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anti hcd19 antibody  (Bio X Cell)


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    Bio X Cell anti hcd19 antibody
    Anti Hcd19 Antibody, supplied by Bio X Cell, used in various techniques. Bioz Stars score: 93/100, based on 6 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/anti+hcd19+antibody/InVivoMAb+anti-human+CD19/pm41162681-249-17-20
    Average 93 stars, based on 6 article reviews
    anti hcd19 antibody - by Bioz Stars, 2026-10
    93/100 stars

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    Related Articles

    Expressing:

    Article Title: Imaging of biphasic signalosomes constructed by checkpoint receptor 2B4 in conventional and chimeric antigen receptor-T cells
    Article Snippet: The mouse MHC class II molecule I-E k with a GPI anchor (I-E k -GPI), the mouse MHC class I molecule H-2K b with a GPI anchor (H-2K b -GPI), and mouse ICAM-1 with a GPI anchor (ICAM-1-GPI) were purified from transfected Chinese hamster ovary (CHO) and baby hamster kidney (BHK) cells, respectively, and were incorporated into dioleoyl phosphatidylcholine liposomes (Avanti Polar Lipids). .. BHK cells (ATCC) highly expressing mCD48-GPI and hCD19-GPI were established. mCD48-GPI and hCD19-GPI were purified from the lysates by affinity column with anti-mCD48 antibody (HM48-1, Bio X cell) and anti-hCD19 antibody (4G7, Bio X cell), respectively. .. The expression level of each GPI-anchored protein on the planar bilayer was quantified using silica beads with a diameter of 5 μm (Bangs Laboratories).

    Article Title: Imaging of biphasic signalosomes constructed by checkpoint receptor 2B4 in conventional and CAR-T cells
    Article Snippet: The mouse MHC class II molecule I-Ek with a GPI anchor (I-Ek-GPI), the mouse MHC class I molecule H-2Kb with a GPI anchor (H-2Kb-GPI), and mouse ICAM-1 with a GPI anchor (ICAM-1-GPI) were purified from transfected Chinese hamster ovary (CHO) and baby hamster kidney (BHK) cells, respectively, and were incorporated into dioleoyl phosphatidylcholine liposomes (Avanti Polar Lipids). .. BHK cells (ATCC) highly expressing mCD48-GPI and hCD19-GPI were established. mCD48-GPI and hCD19-GPI were purified from the lysates by affinity column with antimCD48 antibody (HM48-1, Bio X cell) and anti-hCD19 antibody (4G7, Bio X cell), respectively. .. The expression level of each GPI-anchored protein on the planar bilayer was quantified using silica beads with a diameter of 5 μm (Bangs Laboratories) 28.

    Article Title: CD28-mediated linear and parallel costimulatory signaling cooperatively regulate CAR-T cell functions via CAR-CD28 microclusters
    Article Snippet: The mouse MHC class I molecule H-2K b with a GPI anchor (H-2K b –GPI), mouse ICAM-1 with a GPI anchor (ICAM-1–GPI), and mouse CD80 with a GPI anchor (CD80–GPI) were purified from transfected Chinese hamster ovary and baby hamster kidney cells, respectively, and were incorporated into dioleoyl phosphatidylcholine liposomes (Avanti Polar Lipids). .. BHK cells (ATCC) highly expressing hCD19–GPI were established. hCD19–GPI were purified from the lysates by affinity column with anti-hCD19 antibody (4G7, Bio X cell). .. The expression level of each GPI-anchored protein on the planar bilayer was quantified using silica beads with a diameter of 5 μm (Bangs Laboratories, SS05N) .

    Article Title: CD28-mediated linear and parallel costimulatory signaling cooperatively regulate CAR-T cell functions via CAR-CD28 microclusters.
    Article Snippet: The mouse MHC class I molecule H-2Kb with a GPI anchor (H-2Kb–GPI), mouse ICAM-1 with a GPI anchor (ICAM-1–GPI), and mouse CD80 with a GPI anchor (CD80–GPI) were purified from transfected Chinese hamster ovary and baby hamster kidney cells, respectively, and were incorporated into dioleoyl phosphatidylcholine liposomes (Avanti Polar Lipids). .. BHK cells (ATCC) highly expressing hCD19–GPIwere established. hCD19–GPI were purified from the lysates by affinity column with anti-hCD19 antibody (4G7, Bio X cell). .. The expression level of each GPI-anchored protein on the planar bilayer was quantified using silica beads with a diameter of 5 μm (Bangs Laboratories, SS05N)64.

    Purification:

    Article Title: Imaging of biphasic signalosomes constructed by checkpoint receptor 2B4 in conventional and chimeric antigen receptor-T cells
    Article Snippet: The mouse MHC class II molecule I-E k with a GPI anchor (I-E k -GPI), the mouse MHC class I molecule H-2K b with a GPI anchor (H-2K b -GPI), and mouse ICAM-1 with a GPI anchor (ICAM-1-GPI) were purified from transfected Chinese hamster ovary (CHO) and baby hamster kidney (BHK) cells, respectively, and were incorporated into dioleoyl phosphatidylcholine liposomes (Avanti Polar Lipids). .. BHK cells (ATCC) highly expressing mCD48-GPI and hCD19-GPI were established. mCD48-GPI and hCD19-GPI were purified from the lysates by affinity column with anti-mCD48 antibody (HM48-1, Bio X cell) and anti-hCD19 antibody (4G7, Bio X cell), respectively. .. The expression level of each GPI-anchored protein on the planar bilayer was quantified using silica beads with a diameter of 5 μm (Bangs Laboratories).

