Review



cell sort staining protocol cd4 + -pe ebioscience gk5.1  (Thermo Fisher)


Bioz Verified Symbol Thermo Fisher is a verified supplier
Bioz Manufacturer Symbol Thermo Fisher manufactures this product  
  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
    • 90
      Buy from Supplier

    Structured Review

    Thermo Fisher cell sort staining protocol cd4 + -pe ebioscience gk5.1
    Cell Sort Staining Protocol Cd4 + Pe Ebioscience Gk5.1, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/protocol+cd4/cell+sort+staining+protocol+cd4++++pe+ebioscience+gk5+1/pmc03826814-100-19-22
    Average 90 stars, based on 1 article reviews
    cell sort staining protocol cd4 + -pe ebioscience gk5.1 - by Bioz Stars, 2026-11
    90/100 stars

    Images

    Related Articles

    other:

    Article Title: Hypermethylation of MST1 in IgG4-related autoimmune pancreatitis and rheumatoid arthritis.
    Article Snippet: The serine/threonine kinase Mst1 plays important roles in the control of immune cell trafficking, proliferation, and differentiation.. Previously, we reported that Mst1 was required for thymocyte selection and regulatory T-cell functions, thereby the prevention of autoimmunity in mice.. In humans, MST1 null mutations cause T-cell immunodeficiency and hypergammaglobulinemia with autoantibody production.

    Article Title: Human PBMCs fight or flight response to starvation stress: Increased T-reg, FOXP3, and TGF-β1 with decreased miR-21 and Constant miR-181c levels.
    Article Snippet: Cell surface staining was performed according to manufacturer’s protocol of eBioscience with mAb, anti-CD4 (APC-conjugated, Clone: RPA-T4), anti-CD25 (PE-Cy7-conjugated, Clone: BC96) and anti-CD127 (FITC-conjugated, Clone: eBioRDR5).

    Article Title: Immunization of Mice with a Live Transconjugant Shigella Hybrid Strain Induced Th1 and Th17 Cell-Mediated Immune Responses and Confirmed Passive Protection Against Heterologous Shigellae.
    Article Snippet: An avirulent, live transconjugant Shigella hybrid (LTSHDstx) strain was constructed in our earlier study by introducing a plasmid vector, pPR1347, into a Shiga toxin gene deleted Shigella dysenteriae 1.. Three successive oral administrations of LTSHDstx to female adult mice produced comprehensive passive heterologous protection in their offspring against challenge with wildtype shigellae.. Production of NO and different cytokines such asIL-12p70, IL-1b and IL-23 in peritoneal mice macrophages indicated that LTSHDstx induced innate and adaptive immunity in mice.

    Article Title: HMGB1 regulates T helper 2 and T helper17 cell differentiation both directly and indirectly in asthmatic mice.
    Article Snippet: The Th (T helper) 2 response is characteristic of allergic asthma, and Th17 cells are involved in more severe asthma.. Recent studies demonstrated that HMGB1 (High mobility group box 1 protein) regulates airway inflammation and the Th2, Th17 inflammatory response in asthma.. HMGB1 can interact with Toll-like receptors (TLR) 2 and 4, and the receptor for advanced glycation end products (RAGE), activating the NF-κB (nuclear factor kappa B) signaling pathway and inducing the release of downstream inflammatory mediators.

    Article Title: A population of CD4 + T cells with a naïve phenotype stably polarized to the T H 1 lineage
    Article Snippet: Naïve and effector memory CD4 + T cells were sorted using the protocol, as described above using anti-CD4-PerCPCy5.5 (RM4-5), anti-CD25-PE (PC61), anti-CD62L-PECy7 (MEL-14; Thermo Fisher) and anti-CD44-Pacific Blue (IM1.8.1; Thermo Fisher).

