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STEMCELL Technologies Inc human peripheral blood mononuclear cells pbmcs
Human Peripheral Blood Mononuclear Cells Pbmcs, supplied by STEMCELL Technologies Inc, used in various techniques. Bioz Stars score: 93/100, based on 7 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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Average 93 stars, based on 7 article reviews
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human peripheral blood mononuclear cells pbmcs - by Bioz Stars, 2020-09
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Article Title: Three-Dimensional Explant Platform for Studies on Choroid Plexus Epithelium
Article Snippet: .. In brief, DCs were prepared from human peripheral blood mononuclear cells (PBMCs) isolated from buffy coats (obtained from healthy donors), by centrifugation on a Lymphoprep gradient (STEMCELL technologies). .. Monocytes were isolated from PBMC by plastic adherence.

Article Title: Exploratory examination of inflammation state, immune response and blood cell composition in a human obese cohort to identify potential markers predicting cancer risk
Article Snippet: .. Immunophenotyping Human peripheral blood mononuclear cells (PBMCs) were isolated from heparinized whole blood by density gradient centrifugation with Lymphoprep (StemCell Technologies, Vancouver, BC). .. The PBMCs were stained with GhostDye Violet 450 viability dye (Tonbo Biosciences, San Diego, CA) for 30 min at 4ºC, washed once with PBS containing 2% FBS and 0.05% sodium azide (PFN), and blocked with anti-human CD32 Clone IV.3 (StemCell Technologies, Vancouver, BC) for 15 min at 23°C.

Article Title: Combined DLL3-targeted bispecific antibody with PD-1 inhibition is efficient to suppress small cell lung cancer growth
Article Snippet: .. Human peripheral blood mononuclear cells (PBMC) were isolated from the whole blood of healthy donors (Wuhan Blood Center) by a Ficoll separation method (Stem Cell Technologies, Vancouver, British Columbia, Canada). .. PBMC were cultured in RPMI 1640 medium supplemented with 200 IU/mL human recombinant interleukin (IL)-2 and activated by Dynabeads CD3/CD28 human T-activator (Cat. 11 131D, ThermoFisher, Waltham, Massachusetts) for 3 days according to the manufacturer’s instruction.

Article Title: Systemic Antibiotic Therapy Reduces Circulating Inflammatory Dendritic Cells and Treg-Th17 Plasticity in Periodontitis.
Article Snippet: .. Human peripheral blood mononuclear cells (PBMCs) were isolated from whole blood by density gradient centrifugation using Lymphoprep™ (StemCell Technologies, Vancouver, BC, Canada). .. Isolation and phenotypic analysis of blood DCsCirculating blood DCs were enriched from PBMCs by negative selection using human panDC enrichment kit (StemCell Technologies, Vancouver, BC, Canada).

Centrifugation:

Article Title: Three-Dimensional Explant Platform for Studies on Choroid Plexus Epithelium
Article Snippet: .. In brief, DCs were prepared from human peripheral blood mononuclear cells (PBMCs) isolated from buffy coats (obtained from healthy donors), by centrifugation on a Lymphoprep gradient (STEMCELL technologies). .. Monocytes were isolated from PBMC by plastic adherence.

Gradient Centrifugation:

Article Title: Exploratory examination of inflammation state, immune response and blood cell composition in a human obese cohort to identify potential markers predicting cancer risk
Article Snippet: .. Immunophenotyping Human peripheral blood mononuclear cells (PBMCs) were isolated from heparinized whole blood by density gradient centrifugation with Lymphoprep (StemCell Technologies, Vancouver, BC). .. The PBMCs were stained with GhostDye Violet 450 viability dye (Tonbo Biosciences, San Diego, CA) for 30 min at 4ºC, washed once with PBS containing 2% FBS and 0.05% sodium azide (PFN), and blocked with anti-human CD32 Clone IV.3 (StemCell Technologies, Vancouver, BC) for 15 min at 23°C.

Article Title: Systemic Antibiotic Therapy Reduces Circulating Inflammatory Dendritic Cells and Treg-Th17 Plasticity in Periodontitis.
Article Snippet: .. Human peripheral blood mononuclear cells (PBMCs) were isolated from whole blood by density gradient centrifugation using Lymphoprep™ (StemCell Technologies, Vancouver, BC, Canada). .. Isolation and phenotypic analysis of blood DCsCirculating blood DCs were enriched from PBMCs by negative selection using human panDC enrichment kit (StemCell Technologies, Vancouver, BC, Canada).