    Article Title: Imaging of biphasic signalosomes constructed by checkpoint receptor 2B4 in conventional and CAR-T cells
    Article Snippet: The mouse MHC class II molecule I-Ek with a GPI anchor (I-Ek-GPI), the mouse MHC class I molecule H-2Kb with a GPI anchor (H-2Kb-GPI), and mouse ICAM-1 with a GPI anchor (ICAM-1-GPI) were purified from transfected Chinese hamster ovary (CHO) and baby hamster kidney (BHK) cells, respectively, and were incorporated into dioleoyl phosphatidylcholine liposomes (Avanti Polar Lipids). .. BHK cells (ATCC) highly expressing mCD48-GPI and hCD19-GPI were established. mCD48-GPI and hCD19-GPI were purified from the lysates by affinity column with antimCD48 antibody (HM48-1, Bio X cell) and anti-hCD19 antibody (4G7, Bio X cell), respectively. .. The expression level of each GPI-anchored protein on the planar bilayer was quantified using silica beads with a diameter of 5 μm (Bangs Laboratories) 28.

    Article Title: CD28-mediated linear and parallel costimulatory signaling cooperatively regulate CAR-T cell functions via CAR-CD28 microclusters
    Article Snippet: The mouse MHC class I molecule H-2K b with a GPI anchor (H-2K b –GPI), mouse ICAM-1 with a GPI anchor (ICAM-1–GPI), and mouse CD80 with a GPI anchor (CD80–GPI) were purified from transfected Chinese hamster ovary and baby hamster kidney cells, respectively, and were incorporated into dioleoyl phosphatidylcholine liposomes (Avanti Polar Lipids). .. BHK cells (ATCC) highly expressing hCD19–GPI were established. hCD19–GPI were purified from the lysates by affinity column with anti-hCD19 antibody (4G7, Bio X cell). .. The expression level of each GPI-anchored protein on the planar bilayer was quantified using silica beads with a diameter of 5 μm (Bangs Laboratories, SS05N) .

    Article Title: CD28-mediated linear and parallel costimulatory signaling cooperatively regulate CAR-T cell functions via CAR-CD28 microclusters.
    Article Snippet: The mouse MHC class I molecule H-2Kb with a GPI anchor (H-2Kb–GPI), mouse ICAM-1 with a GPI anchor (ICAM-1–GPI), and mouse CD80 with a GPI anchor (CD80–GPI) were purified from transfected Chinese hamster ovary and baby hamster kidney cells, respectively, and were incorporated into dioleoyl phosphatidylcholine liposomes (Avanti Polar Lipids). .. BHK cells (ATCC) highly expressing hCD19–GPIwere established. hCD19–GPI were purified from the lysates by affinity column with anti-hCD19 antibody (4G7, Bio X cell). .. The expression level of each GPI-anchored protein on the planar bilayer was quantified using silica beads with a diameter of 5 μm (Bangs Laboratories, SS05N)64.

    Affinity Column:

    Article Title: Imaging of biphasic signalosomes constructed by checkpoint receptor 2B4 in conventional and chimeric antigen receptor-T cells
    Article Snippet: The mouse MHC class II molecule I-E k with a GPI anchor (I-E k -GPI), the mouse MHC class I molecule H-2K b with a GPI anchor (H-2K b -GPI), and mouse ICAM-1 with a GPI anchor (ICAM-1-GPI) were purified from transfected Chinese hamster ovary (CHO) and baby hamster kidney (BHK) cells, respectively, and were incorporated into dioleoyl phosphatidylcholine liposomes (Avanti Polar Lipids). .. BHK cells (ATCC) highly expressing mCD48-GPI and hCD19-GPI were established. mCD48-GPI and hCD19-GPI were purified from the lysates by affinity column with anti-mCD48 antibody (HM48-1, Bio X cell) and anti-hCD19 antibody (4G7, Bio X cell), respectively. .. The expression level of each GPI-anchored protein on the planar bilayer was quantified using silica beads with a diameter of 5 μm (Bangs Laboratories).

    Article Title: Imaging of biphasic signalosomes constructed by checkpoint receptor 2B4 in conventional and CAR-T cells
    Article Snippet: The mouse MHC class II molecule I-Ek with a GPI anchor (I-Ek-GPI), the mouse MHC class I molecule H-2Kb with a GPI anchor (H-2Kb-GPI), and mouse ICAM-1 with a GPI anchor (ICAM-1-GPI) were purified from transfected Chinese hamster ovary (CHO) and baby hamster kidney (BHK) cells, respectively, and were incorporated into dioleoyl phosphatidylcholine liposomes (Avanti Polar Lipids). .. BHK cells (ATCC) highly expressing mCD48-GPI and hCD19-GPI were established. mCD48-GPI and hCD19-GPI were purified from the lysates by affinity column with antimCD48 antibody (HM48-1, Bio X cell) and anti-hCD19 antibody (4G7, Bio X cell), respectively. .. The expression level of each GPI-anchored protein on the planar bilayer was quantified using silica beads with a diameter of 5 μm (Bangs Laboratories) 28.