    Article Title: Memory T cells are significantly increased in rejected liver allografts of rhesus monkeys
    Article Snippet: For analysis of forkhead box P3 (FOXP3) expression (eBioscience, San Diego, CA), PBMCs were used, and the staining protocol was essentially the same as surface‐staining with anti‐CD4 and anti‐CD25, except for permeabilization, which was performed using the kit provided with the antibody according to the manufacturer's instructions.

    Isolation:

    Article Title: Treg-Resistant Cytotoxic CD4 + T Cells Dictate T Helper Cells in Their Vicinity: TH17 Skewing and Modulation of Proliferation
    Article Snippet: .. Memory CD4 + T cells were magnetically isolated from human PBMCs according to the manufacturer’s protocol (MagniSort Human CD4 memory T cell Enrichment Kit, Thermo Fisher Scientific, Waltham, MA, USA). .. Cells were plated in 24-wells plates at 5 × 10 5 cells/well, stimulated with plate-bound 2.5 μg/mL anti-CD3 (clone OKT3, Thermo Fisher Scientific, Waltham, MA, USA) and soluble 2 μg/mL anti-CD28 (clone CD28.2, BD Biosciences, Erembodegem, Belgium), supplemented with 50% ( v / v ) CM of CD4 CTL or feeders.

    Staining:

    Article Title: Oocyst-Derived Extract of Toxoplasma Gondii Serves as Potent Immunomodulator in a Mouse Model of Birch Pollen Allergy
    Article Snippet: .. For the analysis of regulatory T cells, splenocytes were stained with the mouse regulatory T cell staining kit according to the manufacturer’s protocol (Mouse regulatory T cell staining Kit, CD4-FITC, CD25-PE, Foxp3-APC, eBioscience). .. Myeloid-derived suppressor cells (MDSC) were identified in lung cells after incubation with purified anti-mouse CD16/32 antibody followed by labelling with 7AAD (10 ng/ml; Sigma), CD11b-PerCP-Cy5.5, Ly6G-FITC, and Ly6C-APC or the relevant isotype controls.