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    STEMCELL Technologies Inc human b lymphocytes peripheral blood mononuclear cells pbmcs
    Endogenous expression of C3 is very low in <t>human</t> B cells. (A) RNA was isolated from malignant B cell lines and blood-derived B cells, reverse-transcribed and analyzed for C3 expression by qPCR. As positive control, blood-derived T cells, <t>PBMCs,</t> and total, liver tissue RNA were used. Data were normalized to the housekeeping HPRT gene and are shown as mean 2-dCt values with SD of three independent experiments. (B) The presence of full length C3 mRNA was confirmed by primer pairs, covering the whole region of human C3 coding sequence. As positive control, liver tissue RNA was used. Data shown are representative of three independent experiments. Numbers indicate DNA length in base pair (bp). The start positions of forward (Fw) and reverse (Rv) primers are shown under the gel picture. (C) Western blot results analyzing endogenous C3 expression of human B cells. Lysates prepared from the human B cell lines, Raji and Namalwa and blood-derived B cells and PBMCs were analyzed for the presence of C3 by Western blot with the goat polyclonal anti-C3 antibody from Quidel under non reducing and reducing conditions. As positive control, lysate of Raji cells incubated with 10% NHS in EDTA-GVB buffer was used. Results shown are representative of five independent experiments.
    Human B Lymphocytes Peripheral Blood Mononuclear Cells Pbmcs, supplied by STEMCELL Technologies Inc, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/human b lymphocytes peripheral blood mononuclear cells pbmcs/product/STEMCELL Technologies Inc
    Average 93 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    human b lymphocytes peripheral blood mononuclear cells pbmcs - by Bioz Stars, 2020-09
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    84
    STEMCELL Technologies Inc hiv infection pbmcs
    <t>HIV</t> egress from the brain into the periphery in the adult astrocyte xenotransplantation model. Adult NSG mice were xenotransplanted with HIV- or HIV+ NHAs as described in Fig 2C and HIV detection outside of the brain was evaluated by Real-time PCR products (HIV DNA, A , or HIV RNA, B ) from brain, cervical lymph node, spleen, peripheral lymph node, splenocytes outgrowth assay and splenocytes outgrowth assay supernatant added to fresh <t>PBMCs</t> analyzed by electrophoresis and human GAPDH. Each column indicates individual animal. HIV- animals are shown to left of ladder, HIV+ animals shown to right for all gels shown. No template control (NTC) is the negative control. ( C ) GFP expression in splenocytes at day 14 day of culturing the cells and ( D ) flow cytometry of cultured splenocytes stained for CD3+/CD4+/GFP; dot blots (left) and cumulative data on right ( p = 0.04, Mann-Whitney U-test). ( E ) Representative image of supernatant from splenocytes cultured for 14 days from neonates from HIV- animal (top) or HIV+ animal (bottom) on fresh PBMCs stimulated with soluble α-CD3 and α-CD28 IL-2. Egress was analyzed in n = 7 (HIV-) and 9 (HIV+) mice; n = 3 (HIV-) and 5 (HIV+) representative mice are shown here.
    Hiv Infection Pbmcs, supplied by STEMCELL Technologies Inc, used in various techniques. Bioz Stars score: 84/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/hiv infection pbmcs/product/STEMCELL Technologies Inc
    Average 84 stars, based on 2 article reviews
    Price from $9.99 to $1999.99
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    94
    STEMCELL Technologies Inc pbmc
    Cytokine release by AFM11-His is strictly dependent on the presence of CD19 + target cells. ( A ) Cytokine release in cultures of <t>PBMC,</t> B cell-depleted PBMC, and enriched human T cells. <t>Unfractionated</t> human PBMC, B cell-depleted PBMC, and enriched T cells were cultured in the presence of: i) 10 µg/mL AFM11-His or OKT3, ii) 100 µg/mL PHA, or iii) without antibodies (w/o). After 48 h incubation IL-2, IL-6, IL-10, TNF, and IFN-γ in the cell-free culture supernatant were quantified by multiplexing and the results of representative experiments were plotted. ( B ) TNF, ( C ) IL-2, ( D ) IL-6, and ( E ) IFN-γ levels in culture supernatants of PBMC from 7 individual donors. Cytokine levels were determined by Luminex upon stimulation with antibodies in solution. PBMC were either stimulated with AFM11 (open circles) or OKT3 (closed triangles) at the indicated concentrations. IL-2 and TNF levels were measured after 24 h, IL-6 and IFN-γ levels were measured after 48 h. Background (asterisks) was measured in cultures supplemented with vehicle (formulation buffer) corresponding to the volume of highest AFM11 concentration used. Horizontal bars indicate mean values across the 7 donors.
    Pbmc, supplied by STEMCELL Technologies Inc, used in various techniques. Bioz Stars score: 94/100, based on 290 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/pbmc/product/STEMCELL Technologies Inc
    Average 94 stars, based on 290 article reviews
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    Endogenous expression of C3 is very low in human B cells. (A) RNA was isolated from malignant B cell lines and blood-derived B cells, reverse-transcribed and analyzed for C3 expression by qPCR. As positive control, blood-derived T cells, PBMCs, and total, liver tissue RNA were used. Data were normalized to the housekeeping HPRT gene and are shown as mean 2-dCt values with SD of three independent experiments. (B) The presence of full length C3 mRNA was confirmed by primer pairs, covering the whole region of human C3 coding sequence. As positive control, liver tissue RNA was used. Data shown are representative of three independent experiments. Numbers indicate DNA length in base pair (bp). The start positions of forward (Fw) and reverse (Rv) primers are shown under the gel picture. (C) Western blot results analyzing endogenous C3 expression of human B cells. Lysates prepared from the human B cell lines, Raji and Namalwa and blood-derived B cells and PBMCs were analyzed for the presence of C3 by Western blot with the goat polyclonal anti-C3 antibody from Quidel under non reducing and reducing conditions. As positive control, lysate of Raji cells incubated with 10% NHS in EDTA-GVB buffer was used. Results shown are representative of five independent experiments.