    Article Title: CD28-mediated linear and parallel costimulatory signaling cooperatively regulate CAR-T cell functions via CAR-CD28 microclusters
    Article Snippet: The mouse MHC class I molecule H-2K b with a GPI anchor (H-2K b –GPI), mouse ICAM-1 with a GPI anchor (ICAM-1–GPI), and mouse CD80 with a GPI anchor (CD80–GPI) were purified from transfected Chinese hamster ovary and baby hamster kidney cells, respectively, and were incorporated into dioleoyl phosphatidylcholine liposomes (Avanti Polar Lipids). .. BHK cells (ATCC) highly expressing hCD19–GPI were established. hCD19–GPI were purified from the lysates by affinity column with anti-hCD19 antibody (4G7, Bio X cell). .. The expression level of each GPI-anchored protein on the planar bilayer was quantified using silica beads with a diameter of 5 μm (Bangs Laboratories, SS05N) .

    Article Title: CD28-mediated linear and parallel costimulatory signaling cooperatively regulate CAR-T cell functions via CAR-CD28 microclusters.
    Article Snippet: The mouse MHC class I molecule H-2Kb with a GPI anchor (H-2Kb–GPI), mouse ICAM-1 with a GPI anchor (ICAM-1–GPI), and mouse CD80 with a GPI anchor (CD80–GPI) were purified from transfected Chinese hamster ovary and baby hamster kidney cells, respectively, and were incorporated into dioleoyl phosphatidylcholine liposomes (Avanti Polar Lipids). .. BHK cells (ATCC) highly expressing hCD19–GPIwere established. hCD19–GPI were purified from the lysates by affinity column with anti-hCD19 antibody (4G7, Bio X cell). .. The expression level of each GPI-anchored protein on the planar bilayer was quantified using silica beads with a diameter of 5 μm (Bangs Laboratories, SS05N)64.



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    Image Search Results


    Reduced activation of FCRL3 + T cells. (A) Surface FCRL3 expression measured by FACS in human naïve and memory CD4 + T helper, Tregs, and CD8 + T lymphocytes freshly isolated from peripheral blood of one representative healthy donor. (B) FCRL3 expression in different T cell populations measured as in A. Each dot represents one donor. N = 6–9; mean ± SD; paired t test, two-tailed. From left to right: **P = 0.0033, *P = 0.0303, *P = 0.0255. (C–E) Surface staining for CD25 (C), OX40 (D), and CD40L (E) expression in sorted FCRL3 + and FCRL3 − CD8 + memory T cells after stimulation with plate-bound anti-CD3/CD28 for 48 h. Each dot represents a different donor, N = 4–10. Mean, paired t test or ratio paired t test, two-tailed. From left to right: (C)***P = 0.0008, ***P = 0.0003, (D)*P = 0.048, **P = 0.0071, (E)**P = 0.0064, **P = 0.0044. Data underlying this figure can be found in .

    Journal: The Journal of Experimental Medicine

    Article Title: FCRL3 is an immunoregulatory receptor that restrains the activation of human memory T lymphocytes

    doi: 10.1084/jem.20242474

    Figure Lengend Snippet: Reduced activation of FCRL3 + T cells. (A) Surface FCRL3 expression measured by FACS in human naïve and memory CD4 + T helper, Tregs, and CD8 + T lymphocytes freshly isolated from peripheral blood of one representative healthy donor. (B) FCRL3 expression in different T cell populations measured as in A. Each dot represents one donor. N = 6–9; mean ± SD; paired t test, two-tailed. From left to right: **P = 0.0033, *P = 0.0303, *P = 0.0255. (C–E) Surface staining for CD25 (C), OX40 (D), and CD40L (E) expression in sorted FCRL3 + and FCRL3 − CD8 + memory T cells after stimulation with plate-bound anti-CD3/CD28 for 48 h. Each dot represents a different donor, N = 4–10. Mean, paired t test or ratio paired t test, two-tailed. From left to right: (C)***P = 0.0008, ***P = 0.0003, (D)*P = 0.048, **P = 0.0071, (E)**P = 0.0064, **P = 0.0044. Data underlying this figure can be found in .

    Article Snippet: Prior to the coculture, JeKo cells were preincubated with the bispecific antibody against the human CD19 and CD3 antigens (InvivoGen) at a concentration of 2–3 ng/ml for 30 min at 37°C.

    Techniques: Activation Assay, Expressing, Isolation, Two Tailed Test, Staining, FACS

    FCRL3 expression is associated with the donors’ genotype. (A) Percentage of surface FCRL3 expression on sorted T cell subsets from genotyped healthy donors. Each dot represents one donor. Mean ± SD; unpaired t test, two-tailed. ***P = 0.0009, **P = 0.003. (B) gMFI of gated FCRL3 + CD8 + T memory cells from genotyped healthy donors. Each dot represents one donor. Results were normalized to the average of “T/T” donors. Mean ± SD; unpaired t test, two-tailed. No difference was statistically significant. (C) Examples of genotyping results by PCR and Sanger sequencing. (D) Examples of FCRL3 expression in donors carrying the indicated SNP variants in the FCRL3 promoter region (−169 bp from the transcription start site). For every donor, FCRL3 expression is shown in CD8 + and CD4 + T memory cells and Tregs. (E) Surface staining for the indicated AIMs (CD25, OX40, CD40L) in unstimulated CD8 + FCRL3 + and FCRL3 − memory T cells. gMFI, geometric mean fluorescence intensity. Data underlying this figure can be found in . Source data are available for this figure: .