    Similar Products

    protocol cd4  (Miltenyi Biotec)
    97
    Miltenyi Biotec protocol cd4
    Protocol Cd4, supplied by Miltenyi Biotec, used in various techniques. Bioz Stars score: 97/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/protocol+cd4/CD4%2B+T+Cell+Isolation+Kit%2C+mouse/pmc13129426-204-12-14
    Average 97 stars, based on 1 article reviews
    protocol cd4 - by Bioz Stars, 2026-11
    97/100 stars
    		  Buy from Supplier
    anti-cd4 (rm4-5)-bv510 (need clone name or described it in star protocol)  (Becton Dickinson)
    90
    Becton Dickinson anti-cd4 (rm4-5)-bv510 (need clone name or described it in star protocol)
    (A) Conventional T cells develop normally in the absence of p53. Upper panel, <t>CD4</t> and CD8 profile of total thymocytes. Lower panel, CD4 and CD8 profile of TCRβ high matured thymocytes. (B) Representative gating strategy for thymic NKT1 , NKT2 and NKT17 subsets. (C) p53 protein expression levels were analyzed by flow cytometry at steady state and after irradiation. For irradiation condition, cells were irradiated (1,000 rad) and incubated for 4 hours at 37°C. Data is pooled result of two independent experiments. (D) γH2AX expression in thymic iNKT subset was analyzed by flow cytometry. Data are mean ± SEM. Statistical differences between groups were analyzed with One-way ANOVA followed by multiple comparison. * * p < 0.005, * * * p < 0.0001.
    Anti Cd4 (Rm4 5) Bv510 (Need Clone Name Or Described It In Star Protocol), supplied by Becton Dickinson, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/protocol+cd4/anti+cd3/bio_rxiv__2024__08__21__608967-69-0-34
    Average 90 stars, based on 1 article reviews
    anti-cd4 (rm4-5)-bv510 (need clone name or described it in star protocol) - by Bioz Stars, 2026-11
    90/100 stars
    		  Buy from Supplier
    protocol 28818 generic cd4  (Jackson Laboratory)
    90
    Jackson Laboratory protocol 28818 generic cd4
    (A) Conventional T cells develop normally in the absence of p53. Upper panel, <t>CD4</t> and CD8 profile of total thymocytes. Lower panel, CD4 and CD8 profile of TCRβ high matured thymocytes. (B) Representative gating strategy for thymic NKT1 , NKT2 and NKT17 subsets. (C) p53 protein expression levels were analyzed by flow cytometry at steady state and after irradiation. For irradiation condition, cells were irradiated (1,000 rad) and incubated for 4 hours at 37°C. Data is pooled result of two independent experiments. (D) γH2AX expression in thymic iNKT subset was analyzed by flow cytometry. Data are mean ± SEM. Statistical differences between groups were analyzed with One-way ANOVA followed by multiple comparison. * * p < 0.005, * * * p < 0.0001.
    Protocol 28818 Generic Cd4, supplied by Jackson Laboratory, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/protocol+cd4/protocol+28818+generic+cd4/pm35373853-269-7-12
    Average 90 stars, based on 1 article reviews
    protocol 28818 generic cd4 - by Bioz Stars, 2026-11
    90/100 stars
    		  Buy from Supplier
    cd4+ t cells isolation protocol  (STEMCELL Technologies Inc)
    90
    STEMCELL Technologies Inc cd4+ t cells isolation protocol
    Poly1 particles increase MHCI-mediated antigen presentation and antigen-specific T cell proliferation. (A) MFI of signal pertaining to SIINFEKL presented in MHCI following treatment of DCs with no SIINFEKL (“Untreated”, gray), buffer with SIINFEKL (“SIIN Only”, gray), free Poly1 with SIINFEKL (blue), or Poly1 particles with SIINFEKL (red) and gated under DAPI− cells. 48 hours after treatment of primary DCs as in (A) but with 0.25ug mL−1 of a different model antigen, MOG35-55,CD4+ cells isolated from <t>2D2</t> mice with T cell receptors specific to MOG35-55 were added to the wells and incubated for 72 hours. (B) Representative flow cytometry traces showing CFSE dilutions after treatment with MOG35-55 and either buffer (“MOG Only”, gray), TLR4a LPS (“TLR4”, black), free Poly1 (“Free”, blue), and Poly1 particles formed (“Particles”, red). (C) MFI of CFSE signal among DAPI−/CD3+/CD4+ cells following treatments as in (B). (D) Percentage of DAPI−/CD3+/CD4+ cells proliferated beyond the 2nd generation following treatment as in (B) as determined by CFSE dilution. Samples were prepared in triplicated and errors bars represent SEM.