    Journal: Frontiers in Immunology

    Article Title: Interaction of Serum-Derived and Internalized C3 With DNA in Human B Cells—A Potential Involvement in Regulation of Gene Transcription

    doi: 10.3389/fimmu.2019.00493

    Figure Lengend Snippet: Endogenous expression of C3 is very low in human B cells. (A) RNA was isolated from malignant B cell lines and blood-derived B cells, reverse-transcribed and analyzed for C3 expression by qPCR. As positive control, blood-derived T cells, PBMCs, and total, liver tissue RNA were used. Data were normalized to the housekeeping HPRT gene and are shown as mean 2-dCt values with SD of three independent experiments. (B) The presence of full length C3 mRNA was confirmed by primer pairs, covering the whole region of human C3 coding sequence. As positive control, liver tissue RNA was used. Data shown are representative of three independent experiments. Numbers indicate DNA length in base pair (bp). The start positions of forward (Fw) and reverse (Rv) primers are shown under the gel picture. (C) Western blot results analyzing endogenous C3 expression of human B cells. Lysates prepared from the human B cell lines, Raji and Namalwa and blood-derived B cells and PBMCs were analyzed for the presence of C3 by Western blot with the goat polyclonal anti-C3 antibody from Quidel under non reducing and reducing conditions. As positive control, lysate of Raji cells incubated with 10% NHS in EDTA-GVB buffer was used. Results shown are representative of five independent experiments.

    Article Snippet: Isolation of Human B Lymphocytes Peripheral blood mononuclear cells (PBMCs) were isolated by Lymphoprep (Stemcell Technologies) density gradient centrifugation from superfluous buffy coat obtained from the Medical Service (Clinical Immunology and Transfusion Medicine, Lund) according to standard procedures ( ) and permit granted by the local ethics committee of Lund.