    Journal: The Journal of Experimental Medicine

    Article Title: FCRL3 is an immunoregulatory receptor that restrains the activation of human memory T lymphocytes

    doi: 10.1084/jem.20242474

    Figure Lengend Snippet: FCRL3 expression is associated with the donors’ genotype. (A) Percentage of surface FCRL3 expression on sorted T cell subsets from genotyped healthy donors. Each dot represents one donor. Mean ± SD; unpaired t test, two-tailed. ***P = 0.0009, **P = 0.003. (B) gMFI of gated FCRL3 + CD8 + T memory cells from genotyped healthy donors. Each dot represents one donor. Results were normalized to the average of “T/T” donors. Mean ± SD; unpaired t test, two-tailed. No difference was statistically significant. (C) Examples of genotyping results by PCR and Sanger sequencing. (D) Examples of FCRL3 expression in donors carrying the indicated SNP variants in the FCRL3 promoter region (−169 bp from the transcription start site). For every donor, FCRL3 expression is shown in CD8 + and CD4 + T memory cells and Tregs. (E) Surface staining for the indicated AIMs (CD25, OX40, CD40L) in unstimulated CD8 + FCRL3 + and FCRL3 − memory T cells. gMFI, geometric mean fluorescence intensity. Data underlying this figure can be found in . Source data are available for this figure: .

    Article Snippet: Prior to the coculture, JeKo cells were preincubated with the bispecific antibody against the human CD19 and CD3 antigens (InvivoGen) at a concentration of 2–3 ng/ml for 30 min at 37°C.

    Techniques: Expressing, Two Tailed Test, Genotyping, DNA Sequencing, Staining, Fluorescence, FACS, Sequencing

    Phenotypic characterization of FCRL3 + T cells. (A) Surface staining for CD25 expression in FCRL3 + and FCRL3 − CD4 + memory T cells. Cells were stimulated with plate-bound anti-CD3/CD28 for 48 h. Each dot represents one donor, N = 5. Mean ± SD, paired t test, two-tailed. From left to right: *P = 0.0174, *P = 0.015. (B) Live/Dead staining showing viability of sorted FCRL3 + and FCRL3 − CD4 + and CD8 + memory T cells, 3 days after activation. Each dot represents a different donor, N = 3–6. Mean ± SD, paired t test, two-tailed. From left to right: *P = 0.0369, *P = 0.0272. (C) BrdU incorporation assay to measure the proliferation of sorted FCRL3 + and FCRL3 − CD4 + and CD8 + memory T cells, 3 days after activation. Each dot represents a different donor, N = 3–4. Mean ± SD, paired t test, two-tailed. *P = 0.0226, **P = 0.0093. (D) Heatmaps showing the differential expression of selected genes from . (E) Surface staining for CXCR5 expression in FCRL3 + and FCRL3 − CD8 + memory T cells. Each dot represents one donor, N = 8. Mean ± SD, paired t test, two-tailed. *P = 0.0221.

    Journal: The Journal of Experimental Medicine

    Article Title: FCRL3 is an immunoregulatory receptor that restrains the activation of human memory T lymphocytes

    doi: 10.1084/jem.20242474

    Figure Lengend Snippet: Phenotypic characterization of FCRL3 + T cells. (A) Surface staining for CD25 expression in FCRL3 + and FCRL3 − CD4 + memory T cells. Cells were stimulated with plate-bound anti-CD3/CD28 for 48 h. Each dot represents one donor, N = 5. Mean ± SD, paired t test, two-tailed. From left to right: *P = 0.0174, *P = 0.015. (B) Live/Dead staining showing viability of sorted FCRL3 + and FCRL3 − CD4 + and CD8 + memory T cells, 3 days after activation. Each dot represents a different donor, N = 3–6. Mean ± SD, paired t test, two-tailed. From left to right: *P = 0.0369, *P = 0.0272. (C) BrdU incorporation assay to measure the proliferation of sorted FCRL3 + and FCRL3 − CD4 + and CD8 + memory T cells, 3 days after activation. Each dot represents a different donor, N = 3–4. Mean ± SD, paired t test, two-tailed. *P = 0.0226, **P = 0.0093. (D) Heatmaps showing the differential expression of selected genes from . (E) Surface staining for CXCR5 expression in FCRL3 + and FCRL3 − CD8 + memory T cells. Each dot represents one donor, N = 8. Mean ± SD, paired t test, two-tailed. *P = 0.0221.

    Article Snippet: Prior to the coculture, JeKo cells were preincubated with the bispecific antibody against the human CD19 and CD3 antigens (InvivoGen) at a concentration of 2–3 ng/ml for 30 min at 37°C.