    Cd4+ T Cells Isolation Protocol, supplied by STEMCELL Technologies Inc, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/protocol+cd4/cd4++t+cell+isolation+cocktail/pmc04754144-111-15-22
    Average 90 stars, based on 1 article reviews
    cd4+ t cells isolation protocol - by Bioz Stars, 2026-11
    90/100 stars
    		  Buy from Supplier
    cd4 enrichment protocol  (STEMCELL Technologies Inc)
    90
    STEMCELL Technologies Inc cd4 enrichment protocol
    Poly1 particles increase MHCI-mediated antigen presentation and antigen-specific T cell proliferation. (A) MFI of signal pertaining to SIINFEKL presented in MHCI following treatment of DCs with no SIINFEKL (“Untreated”, gray), buffer with SIINFEKL (“SIIN Only”, gray), free Poly1 with SIINFEKL (blue), or Poly1 particles with SIINFEKL (red) and gated under DAPI− cells. 48 hours after treatment of primary DCs as in (A) but with 0.25ug mL−1 of a different model antigen, MOG35-55,CD4+ cells isolated from <t>2D2</t> mice with T cell receptors specific to MOG35-55 were added to the wells and incubated for 72 hours. (B) Representative flow cytometry traces showing CFSE dilutions after treatment with MOG35-55 and either buffer (“MOG Only”, gray), TLR4a LPS (“TLR4”, black), free Poly1 (“Free”, blue), and Poly1 particles formed (“Particles”, red). (C) MFI of CFSE signal among DAPI−/CD3+/CD4+ cells following treatments as in (B). (D) Percentage of DAPI−/CD3+/CD4+ cells proliferated beyond the 2nd generation following treatment as in (B) as determined by CFSE dilution. Samples were prepared in triplicated and errors bars represent SEM.
    Cd4 Enrichment Protocol, supplied by STEMCELL Technologies Inc, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/protocol+cd4/cd4+enrichment+protocol/pmc05323111-171-9-19
    Average 90 stars, based on 1 article reviews
    cd4 enrichment protocol - by Bioz Stars, 2026-11
    90/100 stars
    		  Buy from Supplier
    cell sort staining protocol cd4 + -pe ebioscience gk5.1  (Thermo Fisher)
    90
    Thermo Fisher cell sort staining protocol cd4 + -pe ebioscience gk5.1
    Poly1 particles increase MHCI-mediated antigen presentation and antigen-specific T cell proliferation. (A) MFI of signal pertaining to SIINFEKL presented in MHCI following treatment of DCs with no SIINFEKL (“Untreated”, gray), buffer with SIINFEKL (“SIIN Only”, gray), free Poly1 with SIINFEKL (blue), or Poly1 particles with SIINFEKL (red) and gated under DAPI− cells. 48 hours after treatment of primary DCs as in (A) but with 0.25ug mL−1 of a different model antigen, MOG35-55,CD4+ cells isolated from <t>2D2</t> mice with T cell receptors specific to MOG35-55 were added to the wells and incubated for 72 hours. (B) Representative flow cytometry traces showing CFSE dilutions after treatment with MOG35-55 and either buffer (“MOG Only”, gray), TLR4a LPS (“TLR4”, black), free Poly1 (“Free”, blue), and Poly1 particles formed (“Particles”, red). (C) MFI of CFSE signal among DAPI−/CD3+/CD4+ cells following treatments as in (B). (D) Percentage of DAPI−/CD3+/CD4+ cells proliferated beyond the 2nd generation following treatment as in (B) as determined by CFSE dilution. Samples were prepared in triplicated and errors bars represent SEM.
    Cell Sort Staining Protocol Cd4 + Pe Ebioscience Gk5.1, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/protocol+cd4/cell+sort+staining+protocol+cd4++++pe+ebioscience+gk5+1/pmc03826814-100-19-22
    Average 90 stars, based on 1 article reviews
    cell sort staining protocol cd4 + -pe ebioscience gk5.1 - by Bioz Stars, 2026-11
    90/100 stars
    		  Buy from Supplier
    the protocol for the negative retention of the cd4+cd25- cell population  (Miltenyi Biotec)
    90
    Miltenyi Biotec the protocol for the negative retention of the cd4+cd25- cell population
