified as SIG2 + or SIG1 + according to the TOPOSCORE. G, Serum levels of sMAdCAM-1 (ng/mL) in patients with B-cell lymphoma across longitudinal analyses (left) and according to 6-month response status (right). Results are shown as means ± SEM, analyzed using Mann–Whitney tests. H, LEfSe graph generated from baseline (visit 1) metagenomic data, highlighting the most discriminant species between ORR + and ORR − based on linear discriminant analysis scores. MDS, multidimensional scaling.

Article Snippet: Briefly, after obtaining single-cell suspensions, tumors were stained for 30 minutes at 4°C with a mix of antibodies containing anti-CAR T-cell CD19 (Cat. No. 130-127-342, clone REA1297, Miltenyi), anti-CCR6 (Cat. No. 566130, clone 11A9, BD), anti-CCR7 (Cat. No. 566437, clone 3D12 RUO, BD), anti-CD11b (Cat. No. 301325, clone ICRF44), anti-CD11c (Cat. No. 130-128-247, clone REA618, Miltenyi), anti-CD123 (Cat. No. 130-115-272, clone REA918, Miltenyi), anti-CD127 (Cat. No. 351363, clone A019D5, BioLegend), anti-CD141 (Cat. No. 344113, clone M80, BioLegend), anti-CD16 (Cat. No. 130-113-395, clone REA423, Miltenyi), anti-CD163 (Cat. No. 333631, clone GHI/61, BioLegend), anti-CD19 (Cat. No. 612916, clone SJ25C1, BD), anti-CD1c (Cat. No. 331537, clone L161, BioLegend), anti-CD20 (Cat. No. 612906, clone 2H7, BD), anti-CD206 (Cat. No. 130-126-623, clone DCN228, Miltenyi), anti-CD25 (Cat. No. 356145, clone M-A251, BioLegend), anti-CD3 (Cat. No. 130-113-142, clone REA613, Miltenyi), anti-CD38 (Cat. No. 303549, clone HIT2, BioLegend), anti-CD39 (Cat. No. 749967, clone TU66, BD), anti-CD4 (Cat. No. 344669, clone SK3, BioLegend), anti-CD45 (Cat. No. 130-113-120, clone 5B1, Miltenyi), CD45RA (Cat. No. 740298, clone HI100, BD), anti-CD56 (Cat. No. 362513, clone 5.1H11, BioLegend), anti-CD66b (Cat. No. 305111, clone G10F5, BioLegend), CD79b (Cat. No. 612814, clone CB3-1, BD), anti-CD8 (Cat. No. 344759, clone SK1, BioLegend), anti-CXCR3 (Cat. No. 353755, clone G025H7, BioLegend), anti-CXCR5 (Cat. No. 356941, clone J252D4, BioLegend), anti-FOLR2 (Cat. No. 391705, clone 94b/FOLR2, BioLegend), anti–HLA-DR (Cat. No. 307683, clone L243, BioLegend), anti–PD-1 (Cat. No. 329927, clone EH12.2H7, BioLegend), anti-TCF1 (Cat. No. 905115, Cell Signaling Technology), anti–TIM-3 (Cat. No. 345027, clone F38-2E2, BioLegend), anti-TREM2 (Cat. No. FAB17291G, clone 237920, RD), and Viability (Cat. No. 423105, BioLegend).

Techniques: Comparison, MANN-WHITNEY, Generated

Journal: Cancer Discovery

Article Title: Gut Microbiota Modulation through Akkermansia spp . Supplementation Increases CAR T-cell Potency

doi: 10.1158/2159-8290.CD-24-1230

Figure Lengend Snippet: Akkermansia spp. supplementation improves CD19/CD28-ζ CAR T-cell efficacy against B-cell lymphoma. A, Schematic representation of the experimental design for the CD19/CD28-ζ CAR T-cell mouse model. B and C, Tumor growth kinetics and animal survival following CAR T-cell infusion with or without oral live biotherapeutics. B, Tumor sizes monitored by caliper, presented as means ± SEM. A representative experiment from three independent trials is shown, with 15 mice per treatment group. Tumor sizes were analyzed using one-way ANOVA, with specific time points compared using Mann–Whitney tests. C, Survival analysis was performed using the Kaplan–Meier estimator and log-rank test. D, Flow cytometry analysis of the blood compartment on day 14 after CAR T-cell injection. Evaluation of CAR T-cell expansion rate (% CD45.1 EGFR + cells) and percentage of persistent target cells (% CD19 + cells). Data are presented as means ± SEM, with statistical analysis by Mann–Whitney tests. E, Flow cytometry analysis of the tumor compartment on day 24. Representative contour plots and absolute numbers/percentages of viable CAR T cells across treatment groups. F, Phenotype of tumor-infiltrating CAR T cells, including CD4/CD8 ratio and percentage of IFNγ + CD8 + CAR T cells. Data are shown as means ± SEM, with statistical analysis by Mann–Whitney tests.