    Techniques: Staining, Expressing, Two Tailed Test, Activation Assay, BrdU Incorporation Assay, Quantitative Proteomics, FACS

    FCRL3 expression is sufficient to modulate T cell activation. (A) Schematic representation of the experimental setup for repetitive T cell stimulation. Memory CD8 + T cells isolated from peripheral blood were activated on plate-bound anti-CD3 antibody with or without costimulation with an anti-CD28 antibody or restimulation. (B and C) Surface expression of FCRL3 measured 5 days after activation with anti-CD3/CD28 or with two consecutive stimulations (day 0 and day 3) with anti-CD3 antibody. One representative donor (B) and N = 8 independent experiments (C) are shown. Each dot represents one donor. Mean; paired t test, two-tailed. *P = 0.0147. (D) Schematic representation of the FCRL3 locus, with the exons (4 and 7) targeted by the selected sgRNAs. Only the exons targeted by the sgRNAs are shown. (E) Experimental workflow for FCRL3 CRISPR-KO in FCRL3 + CD8 + T cells. (F–H) FCRL3 (F), CD25 (G), and CD137 (H) expression on CD8 + FCRL3 + cells, transfected with RNPs containing sgRNAs targeting the FCRL3 gene, or control. Surface expression was monitored 10 days after reactivation with anti-CD3. N = 6–8 independent experiments. Mean; paired t test, two-tailed. From left to right: (F) ***P = 0.001, (G) *P = 0.0145, (H) *P = 0.0171. (I) Experimental workflow for the ectopic expression of FCRL3 in CD8 + T cells, by lentiviral transduction. After activation with plate-bound anti-CD3/CD28 and transduction, transduced cells are selected by puromycin treatment, followed by recovery and surface staining for FCRL3 and AIMs. (J) Surface expression of FCRL3 in memory CD8 + T cells transduced with FCRL3-encoding lentivirus or empty lentivirus as a control. One representative experiment of N = 4. (K) Surface CD25 expression in FCRL3-transduced memory CD8 + T cells, 6 and 10 days after the initial activation. The left panel shows the result for one representative donors, while the bar graphs represent the results for N = 4 independent donors at day 10. The gMFI and percentage of CD25 + cells are both shown. Mean ± SD; paired t test, two-tailed. *P = 0.0306, **P = 0.0076. Data underlying this figure can be found in .

    Journal: The Journal of Experimental Medicine

    Article Title: FCRL3 is an immunoregulatory receptor that restrains the activation of human memory T lymphocytes

    doi: 10.1084/jem.20242474

    Figure Lengend Snippet: FCRL3 expression is sufficient to modulate T cell activation. (A) Schematic representation of the experimental setup for repetitive T cell stimulation. Memory CD8 + T cells isolated from peripheral blood were activated on plate-bound anti-CD3 antibody with or without costimulation with an anti-CD28 antibody or restimulation. (B and C) Surface expression of FCRL3 measured 5 days after activation with anti-CD3/CD28 or with two consecutive stimulations (day 0 and day 3) with anti-CD3 antibody. One representative donor (B) and N = 8 independent experiments (C) are shown. Each dot represents one donor. Mean; paired t test, two-tailed. *P = 0.0147. (D) Schematic representation of the FCRL3 locus, with the exons (4 and 7) targeted by the selected sgRNAs. Only the exons targeted by the sgRNAs are shown. (E) Experimental workflow for FCRL3 CRISPR-KO in FCRL3 + CD8 + T cells. (F–H) FCRL3 (F), CD25 (G), and CD137 (H) expression on CD8 + FCRL3 + cells, transfected with RNPs containing sgRNAs targeting the FCRL3 gene, or control. Surface expression was monitored 10 days after reactivation with anti-CD3. N = 6–8 independent experiments. Mean; paired t test, two-tailed. From left to right: (F) ***P = 0.001, (G) *P = 0.0145, (H) *P = 0.0171. (I) Experimental workflow for the ectopic expression of FCRL3 in CD8 + T cells, by lentiviral transduction. After activation with plate-bound anti-CD3/CD28 and transduction, transduced cells are selected by puromycin treatment, followed by recovery and surface staining for FCRL3 and AIMs. (J) Surface expression of FCRL3 in memory CD8 + T cells transduced with FCRL3-encoding lentivirus or empty lentivirus as a control. One representative experiment of N = 4. (K) Surface CD25 expression in FCRL3-transduced memory CD8 + T cells, 6 and 10 days after the initial activation. The left panel shows the result for one representative donors, while the bar graphs represent the results for N = 4 independent donors at day 10. The gMFI and percentage of CD25 + cells are both shown. Mean ± SD; paired t test, two-tailed. *P = 0.0306, **P = 0.0076. Data underlying this figure can be found in .

    Article Snippet: Prior to the coculture, JeKo cells were preincubated with the bispecific antibody against the human CD19 and CD3 antigens (InvivoGen) at a concentration of 2–3 ng/ml for 30 min at 37°C.