    Techniques: Expressing, Isolation, Derivative Assay, Real-time Polymerase Chain Reaction, Positive Control, Sequencing, Western Blot, Incubation

    HIV egress from the brain into the periphery in the adult astrocyte xenotransplantation model. Adult NSG mice were xenotransplanted with HIV- or HIV+ NHAs as described in Fig 2C and HIV detection outside of the brain was evaluated by Real-time PCR products (HIV DNA, A , or HIV RNA, B ) from brain, cervical lymph node, spleen, peripheral lymph node, splenocytes outgrowth assay and splenocytes outgrowth assay supernatant added to fresh PBMCs analyzed by electrophoresis and human GAPDH. Each column indicates individual animal. HIV- animals are shown to left of ladder, HIV+ animals shown to right for all gels shown. No template control (NTC) is the negative control. ( C ) GFP expression in splenocytes at day 14 day of culturing the cells and ( D ) flow cytometry of cultured splenocytes stained for CD3+/CD4+/GFP; dot blots (left) and cumulative data on right ( p = 0.04, Mann-Whitney U-test). ( E ) Representative image of supernatant from splenocytes cultured for 14 days from neonates from HIV- animal (top) or HIV+ animal (bottom) on fresh PBMCs stimulated with soluble α-CD3 and α-CD28 IL-2. Egress was analyzed in n = 7 (HIV-) and 9 (HIV+) mice; n = 3 (HIV-) and 5 (HIV+) representative mice are shown here.

    Journal: PLoS Pathogens

    Article Title: HIV infects astrocytes in vivo and egresses from the brain to the periphery

    doi: 10.1371/journal.ppat.1008381

    Figure Lengend Snippet: HIV egress from the brain into the periphery in the adult astrocyte xenotransplantation model. Adult NSG mice were xenotransplanted with HIV- or HIV+ NHAs as described in Fig 2C and HIV detection outside of the brain was evaluated by Real-time PCR products (HIV DNA, A , or HIV RNA, B ) from brain, cervical lymph node, spleen, peripheral lymph node, splenocytes outgrowth assay and splenocytes outgrowth assay supernatant added to fresh PBMCs analyzed by electrophoresis and human GAPDH. Each column indicates individual animal. HIV- animals are shown to left of ladder, HIV+ animals shown to right for all gels shown. No template control (NTC) is the negative control. ( C ) GFP expression in splenocytes at day 14 day of culturing the cells and ( D ) flow cytometry of cultured splenocytes stained for CD3+/CD4+/GFP; dot blots (left) and cumulative data on right ( p = 0.04, Mann-Whitney U-test). ( E ) Representative image of supernatant from splenocytes cultured for 14 days from neonates from HIV- animal (top) or HIV+ animal (bottom) on fresh PBMCs stimulated with soluble α-CD3 and α-CD28 IL-2. Egress was analyzed in n = 7 (HIV-) and 9 (HIV+) mice; n = 3 (HIV-) and 5 (HIV+) representative mice are shown here.

    Article Snippet: PBMC Cell Culture and HIV Infection PBMCs were isolated from HIV sero-negative human donor venous blood and isolated using SepMate PBMC Isolation (STEMCELL Technologies).

    Techniques: Mouse Assay, Real-time Polymerase Chain Reaction, Electrophoresis, Negative Control, Expressing, Flow Cytometry, Cell Culture, Staining, MANN-WHITNEY

    HIV egresses from the brain into the periphery likely through CD4+ T cells. ( A ) Representative image of neonate hippocampus (HPC) immunostained for human T cells (CD3+, magenta), human astrocytes (huGFAP; red), HIV (green) and Nuclei (DAPI, blue) at week 10 post-xenotransplantation and week 4 reconstitution with huPBMCs. Arrows indicates detection of CD3+/HIV+ cell. Scale bar, 10μm. ( B ) Total PBMCs (HIV+) or monocyte/CD4+ T cell depleted PBMCs analyzed by flow cytometry for CD3+/CD4+ positive T cells (top) or CD14 positive or CD14/CD16 positive monocytes before injection into the adult mouse xenotransplanted with HIV VSVg + NHAs. HIV DNA ( C ) and RNA ( D ) from spleen from animals xenotransplanted with HIV VSVg + NHAs and reconstituted with total PBMCs (HIV+) or PBMCs depleted of monocytes and CD4+ T cells (Mono/CD4). No analysis was performed on DNA as Mono/CD4 group had undetectable HIV DNA. RNA is depicted on the right ( p = 0 . 08 , Mann-Whitney U-test). n = 5–7 per group.