    Poly1 particles increase MHCI-mediated antigen presentation and antigen-specific T cell proliferation. (A) MFI of signal pertaining to SIINFEKL presented in MHCI following treatment of DCs with no SIINFEKL (“Untreated”, gray), buffer with SIINFEKL (“SIIN Only”, gray), free Poly1 with SIINFEKL (blue), or Poly1 particles with SIINFEKL (red) and gated under DAPI− cells. 48 hours after treatment of primary DCs as in (A) but with 0.25ug mL−1 of a different model antigen, MOG35-55,CD4+ cells isolated from <t>2D2</t> mice with T cell receptors specific to MOG35-55 were added to the wells and incubated for 72 hours. (B) Representative flow cytometry traces showing CFSE dilutions after treatment with MOG35-55 and either buffer (“MOG Only”, gray), TLR4a LPS (“TLR4”, black), free Poly1 (“Free”, blue), and Poly1 particles formed (“Particles”, red). (C) MFI of CFSE signal among DAPI−/CD3+/CD4+ cells following treatments as in (B). (D) Percentage of DAPI−/CD3+/CD4+ cells proliferated beyond the 2nd generation following treatment as in (B) as determined by CFSE dilution. Samples were prepared in triplicated and errors bars represent SEM.
    The Protocol For The Negative Retention Of The Cd4+Cd25 Cell Population, supplied by Miltenyi Biotec, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/protocol+cd4/cd14/pmc03935776-175-8-14
    Average 90 stars, based on 1 article reviews
    the protocol for the negative retention of the cd4+cd25- cell population - by Bioz Stars, 2026-11
    90/100 stars
    		  Buy from Supplier
    cd4+cd25+ t cells staining protocol  (Thermo Fisher)
    90
    Thermo Fisher cd4+cd25+ t cells staining protocol
    Poly1 particles increase MHCI-mediated antigen presentation and antigen-specific T cell proliferation. (A) MFI of signal pertaining to SIINFEKL presented in MHCI following treatment of DCs with no SIINFEKL (“Untreated”, gray), buffer with SIINFEKL (“SIIN Only”, gray), free Poly1 with SIINFEKL (blue), or Poly1 particles with SIINFEKL (red) and gated under DAPI− cells. 48 hours after treatment of primary DCs as in (A) but with 0.25ug mL−1 of a different model antigen, MOG35-55,CD4+ cells isolated from <t>2D2</t> mice with T cell receptors specific to MOG35-55 were added to the wells and incubated for 72 hours. (B) Representative flow cytometry traces showing CFSE dilutions after treatment with MOG35-55 and either buffer (“MOG Only”, gray), TLR4a LPS (“TLR4”, black), free Poly1 (“Free”, blue), and Poly1 particles formed (“Particles”, red). (C) MFI of CFSE signal among DAPI−/CD3+/CD4+ cells following treatments as in (B). (D) Percentage of DAPI−/CD3+/CD4+ cells proliferated beyond the 2nd generation following treatment as in (B) as determined by CFSE dilution. Samples were prepared in triplicated and errors bars represent SEM.
    Cd4+Cd25+ T Cells Staining Protocol, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/protocol+cd4/anti+cd25/pm23663055-63-0-10
    Average 90 stars, based on 1 article reviews
    cd4+cd25+ t cells staining protocol - by Bioz Stars, 2026-11
    90/100 stars
    		  Buy from Supplier
    reagents and protocol for cd4 t-lymphocyte count  (Partec)
    90
    Partec reagents and protocol for cd4 t-lymphocyte count
    Poly1 particles increase MHCI-mediated antigen presentation and antigen-specific T cell proliferation. (A) MFI of signal pertaining to SIINFEKL presented in MHCI following treatment of DCs with no SIINFEKL (“Untreated”, gray), buffer with SIINFEKL (“SIIN Only”, gray), free Poly1 with SIINFEKL (blue), or Poly1 particles with SIINFEKL (red) and gated under DAPI− cells. 48 hours after treatment of primary DCs as in (A) but with 0.25ug mL−1 of a different model antigen, MOG35-55,CD4+ cells isolated from <t>2D2</t> mice with T cell receptors specific to MOG35-55 were added to the wells and incubated for 72 hours. (B) Representative flow cytometry traces showing CFSE dilutions after treatment with MOG35-55 and either buffer (“MOG Only”, gray), TLR4a LPS (“TLR4”, black), free Poly1 (“Free”, blue), and Poly1 particles formed (“Particles”, red). (C) MFI of CFSE signal among DAPI−/CD3+/CD4+ cells following treatments as in (B). (D) Percentage of DAPI−/CD3+/CD4+ cells proliferated beyond the 2nd generation following treatment as in (B) as determined by CFSE dilution. Samples were prepared in triplicated and errors bars represent SEM.
    Reagents And Protocol For Cd4 T Lymphocyte Count, supplied by Partec, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/protocol+cd4/robust+and+affordable+technique+for+cd4+and+cd4++monitoring/10__1128_slash_cdli__12__1__224___227__2005-26-5-11
    Average 90 stars, based on 1 article reviews
    reagents and protocol for cd4 t-lymphocyte count - by Bioz Stars, 2026-11
    90/100 stars
    		  Buy from Supplier