    Techniques: Expressing, Activation Assay, Cell Stimulation, Isolation, Two Tailed Test, CRISPR, Transfection, Control, Transduction, Staining, FACS, Over Expression

    Overlapping TCR repertoire in FCRL3 + and FCRL3 − cells. (A) Number of unique productive TCR Vβ rearrangements (left) and Simpson’s clonality index (right) in sorted CD8 + FCRL3 + and CD8 + FCRL3 − memory T cell subsets from three healthy donors (D1–D3) (initial input, 5 × 10 5 cells per subset). Each dot represents one donor ( N = 3). Mean ± SD. (B) TCR Vβ gene family usage by CD8 + FCRL3 + (upper panel) and CD8 + FCRL3 − memory T cells (lower panel) in three healthy donors. Slices in the chart represent different Vβ gene families, and their size is proportional to the frequency of clonotypes using that segment. The color-coded legend is reported for the 26 different Vβ gene family. The total number of clonotypes is indicated at the center of the pie chart. (C) Percentage of clonotypes bearing the same CDR3β length defined by the number of nucleotides. The CDR3β length of TCR Vβ clonotypes from CD8 + FCRL3 + and CD8 + FCRL3 − memory T cells is shown in lavender and gray, respectively. Each dot represents one donor ( N = 3). Mean ± SD. (D) Pairwise comparison of TCR Vβ clonotype frequency distribution in CD8 + FCRL3 + memory T cells (x axis) and CD8 + FCRL3 − memory T cells (y axis) from N = 3 donors. Frequencies are shown as a percentage of productive templates. Each dot indicates a unique TCR Vβ clonotype. Dots outside the dashed lines represent clonotypes that were found in only one of the two samples and that were assigned an arbitrary frequency value for graphical purposes. The total number of clonotypes is indicated in the x and y axes. Values in the upper right corner represent the number of clonotypes shared between two samples. (E) Unique and shared TCR Vβ clonotypes between CD8 + FCRL3 + and CD8 + FCRL3 − memory T cells in N = 3 donors. Shown are the percentage of clonotypes based on the Jaccard index (left) and their cumulative frequency (right). Data underlying this figure can be found in .

    Journal: The Journal of Experimental Medicine

    Article Title: FCRL3 is an immunoregulatory receptor that restrains the activation of human memory T lymphocytes

    doi: 10.1084/jem.20242474

    Figure Lengend Snippet: Overlapping TCR repertoire in FCRL3 + and FCRL3 − cells. (A) Number of unique productive TCR Vβ rearrangements (left) and Simpson’s clonality index (right) in sorted CD8 + FCRL3 + and CD8 + FCRL3 − memory T cell subsets from three healthy donors (D1–D3) (initial input, 5 × 10 5 cells per subset). Each dot represents one donor ( N = 3). Mean ± SD. (B) TCR Vβ gene family usage by CD8 + FCRL3 + (upper panel) and CD8 + FCRL3 − memory T cells (lower panel) in three healthy donors. Slices in the chart represent different Vβ gene families, and their size is proportional to the frequency of clonotypes using that segment. The color-coded legend is reported for the 26 different Vβ gene family. The total number of clonotypes is indicated at the center of the pie chart. (C) Percentage of clonotypes bearing the same CDR3β length defined by the number of nucleotides. The CDR3β length of TCR Vβ clonotypes from CD8 + FCRL3 + and CD8 + FCRL3 − memory T cells is shown in lavender and gray, respectively. Each dot represents one donor ( N = 3). Mean ± SD. (D) Pairwise comparison of TCR Vβ clonotype frequency distribution in CD8 + FCRL3 + memory T cells (x axis) and CD8 + FCRL3 − memory T cells (y axis) from N = 3 donors. Frequencies are shown as a percentage of productive templates. Each dot indicates a unique TCR Vβ clonotype. Dots outside the dashed lines represent clonotypes that were found in only one of the two samples and that were assigned an arbitrary frequency value for graphical purposes. The total number of clonotypes is indicated in the x and y axes. Values in the upper right corner represent the number of clonotypes shared between two samples. (E) Unique and shared TCR Vβ clonotypes between CD8 + FCRL3 + and CD8 + FCRL3 − memory T cells in N = 3 donors. Shown are the percentage of clonotypes based on the Jaccard index (left) and their cumulative frequency (right). Data underlying this figure can be found in .

    Article Snippet: Prior to the coculture, JeKo cells were preincubated with the bispecific antibody against the human CD19 and CD3 antigens (InvivoGen) at a concentration of 2–3 ng/ml for 30 min at 37°C.

    Techniques: Comparison

    FCRL3 expression is causally associated with reduced T cell activation. (A) Expression of FCRL3 , TOX , and EOMES transcripts was measured in FCRL3-KO and control cells by RT-qPCR. Transcript expression was normalized to the housekeeping gene UBE2D2 and is shown relative to control samples. N = 5 independent experiments; paired t test, two-tailed. From left to right: ****P < 0.0001, *P = 0.0475, **P = 0.0040. (B) Surface expression of FCRL3 in Jurkat T cells transduced with FCRL3-lentivirus or empty lentivirus as a control. One representative experiment of N = 4 is shown. (C and D) Surface CD25 expression in FCRL3-transduced Jurkat cells. Cells were stimulated for 24 h with anti-CD3/CD28-coated beads. One representative experiment is shown in C, while the bar graphs in D show the compiled results of N = 4 independent experiments. The gMFI and percentage of CD25 + cells are both shown. Mean ± SEM; paired t test, two-tailed. From left to right: **P = 0.0052, **P = 0.0033. (E) Schematic representation of the EGFR-FCRL3 chimeric proteins. (F) Western blot showing the expression of the different truncated chimeric proteins in Jurkat T cells, using an anti-EGFR antibody. Data are representative of N = 2 experiments. (G) Surface expression of the EGFR in Jurkat T cells transduced to ectopically express the EGFR-FCRL3 chimeras. Data are representative of N = 4 independent experiments. (H) Surface expression of CD25 in Jurkat T cells expressing the EGFR-FCRL3 chimeras. Cells were stimulated for 24 h with anti-CD3/CD28-coated beads. One representative experiment is shown. (I) Surface expression of CD25 in stimulated (24 h) Jurkat T cells expressing the EGFR-FCRL3 chimeras. The compiled results of N = 4 independent experiments as in E are shown. Mean ± SEM; paired t test, two-tailed. Left panel, from top to bottom: ***P = 0.0007, *P = 0.0151, **P = 0.0046, ***P = 0.0002, **P = 0.0030, **P = 0.0013. Right panel, from top to bottom: ***P = 0.0001, n.s. (not significant), P = 0.1005, *P = 0.0326, **P = 0.0014, **P = 0.0050, *P = 0.0157. Data underlying this figure can be found in . Source data are available for this figure: .