    Journal: PLoS Pathogens

    Article Title: HIV infects astrocytes in vivo and egresses from the brain to the periphery

    doi: 10.1371/journal.ppat.1008381

    Figure Lengend Snippet: HIV egresses from the brain into the periphery likely through CD4+ T cells. ( A ) Representative image of neonate hippocampus (HPC) immunostained for human T cells (CD3+, magenta), human astrocytes (huGFAP; red), HIV (green) and Nuclei (DAPI, blue) at week 10 post-xenotransplantation and week 4 reconstitution with huPBMCs. Arrows indicates detection of CD3+/HIV+ cell. Scale bar, 10μm. ( B ) Total PBMCs (HIV+) or monocyte/CD4+ T cell depleted PBMCs analyzed by flow cytometry for CD3+/CD4+ positive T cells (top) or CD14 positive or CD14/CD16 positive monocytes before injection into the adult mouse xenotransplanted with HIV VSVg + NHAs. HIV DNA ( C ) and RNA ( D ) from spleen from animals xenotransplanted with HIV VSVg + NHAs and reconstituted with total PBMCs (HIV+) or PBMCs depleted of monocytes and CD4+ T cells (Mono/CD4). No analysis was performed on DNA as Mono/CD4 group had undetectable HIV DNA. RNA is depicted on the right ( p = 0 . 08 , Mann-Whitney U-test). n = 5–7 per group.

    Article Snippet: PBMC Cell Culture and HIV Infection PBMCs were isolated from HIV sero-negative human donor venous blood and isolated using SepMate PBMC Isolation (STEMCELL Technologies).

    Techniques: Flow Cytometry, Injection, MANN-WHITNEY

    HIV egress from the brain into the periphery in the neonate astrocyte xenotransplantation model. Neonate NSG mice were xenotransplanted with HIV- or HIV+ NHAs as described in Fig 2C and HIV detection outside of the brain was evaluated by Real-time PCR products (HIV DNA, A , or HIV RNA, B ) from brain, cervical lymph node, spleen, splenocytes outgrowth assay and splenocytes outgrowth assay supernatant added to fresh PBMCs analyzed by electrophoresis and human GAPDH. Each column indicates individual animal. HIV- animals are shown to left of ladder, HIV+ animals shown to right for all gels shown. No template control (NTC) is the negative control. M/F indicates sex and number is animal number from that group. ( C ) GFP expression in splenocytes at day 14 day of culturing the cells and ( D ) flow cytometry of cultured splenocytes stained for CD3+/CD4+/GFP; dot blots (left) and cumulative data on right ( p = 0.05, Mann-Whitney U-test). ( E ) Representative image of supernatant from splenocytes cultured for 14 days from neonates from HIV- animal (top) or HIV+ animal (bottom) on fresh PBMCs stimulated with soluble α-CD3 and α-CD28 IL-2. Egress for neonates was analyzed in n = 7 (HIV-) and 12 (HIV+) mice; n = 3 (HIV-) and 4 (HIV+) representative mice are shown here; n = 3 (HIV-) and 5 (HIV+) representative mice are shown here.

    Journal: PLoS Pathogens

    Article Title: HIV infects astrocytes in vivo and egresses from the brain to the periphery

    doi: 10.1371/journal.ppat.1008381

    Figure Lengend Snippet: HIV egress from the brain into the periphery in the neonate astrocyte xenotransplantation model. Neonate NSG mice were xenotransplanted with HIV- or HIV+ NHAs as described in Fig 2C and HIV detection outside of the brain was evaluated by Real-time PCR products (HIV DNA, A , or HIV RNA, B ) from brain, cervical lymph node, spleen, splenocytes outgrowth assay and splenocytes outgrowth assay supernatant added to fresh PBMCs analyzed by electrophoresis and human GAPDH. Each column indicates individual animal. HIV- animals are shown to left of ladder, HIV+ animals shown to right for all gels shown. No template control (NTC) is the negative control. M/F indicates sex and number is animal number from that group. ( C ) GFP expression in splenocytes at day 14 day of culturing the cells and ( D ) flow cytometry of cultured splenocytes stained for CD3+/CD4+/GFP; dot blots (left) and cumulative data on right ( p = 0.05, Mann-Whitney U-test). ( E ) Representative image of supernatant from splenocytes cultured for 14 days from neonates from HIV- animal (top) or HIV+ animal (bottom) on fresh PBMCs stimulated with soluble α-CD3 and α-CD28 IL-2. Egress for neonates was analyzed in n = 7 (HIV-) and 12 (HIV+) mice; n = 3 (HIV-) and 4 (HIV+) representative mice are shown here; n = 3 (HIV-) and 5 (HIV+) representative mice are shown here.

