pbmcs  (Millipore)

 
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    Name:
    Human Peripheral Blood Monocytes
    Description:
    Human Peripheral Blood Monocytes HPBM provide a useful in vitro tool for studying various aspects of pathology and biology Our CD14 Monocytes are isolated and purified from human peripheral blood of healthy donors and cryopreserved immediately after isolation HPBM are the precursors of tissue macrophages and dentritic cells and are central to immunological responses including cell mediated cytotoxicity and production of inflammatory mediators The CD14 monocyte subset expresses a component of the lipopolysaccharide LPS receptor complex are large with diameter of 18 um represent 80 90 of circulating monocytes and express higher levels of CD62L L selectin and CD11b also referred to as Mac 1 or CR3 and lower levels of MHC class II than do CD16 monocytes
    Catalog Number:
    6906-50a
    Price:
    None
    Applications:
    Immune system dysfunction. Immunological responses, cell-mediated cytotoxicity, production of inflammatory mediators, hematopoiesis, myeloid differentiation, lipopolysaccharide (LPS), receptor complexes, CD62L (L-selectin), CD11b (Mac-1 or CR3), MHC class II.
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    Structured Review

    Millipore pbmcs
    Human Peripheral Blood Monocytes
    Human Peripheral Blood Monocytes HPBM provide a useful in vitro tool for studying various aspects of pathology and biology Our CD14 Monocytes are isolated and purified from human peripheral blood of healthy donors and cryopreserved immediately after isolation HPBM are the precursors of tissue macrophages and dentritic cells and are central to immunological responses including cell mediated cytotoxicity and production of inflammatory mediators The CD14 monocyte subset expresses a component of the lipopolysaccharide LPS receptor complex are large with diameter of 18 um represent 80 90 of circulating monocytes and express higher levels of CD62L L selectin and CD11b also referred to as Mac 1 or CR3 and lower levels of MHC class II than do CD16 monocytes
    https://www.bioz.com/result/pbmcs/product/Millipore
    Average 86 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    pbmcs - by Bioz Stars, 2021-03
    86/100 stars

    Images

    1) Product Images from "Plasma cytokines, chemokines and cellular immune responses in pre-school Nigerian children infected with Plasmodium falciparum"

    Article Title: Plasma cytokines, chemokines and cellular immune responses in pre-school Nigerian children infected with Plasmodium falciparum

    Journal: Malaria Journal

    doi: 10.1186/1475-2875-12-5

    Ascaris lumbricoides infection did not alter IFN-γ, TNF, IL-10 and IL-2 expression in mitogen-activated T-cell populations. PBMCs were stimulated for 4 h with PMA and ionomycin. Cells were washed and stained for cell surface expression of γδTCR, CD3 and CD4 and were intracellular stained for IFN-γ, IL-2, IL-10 or TNF-a. Appropriate isotype controls were included to define parameter gates. PBMCs were gated on the lymphocyte population and the percentage of positive cells for each group Ascaris infection only (ASC), Malaria infection only (MAL) and co-infected (ASC + MAL) was determined by flow cytometry and compared to uninfected endemic controls (EC) and MAL group was compared to the (ASC + MAL).
    Figure Legend Snippet: Ascaris lumbricoides infection did not alter IFN-γ, TNF, IL-10 and IL-2 expression in mitogen-activated T-cell populations. PBMCs were stimulated for 4 h with PMA and ionomycin. Cells were washed and stained for cell surface expression of γδTCR, CD3 and CD4 and were intracellular stained for IFN-γ, IL-2, IL-10 or TNF-a. Appropriate isotype controls were included to define parameter gates. PBMCs were gated on the lymphocyte population and the percentage of positive cells for each group Ascaris infection only (ASC), Malaria infection only (MAL) and co-infected (ASC + MAL) was determined by flow cytometry and compared to uninfected endemic controls (EC) and MAL group was compared to the (ASC + MAL).

    Techniques Used: Infection, Expressing, Staining, Flow Cytometry, Cytometry

    Malaria infection was not correlated with an increase in the secretion of IFN-γ, TNF, IL-10 and IL-2 in mitogen-activated T-cell populations. PBMCs were stimulated for 4 h with PMA and ionomycin. Cells were washed and stained for cell surface expression of γδTCR, CD3 and CD4 and were intracellular stained for IFN-γ, IL-2, IL-10 or TNF. Appropriate isotype controls were included to define parameter gates. PBMCs were gated on the lymphocyte population and the percentage of positive cells for each group (low (
    Figure Legend Snippet: Malaria infection was not correlated with an increase in the secretion of IFN-γ, TNF, IL-10 and IL-2 in mitogen-activated T-cell populations. PBMCs were stimulated for 4 h with PMA and ionomycin. Cells were washed and stained for cell surface expression of γδTCR, CD3 and CD4 and were intracellular stained for IFN-γ, IL-2, IL-10 or TNF. Appropriate isotype controls were included to define parameter gates. PBMCs were gated on the lymphocyte population and the percentage of positive cells for each group (low (

    Techniques Used: Infection, Staining, Expressing

    PBMCs from infected individuals did not secrete antigen specific immune responses. PBMCs (1x10 6 cells/ml) from malaria infected (n=23) and non-infected (n=7) children were stimulated with medium (Med), mycoplasma free P. falciparum parasites (at a ratio of 1:5) (malaria antigen) and PMA/anti-CD3. After three days IFN-γ, IL-10, and IL-5 were measured in supernatant by commercial assay.
    Figure Legend Snippet: PBMCs from infected individuals did not secrete antigen specific immune responses. PBMCs (1x10 6 cells/ml) from malaria infected (n=23) and non-infected (n=7) children were stimulated with medium (Med), mycoplasma free P. falciparum parasites (at a ratio of 1:5) (malaria antigen) and PMA/anti-CD3. After three days IFN-γ, IL-10, and IL-5 were measured in supernatant by commercial assay.

    Techniques Used: Infection

    2) Product Images from "Functional Inactivation of EBV-Specific T-Lymphocytes in Nasopharyngeal Carcinoma: Implications for Tumor Immunotherapy"

    Article Title: Functional Inactivation of EBV-Specific T-Lymphocytes in Nasopharyngeal Carcinoma: Implications for Tumor Immunotherapy

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0001122

    Immunophenotype of LCL stimulated cultures and TILs from NPC patients (A). Surface markers were detected by directly immunofluorescence and FACS analysis in LCL-stimulated cultures (n = 13) and tumor infiltrating lymphocyte (n = 25) expanded for 1 to 4 weeks in IL-2 medium without antigen stimulation. A relative decrease of CD8+ cells and increase of CD4+ cells was observed in TILs compared to LCL-stimulated cultures; both cultures showed high percentage of CD3+CD45RO+ and CD4+CD25+ cells. Frequency of LMP1 and LMP2 epitope-specific T cells in auto-LCL stimulated cultures and TILs from NPC patients (B). LMP1 and LMP2 epitopes specific T cells were detected by EBV tetramer staining and FACS analysis in autologous LCL-stimulated PBMCs (n = 9) and TILs (n = 6) from NPC patients.
    Figure Legend Snippet: Immunophenotype of LCL stimulated cultures and TILs from NPC patients (A). Surface markers were detected by directly immunofluorescence and FACS analysis in LCL-stimulated cultures (n = 13) and tumor infiltrating lymphocyte (n = 25) expanded for 1 to 4 weeks in IL-2 medium without antigen stimulation. A relative decrease of CD8+ cells and increase of CD4+ cells was observed in TILs compared to LCL-stimulated cultures; both cultures showed high percentage of CD3+CD45RO+ and CD4+CD25+ cells. Frequency of LMP1 and LMP2 epitope-specific T cells in auto-LCL stimulated cultures and TILs from NPC patients (B). LMP1 and LMP2 epitopes specific T cells were detected by EBV tetramer staining and FACS analysis in autologous LCL-stimulated PBMCs (n = 9) and TILs (n = 6) from NPC patients.

    Techniques Used: Immunofluorescence, FACS, Staining

    Cytotoxic activity of auto-LCL activated cultures from PBMCs of NPC patients. Polyclonal CTL cultures were test for cytotoxic activity against a panel of targets including the autologous EBV transformed LCL, PHA blasts, allogenic HLA class I matched or mismatched LCL. A. Representative experiments illustrating three pattern of cytotoxic activity. Pattern I: Lysis of the auto-LCL ≥25% and HLA class I mismatched LCL ≤10%; Pattern II: lysis to both HLA class I matched and mismatched LCL; Pattern III: less than 10% lysis against autologous or allogenic EBV positive or negative targets B. EBV-specific CTL cultures from NPC patients lysed freshly isolated autologous NPC tumor cells. Representative 51 Cr release assays performed with CTL cultures from two NPC patients are shown in the figure.
    Figure Legend Snippet: Cytotoxic activity of auto-LCL activated cultures from PBMCs of NPC patients. Polyclonal CTL cultures were test for cytotoxic activity against a panel of targets including the autologous EBV transformed LCL, PHA blasts, allogenic HLA class I matched or mismatched LCL. A. Representative experiments illustrating three pattern of cytotoxic activity. Pattern I: Lysis of the auto-LCL ≥25% and HLA class I mismatched LCL ≤10%; Pattern II: lysis to both HLA class I matched and mismatched LCL; Pattern III: less than 10% lysis against autologous or allogenic EBV positive or negative targets B. EBV-specific CTL cultures from NPC patients lysed freshly isolated autologous NPC tumor cells. Representative 51 Cr release assays performed with CTL cultures from two NPC patients are shown in the figure.

    Techniques Used: Activity Assay, CTL Assay, Transformation Assay, Lysis, Isolation

    Frequency of LMP1 and LMP2 epitope- specific T cells in PBMCs from healthy donors and NPC patients. The frequency of T cells specific for HLA A2 restricted on LMP1 and LMP2 was detected by EBV tetramer staining and FACS analysis in freshly isolated PBMCs from healthy donors (n = 7) and NPC patients (n = 9). The number of tetramer positive cells in 100 lymphocytes are shown in the figure.
    Figure Legend Snippet: Frequency of LMP1 and LMP2 epitope- specific T cells in PBMCs from healthy donors and NPC patients. The frequency of T cells specific for HLA A2 restricted on LMP1 and LMP2 was detected by EBV tetramer staining and FACS analysis in freshly isolated PBMCs from healthy donors (n = 7) and NPC patients (n = 9). The number of tetramer positive cells in 100 lymphocytes are shown in the figure.

    Techniques Used: Staining, FACS, Isolation

    3) Product Images from "Small molecule MIRA-1 induces in vitro and in vivo anti-myeloma activity and synergizes with current anti-myeloma agents"

    Article Title: Small molecule MIRA-1 induces in vitro and in vivo anti-myeloma activity and synergizes with current anti-myeloma agents

    Journal: British Journal of Cancer

    doi: 10.1038/bjc.2014.164

    MIRA-1 demonstrated potent anti-myeloma activity in vitro . ( A ) Chemical structures of MIRA-1; Chemical name: 1-[(1-Oxopropoxy)methyl]-1 H -pyrrole-2,5-dione. Molecular formula: C 8 H 9 NO 4 ( B – E ). ( B ) MM cell lines expressing wild-type p53 (MM.1S, H929) or mutant p53 (LP1, U266, and 8226) were incubated with MIRA-1 (0–20 μ mol l −1 ), and viability was determined at 48 h using MTT assay. ( C ) MIRA-1-induced cytotoxicity in primary MM samples from newly diagnosed MM patients. Myeloma cells isolated from the bone marrow via CD138+ selection were cultured with MIRA-1 (0–20 μ mol l −1 ), and viability was assessed by MTT at 48 h. ( D ) PBMCs and ( E ) BMMNCs were treated similarly with MIRA-1 and cyototoxicity was assessed by MTT assay. Viability of the cells was expressed as percentage of the DMSO-treated control. Data represents means (±s.d.) of triplicate cultures.
    Figure Legend Snippet: MIRA-1 demonstrated potent anti-myeloma activity in vitro . ( A ) Chemical structures of MIRA-1; Chemical name: 1-[(1-Oxopropoxy)methyl]-1 H -pyrrole-2,5-dione. Molecular formula: C 8 H 9 NO 4 ( B – E ). ( B ) MM cell lines expressing wild-type p53 (MM.1S, H929) or mutant p53 (LP1, U266, and 8226) were incubated with MIRA-1 (0–20 μ mol l −1 ), and viability was determined at 48 h using MTT assay. ( C ) MIRA-1-induced cytotoxicity in primary MM samples from newly diagnosed MM patients. Myeloma cells isolated from the bone marrow via CD138+ selection were cultured with MIRA-1 (0–20 μ mol l −1 ), and viability was assessed by MTT at 48 h. ( D ) PBMCs and ( E ) BMMNCs were treated similarly with MIRA-1 and cyototoxicity was assessed by MTT assay. Viability of the cells was expressed as percentage of the DMSO-treated control. Data represents means (±s.d.) of triplicate cultures.

    Techniques Used: Activity Assay, In Vitro, Expressing, Mutagenesis, Incubation, MTT Assay, Isolation, Selection, Cell Culture

    4) Product Images from "Ascorbic Acid Has Superior Ex Vivo Antiproliferative, Cell Death-Inducing and Immunomodulatory Effects over IFN-? in HTLV-1-Associated Myelopathy"

    Article Title: Ascorbic Acid Has Superior Ex Vivo Antiproliferative, Cell Death-Inducing and Immunomodulatory Effects over IFN-? in HTLV-1-Associated Myelopathy

    Journal: PLoS Neglected Tropical Diseases

    doi: 10.1371/journal.pntd.0001729

    High-dose AA, but not IFN-α, exerts strong antiproliferative effects in HAM/TSP PBMCs. (A) Lymphoproliferation of normal donors (NDs) PBMCs (n = 4) and HAM/TSP PBMCs (n = 3) was quantified by [ 3 H]thymidine incorporation (cpm), performed in triplicate. NDs and HAM/TSP PBMCs were cultured for 5 days with no treatment (NT), high-dose AA (100 µg/ml) or IFN-α (1000 IU/ml). Thymidine incorporation is shown in the y -axis, whereas the treatment conditions are shown in the x -axis. ANOVA with Bonferroni post-test for multiple testing was used and the p-values are indicated by asterisks ( *
    Figure Legend Snippet: High-dose AA, but not IFN-α, exerts strong antiproliferative effects in HAM/TSP PBMCs. (A) Lymphoproliferation of normal donors (NDs) PBMCs (n = 4) and HAM/TSP PBMCs (n = 3) was quantified by [ 3 H]thymidine incorporation (cpm), performed in triplicate. NDs and HAM/TSP PBMCs were cultured for 5 days with no treatment (NT), high-dose AA (100 µg/ml) or IFN-α (1000 IU/ml). Thymidine incorporation is shown in the y -axis, whereas the treatment conditions are shown in the x -axis. ANOVA with Bonferroni post-test for multiple testing was used and the p-values are indicated by asterisks ( *

    Techniques Used: Cell Culture

    5) Product Images from "Genetic analysis of innate immunity in Behcet’s disease identifies an association with IL-37 and IL-18RAP"

    Article Title: Genetic analysis of innate immunity in Behcet’s disease identifies an association with IL-37 and IL-18RAP

    Journal: Scientific Reports

    doi: 10.1038/srep35802

    Effect of IL37/rs3811047 genotype on cytokine production by LPS stimulated PBMCs from healthy genotyped individuals (GG: N = 10, and AG: N = 10). Data are expressed as the mean ± SD. **P
    Figure Legend Snippet: Effect of IL37/rs3811047 genotype on cytokine production by LPS stimulated PBMCs from healthy genotyped individuals (GG: N = 10, and AG: N = 10). Data are expressed as the mean ± SD. **P

    Techniques Used:

    Related Articles

    Isolation:

    Article Title: Disruption of Protein Mannosylation Affects Candida guilliermondii Cell Wall, Immune Sensing, and Virulence
    Article Snippet: .. Isolation and stimulation of human PBMCs with C. guilliermondii cells Human PBMCs were isolated using Histopaque-1077 (Sigma) as reported (Endres et al., ). .. To stimulate cytokine production, 100 μL containing 5 × 105 PBMCs in RPMI 1640 Dutch modification (added with 2 mM glutamine, 0.1 mM pyruvate and 0.05 mg/mL gentamycin; all reagents from Sigma) were plated onto round-bottom 96-well microplates, and 100 μL with 1 × 105 fungal cells freshly harvested or treated were added to each well.

    Article Title: The Kruppel-like factor KLF4 is a critical regulator of monocyte differentiation
    Article Snippet: .. Cell culture and reagents Human peripheral blood monocytes were isolated from healthy donors (Blood Bank, Children's Hospital, Boston) by the Ficoll–Hypaque centrifugation technique, as described previously ( ). .. HL-60, U-937, THP-1, Jurkat, Raji, and U-266 cells were obtained from the American Type Culture Collection (ATCC) and cultured as recommended.

    Article Title: Acute exposure to apolipoprotein A1 inhibits macrophage chemotaxis in vitro and monocyte recruitment in vivo
    Article Snippet: The total number of interacting PBMCs was quantified as captured and further classified as rolling (cells that moved in the direction of flow over the 10 s analysis period), adherent (cells that remained stationary over the 10 s analysis period) or transmigrated (cells that migrated through the endothelial monolayer) using Image-Pro Plus software (MediaCybernetics, Buckinghamshire, UK). .. Human monocyte flow cytometry PBMCs were freshly isolated as previously described ( ) from healthy volunteers using a double density gradient of histopaque (Sigma-Aldrich). .. PBMCs were pre-incubated with apoA1 for 45 min at 37°C, diluted to 1 × 106 cells/ml in PBS-BSA (5%) and stimulated with C5a (10 nM) for 5 min and then stained with active CD11b (Clone CBRM 1/5; eBioscience), total CD11b (Clone ICRF44; Biolegend), CD49b (Clone P1E6-C5; Biolegend), CD14 (Clone 63D3; Biolegend), C5aR1 (Clone S5/1; Biolegend) and CD62L (Clone DREG-56; Biolegend) and analysed by flow cytometry.

    Irradiation:

    Article Title: Enhanced immunogenicity of CTL antigens through mutation of the CD8 binding MHC class I invariant region
    Article Snippet: CTL priming C1R-A2 cells were pulsed with 1 μM Melan A26–35 (ELAGIGILTV) peptide for 90 min, irradiated and washed once in RPMI 1640 medium. .. Pulsed irradiated C1R-A2 cells (2×105 ) were incubated with 106 fresh HLA-A2+ human PBMC in RPMI 1640 supplemented with 10% fetal calf serum, 100 U/mL penicillin, 100 μg/mL streptomycin and 2 mM glutamine (Sigma) (R10 medium); 200 U/mL IL-2 was added on day 3. ..

    Incubation:

    Article Title: Enhanced immunogenicity of CTL antigens through mutation of the CD8 binding MHC class I invariant region
    Article Snippet: CTL priming C1R-A2 cells were pulsed with 1 μM Melan A26–35 (ELAGIGILTV) peptide for 90 min, irradiated and washed once in RPMI 1640 medium. .. Pulsed irradiated C1R-A2 cells (2×105 ) were incubated with 106 fresh HLA-A2+ human PBMC in RPMI 1640 supplemented with 10% fetal calf serum, 100 U/mL penicillin, 100 μg/mL streptomycin and 2 mM glutamine (Sigma) (R10 medium); 200 U/mL IL-2 was added on day 3. ..

    Cell Culture:

    Article Title: The Kruppel-like factor KLF4 is a critical regulator of monocyte differentiation
    Article Snippet: .. Cell culture and reagents Human peripheral blood monocytes were isolated from healthy donors (Blood Bank, Children's Hospital, Boston) by the Ficoll–Hypaque centrifugation technique, as described previously ( ). .. HL-60, U-937, THP-1, Jurkat, Raji, and U-266 cells were obtained from the American Type Culture Collection (ATCC) and cultured as recommended.

    Article Title: Oligomerized CARD16 promotes caspase-1 assembly and IL-1β processing
    Article Snippet: 2.3 Isolation of human PBMCs and cell culture HEK-293T and HeLa cells were cultured in Dulbecco’s modified Eagle’s medium (Wako, Osaka, Japan) supplemented with 10% fetal calf serum (FCS) and antibiotics. .. THP-1 cells, HL60 cells, and human PBMCs were cultured in RPMI1640 (Sigma, St Louis, MO, USA) supplemented with 10% FCS and antibiotics. .. PBMCs were isolated from five healthy male volunteers (25–40 years old) with Ficoll-Paque PLUS (GE Healthcare, Little Chalfont, UK).

    Centrifugation:

    Article Title: The Kruppel-like factor KLF4 is a critical regulator of monocyte differentiation
    Article Snippet: .. Cell culture and reagents Human peripheral blood monocytes were isolated from healthy donors (Blood Bank, Children's Hospital, Boston) by the Ficoll–Hypaque centrifugation technique, as described previously ( ). .. HL-60, U-937, THP-1, Jurkat, Raji, and U-266 cells were obtained from the American Type Culture Collection (ATCC) and cultured as recommended.

    Ex Vivo:

    Article Title: Identification of Cryptic MHC I-restricted Epitopes Encoded by HIV-1 Alternative Reading Frames
    Article Snippet: Human CTL Activity Assays. .. Ex vivo peptide stimulation of human PBMCs was performed using HCV-derived HLA-B7–restricted epitope G9AT (GPRLGVRAT; reference ) as negative control, phytohemagglutinin (Sigma-Aldrich) and CMV-derived epitope T10AM (pp65417 TPRVTGGGAM426 ; reference ) as positive controls, and known HIV-1–derived epitopes described in the Los Alamos Database as follows ( http://www.hiv.lanl.gov/content/hiv-db/mainpage.html ): S9WV, p2416 SPRTLNAWV24 ; T9ML, p2448 TPQDLNTML56 ; Y10LF, p636 YPLASLRSLF45 ; R10SI, gp160298 RPNNNTRKSI307 ; I9GL, gp160843 IPRRIRQGL851 ; F10LR, T10PL, Nef128 TPGPGVRYPL137 ; R9AL, Nef77 RPMTYKAAL85 ; and F9GL, Vpr34 FPRIWLHGL42 . .. For ELISPOT assay, 2 × 105 PBMCs were incubated for 16 h at 37°C with individual peptides (5 × 10−6 M), and IFN-γ release was measured as described previously ( ).

    Negative Control:

    Article Title: Identification of Cryptic MHC I-restricted Epitopes Encoded by HIV-1 Alternative Reading Frames
    Article Snippet: Human CTL Activity Assays. .. Ex vivo peptide stimulation of human PBMCs was performed using HCV-derived HLA-B7–restricted epitope G9AT (GPRLGVRAT; reference ) as negative control, phytohemagglutinin (Sigma-Aldrich) and CMV-derived epitope T10AM (pp65417 TPRVTGGGAM426 ; reference ) as positive controls, and known HIV-1–derived epitopes described in the Los Alamos Database as follows ( http://www.hiv.lanl.gov/content/hiv-db/mainpage.html ): S9WV, p2416 SPRTLNAWV24 ; T9ML, p2448 TPQDLNTML56 ; Y10LF, p636 YPLASLRSLF45 ; R10SI, gp160298 RPNNNTRKSI307 ; I9GL, gp160843 IPRRIRQGL851 ; F10LR, T10PL, Nef128 TPGPGVRYPL137 ; R9AL, Nef77 RPMTYKAAL85 ; and F9GL, Vpr34 FPRIWLHGL42 . .. For ELISPOT assay, 2 × 105 PBMCs were incubated for 16 h at 37°C with individual peptides (5 × 10−6 M), and IFN-γ release was measured as described previously ( ).

    Flow Cytometry:

    Article Title: Acute exposure to apolipoprotein A1 inhibits macrophage chemotaxis in vitro and monocyte recruitment in vivo
    Article Snippet: The total number of interacting PBMCs was quantified as captured and further classified as rolling (cells that moved in the direction of flow over the 10 s analysis period), adherent (cells that remained stationary over the 10 s analysis period) or transmigrated (cells that migrated through the endothelial monolayer) using Image-Pro Plus software (MediaCybernetics, Buckinghamshire, UK). .. Human monocyte flow cytometry PBMCs were freshly isolated as previously described ( ) from healthy volunteers using a double density gradient of histopaque (Sigma-Aldrich). .. PBMCs were pre-incubated with apoA1 for 45 min at 37°C, diluted to 1 × 106 cells/ml in PBS-BSA (5%) and stimulated with C5a (10 nM) for 5 min and then stained with active CD11b (Clone CBRM 1/5; eBioscience), total CD11b (Clone ICRF44; Biolegend), CD49b (Clone P1E6-C5; Biolegend), CD14 (Clone 63D3; Biolegend), C5aR1 (Clone S5/1; Biolegend) and CD62L (Clone DREG-56; Biolegend) and analysed by flow cytometry.

    Concentration Assay:

    Article Title: The kinase inhibitor imatinib mesylate inhibits TNF-? production in vitro and prevents TNF-dependent acute hepatic inflammation
    Article Snippet: Pharmacologic Kinase Inhibition. .. Human PBMCs or CD14-selected monocytes were stimulated with LPS at a concentration of 100 ng/ml for 12 h in the presence of either solvent or the indicated kinase inhibitor ( , SB203580, PD98059, and JNK-Inhibitor II, all used in a final concentration of 35 μM and purchased from Calbiochem). ..

    Diagnostic Assay:

    Article Title: Trypanosoma cruzi SSP4 Amastigote Protein Induces Expression of Immunoregulatory and Immunosuppressive Molecules in Peripheral Blood Mononuclear Cells
    Article Snippet: .. In brief, human PBMC were stimulated with either rMBP::SSP4 (10 μ g/mL), MBP (10 μ g/mL), LPS from Escherichia coli (0.0111:B4, 4 ng/mL) (Sigma Chemical Co), IFN-γ (100 U/mL) (Genzyme Diagnostic), or LPS plus IFN-γ , respectively. ..

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  • 98
    Millipore pbmcs proliferation
    <t>MSCs</t> from pericardial and subcutaneous adipose tissue equally suppress T‐cell proliferation. (A) : Representative example of a flow cytometry proliferation analysis of monocyte depleted peripheral blood mononuclear cells in coculture with subcutaneous or pericardial MSCs. MSCs from subcutaneous and pericardial fat have similar ability to suppress activated T‐cells' proliferation (B) and to support T‐cell viability (C) ( n = 8). Abbreviations: 7 AAD, 7‐aminoactinomycin D; adMSCs, adipose tissue‐derived MSCs; EI, expansion index; CFSE, carboxyfluorescein succinimidyl ester; FSC‐A: forward scatter area; SSC‐A: side scatter area; SSC‐H: side scatter height; SSC‐W: side scatter width.
    Pbmcs Proliferation, supplied by Millipore, used in various techniques. Bioz Stars score: 98/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/pbmcs proliferation/product/Millipore
    Average 98 stars, based on 1 article reviews
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    99
    Millipore peripheral blood mononuclear cells
    Raman fingerprints of extracellular vesicles. Raman spectra of trophoblast (a) and <t>peripheral</t> <t>mononuclear</t> <t>blood</t> cell (PBMC) (b) extracellular vesicles, n = 30–35 . The spectral shift positions are highlighted in cyan color and indicated by red numbers.
    Peripheral Blood Mononuclear Cells, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/peripheral blood mononuclear cells/product/Millipore
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    94
    Millipore shiv rt infected pbmcs
    Infection of macaque vaginal tissue after challenge with cell-associated <t>SHIV-RT.</t> Explants were prepared from vaginal biopsy specimens of SHIV-exposed uninfected animals ( n = 5). PHA/IL-2-stimulated explants were incubated with mitomycin-C-treated SHIV-RT-infected <t>PBMCs</t> (50 to 5 × 10 4 cells/explant; 3 to 4 explants/donor/condition) in 96-well plates for 18 h. Then the tissues were washed and cultured for 11 days in the presence of IL-2. PBMCs (5 × 10 4 ) cultured alone (PBMCs only) in the presence of IL-2 served as controls. SIV p27 release was measured at 0, 3, 7, and 11 days of culture by p27 ELISA. (A) Shown is the summary from 3 to 5 independent experiments using 10 2 to 10 4 infected PBMCs (means ± SEM) and an individual experiment using 50 and 5 × 10 4 cells (means ± SEM of replicate wells under each condition). (B) Shown are log 10 -transformed SOFT and CUM analyses (days 3 to 11 of culture) of the experiments presented in panel A. The LLOQ of the assay is denoted for both SOFT (solid line) and CUM (dotted line).
    Shiv Rt Infected Pbmcs, supplied by Millipore, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    MSCs from pericardial and subcutaneous adipose tissue equally suppress T‐cell proliferation. (A) : Representative example of a flow cytometry proliferation analysis of monocyte depleted peripheral blood mononuclear cells in coculture with subcutaneous or pericardial MSCs. MSCs from subcutaneous and pericardial fat have similar ability to suppress activated T‐cells' proliferation (B) and to support T‐cell viability (C) ( n = 8). Abbreviations: 7 AAD, 7‐aminoactinomycin D; adMSCs, adipose tissue‐derived MSCs; EI, expansion index; CFSE, carboxyfluorescein succinimidyl ester; FSC‐A: forward scatter area; SSC‐A: side scatter area; SSC‐H: side scatter height; SSC‐W: side scatter width.

    Journal: Stem Cells Translational Medicine

    Article Title: A Proinflammatory Secretome Mediates the Impaired Immunopotency of Human Mesenchymal Stromal Cells in Elderly Patients with Atherosclerosis

    doi: 10.1002/sctm.16-0221

    Figure Lengend Snippet: MSCs from pericardial and subcutaneous adipose tissue equally suppress T‐cell proliferation. (A) : Representative example of a flow cytometry proliferation analysis of monocyte depleted peripheral blood mononuclear cells in coculture with subcutaneous or pericardial MSCs. MSCs from subcutaneous and pericardial fat have similar ability to suppress activated T‐cells' proliferation (B) and to support T‐cell viability (C) ( n = 8). Abbreviations: 7 AAD, 7‐aminoactinomycin D; adMSCs, adipose tissue‐derived MSCs; EI, expansion index; CFSE, carboxyfluorescein succinimidyl ester; FSC‐A: forward scatter area; SSC‐A: side scatter area; SSC‐H: side scatter height; SSC‐W: side scatter width.

    Article Snippet: To assess the effect of MSCs on suppressing monocyte‐depleted PBMCs proliferation, PBMCs were labeled with 10 uM carboxyfluorescein succinimidyl ester (CFSE) (Sigma), stimulated with anti‐CD3/CD28 beads (1 bead per cell) (Dynabeads Human T‐Activator CD3/CD28, Life Technologies) and cultured for 4 days with MSCs.

    Techniques: Flow Cytometry, Cytometry, Derivative Assay

    Raman fingerprints of extracellular vesicles. Raman spectra of trophoblast (a) and peripheral mononuclear blood cell (PBMC) (b) extracellular vesicles, n = 30–35 . The spectral shift positions are highlighted in cyan color and indicated by red numbers.

    Journal: PLoS ONE

    Article Title: Raman Spectroscopy characterization extracellular vesicles from bovine placenta and peripheral blood mononuclear cells

    doi: 10.1371/journal.pone.0235214

    Figure Lengend Snippet: Raman fingerprints of extracellular vesicles. Raman spectra of trophoblast (a) and peripheral mononuclear blood cell (PBMC) (b) extracellular vesicles, n = 30–35 . The spectral shift positions are highlighted in cyan color and indicated by red numbers.

    Article Snippet: Peripheral blood mononuclear cells were isolated by density gradient centrifugation (Accu-Paque, Accurate Chem.

    Techniques:

    PCA and LDA analyses of the extracellular vesicles derived from peripheral mononuclear blood cells (P001, P002 and P003) and trophoblast cells (T001, T002 and T003). Scatter plot of the PCA results show the PC1 and PC2 scores assigned to each spectrum ( N = 90 ) (a). Linear discriminant analysis, the first 10 PC loadings calculated by means of PCA were used for LDA. Each spot represents a single spectrum. The crosses indicate the mean canonical observation score obtained for each group ( N = 90 ) (b). 2D PCA (c) and LDA (d) plots of PBMC-derived vesicles from three different cows, and 2D PCA (e) and LDA (f) plots of trophoblast-derived vesicles from three different animal individuals.

    Journal: PLoS ONE

    Article Title: Raman Spectroscopy characterization extracellular vesicles from bovine placenta and peripheral blood mononuclear cells

    doi: 10.1371/journal.pone.0235214

    Figure Lengend Snippet: PCA and LDA analyses of the extracellular vesicles derived from peripheral mononuclear blood cells (P001, P002 and P003) and trophoblast cells (T001, T002 and T003). Scatter plot of the PCA results show the PC1 and PC2 scores assigned to each spectrum ( N = 90 ) (a). Linear discriminant analysis, the first 10 PC loadings calculated by means of PCA were used for LDA. Each spot represents a single spectrum. The crosses indicate the mean canonical observation score obtained for each group ( N = 90 ) (b). 2D PCA (c) and LDA (d) plots of PBMC-derived vesicles from three different cows, and 2D PCA (e) and LDA (f) plots of trophoblast-derived vesicles from three different animal individuals.

    Article Snippet: Peripheral blood mononuclear cells were isolated by density gradient centrifugation (Accu-Paque, Accurate Chem.

    Techniques: Derivative Assay

    Principal component analysis (PCA) loading plots corresponding to PC1, PC2, and PC3 from the data of Fig 6A for extracellular vesicles derived from peripheral mononuclear blood cells and trophoblast cells.

    Journal: PLoS ONE

    Article Title: Raman Spectroscopy characterization extracellular vesicles from bovine placenta and peripheral blood mononuclear cells

    doi: 10.1371/journal.pone.0235214

    Figure Lengend Snippet: Principal component analysis (PCA) loading plots corresponding to PC1, PC2, and PC3 from the data of Fig 6A for extracellular vesicles derived from peripheral mononuclear blood cells and trophoblast cells.

    Article Snippet: Peripheral blood mononuclear cells were isolated by density gradient centrifugation (Accu-Paque, Accurate Chem.

    Techniques: Derivative Assay

    Raman fingerprint of extracellular vesicles (EVs) isolated from peripheral mononuclear blood cells (PBMC) and trophoblast cells. The solid line indicates the average of 90 spectra ± 1 standard deviation (shaded grey areas). The areas highlighted in cyan color showed the intensity differences. The peaks marked in red only exist in PBMC-derived EVs, while the peaks in blue only exist in trophoblast-derived EVs (a). Mean Raman peak intensity analysis of PBMC and trophoblast-derived EVs, N = 90 , * means P

    Journal: PLoS ONE

    Article Title: Raman Spectroscopy characterization extracellular vesicles from bovine placenta and peripheral blood mononuclear cells

    doi: 10.1371/journal.pone.0235214

    Figure Lengend Snippet: Raman fingerprint of extracellular vesicles (EVs) isolated from peripheral mononuclear blood cells (PBMC) and trophoblast cells. The solid line indicates the average of 90 spectra ± 1 standard deviation (shaded grey areas). The areas highlighted in cyan color showed the intensity differences. The peaks marked in red only exist in PBMC-derived EVs, while the peaks in blue only exist in trophoblast-derived EVs (a). Mean Raman peak intensity analysis of PBMC and trophoblast-derived EVs, N = 90 , * means P

    Article Snippet: Peripheral blood mononuclear cells were isolated by density gradient centrifugation (Accu-Paque, Accurate Chem.

    Techniques: Isolation, Standard Deviation, Derivative Assay

    Generation, isolation, and characterization of epidermal growth factor receptor (EGFR)‐specific chimeric antigen receptor (CAR)‐engineered natural killer (NK) cells (EGFR‐CAR NK cells). (A) Flow cytometric analysis of phenotypic and subset composition of peripheral blood mononuclear cells (PBMCs) labeled with anti‐CD3‐PE‐Cy7, anti‐CD56‐PE. (B‐C) Flow cytometric analysis of phenotypic and subset composition of NK cells labeled with anti‐CD3‐PE‐Cy7, anti‐CD56‐PE, and anti‐CD69‐APC‐Cy7. (D) The percentage of CD3‐/CD56 + cells in day 0 PBMCs and day14 PBMC culture. (E) Real‐time PCR and (F) Western blotting analyses of the expression of exogenous CD3ζ in the non‐transduced NK cells, Con‐CAR NK cells, EGFR‐CAR‐1 NK cells, and EGFR‐CAR‐2 NK cells. β‐actin was used as an endogenous control. (G) The transduced NK cells stained with IgG‐FITC and EGFR‐FITC antibodies were detected by flow cytometry

    Journal: Cell Proliferation

    Article Title: Targeting epidermal growth factor‐overexpressing triple‐negative breast cancer by natural killer cells expressing a specific chimeric antigen receptor, et al. Targeting epidermal growth factor‐overexpressing triple‐negative breast cancer by natural killer cells expressing a specific chimeric antigen receptor

    doi: 10.1111/cpr.12858

    Figure Lengend Snippet: Generation, isolation, and characterization of epidermal growth factor receptor (EGFR)‐specific chimeric antigen receptor (CAR)‐engineered natural killer (NK) cells (EGFR‐CAR NK cells). (A) Flow cytometric analysis of phenotypic and subset composition of peripheral blood mononuclear cells (PBMCs) labeled with anti‐CD3‐PE‐Cy7, anti‐CD56‐PE. (B‐C) Flow cytometric analysis of phenotypic and subset composition of NK cells labeled with anti‐CD3‐PE‐Cy7, anti‐CD56‐PE, and anti‐CD69‐APC‐Cy7. (D) The percentage of CD3‐/CD56 + cells in day 0 PBMCs and day14 PBMC culture. (E) Real‐time PCR and (F) Western blotting analyses of the expression of exogenous CD3ζ in the non‐transduced NK cells, Con‐CAR NK cells, EGFR‐CAR‐1 NK cells, and EGFR‐CAR‐2 NK cells. β‐actin was used as an endogenous control. (G) The transduced NK cells stained with IgG‐FITC and EGFR‐FITC antibodies were detected by flow cytometry

    Article Snippet: Peripheral blood mononuclear cells were isolated from the whole blood of healthy donors by Ficoll density gradient centrifugation.

    Techniques: Isolation, Labeling, Real-time Polymerase Chain Reaction, Western Blot, Expressing, Staining, Flow Cytometry

    Infection of macaque vaginal tissue after challenge with cell-associated SHIV-RT. Explants were prepared from vaginal biopsy specimens of SHIV-exposed uninfected animals ( n = 5). PHA/IL-2-stimulated explants were incubated with mitomycin-C-treated SHIV-RT-infected PBMCs (50 to 5 × 10 4 cells/explant; 3 to 4 explants/donor/condition) in 96-well plates for 18 h. Then the tissues were washed and cultured for 11 days in the presence of IL-2. PBMCs (5 × 10 4 ) cultured alone (PBMCs only) in the presence of IL-2 served as controls. SIV p27 release was measured at 0, 3, 7, and 11 days of culture by p27 ELISA. (A) Shown is the summary from 3 to 5 independent experiments using 10 2 to 10 4 infected PBMCs (means ± SEM) and an individual experiment using 50 and 5 × 10 4 cells (means ± SEM of replicate wells under each condition). (B) Shown are log 10 -transformed SOFT and CUM analyses (days 3 to 11 of culture) of the experiments presented in panel A. The LLOQ of the assay is denoted for both SOFT (solid line) and CUM (dotted line).

    Journal: Antimicrobial Agents and Chemotherapy

    Article Title: MIV-150/Zinc Acetate Gel Inhibits Cell-Associated Simian-Human Immunodeficiency Virus Reverse Transcriptase Infection in a Macaque Vaginal Explant Model

    doi: 10.1128/AAC.00073-15

    Figure Lengend Snippet: Infection of macaque vaginal tissue after challenge with cell-associated SHIV-RT. Explants were prepared from vaginal biopsy specimens of SHIV-exposed uninfected animals ( n = 5). PHA/IL-2-stimulated explants were incubated with mitomycin-C-treated SHIV-RT-infected PBMCs (50 to 5 × 10 4 cells/explant; 3 to 4 explants/donor/condition) in 96-well plates for 18 h. Then the tissues were washed and cultured for 11 days in the presence of IL-2. PBMCs (5 × 10 4 ) cultured alone (PBMCs only) in the presence of IL-2 served as controls. SIV p27 release was measured at 0, 3, 7, and 11 days of culture by p27 ELISA. (A) Shown is the summary from 3 to 5 independent experiments using 10 2 to 10 4 infected PBMCs (means ± SEM) and an individual experiment using 50 and 5 × 10 4 cells (means ± SEM of replicate wells under each condition). (B) Shown are log 10 -transformed SOFT and CUM analyses (days 3 to 11 of culture) of the experiments presented in panel A. The LLOQ of the assay is denoted for both SOFT (solid line) and CUM (dotted line).

    Article Snippet: Tissues were washed twice in PBS, cut using an Acu-Punch (Acuderm, Fort Lauderdale, FL) to ∼3- by 3-mm pieces, and cultured in 96-well plates in the presence of 5 μg/ml PHA and 100 U/ml IL-2 in cDMEM (Dulbecco's modified Eagle's medium [DMEM] [Cellgro Mediatech] supplemented with 10% FBS, 100 U/ml penicillin, 100 μg/ml streptomycin, and 0.1 mM minimal essential medium [MEM] nonessential amino acids [Irvine Scientific, Santa Ana, CA]) for 48 h. Thawed SHIV-RT-infected PBMCs were cultured overnight with 50 U/ml IL-2 in R10, incubated with 200 μg/ml mitomycin C (Sigma-Aldrich) for 1 h, and washed (3 times) in PBS before the coculture was set up.

    Techniques: Infection, Incubation, Cell Culture, Enzyme-linked Immunosorbent Assay, Transformation Assay

    De novo viral production in SHIV-RT-infected PBMCs is inhibited by mitomycin-C. Infected PBMCs were thawed, cultured overnight in the presence of IL-2, and then treated with mitomycin-C (versus untreated control PBMCs) and cultured for 14 days in the presence of IL-2 ( n = 3 replicates per condition). Supernatants were collected at days 0 (18 h after culture setup), 3, 7, 11, and 14, and replicates were pooled for analysis by qRT-PCR. Shown are the SIV gag copies per milliliter from two independent experiments.

    Journal: Antimicrobial Agents and Chemotherapy

    Article Title: MIV-150/Zinc Acetate Gel Inhibits Cell-Associated Simian-Human Immunodeficiency Virus Reverse Transcriptase Infection in a Macaque Vaginal Explant Model

    doi: 10.1128/AAC.00073-15

    Figure Lengend Snippet: De novo viral production in SHIV-RT-infected PBMCs is inhibited by mitomycin-C. Infected PBMCs were thawed, cultured overnight in the presence of IL-2, and then treated with mitomycin-C (versus untreated control PBMCs) and cultured for 14 days in the presence of IL-2 ( n = 3 replicates per condition). Supernatants were collected at days 0 (18 h after culture setup), 3, 7, 11, and 14, and replicates were pooled for analysis by qRT-PCR. Shown are the SIV gag copies per milliliter from two independent experiments.

    Article Snippet: Tissues were washed twice in PBS, cut using an Acu-Punch (Acuderm, Fort Lauderdale, FL) to ∼3- by 3-mm pieces, and cultured in 96-well plates in the presence of 5 μg/ml PHA and 100 U/ml IL-2 in cDMEM (Dulbecco's modified Eagle's medium [DMEM] [Cellgro Mediatech] supplemented with 10% FBS, 100 U/ml penicillin, 100 μg/ml streptomycin, and 0.1 mM minimal essential medium [MEM] nonessential amino acids [Irvine Scientific, Santa Ana, CA]) for 48 h. Thawed SHIV-RT-infected PBMCs were cultured overnight with 50 U/ml IL-2 in R10, incubated with 200 μg/ml mitomycin C (Sigma-Aldrich) for 1 h, and washed (3 times) in PBS before the coculture was set up.

    Techniques: Infection, Cell Culture, Quantitative RT-PCR

    Comparison of vaginal tissue infection after challenge with cell-associated SHIV-RT in contact-dependent and -independent models. Explants were prepared from vaginal biopsy or necropsy tissues of naive ( n = 2) and SHIV-exposed uninfected ( n = 2) animals. PHA/IL-2-stimulated explants were placed in the bottom chamber of a 12-well Transwell plate with 0.4-μm-pore inserts ( n = 4 explants per well/donor/condition). SHIV-RT-infected, mitomycin C-treated PBMCs (10 3 cells per explant) were added to the bottom chamber of the Transwell plate with tissues (Contact) or on the top of the insert (No Contact). Cultures were maintained for 14 days in the presence of IL-2. SIV p27 release in the bottom chamber was measured at 0 (18 h after culture setup), 3, 7, 11, and 14 days of culture by p27 ELISA. Shown are individual experiments (A) and the summaries from the SOFT and CUM analyses (B) (means ± SEM) of the experiments presented in panel A. The LLOQs of the assay are denoted as dotted lines in panel A. The LLOQs for both SOFT (solid line) and CUM (dotted line) are indicated in panel B.

    Journal: Antimicrobial Agents and Chemotherapy

    Article Title: MIV-150/Zinc Acetate Gel Inhibits Cell-Associated Simian-Human Immunodeficiency Virus Reverse Transcriptase Infection in a Macaque Vaginal Explant Model

    doi: 10.1128/AAC.00073-15

    Figure Lengend Snippet: Comparison of vaginal tissue infection after challenge with cell-associated SHIV-RT in contact-dependent and -independent models. Explants were prepared from vaginal biopsy or necropsy tissues of naive ( n = 2) and SHIV-exposed uninfected ( n = 2) animals. PHA/IL-2-stimulated explants were placed in the bottom chamber of a 12-well Transwell plate with 0.4-μm-pore inserts ( n = 4 explants per well/donor/condition). SHIV-RT-infected, mitomycin C-treated PBMCs (10 3 cells per explant) were added to the bottom chamber of the Transwell plate with tissues (Contact) or on the top of the insert (No Contact). Cultures were maintained for 14 days in the presence of IL-2. SIV p27 release in the bottom chamber was measured at 0 (18 h after culture setup), 3, 7, 11, and 14 days of culture by p27 ELISA. Shown are individual experiments (A) and the summaries from the SOFT and CUM analyses (B) (means ± SEM) of the experiments presented in panel A. The LLOQs of the assay are denoted as dotted lines in panel A. The LLOQs for both SOFT (solid line) and CUM (dotted line) are indicated in panel B.

    Article Snippet: Tissues were washed twice in PBS, cut using an Acu-Punch (Acuderm, Fort Lauderdale, FL) to ∼3- by 3-mm pieces, and cultured in 96-well plates in the presence of 5 μg/ml PHA and 100 U/ml IL-2 in cDMEM (Dulbecco's modified Eagle's medium [DMEM] [Cellgro Mediatech] supplemented with 10% FBS, 100 U/ml penicillin, 100 μg/ml streptomycin, and 0.1 mM minimal essential medium [MEM] nonessential amino acids [Irvine Scientific, Santa Ana, CA]) for 48 h. Thawed SHIV-RT-infected PBMCs were cultured overnight with 50 U/ml IL-2 in R10, incubated with 200 μg/ml mitomycin C (Sigma-Aldrich) for 1 h, and washed (3 times) in PBS before the coculture was set up.

    Techniques: Infection, Enzyme-linked Immunosorbent Assay

    Phenotype of SHIV-RT-infected PBMCs. Uninfected control (A) and SHIV-RT-infected (B to D) PBMCs were thawed and cultured overnight in the presence of IL-2. Then PBMCs were incubated with Live/Dead Aqua stain and anti-CD3 (Alexa Fluor 700), anti-CD4 (PerCP-Cy5.5), anti-CD95 (V450), and anti-CD28 APC antibodies followed by intracellular staining with anti-p27 RPE antibody. Mononuclear cells were gated on the total live (Aqua − ) population. The gating strategy is presented in B to D. First, CD3 − p27 + and CD3 + p27 + T cells were gated (B). Next, CD4 + T cells (C) were gated within CD3 + p27 + T cells. Then, infected T cell subsets were identified within CD3 + CD4 + p27 + T cells (D).

    Journal: Antimicrobial Agents and Chemotherapy

    Article Title: MIV-150/Zinc Acetate Gel Inhibits Cell-Associated Simian-Human Immunodeficiency Virus Reverse Transcriptase Infection in a Macaque Vaginal Explant Model

    doi: 10.1128/AAC.00073-15

    Figure Lengend Snippet: Phenotype of SHIV-RT-infected PBMCs. Uninfected control (A) and SHIV-RT-infected (B to D) PBMCs were thawed and cultured overnight in the presence of IL-2. Then PBMCs were incubated with Live/Dead Aqua stain and anti-CD3 (Alexa Fluor 700), anti-CD4 (PerCP-Cy5.5), anti-CD95 (V450), and anti-CD28 APC antibodies followed by intracellular staining with anti-p27 RPE antibody. Mononuclear cells were gated on the total live (Aqua − ) population. The gating strategy is presented in B to D. First, CD3 − p27 + and CD3 + p27 + T cells were gated (B). Next, CD4 + T cells (C) were gated within CD3 + p27 + T cells. Then, infected T cell subsets were identified within CD3 + CD4 + p27 + T cells (D).

    Article Snippet: Tissues were washed twice in PBS, cut using an Acu-Punch (Acuderm, Fort Lauderdale, FL) to ∼3- by 3-mm pieces, and cultured in 96-well plates in the presence of 5 μg/ml PHA and 100 U/ml IL-2 in cDMEM (Dulbecco's modified Eagle's medium [DMEM] [Cellgro Mediatech] supplemented with 10% FBS, 100 U/ml penicillin, 100 μg/ml streptomycin, and 0.1 mM minimal essential medium [MEM] nonessential amino acids [Irvine Scientific, Santa Ana, CA]) for 48 h. Thawed SHIV-RT-infected PBMCs were cultured overnight with 50 U/ml IL-2 in R10, incubated with 200 μg/ml mitomycin C (Sigma-Aldrich) for 1 h, and washed (3 times) in PBS before the coculture was set up.

    Techniques: Infection, Cell Culture, Incubation, Staining

    Characterization of cell-associated SHIV-RT stock. (A) Total DNA and RNA were isolated from 10 6 infected PBMCs. SIV gag qPCR and qRT-PCR were run using 50 μg of DNA and RNA as the templates, respectively. Shown are calculated per cell SIV gag copies (single experiment). (B) Infected PBMCs were thawed and immediately used in the p27 ELISA. Shown is p27 content in lysed PBMCs (single experiment).

    Journal: Antimicrobial Agents and Chemotherapy

    Article Title: MIV-150/Zinc Acetate Gel Inhibits Cell-Associated Simian-Human Immunodeficiency Virus Reverse Transcriptase Infection in a Macaque Vaginal Explant Model

    doi: 10.1128/AAC.00073-15

    Figure Lengend Snippet: Characterization of cell-associated SHIV-RT stock. (A) Total DNA and RNA were isolated from 10 6 infected PBMCs. SIV gag qPCR and qRT-PCR were run using 50 μg of DNA and RNA as the templates, respectively. Shown are calculated per cell SIV gag copies (single experiment). (B) Infected PBMCs were thawed and immediately used in the p27 ELISA. Shown is p27 content in lysed PBMCs (single experiment).

    Article Snippet: Tissues were washed twice in PBS, cut using an Acu-Punch (Acuderm, Fort Lauderdale, FL) to ∼3- by 3-mm pieces, and cultured in 96-well plates in the presence of 5 μg/ml PHA and 100 U/ml IL-2 in cDMEM (Dulbecco's modified Eagle's medium [DMEM] [Cellgro Mediatech] supplemented with 10% FBS, 100 U/ml penicillin, 100 μg/ml streptomycin, and 0.1 mM minimal essential medium [MEM] nonessential amino acids [Irvine Scientific, Santa Ana, CA]) for 48 h. Thawed SHIV-RT-infected PBMCs were cultured overnight with 50 U/ml IL-2 in R10, incubated with 200 μg/ml mitomycin C (Sigma-Aldrich) for 1 h, and washed (3 times) in PBS before the coculture was set up.

    Techniques: Isolation, Infection, Real-time Polymerase Chain Reaction, Quantitative RT-PCR, Enzyme-linked Immunosorbent Assay

    MZC inhibits ex vivo cell-associated SHIV-RT infection up to 4 days after gel exposure. Explants were prepared from vaginal biopsy or necropsy tissues of SHIV-RT-exposed uninfected ( n = 5, 24-h group), naive ( n = 3, 4-day group), and SHIV-RT-infected animals ( n = 2, 24-h group and n = 2, 4-day group). PHA/IL-2-stimulated explants were challenged with cell-associated SHIV-RT (10 3 infected PBMCs per explant; 3 to 4 explants/donor/condition) in 96-well plates 24 h or 4 days after exposure to diluted gels. Shown are log 10 -transformed SOFT and CUM analyses (means ± SEM; days 3 to 14 of culture). Summaries from 5 to 7 independent experiments (24 h) and 5 independent experiments (4 days) are shown. The LLOQs of the assay are denoted for both SOFT (solid line) and CUM (dotted line). *, P

    Journal: Antimicrobial Agents and Chemotherapy

    Article Title: MIV-150/Zinc Acetate Gel Inhibits Cell-Associated Simian-Human Immunodeficiency Virus Reverse Transcriptase Infection in a Macaque Vaginal Explant Model

    doi: 10.1128/AAC.00073-15

    Figure Lengend Snippet: MZC inhibits ex vivo cell-associated SHIV-RT infection up to 4 days after gel exposure. Explants were prepared from vaginal biopsy or necropsy tissues of SHIV-RT-exposed uninfected ( n = 5, 24-h group), naive ( n = 3, 4-day group), and SHIV-RT-infected animals ( n = 2, 24-h group and n = 2, 4-day group). PHA/IL-2-stimulated explants were challenged with cell-associated SHIV-RT (10 3 infected PBMCs per explant; 3 to 4 explants/donor/condition) in 96-well plates 24 h or 4 days after exposure to diluted gels. Shown are log 10 -transformed SOFT and CUM analyses (means ± SEM; days 3 to 14 of culture). Summaries from 5 to 7 independent experiments (24 h) and 5 independent experiments (4 days) are shown. The LLOQs of the assay are denoted for both SOFT (solid line) and CUM (dotted line). *, P

    Article Snippet: Tissues were washed twice in PBS, cut using an Acu-Punch (Acuderm, Fort Lauderdale, FL) to ∼3- by 3-mm pieces, and cultured in 96-well plates in the presence of 5 μg/ml PHA and 100 U/ml IL-2 in cDMEM (Dulbecco's modified Eagle's medium [DMEM] [Cellgro Mediatech] supplemented with 10% FBS, 100 U/ml penicillin, 100 μg/ml streptomycin, and 0.1 mM minimal essential medium [MEM] nonessential amino acids [Irvine Scientific, Santa Ana, CA]) for 48 h. Thawed SHIV-RT-infected PBMCs were cultured overnight with 50 U/ml IL-2 in R10, incubated with 200 μg/ml mitomycin C (Sigma-Aldrich) for 1 h, and washed (3 times) in PBS before the coculture was set up.

    Techniques: Ex Vivo, Infection, Transformation Assay