    Journal: The Journal of Experimental Medicine

    Article Title: FCRL3 is an immunoregulatory receptor that restrains the activation of human memory T lymphocytes

    doi: 10.1084/jem.20242474

    Figure Lengend Snippet: FCRL3 expression is causally associated with reduced T cell activation. (A) Expression of FCRL3 , TOX , and EOMES transcripts was measured in FCRL3-KO and control cells by RT-qPCR. Transcript expression was normalized to the housekeeping gene UBE2D2 and is shown relative to control samples. N = 5 independent experiments; paired t test, two-tailed. From left to right: ****P < 0.0001, *P = 0.0475, **P = 0.0040. (B) Surface expression of FCRL3 in Jurkat T cells transduced with FCRL3-lentivirus or empty lentivirus as a control. One representative experiment of N = 4 is shown. (C and D) Surface CD25 expression in FCRL3-transduced Jurkat cells. Cells were stimulated for 24 h with anti-CD3/CD28-coated beads. One representative experiment is shown in C, while the bar graphs in D show the compiled results of N = 4 independent experiments. The gMFI and percentage of CD25 + cells are both shown. Mean ± SEM; paired t test, two-tailed. From left to right: **P = 0.0052, **P = 0.0033. (E) Schematic representation of the EGFR-FCRL3 chimeric proteins. (F) Western blot showing the expression of the different truncated chimeric proteins in Jurkat T cells, using an anti-EGFR antibody. Data are representative of N = 2 experiments. (G) Surface expression of the EGFR in Jurkat T cells transduced to ectopically express the EGFR-FCRL3 chimeras. Data are representative of N = 4 independent experiments. (H) Surface expression of CD25 in Jurkat T cells expressing the EGFR-FCRL3 chimeras. Cells were stimulated for 24 h with anti-CD3/CD28-coated beads. One representative experiment is shown. (I) Surface expression of CD25 in stimulated (24 h) Jurkat T cells expressing the EGFR-FCRL3 chimeras. The compiled results of N = 4 independent experiments as in E are shown. Mean ± SEM; paired t test, two-tailed. Left panel, from top to bottom: ***P = 0.0007, *P = 0.0151, **P = 0.0046, ***P = 0.0002, **P = 0.0030, **P = 0.0013. Right panel, from top to bottom: ***P = 0.0001, n.s. (not significant), P = 0.1005, *P = 0.0326, **P = 0.0014, **P = 0.0050, *P = 0.0157. Data underlying this figure can be found in . Source data are available for this figure: .

    Article Snippet: Prior to the coculture, JeKo cells were preincubated with the bispecific antibody against the human CD19 and CD3 antigens (InvivoGen) at a concentration of 2–3 ng/ml for 30 min at 37°C.

    Techniques: Expressing, Activation Assay, Control, Quantitative RT-PCR, Two Tailed Test, Transduction, Western Blot, FACS