    Article Snippet: PBMC Cell Culture and HIV Infection PBMCs were isolated from HIV sero-negative human donor venous blood and isolated using SepMate PBMC Isolation (STEMCELL Technologies).

    Techniques: Mouse Assay, Real-time Polymerase Chain Reaction, Electrophoresis, Negative Control, Expressing, Flow Cytometry, Cell Culture, Staining, MANN-WHITNEY

    STING ligands increase the amount of cellular SIV-RNA/latently-infected cells. Effects of STING ligands and R848 on SIV Gag cellular/supernatant RNA copy numbers are indicated. PBMCs isolated 40 weeks after SIV-infection were treated with indicated ligands for 4 days; then, RNA from cells and supernatants were analysed by qRT-PCR. ( a ) SIV Gag cellular RNA copies normalized to infected cell numbers. ( b ) Fold-induction of SIV Gag cellular RNA copies/infected cell. ( c ) SIV Gag RNA copies in culture supernatant (sup) normalized to infected cell numbers. ( d ) Fold-induction of SIV Gag RNA copies in supernatant/infected cell. ( e ) HIV Gag cellular RNA copies normalized to infected EGFP + cell numbers. ( f ) Fold-induction of HIV Gag cellular RNA copies/infected EGFP + cell were plotted. Statistical significances were indicated by stars based on p-values (*P

    Journal: Scientific Reports

    Article Title: STING agonists activate latently infected cells and enhance SIV-specific responses ex vivo in naturally SIV controlled cynomolgus macaques

    doi: 10.1038/s41598-019-42253-3

    Figure Lengend Snippet: STING ligands increase the amount of cellular SIV-RNA/latently-infected cells. Effects of STING ligands and R848 on SIV Gag cellular/supernatant RNA copy numbers are indicated. PBMCs isolated 40 weeks after SIV-infection were treated with indicated ligands for 4 days; then, RNA from cells and supernatants were analysed by qRT-PCR. ( a ) SIV Gag cellular RNA copies normalized to infected cell numbers. ( b ) Fold-induction of SIV Gag cellular RNA copies/infected cell. ( c ) SIV Gag RNA copies in culture supernatant (sup) normalized to infected cell numbers. ( d ) Fold-induction of SIV Gag RNA copies in supernatant/infected cell. ( e ) HIV Gag cellular RNA copies normalized to infected EGFP + cell numbers. ( f ) Fold-induction of HIV Gag cellular RNA copies/infected EGFP + cell were plotted. Statistical significances were indicated by stars based on p-values (*P

    Article Snippet: Human primary CD4+ T cells were prepared with HIV− PBMCs using the EasySep Human CD4+ T Cell Enrichment Kit (StemCell Technologies, Vancouver, BC, Canada).

    Techniques: Infection, Isolation, Quantitative RT-PCR

    Cytokine release by AFM11-His is strictly dependent on the presence of CD19 + target cells. ( A ) Cytokine release in cultures of PBMC, B cell-depleted PBMC, and enriched human T cells. Unfractionated human PBMC, B cell-depleted PBMC, and enriched T cells were cultured in the presence of: i) 10 µg/mL AFM11-His or OKT3, ii) 100 µg/mL PHA, or iii) without antibodies (w/o). After 48 h incubation IL-2, IL-6, IL-10, TNF, and IFN-γ in the cell-free culture supernatant were quantified by multiplexing and the results of representative experiments were plotted. ( B ) TNF, ( C ) IL-2, ( D ) IL-6, and ( E ) IFN-γ levels in culture supernatants of PBMC from 7 individual donors. Cytokine levels were determined by Luminex upon stimulation with antibodies in solution. PBMC were either stimulated with AFM11 (open circles) or OKT3 (closed triangles) at the indicated concentrations. IL-2 and TNF levels were measured after 24 h, IL-6 and IFN-γ levels were measured after 48 h. Background (asterisks) was measured in cultures supplemented with vehicle (formulation buffer) corresponding to the volume of highest AFM11 concentration used. Horizontal bars indicate mean values across the 7 donors.

    Journal: mAbs

    Article Title: A tetravalent bispecific TandAb (CD19/CD3), AFM11, efficiently recruits T cells for the potent lysis of CD19+ tumor cells

    doi: 10.1080/19420862.2015.1029216

    Figure Lengend Snippet: Cytokine release by AFM11-His is strictly dependent on the presence of CD19 + target cells. ( A ) Cytokine release in cultures of PBMC, B cell-depleted PBMC, and enriched human T cells. Unfractionated human PBMC, B cell-depleted PBMC, and enriched T cells were cultured in the presence of: i) 10 µg/mL AFM11-His or OKT3, ii) 100 µg/mL PHA, or iii) without antibodies (w/o). After 48 h incubation IL-2, IL-6, IL-10, TNF, and IFN-γ in the cell-free culture supernatant were quantified by multiplexing and the results of representative experiments were plotted. ( B ) TNF, ( C ) IL-2, ( D ) IL-6, and ( E ) IFN-γ levels in culture supernatants of PBMC from 7 individual donors. Cytokine levels were determined by Luminex upon stimulation with antibodies in solution. PBMC were either stimulated with AFM11 (open circles) or OKT3 (closed triangles) at the indicated concentrations. IL-2 and TNF levels were measured after 24 h, IL-6 and IFN-γ levels were measured after 48 h. Background (asterisks) was measured in cultures supplemented with vehicle (formulation buffer) corresponding to the volume of highest AFM11 concentration used. Horizontal bars indicate mean values across the 7 donors.

    Article Snippet: Negatively-selected unfractionated human T cells were immunomagnetically isolated from the PBMC (EasySep™ Human T Cell Enrichment Kit and the Big Easy EasySep™ Magnet, Stem Cell Technologies, Grenoble, France, cat: 19051) according to manufacturer's instructions.

    Techniques: Cell Culture, Incubation, Multiplexing, Luminex, Concentration Assay

    AFM11 does not facilitate activation of human T cells in the absence of CD19 + target cells. Dose-responsive induction of CD25 by AFM11-His ( A ) and CD69 ( B ) expression on human T cells was assayed in cultures of human PBMC, B cell-depleted PBMC, and enriched T cells after 48 h incubation. Unfractionated PBMC contained 3.8% CD19 + cells. B cell-depleted cultures possessed 0.6% CD19 + cells, and the enriched T cell cultures contained 0.1% CD19 + cells. The kinetics of CD25 ( C ) and CD69 ( D ) expression induced by AFM11-His (10 ng/mL) were determined in human PBMC cultures containing 3.5% CD19 + cells and in B cell-depleted PBMC cultures containing

    Journal: mAbs

    Article Title: A tetravalent bispecific TandAb (CD19/CD3), AFM11, efficiently recruits T cells for the potent lysis of CD19+ tumor cells

    doi: 10.1080/19420862.2015.1029216

    Figure Lengend Snippet: AFM11 does not facilitate activation of human T cells in the absence of CD19 + target cells. Dose-responsive induction of CD25 by AFM11-His ( A ) and CD69 ( B ) expression on human T cells was assayed in cultures of human PBMC, B cell-depleted PBMC, and enriched T cells after 48 h incubation. Unfractionated PBMC contained 3.8% CD19 + cells. B cell-depleted cultures possessed 0.6% CD19 + cells, and the enriched T cell cultures contained 0.1% CD19 + cells. The kinetics of CD25 ( C ) and CD69 ( D ) expression induced by AFM11-His (10 ng/mL) were determined in human PBMC cultures containing 3.5% CD19 + cells and in B cell-depleted PBMC cultures containing

    Article Snippet: Negatively-selected unfractionated human T cells were immunomagnetically isolated from the PBMC (EasySep™ Human T Cell Enrichment Kit and the Big Easy EasySep™ Magnet, Stem Cell Technologies, Grenoble, France, cat: 19051) according to manufacturer's instructions.

    Techniques: Activation Assay, Expressing, Incubation