Article Snippet: Briefly, after obtaining single-cell suspensions, tumors were stained for 30 minutes at 4°C with a mix of antibodies containing anti-CAR T-cell CD19 (Cat. No. 130-127-342, clone REA1297, Miltenyi), anti-CCR6 (Cat. No. 566130, clone 11A9, BD), anti-CCR7 (Cat. No. 566437, clone 3D12 RUO, BD), anti-CD11b (Cat. No. 301325, clone ICRF44), anti-CD11c (Cat. No. 130-128-247, clone REA618, Miltenyi), anti-CD123 (Cat. No. 130-115-272, clone REA918, Miltenyi), anti-CD127 (Cat. No. 351363, clone A019D5, BioLegend), anti-CD141 (Cat. No. 344113, clone M80, BioLegend), anti-CD16 (Cat. No. 130-113-395, clone REA423, Miltenyi), anti-CD163 (Cat. No. 333631, clone GHI/61, BioLegend), anti-CD19 (Cat. No. 612916, clone SJ25C1, BD), anti-CD1c (Cat. No. 331537, clone L161, BioLegend), anti-CD20 (Cat. No. 612906, clone 2H7, BD), anti-CD206 (Cat. No. 130-126-623, clone DCN228, Miltenyi), anti-CD25 (Cat. No. 356145, clone M-A251, BioLegend), anti-CD3 (Cat. No. 130-113-142, clone REA613, Miltenyi), anti-CD38 (Cat. No. 303549, clone HIT2, BioLegend), anti-CD39 (Cat. No. 749967, clone TU66, BD), anti-CD4 (Cat. No. 344669, clone SK3, BioLegend), anti-CD45 (Cat. No. 130-113-120, clone 5B1, Miltenyi), CD45RA (Cat. No. 740298, clone HI100, BD), anti-CD56 (Cat. No. 362513, clone 5.1H11, BioLegend), anti-CD66b (Cat. No. 305111, clone G10F5, BioLegend), CD79b (Cat. No. 612814, clone CB3-1, BD), anti-CD8 (Cat. No. 344759, clone SK1, BioLegend), anti-CXCR3 (Cat. No. 353755, clone G025H7, BioLegend), anti-CXCR5 (Cat. No. 356941, clone J252D4, BioLegend), anti-FOLR2 (Cat. No. 391705, clone 94b/FOLR2, BioLegend), anti–HLA-DR (Cat. No. 307683, clone L243, BioLegend), anti–PD-1 (Cat. No. 329927, clone EH12.2H7, BioLegend), anti-TCF1 (Cat. No. 905115, Cell Signaling Technology), anti–TIM-3 (Cat. No. 345027, clone F38-2E2, BioLegend), anti-TREM2 (Cat. No. FAB17291G, clone 237920, RD), and Viability (Cat. No. 423105, BioLegend).

Techniques: MANN-WHITNEY, Flow Cytometry, Injection

Journal: Cancer Discovery

Article Title: Gut Microbiota Modulation through Akkermansia spp . Supplementation Increases CAR T-cell Potency

doi: 10.1158/2159-8290.CD-24-1230

Figure Lengend Snippet: Prevalence of intestinal Akkermansia spp. and lymphoma TME. A, CAR T-cell expansion in the peripheral blood of patients receiving anti-CD19 CAR T cells, stratified by the presence of Akkermansia spp. based on MGS data ( n = 6 Akk + ; n = 17 Akk − ). CAR T-cell expansion is represented as a percentage (left) and absolute numbers per mL (right). Data are shown as means ± SEM, with statistical significance determined by Mann–Whitney tests. B, Schematic representation of the experimental workflow. Tumor biopsies obtained shortly after CAR T-cell infusion were analyzed by spectral flow cytometry in patients with available MGS data ( n = 3 Akk + ; n = 3 Akk − ). Tumor supernatants were further analyzed using Meso Scale Discovery (MSD) multiplex assays. C, t-Distributed stochastic neighbor embedding (t-SNE) visualization of unsupervised spectral flow cytometry analysis. D, Comparisons of CAR T cells and T-cell frequencies between Akk + and Akk − patients. Data are presented as means ± SEM, with statistical comparisons performed using Mann–Whitney tests. E, Heatmap representation of expression levels [mean fluorescence intensity (MFI)] of CD39, CD38, and PD-1 within the T-cell population. F, Tumor secretome analysis in patients with available metagenomic data ( n = 6), focusing on levels of GM-CSF, IFNγ, and LAG-3. Results are shown as means ± SEM, with statistical significance assessed by Mann–Whitney tests.

Article Snippet: Briefly, after obtaining single-cell suspensions, tumors were stained for 30 minutes at 4°C with a mix of antibodies containing anti-CAR T-cell CD19 (Cat. No. 130-127-342, clone REA1297, Miltenyi), anti-CCR6 (Cat. No. 566130, clone 11A9, BD), anti-CCR7 (Cat. No. 566437, clone 3D12 RUO, BD), anti-CD11b (Cat. No. 301325, clone ICRF44), anti-CD11c (Cat. No. 130-128-247, clone REA618, Miltenyi), anti-CD123 (Cat. No. 130-115-272, clone REA918, Miltenyi), anti-CD127 (Cat. No. 351363, clone A019D5, BioLegend), anti-CD141 (Cat. No. 344113, clone M80, BioLegend), anti-CD16 (Cat. No. 130-113-395, clone REA423, Miltenyi), anti-CD163 (Cat. No. 333631, clone GHI/61, BioLegend), anti-CD19 (Cat. No. 612916, clone SJ25C1, BD), anti-CD1c (Cat. No. 331537, clone L161, BioLegend), anti-CD20 (Cat. No. 612906, clone 2H7, BD), anti-CD206 (Cat. No. 130-126-623, clone DCN228, Miltenyi), anti-CD25 (Cat. No. 356145, clone M-A251, BioLegend), anti-CD3 (Cat. No. 130-113-142, clone REA613, Miltenyi), anti-CD38 (Cat. No. 303549, clone HIT2, BioLegend), anti-CD39 (Cat. No. 749967, clone TU66, BD), anti-CD4 (Cat. No. 344669, clone SK3, BioLegend), anti-CD45 (Cat. No. 130-113-120, clone 5B1, Miltenyi), CD45RA (Cat. No. 740298, clone HI100, BD), anti-CD56 (Cat. No. 362513, clone 5.1H11, BioLegend), anti-CD66b (Cat. No. 305111, clone G10F5, BioLegend), CD79b (Cat. No. 612814, clone CB3-1, BD), anti-CD8 (Cat. No. 344759, clone SK1, BioLegend), anti-CXCR3 (Cat. No. 353755, clone G025H7, BioLegend), anti-CXCR5 (Cat. No. 356941, clone J252D4, BioLegend), anti-FOLR2 (Cat. No. 391705, clone 94b/FOLR2, BioLegend), anti–HLA-DR (Cat. No. 307683, clone L243, BioLegend), anti–PD-1 (Cat. No. 329927, clone EH12.2H7, BioLegend), anti-TCF1 (Cat. No. 905115, Cell Signaling Technology), anti–TIM-3 (Cat. No. 345027, clone F38-2E2, BioLegend), anti-TREM2 (Cat. No. FAB17291G, clone 237920, RD), and Viability (Cat. No. 423105, BioLegend).

Techniques: MANN-WHITNEY, Flow Cytometry, Multiplex Assay, Expressing, Fluorescence

Journal: Cancer Discovery

Article Title: Gut Microbiota Modulation through Akkermansia spp . Supplementation Increases CAR T-cell Potency

doi: 10.1158/2159-8290.CD-24-1230

Figure Lengend Snippet: Akkermansia spp. systemically releases immunogenic indole metabolites, increasing CAR T-cell efficacy via AhR activation. A, Targeted mass spectrometry-based metabolomic analyses of plasma from patients receiving anti-CD19 CAR T cells at three visits ( n = 43 for visit 1; n = 38 for visit 2; n = 29 for visit 3). Heatmaps represent the log 2 fold change of normalized metabolite values. B, Correlation between plasma levels of IPA and Akkermansia spp. presence in patients with available MGS data, analyzed longitudinally. Results are shown as means of log 2 fold change values ±SEM, with Wilcoxon signed-rank tests applied. C, Spearman correlation between bacteria with the highest relative abundance in ORR + or ORR − patients and metabolites from the indole pathway. Statistical significance was determined using Benjamini–Hochberg FDR–corrected P values for each microorganism across all indoles. D, Schematic representation of CAR T-cell experimental design comparing wild-type and AhR-deficient CAR T cells with or without Akk. p2261 supplementation. E and F, Tumor growth kinetics ( E ) and cross-sectional tumor size comparisons between treatment arms at day 13 ( F ). G, Phenotypic characterization comparison of wild-type and AhR-deficient CAR T cells supplemented with supernatant from anaerobic cultures of Akkermansia spp. at 10% (or control media). H, Tumor growth kinetics of lymphoma following CAR T-cell infusion with or without daily oral gavage administration of IPA( n = 10 mice per group). One-way ANOVA was used for tumor growth curve analyses. Data are presented as mean ± SEM; Mann–Whitney tests were performed for specific comparisons. KO, knockout.

Article Snippet: Briefly, after obtaining single-cell suspensions, tumors were stained for 30 minutes at 4°C with a mix of antibodies containing anti-CAR T-cell CD19 (Cat. No. 130-127-342, clone REA1297, Miltenyi), anti-CCR6 (Cat. No. 566130, clone 11A9, BD), anti-CCR7 (Cat. No. 566437, clone 3D12 RUO, BD), anti-CD11b (Cat. No. 301325, clone ICRF44), anti-CD11c (Cat. No. 130-128-247, clone REA618, Miltenyi), anti-CD123 (Cat. No. 130-115-272, clone REA918, Miltenyi), anti-CD127 (Cat. No. 351363, clone A019D5, BioLegend), anti-CD141 (Cat. No. 344113, clone M80, BioLegend), anti-CD16 (Cat. No. 130-113-395, clone REA423, Miltenyi), anti-CD163 (Cat. No. 333631, clone GHI/61, BioLegend), anti-CD19 (Cat. No. 612916, clone SJ25C1, BD), anti-CD1c (Cat. No. 331537, clone L161, BioLegend), anti-CD20 (Cat. No. 612906, clone 2H7, BD), anti-CD206 (Cat. No. 130-126-623, clone DCN228, Miltenyi), anti-CD25 (Cat. No. 356145, clone M-A251, BioLegend), anti-CD3 (Cat. No. 130-113-142, clone REA613, Miltenyi), anti-CD38 (Cat. No. 303549, clone HIT2, BioLegend), anti-CD39 (Cat. No. 749967, clone TU66, BD), anti-CD4 (Cat. No. 344669, clone SK3, BioLegend), anti-CD45 (Cat. No. 130-113-120, clone 5B1, Miltenyi), CD45RA (Cat. No. 740298, clone HI100, BD), anti-CD56 (Cat. No. 362513, clone 5.1H11, BioLegend), anti-CD66b (Cat. No. 305111, clone G10F5, BioLegend), CD79b (Cat. No. 612814, clone CB3-1, BD), anti-CD8 (Cat. No. 344759, clone SK1, BioLegend), anti-CXCR3 (Cat. No. 353755, clone G025H7, BioLegend), anti-CXCR5 (Cat. No. 356941, clone J252D4, BioLegend), anti-FOLR2 (Cat. No. 391705, clone 94b/FOLR2, BioLegend), anti–HLA-DR (Cat. No. 307683, clone L243, BioLegend), anti–PD-1 (Cat. No. 329927, clone EH12.2H7, BioLegend), anti-TCF1 (Cat. No. 905115, Cell Signaling Technology), anti–TIM-3 (Cat. No. 345027, clone F38-2E2, BioLegend), anti-TREM2 (Cat. No. FAB17291G, clone 237920, RD), and Viability (Cat. No. 423105, BioLegend).

Techniques: Activation Assay, Mass Spectrometry, Clinical Proteomics, Bacteria, Comparison, Control, MANN-WHITNEY, Knock-Out