pbmcs (Cellular Technology Ltd)
86
Structured Review
Cellular Technology Ltd
pbmcs
Pbmcs, supplied by Cellular Technology Ltd, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/pbmcs/product/Cellular Technology Ltd
Average 86 stars, based on 1 article reviews
Price from $9.99 to $1999.99
Pbmcs, supplied by Cellular Technology Ltd, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/pbmcs/product/Cellular Technology Ltd
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 "Fludarabine Modulates Immune Response and Extends In Vivo Survival of Adoptively Transferred CD8 T Cells in Patients with Metastatic Melanoma"
Article Title: Fludarabine Modulates Immune Response and Extends In Vivo Survival of Adoptively Transferred CD8 T Cells in Patients with Metastatic Melanoma
Journal: PLoS ONE
doi: 10.1371/journal.pone.0004749
Figure Legend Snippet: A. In vivo persistence of CTL clones as measured by tetramer+ CD8+ T cells from post-infusion, cryopreserved PBMCs. Superimposed are the correlative IFN-γ ELISPOT data. ELISPOT results are presented as the mean number of spot forming cells/10 5 PBMCs. Shown is patient #8 who demonstrated prolonged persistence of Mart-1 27–35 specific CTL. B. In vivo persistence and ELIspot data for patient #7 who received only one infusion of Gp100 154–162 specific CTL clones. The long arrow indicates initiation of high dose dexamethasone (dex).
Techniques Used: In Vivo, CTL Assay, Clone Assay, Enzyme-linked Immunospot
2) Product Images from "Nuclease-resistant immunostimulatory phosphodiester CpG oligodeoxynucleotides as human Toll-like receptor 9 agonists"
Article Title: Nuclease-resistant immunostimulatory phosphodiester CpG oligodeoxynucleotides as human Toll-like receptor 9 agonists
Journal: BMC Biotechnology
doi: 10.1186/1472-6750-11-88
Figure Legend Snippet: TLR9 activation by B lymphocytes, PBMC, and CAL-1 cells . (A) Activation of NF-κB in human B lymphocyte cell lines, Ramos-Blue cells, stably transfected with NF-kB/AP-1-inducible SEAP reporter gene. n = 3, mean (sd). All P values are shown relative to the PD-ODN2006: * P
Techniques Used: Activation Assay, Stable Transfection, Transfection
3) Product Images from "ChAdOx1 nCoV-19 vaccination prevents SARS-CoV-2 pneumonia in rhesus macaques"
Article Title: ChAdOx1 nCoV-19 vaccination prevents SARS-CoV-2 pneumonia in rhesus macaques
Journal: bioRxiv
doi: 10.1101/2020.05.13.093195
Figure Legend Snippet: Humoral and cellular immune responses to ChAdOx1 nCoV-19 vaccination in rhesus macaques. a. Study schedule for NHPs. V = vaccination with ChAdOx1 nCoV-19; G = vaccination with ChAdOx1 GFP; E = exam; N = exam and necropsy. b. Virus neutralizing titer in serum. d. Summed IFN-γ ELISpot responses in PBMCs toward peptides spanning the spike protein Vaccinated animals = red circles; control animals = blue squares; dotted line = limit of detection; line – median; SFU = spot-forming units.
Techniques Used: Enzyme-linked Immunospot
4) Product Images from "Novel HLA-A2-restricted human metapneumovirus epitopes reduce viral titers in mice and are recognized by human T cells"
Article Title: Novel HLA-A2-restricted human metapneumovirus epitopes reduce viral titers in mice and are recognized by human T cells
Journal: Vaccine
doi: 10.1016/j.vaccine.2016.04.034
Figure Legend Snippet: Human HLA-A*0201 PBMCs recognize HMPV N39 and M39 peptides
Techniques Used:
Figure Legend Snippet: Frequency of immune T CD8 in human HLA-A*0201 PBMCs recognizing the HMPV M39 epitope
Techniques Used:
5) Product Images from "Severe immunosuppression and not a cytokine storm characterizes COVID-19 infections"
Article Title: Severe immunosuppression and not a cytokine storm characterizes COVID-19 infections
Journal: JCI Insight
doi: 10.1172/jci.insight.140329
Figure Legend Snippet: Suppressed innate immune TNF-α response in COVID-19. Representative ELISpot photomicrographs displaying baseline innate immune (monocyte) function with LPS-stimulated TNF-α production in PBMCs. Comparison between different donor types, including ( A ) healthy control volunteers and ( B ) CINS, ( C ) septic, and ( D ) COVID-19 patients. Number of spots demonstrates the number of cytokine-producing monocytes, and counts are presented as corrected number of spots per thousand monocytes plated as fraction of the 2.5 × 10 3 PBMCs plated in each well. COVID-19 patients had suppressed TNF-α production when compared with controls. Each photomicrograph was captured with the same magnification, and each image is to scale. ELISpot assays were performed using the PBMC fraction from freshly drawn whole blood. Each condition was run in duplicate for control samples and triplicate for COVID-19 samples.
