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GE Healthcare mononuclear cells mnc
Mononuclear Cells Mnc, supplied by GE Healthcare, used in various techniques. Bioz Stars score: 99/100, based on 51 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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Average 99 stars, based on 51 article reviews
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mononuclear cells mnc - by Bioz Stars, 2020-09
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DNA Extraction:

Article Title: Detection and Correlation of Single and Concomitant TP53, PTEN, and CDKN2A Alterations in Gliomas
Article Snippet: .. DNA Extraction High molecular weight DNA was isolated from the tumor samples using the Illustra Tissue and Cells GenomicPrep Mini Spin Kit (GE Healthcare Life Science, (Little Chalfont, Buckinghamshire, UK). .. The concentration and purity of DNA was evaluated by agarose gel electrophoresis and by Nanodrop ND-2000 Spectrophotometer (Thermo Scientific, Waltham, Massachusetts, USA).

Article Title: Molecular detection of Bartonella spp. and Rickettsia spp. in bat ectoparasites in Brazil
Article Snippet: .. DNA extraction and quality assessment DNA was extracted individually from each fly specimen and from pools comprising 10 mites of the Spintunicidae and Macronyssidae specimens, grouped according the species and host from where they were collected, using the Illustra Tissue and Cells Genomic Prep Mini Spin Kit (GE Healthcare Life Sciences), following manufacturer’s instructions. ..

Article Title: Identification of Different Bartonella Species in the Cattle Tail Louse ( Haematopinus quadripertusus) and in Cattle Blood
Article Snippet: .. Finally, the DNA was extracted using a DNA extraction kit (Illustra Tissue & Cells GenomicPrep Mini Spin kit; GE Healthcare, Buckinghamshire, United Kingdom) according to the manufacturer's instructions. .. DNA was extracted from 50 μl of EDTA-blood of each cow using a DNA extraction kit (BiOstic bacteremia DNA isolation kit; MO BIO Laboratories, Inc., Carlsbad, CA) according to the manufacturer's instructions.

Article Title: A shotgun antisense approach to the identification of novel essential genes in Pseudomonas aeruginosa
Article Snippet: .. Construction and screening of PAO1 shotgun antisense libraries Genomic DNA was isolated from P. aeruginosa PAO1 using an illustra GenomicPrep Cells and Tissue DNA Isolation Kit (GE Healthcare). ..

Article Title: Evaluation of PCR for Diagnosis of American Cutaneous Leishmaniasis in an Area of Endemicity in Northeastern Brazil
Article Snippet: .. DNA was purified for PCR with the GenomicPrep Cells and Tissue DNA isolation kit (Amersham Pharmacia Biotech) according to the instructions of the supplier. ..

Isolation:

Article Title: Detection and Correlation of Single and Concomitant TP53, PTEN, and CDKN2A Alterations in Gliomas
Article Snippet: .. DNA Extraction High molecular weight DNA was isolated from the tumor samples using the Illustra Tissue and Cells GenomicPrep Mini Spin Kit (GE Healthcare Life Science, (Little Chalfont, Buckinghamshire, UK). .. The concentration and purity of DNA was evaluated by agarose gel electrophoresis and by Nanodrop ND-2000 Spectrophotometer (Thermo Scientific, Waltham, Massachusetts, USA).

Article Title: Intracytoplasmic Copper Homeostasis Controls Cytochrome c Oxidase Production
Article Snippet: .. Total RNA was isolated using the GE Healthcare Mini Spin kit, according to the manufacturer’s instructions. ..

Article Title: A shotgun antisense approach to the identification of novel essential genes in Pseudomonas aeruginosa
Article Snippet: .. Construction and screening of PAO1 shotgun antisense libraries Genomic DNA was isolated from P. aeruginosa PAO1 using an illustra GenomicPrep Cells and Tissue DNA Isolation Kit (GE Healthcare). ..

Article Title: The ScoI homologue SenC is a copper binding protein that interacts directly with the cbb3-type cytochrome oxidase in Rhodobacter capsulatus
Article Snippet: .. Total RNA was isolated using GE Healthcare Mini Spin kit following the manufacturer’s protocol, and 2μg of total RNA were digested with DNaseI for 30 min at 25° C in the presence of RNase inhibitor RNasin. .. 50 ng of DNaseI treated RNA was used for RT-PCR reactions with the One Step RT-PCR kit (Qiagen, Hilden, Germany).

Purification:

Article Title: Evaluation of PCR for Diagnosis of American Cutaneous Leishmaniasis in an Area of Endemicity in Northeastern Brazil
Article Snippet: .. DNA was purified for PCR with the GenomicPrep Cells and Tissue DNA isolation kit (Amersham Pharmacia Biotech) according to the instructions of the supplier. ..

Molecular Weight:

Article Title: Detection and Correlation of Single and Concomitant TP53, PTEN, and CDKN2A Alterations in Gliomas
Article Snippet: .. DNA Extraction High molecular weight DNA was isolated from the tumor samples using the Illustra Tissue and Cells GenomicPrep Mini Spin Kit (GE Healthcare Life Science, (Little Chalfont, Buckinghamshire, UK). .. The concentration and purity of DNA was evaluated by agarose gel electrophoresis and by Nanodrop ND-2000 Spectrophotometer (Thermo Scientific, Waltham, Massachusetts, USA).

Polymerase Chain Reaction:

Article Title: Evaluation of PCR for Diagnosis of American Cutaneous Leishmaniasis in an Area of Endemicity in Northeastern Brazil
Article Snippet: .. DNA was purified for PCR with the GenomicPrep Cells and Tissue DNA isolation kit (Amersham Pharmacia Biotech) according to the instructions of the supplier. ..

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  • 85
    GE Healthcare nucleofection peripheral blood mononuclear cells pbmcs
    Genetic analysis of GPM6A . ( a ) Sequencing strategy and overview of the detected variants. Displayed are the coding exons (filled boxes) and the noncoding region of GPM6A (empty box). Arrows indicate rare variants found. Frequencies of rare variants in cases (black) versus controls (gray) are given. ( b) Pedigrees of two claustrophobic individuals (SIWO and THKA), carrying the mutation at locus c.*1834 (position 2882 in human GPM6A transcript variant 1, mRNA; NM_005277.3), suggesting an association between this mutation and the claustrophobic phenotype. ( c ) Highly phylogenetically conserved genomic structure surrounding c.*1834T > C within the seed sequence of miR124 in the 3′untranslated region of GPM6A . ( d ) Expression analysis after miR124 <t>nucleofection.</t> Shown are the results of GPM6A RNA expression in peripheral blood mononuclear cells <t>(PBMCs)</t> after nucleofection with miR124 from two patients and six controls (that is, not carrying the variant; age, gender and disease matched; three controls per patient). Results were standardized to the results after just a pulse. ( e ) Restraint stress induces upregulation of miR124 in the amygdala of male mice, identifying this miR as a stress-regulated transcript ( N =22 per group).
    Nucleofection Peripheral Blood Mononuclear Cells Pbmcs, supplied by GE Healthcare, used in various techniques. Bioz Stars score: 85/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/nucleofection peripheral blood mononuclear cells pbmcs/product/GE Healthcare
    Average 85 stars, based on 1 article reviews
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    89
    GE Healthcare supernatant collection mononuclear cells mnc
    A proinflammatory profile is associated with activation of NF κ B and STAT3. Immunofluorescence microscopy was used to demonstrate pNF κ B p65 and pSTAT3 transcription factors within <t>MNC</t> fractions from <t>ALL</t> patients showing a proinflammatory profile (Infl BM) and a noninflammatory profile (Non Infl) or from normal individuals (Control BM). Propidium iodide was used for nuclear staining. A representative figure of four independent experiments is shown.
    Supernatant Collection Mononuclear Cells Mnc, supplied by GE Healthcare, used in various techniques. Bioz Stars score: 89/100, based on 9 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/supernatant collection mononuclear cells mnc/product/GE Healthcare
    Average 89 stars, based on 9 article reviews
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    85
    GE Healthcare fiv specific pbmc proliferative activity
    Virus-specific lymphoproliferative activity in the study cats. At the times indicated relative to the first <t>FIV-MDC</t> inoculum, <t>PBMCs</t> were exposed to intact FIV-M2 virions (a) or to pooled FIV-M2 Gag oligopeptides (b) and then examined for 3 H-thymidine incorporation. Columns represent the stimulation index (S.I.) of individual animals, which was considered positive when above threshold dotted line (S.I. > 2). All cats responded to the nonspecific stimulus Concavalin A with counts per minute ranging from 10,000 to 38,000 counts per minute, while background proliferation in medium alone ranged between 100 and 1,100. At each sampling point, the animals are represented in the same order as shown in Figure 1.
    Fiv Specific Pbmc Proliferative Activity, supplied by GE Healthcare, used in various techniques. Bioz Stars score: 85/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/fiv specific pbmc proliferative activity/product/GE Healthcare
    Average 85 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
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    94
    GE Healthcare pbmc
    Immune suppressive MDSC and M2 persistence in <t>PDAC-SIS-PDX</t> mice. Frequency of ( A ) MDSC (human CD45 + CD3 − CD56 − CD16 − HLADR low/− CD11b + CD33 + ), ( B ) IL-10 producing MDSC and ( C ) TGF-beta-producing MDSC, as well as ( D ) PDAC-SIS-PDX M2 macrophages (CD45 + CD3 − CD56 − CD14 + CD163 + ), ( E ) IL-10 producing M2 macrophages and ( F ) TGF-beta-producing M2 monocytes in <t>PBMC,</t> spleens and PDAC of PDAC-SIS-PDX mice 4-months post-transplant as determined by flow cytometry. N = 7 SIS-PDX mice from three genetically unrelated donor cohorts were analyzed.
    Pbmc, supplied by GE Healthcare, used in various techniques. Bioz Stars score: 94/100, based on 426 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Image Search Results