    Image Search Results


    (A) Conventional T cells develop normally in the absence of p53. Upper panel, CD4 and CD8 profile of total thymocytes. Lower panel, CD4 and CD8 profile of TCRβ high matured thymocytes. (B) Representative gating strategy for thymic NKT1 , NKT2 and NKT17 subsets. (C) p53 protein expression levels were analyzed by flow cytometry at steady state and after irradiation. For irradiation condition, cells were irradiated (1,000 rad) and incubated for 4 hours at 37°C. Data is pooled result of two independent experiments. (D) γH2AX expression in thymic iNKT subset was analyzed by flow cytometry. Data are mean ± SEM. Statistical differences between groups were analyzed with One-way ANOVA followed by multiple comparison. * * p < 0.005, * * * p < 0.0001.

    Journal: bioRxiv

    Article Title: Tumor suppressor p53 controls thymic NKT17 development

    doi: 10.1101/2024.08.21.608967

    Figure Lengend Snippet: (A) Conventional T cells develop normally in the absence of p53. Upper panel, CD4 and CD8 profile of total thymocytes. Lower panel, CD4 and CD8 profile of TCRβ high matured thymocytes. (B) Representative gating strategy for thymic NKT1 , NKT2 and NKT17 subsets. (C) p53 protein expression levels were analyzed by flow cytometry at steady state and after irradiation. For irradiation condition, cells were irradiated (1,000 rad) and incubated for 4 hours at 37°C. Data is pooled result of two independent experiments. (D) γH2AX expression in thymic iNKT subset was analyzed by flow cytometry. Data are mean ± SEM. Statistical differences between groups were analyzed with One-way ANOVA followed by multiple comparison. * * p < 0.005, * * * p < 0.0001.

    Article Snippet: Anti-CD4 (RM4-5)-BV510 (Need clone name or described it in Star Protocol), anti-B220 (RA3-6B2)-AF700, anti-CD44 (IM7)-FITC, anti-TCRβ (H57-597)-PE/Cy7, anti-RORγt (Q31-378)-BV786, anti-T-bet (O4-46)-BV421, anti-PLZF (R17-809)-AF647, anti-CCR7 (4B12)-BV786, anti-IL-17A (TC11-18H10)-FITC, anti-IFNγ (XMG1.2)-FITC, anti-IL-4 (11B11)-BV421 were purchased from BD Biosciences.

    Techniques: Expressing, Flow Cytometry, Irradiation, Incubation, Comparison

    (A) Ki67 expression in thymic iNKT subset was analyzed by flow cytometry. Data is pooled result of two independent experiments. WT, n=2; cKO, n=2. (B) γH2AX expression in thymic iNKT subset was analyzed by flow cytometry. Data is pooled result of three independent experiments. SP4; CD4 single positive thymocytes. WT, n=3; cKO, n=4. Data are mean ± SEM. Statistical differences were analyzed with student t-test. * p < 0.05.

    Journal: bioRxiv

    Article Title: Tumor suppressor p53 controls thymic NKT17 development

    doi: 10.1101/2024.08.21.608967

    Figure Lengend Snippet: (A) Ki67 expression in thymic iNKT subset was analyzed by flow cytometry. Data is pooled result of two independent experiments. WT, n=2; cKO, n=2. (B) γH2AX expression in thymic iNKT subset was analyzed by flow cytometry. Data is pooled result of three independent experiments. SP4; CD4 single positive thymocytes. WT, n=3; cKO, n=4. Data are mean ± SEM. Statistical differences were analyzed with student t-test. * p < 0.05.

    Article Snippet: Anti-CD4 (RM4-5)-BV510 (Need clone name or described it in Star Protocol), anti-B220 (RA3-6B2)-AF700, anti-CD44 (IM7)-FITC, anti-TCRβ (H57-597)-PE/Cy7, anti-RORγt (Q31-378)-BV786, anti-T-bet (O4-46)-BV421, anti-PLZF (R17-809)-AF647, anti-CCR7 (4B12)-BV786, anti-IL-17A (TC11-18H10)-FITC, anti-IFNγ (XMG1.2)-FITC, anti-IL-4 (11B11)-BV421 were purchased from BD Biosciences.

    Techniques: Expressing, Flow Cytometry

    Poly1 particles increase MHCI-mediated antigen presentation and antigen-specific T cell proliferation. (A) MFI of signal pertaining to SIINFEKL presented in MHCI following treatment of DCs with no SIINFEKL (“Untreated”, gray), buffer with SIINFEKL (“SIIN Only”, gray), free Poly1 with SIINFEKL (blue), or Poly1 particles with SIINFEKL (red) and gated under DAPI− cells. 48 hours after treatment of primary DCs as in (A) but with 0.25ug mL−1 of a different model antigen, MOG35-55,CD4+ cells isolated from 2D2 mice with T cell receptors specific to MOG35-55 were added to the wells and incubated for 72 hours. (B) Representative flow cytometry traces showing CFSE dilutions after treatment with MOG35-55 and either buffer (“MOG Only”, gray), TLR4a LPS (“TLR4”, black), free Poly1 (“Free”, blue), and Poly1 particles formed (“Particles”, red). (C) MFI of CFSE signal among DAPI−/CD3+/CD4+ cells following treatments as in (B). (D) Percentage of DAPI−/CD3+/CD4+ cells proliferated beyond the 2nd generation following treatment as in (B) as determined by CFSE dilution. Samples were prepared in triplicated and errors bars represent SEM.

    Journal: Acta biomaterialia

    Article Title: Intrinsic immunogenicity of rapidly-degradable polymers evolves during degradation

    doi: 10.1016/j.actbio.2015.12.026

    Figure Lengend Snippet: Poly1 particles increase MHCI-mediated antigen presentation and antigen-specific T cell proliferation. (A) MFI of signal pertaining to SIINFEKL presented in MHCI following treatment of DCs with no SIINFEKL (“Untreated”, gray), buffer with SIINFEKL (“SIIN Only”, gray), free Poly1 with SIINFEKL (blue), or Poly1 particles with SIINFEKL (red) and gated under DAPI− cells. 48 hours after treatment of primary DCs as in (A) but with 0.25ug mL−1 of a different model antigen, MOG35-55,CD4+ cells isolated from 2D2 mice with T cell receptors specific to MOG35-55 were added to the wells and incubated for 72 hours. (B) Representative flow cytometry traces showing CFSE dilutions after treatment with MOG35-55 and either buffer (“MOG Only”, gray), TLR4a LPS (“TLR4”, black), free Poly1 (“Free”, blue), and Poly1 particles formed (“Particles”, red). (C) MFI of CFSE signal among DAPI−/CD3+/CD4+ cells following treatments as in (B). (D) Percentage of DAPI−/CD3+/CD4+ cells proliferated beyond the 2nd generation following treatment as in (B) as determined by CFSE dilution. Samples were prepared in triplicated and errors bars represent SEM.

    Article Snippet: After 48 hours in culture, CD4 + T cells were isolated from the spleens of 2D2 mice using the manufacturer’s recommended protocol (STEMCELL).

    Techniques: Immunopeptidomics, Isolation, Incubation, Flow Cytometry