Techniques Used: Enzyme-linked Immunospot
Figure Legend Snippet: Adaptive immune suppression in COVID-19 patients. Representative ELISpot photomicrographs displaying IFN-ɣ production following overnight stimulation with anti-CD3/anti-CD28 antibodies for ( A ) healthy volunteers, ( B ) CINS patients, and ( C ) septic non–COVID-19 patients. ( D ) Three representative COVID-19–positive samples. Number of spots demonstrates the number of cytokine-producing T cells. Counts are presented as the corrected number of spots per thousand lymphocytes plated as fraction of the 2.5 × 10 4 PBMCs plated in each well. Note the reduction in IFN-ɣ production in both septic and COVID-19 patients compared with CINS patients. Note also a degree of heterogeneity in IFN-ɣ production in COVID-19 and septic patients. Each photomicrograph was captured with the same magnification, and each image is to scale. ELISpot assays were performed using the PBMC fraction from freshly drawn whole blood. Each condition was run in duplicate for control samples and triplicate for COVID-19 samples.
Techniques Used: Enzyme-linked Immunospot
6) Product Images from "Evaluation of a DNA Aβ42 vaccine in adult rhesus monkeys (Macaca mulatta): antibody kinetics and immune profile after intradermal immunization with full-length DNA Aβ42 trimer"
Article Title: Evaluation of a DNA Aβ42 vaccine in adult rhesus monkeys (Macaca mulatta): antibody kinetics and immune profile after intradermal immunization with full-length DNA Aβ42 trimer
Journal: Alzheimer's Research & Therapy
doi: 10.1186/s13195-017-0257-7
Figure Legend Snippet: Interleukin (IL)-17 and IL-4 Enzyme-Linked ImmunoSpot (ELISPOT) assays, as well as interferon (IFN)-γ enzyme-linked immunosorbent assay (ELISA) from amyloid-β peptide 1–42 (Aβ42) peptide-restimulated peripheral blood mononuclear cell cultures of DNA Aβ42-immunized rhesus monkeys after a 3-month rest period. IL-17- and IL-4-secreting cells were analyzed in an ELISPOT assay ( a–c ). IFN-γ secretion was tested with a cytokine ELISA ( d ). The individual rhesus monkeys are indicated with numbers on the x -axis of all graphs. a–c The y -axis shows the number of cytokine-secreting cells (spots) per 10 6 cells. d The y -axis shows the amount of IFN-γ found (expressed in picograms per milliliter of culture supernatant)
Techniques Used: Enzyme-linked Immunospot, Enzyme-linked Immunosorbent Assay
Figure Legend Snippet: Interferon (IFN)-γ, interleukin (IL)-17, and IL-4 Enzyme-Linked ImmunoSpot assays for amyloid-β peptide 1–42 (Aβ42) peptide restimulated peripheral blood mononuclear cell cultures of DNA Aβ42-immunized rhesus monkeys. The first column shows the number of IFN-γ-, IL-17-, and IL-4-secreting cells after three immunization time points (48 h in cell culture, medium controls, and Aβ42 peptide restimulation). The second column provides results from the same analyses after the sixth immunization time point. Individual rhesus monkeys are indicated with numbers on the x -axis of all graphs. The y -axis of all graphs shows the number of cytokine-secreting cells (spots) per 10 6 cells. Increased numbers for all three cytokines were found in the immunized animals as well as in the nontreated control animals. Therefore, the cytokine secretion was considered as nonspecific and not due to the DNA Aβ42 immunization. a IL-17-secreting cells. b IFN-γ-secreting cells. c The number of IL-4-secreting cells per 10 6 splenocytes. * p
Techniques Used: Enzyme-linked Immunospot, Cell Culture
7) Product Images from "Human Immunodeficiency Virus Type-1 Myeloid Derived Suppressor Cells Inhibit Cytomegalovirus Inflammation through Interleukin-27 and B7-H4"
Article Title: Human Immunodeficiency Virus Type-1 Myeloid Derived Suppressor Cells Inhibit Cytomegalovirus Inflammation through Interleukin-27 and B7-H4
Journal: Scientific Reports
doi: 10.1038/srep44485
Figure Legend Snippet: HIV MDSC control CMV specific IFNγ production from CD4 + CX3CR1 + T cells. HIV MDSC were expanded, sorted and cultured with autologous freshly isolated PBMCs as in Fig. 2 . Cells were sorted and cultured for 48 hrs, Brefeldin A was added for the last 5 hrs of culture. Cells were surface stained with anti-CD14, -CD3, -CD4 and –CX3CR1, fixed, permeabilized and stained using anti-IFNγ for intracellular IFNγ. (a) Representative flow cytometry histograms are shown. (b) Percentage of CD14 − CD3 + CD4 + CX3CR1 + IFNγ + cells was determined by flow cytometry. (c) Supernatants were collected and the quantity of IFNγ determined by ELISA. (d) Blood obtained from HIV(−) (n = 7) and HIV(+) individuals on ART (HIV+) (n = 12) was stained with anti-CD3,-CD19,-CD11b,-CD33,-CD14, -HLA DR Abs, cells were analyzed as CD3 − CD19 − CD11b + CD33 + CD14 + HLA DR −/lo by flow cytometry. Percentages of MDSCs are shown. (e) Representative dot plot with HLA DR −/lo region is shown. For ( d ) Each dot in the plots depict data of each individual donor, the plots include observations from 25 th to 75 th percentile. The horizontal line represents the median value. For ( b and c ), histograms are presented as mean+/−SD; n = 3 donors; *p
Techniques Used: Cell Culture, Isolation, Staining, Flow Cytometry, Cytometry, Enzyme-linked Immunosorbent Assay
Figure Legend Snippet: IL-27 regulates B7-H4 expression on HIV expanded MDSC. (a) To determine if IL-27 regulates B7-H4 expression, PBMCs from HIV(−) donors were cultured with or without rIL-27 (5 ng/ml) for 72 hrs. Cells cultured in the presence of non-infectious HIV BaL (p24, 12 ng/10 7 cells) or rIL-10 (5 ng/ml) served as additional controls. Whole cell lysates of PBMCs were prepared and immunoblotted with Abs to GAPDH as loading control and B7-H4. (ai) Immunoblot shown is representative of 4 donors; cells cultured with rIL10 and rIL-27 were run in duplicates. (aii) The histogram bar graph shows mean+/−SD of the densitometric analysis for B7-H4 (n = 4). (b–d) Effect of IL-6, IL-10 and IL-27 on MDSC and B7-H4 + HIV MDSC was determined. PBMCs of healthy donors were cultured with or without non-infectious HIV BaL (p24, 12 ng/10 7 cells) in the presence or absence of neutralizing anti-IL6, -IL-10 or –IL-27 or respective isotype antibodies for 5 days. Cells were stained with anti-CD11b, -CD33, -CD14, -HLA DR and –B7-H4 Abs and analyzed using flow cytometry. (b) %B7-H4 + MDSC (bi) and MFI of B7-H4 on MDSC (bii) is shown in presence of anti-IL-6 or-IL-10 antibodies. (c) %B7-H4 + MDSC (ci) and MFI of B7-H4 on MDSC (cii) is shown in presence of anti-IL-27 antibody. (d) %MDSC is shown in presence of anti-IL-6 and –IL-10 (di) and anti-IL-27 (dii) antibodies. Histograms are presented as mean+/−SD; n = 4 donors. (e) Regulation of immunity in HIV/CMV co-infection: HIV/CMV co-infected individuals on ART with suppressed HIV replication and recovered CD4 + T cell numbers have increased IL-27 and reduced MDSC, IL-27 induces (1) B7-H4 on MDSC which binds to CMV-CD4 + T cells and (2) mediates CD4 + IL-10 + T cells, these collectively regulates T cell activation such that IFNγ is produced that is sufficient to control CMV replication without inducing inflammation (right side of straight line) . HIV/CMV co-infected ART naïve individuals with replicating virus have increased IL-6 that induces MDSC expansion and MDSC contributes to immune suppression (left side of straight line) observed during chronic disease stage. *p
Techniques Used: Expressing, Cell Culture, Staining, Flow Cytometry, Cytometry, Infection, Activation Assay, Produced
Figure Legend Snippet: In vitro expanded HIV MDSC are functionally similar to MDSC in HIV-infected individuals and modulate CMV specific T cell response. PBMCs of CMV(+) HIV(−) donors were cultured without (PBMCs) or with inactivated HIV BaL (HIV PBMCs) (p24 Ag, 12 ng/10 7 cells). After 5 days, cells were stained with anti-CD11b, -CD33, CD14, HLA DR. ( a and b ) CD14 + HLA DR −/lo MDSC were depleted from HIV PBMCs. HIV PBMCs, HIV PBMCs and MDSC depleted HIV PBMCs (HIV MDSC dep) were cultured with or without CMVpp65 for 24 and 48 hrs. The amounts of IL-2 ( a ) and IFNγ ( b ) were determined in culture supernatants at 24 and 48 hrs, respectively. Net IL-2 or IFNγ was calculated as CMVpp65 – Control. ( c and d ) PBMCs were cultured and stained as above, DR hi and MDSC were sorted. (ci and cii) Representative dot plots show HLA DR vs CD14 in control cells before sort and after sort (Left panel) and in HIV BaL treated cells before sort and after sort (Right panel) . Cells isolated are > 98% pure. ( d and e ) Isolated HIV DR hi and MDSC (5 × 10 4 ) were cultured overnight with autologous freshly isolated PBMCs (1 × 10 5 ) on ELISPOT plates with or without CMVpp65 to determine frequency of ( d ) IFNγ producing cells and ( e ) IL-10 producing cells. Histograms are presented as mean+/−SD; n = 3 donors; *p
Techniques Used: In Vitro, Infection, Cell Culture, Staining, Isolation, Enzyme-linked Immunospot
Figure Legend Snippet: B7-H4 down regulates pAkt to regulate CMV induced T cell activation. To determine the effect of HIV MDSC on CMVpp65 induced pZap70 or pAkt, PBMCs of CMV(+) HIV(−) donors were cultured with or without non-infectious HIV BaL (p24, 12 ng/10 7 cells). After 5 days, cells were stained with anti -CD11b, -CD33, CD14, HLA DR; DR hi and MDSC were sorted. (a–c) Sorted cells and freshly isolated autologous PBMCs were cultured in a ratio of 1:2 (1 sorted cell: 2 PBMCs) with or without CMVpp65 for 15 min. pZap70 was determined as detailed in Methods section. (a) Representative flow cytometry histogram plot is shown, (b) pZap70 expressed as %Net CD3 + CD4 + pZap70 + cells and (c) MFI of pZap70 in CD3 + CD4 + cells was determined. (d–f) Sorted MDSC were transfected with control or B7-H4 siRNA (75 nM) using Lipofectamine RNAiMAX reagent (Invitrogen); HIV DR hi cells were cultured in presence of Lipofectamine RNAiMAX reagent used for transfecting MDSC. After 48–72 hrs, transfected cells and freshly isolated autologous PBMCs were cultured in a ratio of 1:2 (1 sorted cell: 2 PBMCs) with or without CMVpp65 peptide pool for 15–30 min pAkt was determined as above. (d) Representative flow cytometry histogram plot is shown, (e) pAkt expressed as %Net CD3 + CD4 + pAkt + cells and (f) MFI of pAkt in CD3 + CD4 + cells was determined. Histograms are presented as mean+/−SD; n = 4 donors. *p
Techniques Used: Activation Assay, Cell Culture, Staining, Isolation, Flow Cytometry, Cytometry, Transfection
Figure Legend Snippet: HIV MDSC overexpress the inhibitory ligand B7-H4 and regulate CMV T cell activation. PBMCs from healthy donors were cultured in the presence or absence of HIV BaL . After 5 days, cells were stained using anti-CD11b, -CD33, -CD14, -HLA DR and -B7-H1/-B7-H3/-PD-L2/-ICOS-L or –B7-H4. (a and b ) Expression of B7-H1, B7-H3, PD-L2, ICOS-L or B7-H4 was determined by flow cytometry and fold expression of each ligand calculated: MDSC (Control cells - HIV BaL stimulated cells)/DR hi (Control cell - HIV BaL stimulated cells). (a) Mean values ± SD from 5 donors are shown (b) Representative flow cytometry histogram plot is shown. (c) Net Mean Fluorescence Intensity (MFI) of respective inhibitory ligands in DR hi and MDSC was determined: MFI in Control cells – MFI in HIV BaL stimulated. Mean ± SD from 5 donors are shown. ( d and e ) PBMCs of CMV(+) HIV(−) healthy donors were cultured with or without non-infectious HIV BaL (p24, 12 ng/10 7 cells). After 5 days, cells were stained with anti- CD11b, -CD33, CD14, HLA DR; DR hi and MDSC were sorted. MDSC were transfected with control or B7-H4 siRNA (75 nM) using Lipofectamine RNAiMAX reagent (Invitrogen). HIV DR hi cells were cultured in the presence of Lipofectamine RNAiMAX reagent used for transfecting MDSC. After 48–72 hrs, cell lysates of control and B7-H4 siRNA transfected cells were prepared and immunoblotted using anti-GAPDH and –B7-H4 Ab to determine expression of B7-H4. 2.5 × 10 4 transfected cells were cultured with 5 × 10 4 freshly isolated autologous PBMCs in the presence or absence of CMVpp65. (d) After 18 hrs, IL-2 and (e) after 48 hrs IFNγ was determined in culture supernatants by ELISA; partial knockdown of B7-H4 is shown in d. (f and g) PBMCs of CMV(+) HIV(−) healthy donors were cultured with or without non-infectious HIV BaL (p24, 12 ng/10 7 cells) and MDSC isolated as above. 0.05 × 10 6 MDSC were cultured with 0.1 × 10 6 autologous PBMCs in presence or absence of CMVpp65 with or without rB7-H4 (10 ng/ml). (f) After 18 hrs, IL-2 and (g) after 48 hrs IFNγ was determined in culture supernatants by ELISA. Mean+/−SD are shown for 3 healthy donors. *p
Techniques Used: Activation Assay, Cell Culture, Staining, Expressing, Flow Cytometry, Cytometry, Fluorescence, Transfection, Isolation, Enzyme-linked Immunosorbent Assay
Figure Legend Snippet: MDSC regulate CD4 + CX3CR1 + IFNγ + cells in HIV/CMV co-infected individuals. (a–c) All donors were CMV(+) and either HIV-infected (HIV+) or HIV–uninfected (HIV−). PBMCs of HIV(−) and HIV(+) donors were cultured with or without CMVpp65 for 72 hrs. Brefeldin A was added for the last 5 hrs of culture. Cells were surface stained with anti-CD14, -CD3, -CD4 and –CX3CR1, fixed, permeabilized and stained using anti-IFNγ for intracellular IFNγ. ( a ) Percentage of CD14 − CD3 + CD4 + CX3CR1 + IFNγ + cells was determined by flow cytometry. The plots include observations from 25th to 75th percentile; the horizontal line represents the median value. ( b ) Percentage of IFNγ + cells were determined in CD14 − CD3 + CD4 + CX3CR1 + (open histograms) and CD14 − CD3 + CD4 + CX3CR1 − (shaded histograms) subsets. ( c ) Representative dot plots showing CD4 + IFNγ + cells gated on CD14 − CD3 + CD4 + CX3CR1 + cells are shown from HIV-uninfected (ci HIV-ve) and HIV-infected (cii HIV+ve) individual. (d and e) Freshly isolated PBMCs from CMV(+) HIV-infected individuals on ART and with suppressed viral replication were stained with anti-CD14 and –HLA DR antibodies; CD14 + HLA DR −/lo MDSC were depleted from PBMCs by flow cytometry. Whole PBMC (PBMC) and MDSC depleted PBMC (PBMC-MDSC) were cultured with or without CMVpp65 for 72 hrs as above. Culture supernatant was stored and cells were stained as detailed above. ( d ) Percentage of CD14 − CD3 + CD4 + CX3CR1 + IFNγ + cells was determined by flow cytometry. ( e ) Amount of IFNγ in the culture supernatants was determined by ELISA. The histograms in b show mean values+/−SD; n = 8 donors. For all other graphs each dot represents an individual donor; *p
Techniques Used: Infection, Cell Culture, Staining, Flow Cytometry, Cytometry, Isolation, Enzyme-linked Immunosorbent Assay
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