    Genetic analysis of GPM6A . ( a ) Sequencing strategy and overview of the detected variants. Displayed are the coding exons (filled boxes) and the noncoding region of GPM6A (empty box). Arrows indicate rare variants found. Frequencies of rare variants in cases (black) versus controls (gray) are given. ( b) Pedigrees of two claustrophobic individuals (SIWO and THKA), carrying the mutation at locus c.*1834 (position 2882 in human GPM6A transcript variant 1, mRNA; NM_005277.3), suggesting an association between this mutation and the claustrophobic phenotype. ( c ) Highly phylogenetically conserved genomic structure surrounding c.*1834T > C within the seed sequence of miR124 in the 3′untranslated region of GPM6A . ( d ) Expression analysis after miR124 nucleofection. Shown are the results of GPM6A RNA expression in peripheral blood mononuclear cells (PBMCs) after nucleofection with miR124 from two patients and six controls (that is, not carrying the variant; age, gender and disease matched; three controls per patient). Results were standardized to the results after just a pulse. ( e ) Restraint stress induces upregulation of miR124 in the amygdala of male mice, identifying this miR as a stress-regulated transcript ( N =22 per group).

    Journal: Translational Psychiatry

    Article Title: A single gene defect causing claustrophobia

    doi: 10.1038/tp.2013.28

    Figure Lengend Snippet: Genetic analysis of GPM6A . ( a ) Sequencing strategy and overview of the detected variants. Displayed are the coding exons (filled boxes) and the noncoding region of GPM6A (empty box). Arrows indicate rare variants found. Frequencies of rare variants in cases (black) versus controls (gray) are given. ( b) Pedigrees of two claustrophobic individuals (SIWO and THKA), carrying the mutation at locus c.*1834 (position 2882 in human GPM6A transcript variant 1, mRNA; NM_005277.3), suggesting an association between this mutation and the claustrophobic phenotype. ( c ) Highly phylogenetically conserved genomic structure surrounding c.*1834T > C within the seed sequence of miR124 in the 3′untranslated region of GPM6A . ( d ) Expression analysis after miR124 nucleofection. Shown are the results of GPM6A RNA expression in peripheral blood mononuclear cells (PBMCs) after nucleofection with miR124 from two patients and six controls (that is, not carrying the variant; age, gender and disease matched; three controls per patient). Results were standardized to the results after just a pulse. ( e ) Restraint stress induces upregulation of miR124 in the amygdala of male mice, identifying this miR as a stress-regulated transcript ( N =22 per group).

    Article Snippet: Expression analysis after nucleofection Peripheral blood mononuclear cells (PBMCs) of claustrophobic patients with the mutation in the 3′UTR (N =2) and three matches per subject were freshly isolated using the standard Ficoll-Paque Plus isolation procedure (GE Healthcare, Munich, Germany).

    Techniques: Sequencing, Mutagenesis, Variant Assay, Expressing, RNA Expression, Mouse Assay

    A proinflammatory profile is associated with activation of NF κ B and STAT3. Immunofluorescence microscopy was used to demonstrate pNF κ B p65 and pSTAT3 transcription factors within MNC fractions from ALL patients showing a proinflammatory profile (Infl BM) and a noninflammatory profile (Non Infl) or from normal individuals (Control BM). Propidium iodide was used for nuclear staining. A representative figure of four independent experiments is shown.

    Journal: BioMed Research International

    Article Title: Bone Marrow Cells in Acute Lymphoblastic Leukemia Create a Proinflammatory Microenvironment Influencing Normal Hematopoietic Differentiation Fates

    doi: 10.1155/2015/386165

    Figure Lengend Snippet: A proinflammatory profile is associated with activation of NF κ B and STAT3. Immunofluorescence microscopy was used to demonstrate pNF κ B p65 and pSTAT3 transcription factors within MNC fractions from ALL patients showing a proinflammatory profile (Infl BM) and a noninflammatory profile (Non Infl) or from normal individuals (Control BM). Propidium iodide was used for nuclear staining. A representative figure of four independent experiments is shown.

    Article Snippet: Cell Isolation and Supernatant Collection Mononuclear cells (MNC) from B-cell precursor ALL patients were prepared by Ficoll-Paque Plus (GE Healthcare Bioscience) gradient separation and placed in culture with Alpha MEM medium (Gibco by Life Technologies) (200,000 MNC per 200 μ L medium).

    Techniques: Activation Assay, Immunofluorescence, Microscopy, Staining

    Virus-specific lymphoproliferative activity in the study cats. At the times indicated relative to the first FIV-MDC inoculum, PBMCs were exposed to intact FIV-M2 virions (a) or to pooled FIV-M2 Gag oligopeptides (b) and then examined for 3 H-thymidine incorporation. Columns represent the stimulation index (S.I.) of individual animals, which was considered positive when above threshold dotted line (S.I. > 2). All cats responded to the nonspecific stimulus Concavalin A with counts per minute ranging from 10,000 to 38,000 counts per minute, while background proliferation in medium alone ranged between 100 and 1,100. At each sampling point, the animals are represented in the same order as shown in Figure 1.

    Journal: Retrovirology

    Article Title: Immunotherapy with internally inactivated virus loaded dendritic cells boosts cellular immunity but does not affect feline immunodeficiency virus infection course

    doi: 10.1186/1742-4690-5-33

    Figure Lengend Snippet: Virus-specific lymphoproliferative activity in the study cats. At the times indicated relative to the first FIV-MDC inoculum, PBMCs were exposed to intact FIV-M2 virions (a) or to pooled FIV-M2 Gag oligopeptides (b) and then examined for 3 H-thymidine incorporation. Columns represent the stimulation index (S.I.) of individual animals, which was considered positive when above threshold dotted line (S.I. > 2). All cats responded to the nonspecific stimulus Concavalin A with counts per minute ranging from 10,000 to 38,000 counts per minute, while background proliferation in medium alone ranged between 100 and 1,100. At each sampling point, the animals are represented in the same order as shown in Figure 1.

    Article Snippet: FIV-specific PBMC proliferative activity was determined by incubating 1.5 × 105 PBMCs in 96-well plates in 200 μl medium supplemented with 10% human serum with either 1 μg/well of purified sonicated FIV-M2 or no stimulus for 4 days, and counting after addition of 3 H-thymidine (GE Healthcare, Milan, Italy) for 18 h. Results are reported as stimulation index (S.I.), that is the ratio of radioactivity incorporated by test PBMCs in the presence or absence of antigen, which was considered positive when > 2 [ ].

    Techniques: Activity Assay, Sampling

    Plasma viremia (a), proviral load in the PBMCs (b), and circulating CD4 + T lymphocyte percentages (c) of the study cats at the times indicated relative to the first FIV-MDC inoculum. CD4 + T cells were monitored by flow cytometry in peripheral blood by direct staining with anti-feline CD4-PE (clone vpg34, AbD Serotec, Raleigh, NC) for 30 min as previously described (8). Symbols represent individual animals. Arrows indicate the times of FIV-MDC inoculation.

    Journal: Retrovirology

    Article Title: Immunotherapy with internally inactivated virus loaded dendritic cells boosts cellular immunity but does not affect feline immunodeficiency virus infection course

    doi: 10.1186/1742-4690-5-33

    Figure Lengend Snippet: Plasma viremia (a), proviral load in the PBMCs (b), and circulating CD4 + T lymphocyte percentages (c) of the study cats at the times indicated relative to the first FIV-MDC inoculum. CD4 + T cells were monitored by flow cytometry in peripheral blood by direct staining with anti-feline CD4-PE (clone vpg34, AbD Serotec, Raleigh, NC) for 30 min as previously described (8). Symbols represent individual animals. Arrows indicate the times of FIV-MDC inoculation.

    Article Snippet: FIV-specific PBMC proliferative activity was determined by incubating 1.5 × 105 PBMCs in 96-well plates in 200 μl medium supplemented with 10% human serum with either 1 μg/well of purified sonicated FIV-M2 or no stimulus for 4 days, and counting after addition of 3 H-thymidine (GE Healthcare, Milan, Italy) for 18 h. Results are reported as stimulation index (S.I.), that is the ratio of radioactivity incorporated by test PBMCs in the presence or absence of antigen, which was considered positive when > 2 [ ].

    Techniques: Flow Cytometry, Cytometry, Staining

    Immune suppressive MDSC and M2 persistence in PDAC-SIS-PDX mice. Frequency of ( A ) MDSC (human CD45 + CD3 − CD56 − CD16 − HLADR low/− CD11b + CD33 + ), ( B ) IL-10 producing MDSC and ( C ) TGF-beta-producing MDSC, as well as ( D ) PDAC-SIS-PDX M2 macrophages (CD45 + CD3 − CD56 − CD14 + CD163 + ), ( E ) IL-10 producing M2 macrophages and ( F ) TGF-beta-producing M2 monocytes in PBMC, spleens and PDAC of PDAC-SIS-PDX mice 4-months post-transplant as determined by flow cytometry. N = 7 SIS-PDX mice from three genetically unrelated donor cohorts were analyzed.

    Journal: bioRxiv

    Article Title: NK cells and CTLs are required to clear solid tumor in a novel model of patient-derived-xenograft

    doi: 10.1101/2020.05.24.112722

    Figure Lengend Snippet: Immune suppressive MDSC and M2 persistence in PDAC-SIS-PDX mice. Frequency of ( A ) MDSC (human CD45 + CD3 − CD56 − CD16 − HLADR low/− CD11b + CD33 + ), ( B ) IL-10 producing MDSC and ( C ) TGF-beta-producing MDSC, as well as ( D ) PDAC-SIS-PDX M2 macrophages (CD45 + CD3 − CD56 − CD14 + CD163 + ), ( E ) IL-10 producing M2 macrophages and ( F ) TGF-beta-producing M2 monocytes in PBMC, spleens and PDAC of PDAC-SIS-PDX mice 4-months post-transplant as determined by flow cytometry. N = 7 SIS-PDX mice from three genetically unrelated donor cohorts were analyzed.

    Article Snippet: PBMC from LUAD and PDAC donors To isolate PBMC, peripheral blood was diluted 1:1 with Ca++ , Mg++ free phosphate buffered saline (PBS) and layered over Ficoll-Pacque (GE Healthcare), centrifuged at 2,000 rpm with no brake for 20 minutes at room temperature (RT) according to the manufacturer’s instructions.

    Techniques: Mouse Assay, Flow Cytometry

    Long-term reconstitution of LUAD-SIS-PDX mice with human B cells and dendritic cells (DC). Frequencies of Human B cells (CD45+CD3−CD56−CD19+CD20+) and DC (CD45+CD3−CD56−CD19−CD11c+) in PBMC (“blood”), spleen and transplanted tumor of LUAD-SIS-PDX mice ( A ) 2- and ( B ) 6-months post tumor-engraftment, as determined by flow cytometry. Each dot represents one LUAD-SIS-PDX mouse, N= 10-15 animals per tissue.

    Journal: bioRxiv

    Article Title: NK cells and CTLs are required to clear solid tumor in a novel model of patient-derived-xenograft

    doi: 10.1101/2020.05.24.112722

    Figure Lengend Snippet: Long-term reconstitution of LUAD-SIS-PDX mice with human B cells and dendritic cells (DC). Frequencies of Human B cells (CD45+CD3−CD56−CD19+CD20+) and DC (CD45+CD3−CD56−CD19−CD11c+) in PBMC (“blood”), spleen and transplanted tumor of LUAD-SIS-PDX mice ( A ) 2- and ( B ) 6-months post tumor-engraftment, as determined by flow cytometry. Each dot represents one LUAD-SIS-PDX mouse, N= 10-15 animals per tissue.

    Article Snippet: PBMC from LUAD and PDAC donors To isolate PBMC, peripheral blood was diluted 1:1 with Ca++ , Mg++ free phosphate buffered saline (PBS) and layered over Ficoll-Pacque (GE Healthcare), centrifuged at 2,000 rpm with no brake for 20 minutes at room temperature (RT) according to the manufacturer’s instructions.

    Techniques: Mouse Assay, Flow Cytometry

    Human T cells are exhausted in phenotype and enriched in T regulatory cells in NSCLC (LUAD). Immune phenotyping of human immune cells and T cell subsets in indicated tissues by flow cytometry. ( A, F ) Frequencies of human hematopoietic cells (human CD45 + ), ( B, G ) conventional T cells (CD45 + CD3 + CD56 − ), ( C, H ) PD-1, PD-L1 and/or PD-L2 expressing conventional T cells, ( D, I ) T helper (CD45 + CD3 + CD4 + CD8 − CD56 − ), and CTL CD45 + CD3 + CD4 − CD8 + CD56 − subsets, ( E, J ) T-regulatory cell (CD45 + CD3 + CD4 + Foxp-3 + CD8 − CD56 − ) subset in donor-matched lung vs. NSCLC (LUAD) ( A-E, top ), or patient derived PBMC, lung tissue and LUAD ( F-J, bottom ). Each data point represents one genetically unrelated human donor (N= 8-31 depending on tissue, same donors as for Figure 1 ). Paired t-test for donor-matched lung tissue vs. LUAD, one-way ANOVA with post-hoc Tukey test for comparisons between LUAD-patient derived PBMC, lung tissue and LUAD (not always donor matched between all three tissues). *p

    Journal: bioRxiv

    Article Title: NK cells and CTLs are required to clear solid tumor in a novel model of patient-derived-xenograft

    doi: 10.1101/2020.05.24.112722

    Figure Lengend Snippet: Human T cells are exhausted in phenotype and enriched in T regulatory cells in NSCLC (LUAD). Immune phenotyping of human immune cells and T cell subsets in indicated tissues by flow cytometry. ( A, F ) Frequencies of human hematopoietic cells (human CD45 + ), ( B, G ) conventional T cells (CD45 + CD3 + CD56 − ), ( C, H ) PD-1, PD-L1 and/or PD-L2 expressing conventional T cells, ( D, I ) T helper (CD45 + CD3 + CD4 + CD8 − CD56 − ), and CTL CD45 + CD3 + CD4 − CD8 + CD56 − subsets, ( E, J ) T-regulatory cell (CD45 + CD3 + CD4 + Foxp-3 + CD8 − CD56 − ) subset in donor-matched lung vs. NSCLC (LUAD) ( A-E, top ), or patient derived PBMC, lung tissue and LUAD ( F-J, bottom ). Each data point represents one genetically unrelated human donor (N= 8-31 depending on tissue, same donors as for Figure 1 ). Paired t-test for donor-matched lung tissue vs. LUAD, one-way ANOVA with post-hoc Tukey test for comparisons between LUAD-patient derived PBMC, lung tissue and LUAD (not always donor matched between all three tissues). *p

    Article Snippet: PBMC from LUAD and PDAC donors To isolate PBMC, peripheral blood was diluted 1:1 with Ca++ , Mg++ free phosphate buffered saline (PBS) and layered over Ficoll-Pacque (GE Healthcare), centrifuged at 2,000 rpm with no brake for 20 minutes at room temperature (RT) according to the manufacturer’s instructions.

    Techniques: Flow Cytometry, Expressing, Derivative Assay

    Immune cells in PDAC-SIS-PDX mice resemble exhausted TILs of PDAC donors. ( A-H ) Flow cytometry of human immune cells from PBMC, spleen, and transplanted PDAC of PDX mice analyzed 4-6 months post-transplant. ( A ) Frequency of human hematopoietic cells, ( B ) human NK cells (CD45 + CD56 + CD3 − ), and ( C ) expression of activating receptors CD16 and NKG2D as well as ( D ) markers of exhaustion PD-1, PD-L1 and PD-L2 on NK cells in PDX derived PBMC, spleen and transplanted PDAC. ( E ) Frequency of conventional T cells (CD45 + CD56 − CD3 + ) and ( E ) expression of PD-1, PD-L1 and PD-L2 on conventional T cells, ( G ) frequencies of Th (CD45 + CD3 + CD4 + CD8 − ), and CTL (CD45 + CD3 + CD4 − CD8 + ) as percent of T cells, and ( H ) Treg (CD45 + CD3 + CD4 − CD8 + Foxp3 + ) as percent of Th cell subsets in PDX derived PBMC, spleen, and transplanted PDAC. Each data point represents one PDAC-SIS-PDX mouse tissue (n= 12-33 PDX mice per group, seven donor-cohorts). Data are represented as mean SEM; Paired t-test for donor-matched lung tissue vs. PDAC, one-way ANOVA with post-hoc Tukey test *p

    Journal: bioRxiv

    Article Title: NK cells and CTLs are required to clear solid tumor in a novel model of patient-derived-xenograft

    doi: 10.1101/2020.05.24.112722

    Figure Lengend Snippet: Immune cells in PDAC-SIS-PDX mice resemble exhausted TILs of PDAC donors. ( A-H ) Flow cytometry of human immune cells from PBMC, spleen, and transplanted PDAC of PDX mice analyzed 4-6 months post-transplant. ( A ) Frequency of human hematopoietic cells, ( B ) human NK cells (CD45 + CD56 + CD3 − ), and ( C ) expression of activating receptors CD16 and NKG2D as well as ( D ) markers of exhaustion PD-1, PD-L1 and PD-L2 on NK cells in PDX derived PBMC, spleen and transplanted PDAC. ( E ) Frequency of conventional T cells (CD45 + CD56 − CD3 + ) and ( E ) expression of PD-1, PD-L1 and PD-L2 on conventional T cells, ( G ) frequencies of Th (CD45 + CD3 + CD4 + CD8 − ), and CTL (CD45 + CD3 + CD4 − CD8 + ) as percent of T cells, and ( H ) Treg (CD45 + CD3 + CD4 − CD8 + Foxp3 + ) as percent of Th cell subsets in PDX derived PBMC, spleen, and transplanted PDAC. Each data point represents one PDAC-SIS-PDX mouse tissue (n= 12-33 PDX mice per group, seven donor-cohorts). Data are represented as mean SEM; Paired t-test for donor-matched lung tissue vs. PDAC, one-way ANOVA with post-hoc Tukey test *p

    Article Snippet: PBMC from LUAD and PDAC donors To isolate PBMC, peripheral blood was diluted 1:1 with Ca++ , Mg++ free phosphate buffered saline (PBS) and layered over Ficoll-Pacque (GE Healthcare), centrifuged at 2,000 rpm with no brake for 20 minutes at room temperature (RT) according to the manufacturer’s instructions.

    Techniques: Mouse Assay, Flow Cytometry, Expressing, Derivative Assay

    Immune cells in LUAD-SIS-PDX mice resemble exhausted Tumor Infiltrating Immune cells (TILs) cells of LUAD donors. ( A-H ) Flow cytometry of human immune cells from PBMC, spleen, and transplanted LUAD of PDX mice (“tumor”) analyzed 2-months post-transplant. ( A ) Frequency of human hematopoietic cells (CD45 + ), ( B ) NK cells (CD45 + CD3 − CD56 + ), and ( C ) expression of activating receptors CD16 and NKG2D on NK cells, and ( D ) PD-1, PD-L1 and PD-L2 on NK cells in PDX mouse derived PBMC, spleen and tumor. ( E ) Frequency of conventional T cells (CD45 + CD3 + CD56 − ), and ( F ) expression of PD-1, PD-L1 and PD-L2 on conventional T cells, as well as ( G ) frequencies of T helper (CD45 + CD3 + CD4 + CD8 − CD56 − ) and CTL CD45 + CD3 + CD4 − CD8 + CD56 − ), and ( H ) T-reg (CD45 + CD3 + CD4 + Foxp-3 + CD8 − CD56 − ) subsets in PDX mouse derived PBMC, spleen and tumor. (N = 10 - 23 PDX mice per group from five genetically unrelated human donor cohorts). Data are represented as mean SEM; one-way ANOVA with post-hoc Tukey test *p

    Journal: bioRxiv

    Article Title: NK cells and CTLs are required to clear solid tumor in a novel model of patient-derived-xenograft

    doi: 10.1101/2020.05.24.112722

    Figure Lengend Snippet: Immune cells in LUAD-SIS-PDX mice resemble exhausted Tumor Infiltrating Immune cells (TILs) cells of LUAD donors. ( A-H ) Flow cytometry of human immune cells from PBMC, spleen, and transplanted LUAD of PDX mice (“tumor”) analyzed 2-months post-transplant. ( A ) Frequency of human hematopoietic cells (CD45 + ), ( B ) NK cells (CD45 + CD3 − CD56 + ), and ( C ) expression of activating receptors CD16 and NKG2D on NK cells, and ( D ) PD-1, PD-L1 and PD-L2 on NK cells in PDX mouse derived PBMC, spleen and tumor. ( E ) Frequency of conventional T cells (CD45 + CD3 + CD56 − ), and ( F ) expression of PD-1, PD-L1 and PD-L2 on conventional T cells, as well as ( G ) frequencies of T helper (CD45 + CD3 + CD4 + CD8 − CD56 − ) and CTL CD45 + CD3 + CD4 − CD8 + CD56 − ), and ( H ) T-reg (CD45 + CD3 + CD4 + Foxp-3 + CD8 − CD56 − ) subsets in PDX mouse derived PBMC, spleen and tumor. (N = 10 - 23 PDX mice per group from five genetically unrelated human donor cohorts). Data are represented as mean SEM; one-way ANOVA with post-hoc Tukey test *p

    Article Snippet: PBMC from LUAD and PDAC donors To isolate PBMC, peripheral blood was diluted 1:1 with Ca++ , Mg++ free phosphate buffered saline (PBS) and layered over Ficoll-Pacque (GE Healthcare), centrifuged at 2,000 rpm with no brake for 20 minutes at room temperature (RT) according to the manufacturer’s instructions.

    Techniques: Mouse Assay, Flow Cytometry, Expressing, Derivative Assay

    Immune cell reconstitution across a cohort of LUAD-SIS-PDX mice 4 months post engraftment. Flow cytometry of human immune cells from PBMC of single LUAD-PDX cohort of 15 mice analyzed 4-months post-transplant. Frequency of human hematopoietic cells (human CD45 + of total CD45 + ), NK cells (CD45 + CD56 + CD3 − ), NKT cells (CD45 + CD56 + CD3 + ), conventional T cells (CD45 + CD56 − CD3 + ), T helper cells (CD45 + CD56 − CD3 + CD4 + CD8 − ), and Cytotoxic T cells (CD45 + CD56 − CD3 + CD8 + CD4 − ). Each data point represents one LUAD-SIS-PDX mouse of a single human donor cohort, N=15 animals.

    Journal: bioRxiv

    Article Title: NK cells and CTLs are required to clear solid tumor in a novel model of patient-derived-xenograft

    doi: 10.1101/2020.05.24.112722

    Figure Lengend Snippet: Immune cell reconstitution across a cohort of LUAD-SIS-PDX mice 4 months post engraftment. Flow cytometry of human immune cells from PBMC of single LUAD-PDX cohort of 15 mice analyzed 4-months post-transplant. Frequency of human hematopoietic cells (human CD45 + of total CD45 + ), NK cells (CD45 + CD56 + CD3 − ), NKT cells (CD45 + CD56 + CD3 + ), conventional T cells (CD45 + CD56 − CD3 + ), T helper cells (CD45 + CD56 − CD3 + CD4 + CD8 − ), and Cytotoxic T cells (CD45 + CD56 − CD3 + CD8 + CD4 − ). Each data point represents one LUAD-SIS-PDX mouse of a single human donor cohort, N=15 animals.

    Article Snippet: PBMC from LUAD and PDAC donors To isolate PBMC, peripheral blood was diluted 1:1 with Ca++ , Mg++ free phosphate buffered saline (PBS) and layered over Ficoll-Pacque (GE Healthcare), centrifuged at 2,000 rpm with no brake for 20 minutes at room temperature (RT) according to the manufacturer’s instructions.

    Techniques: Mouse Assay, Flow Cytometry

    Human LUAD associated NK cells are exhausted in phenotype. Immune phenotyping of human NK cells in indicated tissues by flow cytometry. ( A, D ) Frequency of human NK cells (CD45 + CD3 − CD56 + ) in donor-matched lung vs. NSCLC (LUAD) tissue. ( B, E ) Frequency of CD16 and/or NKG2D expressing human NK cells, ( C, F ) Frequency of PD-1, PDL-1 and/or PDL-2-expressing human NK cells in donor-matched lung-tissue vs. LUAD ( A-C, top ), or LUAD-patient derived PBMC, lung tissue and LUAD ( D-F, bottom ). Each data point represents one genetically unrelated human donor (N= eight to 31 depending on tissue). Paired t-test for donor-matched lung tissue vs. LUAD, one-way ANOVA with post-hoc Tukey test for comparisons between LUAD-patient derived PBMC, lung tissue and LUAD (not always donor matched between all three tissues). *p

    Journal: bioRxiv

    Article Title: NK cells and CTLs are required to clear solid tumor in a novel model of patient-derived-xenograft

    doi: 10.1101/2020.05.24.112722

    Figure Lengend Snippet: Human LUAD associated NK cells are exhausted in phenotype. Immune phenotyping of human NK cells in indicated tissues by flow cytometry. ( A, D ) Frequency of human NK cells (CD45 + CD3 − CD56 + ) in donor-matched lung vs. NSCLC (LUAD) tissue. ( B, E ) Frequency of CD16 and/or NKG2D expressing human NK cells, ( C, F ) Frequency of PD-1, PDL-1 and/or PDL-2-expressing human NK cells in donor-matched lung-tissue vs. LUAD ( A-C, top ), or LUAD-patient derived PBMC, lung tissue and LUAD ( D-F, bottom ). Each data point represents one genetically unrelated human donor (N= eight to 31 depending on tissue). Paired t-test for donor-matched lung tissue vs. LUAD, one-way ANOVA with post-hoc Tukey test for comparisons between LUAD-patient derived PBMC, lung tissue and LUAD (not always donor matched between all three tissues). *p

    Article Snippet: PBMC from LUAD and PDAC donors To isolate PBMC, peripheral blood was diluted 1:1 with Ca++ , Mg++ free phosphate buffered saline (PBS) and layered over Ficoll-Pacque (GE Healthcare), centrifuged at 2,000 rpm with no brake for 20 minutes at room temperature (RT) according to the manufacturer’s instructions.

    Techniques: Flow Cytometry, Expressing, Derivative Assay

    Human PDAC associated NK and T cells are exhausted in phenotype. Frequency of ( A ) human hematopoietic cells (human CD45 + ), ( B ) NK cells (CD45 + CD56 + CD3 − ), ( C ) CD16 and/or NKG2D expressing human NK cells, ( D ) PD-1, PDL-1 and/or PDL-2-expressing human NK cells in in indicated freshly excised tissues of PDAC patients. Frequency of ( E ) human conventional T cells (CD45 + CD56 − CD3 + ), ( F ) PD-1, PDL-1 and/or PDL-2-expressing human T cells in PDAC-patient derived PBMC, pancreas and PDAC. Frequency of ( G ) Th (CD45 + CD3 + CD4 + CD8 − ) and CTL (CD45 + CD3 + CD4 − CD8 + ) as percent of human T cells, and ( H ) Treg (CD45 + CD3 + CD4 − CD8 + Foxp3 + ) as percent of Th cells in PDAC-patient derived PBMC, pancreas and PDAC. Each data point represents one genetically unrelated human donor. Paired t-test for donor-matched lung tissue vs. PDAC, one-way ANOVA with post-hoc Tukey test for comparisons between PDAC-patient derived PBMC, pancreas tissue and PDAC. *p

    Journal: bioRxiv

    Article Title: NK cells and CTLs are required to clear solid tumor in a novel model of patient-derived-xenograft

    doi: 10.1101/2020.05.24.112722

    Figure Lengend Snippet: Human PDAC associated NK and T cells are exhausted in phenotype. Frequency of ( A ) human hematopoietic cells (human CD45 + ), ( B ) NK cells (CD45 + CD56 + CD3 − ), ( C ) CD16 and/or NKG2D expressing human NK cells, ( D ) PD-1, PDL-1 and/or PDL-2-expressing human NK cells in in indicated freshly excised tissues of PDAC patients. Frequency of ( E ) human conventional T cells (CD45 + CD56 − CD3 + ), ( F ) PD-1, PDL-1 and/or PDL-2-expressing human T cells in PDAC-patient derived PBMC, pancreas and PDAC. Frequency of ( G ) Th (CD45 + CD3 + CD4 + CD8 − ) and CTL (CD45 + CD3 + CD4 − CD8 + ) as percent of human T cells, and ( H ) Treg (CD45 + CD3 + CD4 − CD8 + Foxp3 + ) as percent of Th cells in PDAC-patient derived PBMC, pancreas and PDAC. Each data point represents one genetically unrelated human donor. Paired t-test for donor-matched lung tissue vs. PDAC, one-way ANOVA with post-hoc Tukey test for comparisons between PDAC-patient derived PBMC, pancreas tissue and PDAC. *p

    Article Snippet: PBMC from LUAD and PDAC donors To isolate PBMC, peripheral blood was diluted 1:1 with Ca++ , Mg++ free phosphate buffered saline (PBS) and layered over Ficoll-Pacque (GE Healthcare), centrifuged at 2,000 rpm with no brake for 20 minutes at room temperature (RT) according to the manufacturer’s instructions.

    Techniques: Expressing, Derivative Assay