    Intracellular portion of FCRL3 interacts with UBASH3A. (A) Schematic representation of the Protein A-TurboID experimental workflow. An anti-FCRL3 antibody was added to FCRL3-expressing or nonexpressing Jurkat cells followed by the addition of Protein A-TurboID fusion protein and biotin. After biotinylation and extensive washing, biotinylated proteins were recovered using streptavidin-conjugated beads and subjected to mass spectrometry analysis. (B) Example of Protein A-TurboID experiment. After incubation of FCRL3-expressing Jurkat cells (or empty vector control cells) with a mouse monoclonal anti-FCRL3 antibody together with recombinant Protein A-TurboID, biotinylated proteins were recovered by streptavidin pull-down, followed by western blot to confirm enrichment of FCRL3. The antibody used for immunoblot was a rabbit polyclonal anti-FCRL3. (C) Differentially retrieved proteins following FCRL3 Protein A-TurboID and mass spectrometry of FCRL3-expressing versus nonexpressing cells (log 2 FC ≥ |2|; P ≤0.01; N = 5 replicates). (D) Example of FCRL3 IP. Jurkat cells expressing FCRL3 (or empty vector control) were lysed, and 1.5 mg of protein extract was used for IP with a rabbit polyclonal anti-FCRL3 antibody, followed by western blot with the same antibody. Two independent representative FCRL3 IPs are shown. (E) Differentially retrieved proteins following FCRL3 IP-MS of FCRL3-expressing versus nonexpressing cells (log 2 FC ≥ |2|; P ≤0.01; N = 4 replicates). (F) Co-IP of FCRL3 with UBASH3A. HEK cells were transfected with the indicated plasmids, followed by IP of FCRL3 using a rabbit polyclonal anti-FCRL3 antibody and immunoblot for either UBASH3A (top) or FCRL3 itself (bottom). Data are representative of N = 2 independent experiments. (G) Schematic representation of the FCRL3 protein and C-terminal truncations (top). All proteins were efficiently expressed, as shown by western blot (bottom). (H) Heatmap showing the differentially enriched proteins by IP-MS of Jurkat cells transduced with full-length FCRL3 and its truncations. N = 3–4 independent samples. Significant differences between multiple experimental conditions were assessed with an ANOVA multiple-sample test (0.01 permutation-based FDR cut-off, 250 randomizations). Significant proteins were filtered, and Z-score normalization was applied to each protein across all samples. Unsupervised hierarchical clustering was employed to visualize on a heatmap proteins with a positive Z-score value for all replicates of the conditions LV-FCRL3 and LV-FCRL3-93 aa. IP, immunoprecipitation. Data underlying this figure can be found in . Source data are available for this figure: .

    Journal: The Journal of Experimental Medicine

    Article Title: FCRL3 is an immunoregulatory receptor that restrains the activation of human memory T lymphocytes

    doi: 10.1084/jem.20242474

    Figure Lengend Snippet: Intracellular portion of FCRL3 interacts with UBASH3A. (A) Schematic representation of the Protein A-TurboID experimental workflow. An anti-FCRL3 antibody was added to FCRL3-expressing or nonexpressing Jurkat cells followed by the addition of Protein A-TurboID fusion protein and biotin. After biotinylation and extensive washing, biotinylated proteins were recovered using streptavidin-conjugated beads and subjected to mass spectrometry analysis. (B) Example of Protein A-TurboID experiment. After incubation of FCRL3-expressing Jurkat cells (or empty vector control cells) with a mouse monoclonal anti-FCRL3 antibody together with recombinant Protein A-TurboID, biotinylated proteins were recovered by streptavidin pull-down, followed by western blot to confirm enrichment of FCRL3. The antibody used for immunoblot was a rabbit polyclonal anti-FCRL3. (C) Differentially retrieved proteins following FCRL3 Protein A-TurboID and mass spectrometry of FCRL3-expressing versus nonexpressing cells (log 2 FC ≥ |2|; P ≤0.01; N = 5 replicates). (D) Example of FCRL3 IP. Jurkat cells expressing FCRL3 (or empty vector control) were lysed, and 1.5 mg of protein extract was used for IP with a rabbit polyclonal anti-FCRL3 antibody, followed by western blot with the same antibody. Two independent representative FCRL3 IPs are shown. (E) Differentially retrieved proteins following FCRL3 IP-MS of FCRL3-expressing versus nonexpressing cells (log 2 FC ≥ |2|; P ≤0.01; N = 4 replicates). (F) Co-IP of FCRL3 with UBASH3A. HEK cells were transfected with the indicated plasmids, followed by IP of FCRL3 using a rabbit polyclonal anti-FCRL3 antibody and immunoblot for either UBASH3A (top) or FCRL3 itself (bottom). Data are representative of N = 2 independent experiments. (G) Schematic representation of the FCRL3 protein and C-terminal truncations (top). All proteins were efficiently expressed, as shown by western blot (bottom). (H) Heatmap showing the differentially enriched proteins by IP-MS of Jurkat cells transduced with full-length FCRL3 and its truncations. N = 3–4 independent samples. Significant differences between multiple experimental conditions were assessed with an ANOVA multiple-sample test (0.01 permutation-based FDR cut-off, 250 randomizations). Significant proteins were filtered, and Z-score normalization was applied to each protein across all samples. Unsupervised hierarchical clustering was employed to visualize on a heatmap proteins with a positive Z-score value for all replicates of the conditions LV-FCRL3 and LV-FCRL3-93 aa. IP, immunoprecipitation. Data underlying this figure can be found in . Source data are available for this figure: .

    Article Snippet: Prior to the coculture, JeKo cells were preincubated with the bispecific antibody against the human CD19 and CD3 antigens (InvivoGen) at a concentration of 2–3 ng/ml for 30 min at 37°C.

    Techniques: Expressing, Biotinylation, Mass Spectrometry, Analysis, Incubation, Plasmid Preparation, Control, Recombinant, Pull Down Assay, Western Blot, Protein-Protein interactions, Co-Immunoprecipitation Assay, Transfection, Transduction, Immunoprecipitation, Significance Assay

    Journal: The Journal of Experimental Medicine

    Article Title: FCRL3 is an immunoregulatory receptor that restrains the activation of human memory T lymphocytes

    doi: 10.1084/jem.20242474

    Figure Lengend Snippet:

    Article Snippet: Prior to the coculture, JeKo cells were preincubated with the bispecific antibody against the human CD19 and CD3 antigens (InvivoGen) at a concentration of 2–3 ng/ml for 30 min at 37°C.

    Techniques: