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cd27 positive selection beads  (Miltenyi Biotec)


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    Miltenyi Biotec cd27 positive selection beads
    Cd27 Positive Selection Beads, supplied by Miltenyi Biotec, used in various techniques. Bioz Stars score: 93/100, based on 63 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/cd27+microbeads/CD27+MicroBeads%2C+human/us12590386-1166-6-10
    Average 93 stars, based on 63 article reviews
    cd27 positive selection beads - by Bioz Stars, 2026-09
    93/100 stars

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    Related Articles

    Isolation:

    Article Title: HIV-1 diversity in viral reservoirs obtained from circulating T-cell subsets during early ART and beyond.
    Article Snippet: Up to 40 mL of whole blood was drawn from HIV-1 positive individuals on ART, PBMC were isolated by density gradient centrifugation using Histopaque 1.077 g/mol (Sigma). .. Subsequently, untouched CD4+ T cells were isolated by negative selection (CD4 T Cell Isolation Kit, Miltenyi), The individual T cell memory compartments of CD4 +T cells were separated by MACS using CD45RA microbeads (CD45RA MicroBeads human, Miltenyi), CD62L microbeads (CD62L MicroBeads human, Miltenyi), CD27 microbeads (CD27 MicroBeads human, Miltenyi) to separate T naïve (TN: CD45RA+CD62L+CD27+), T central memory (TCM: CD45RA-CD62L+CD27+), T translational memory (TTM: CD45RA-CD62L-CD27+)), and T effector memory (TEM: CD45RA-CD62L-CD27-) cells. ..

    Article Title: HIV-1 diversity in viral reservoirs obtained from circulating T-cell subsets during early ART and beyond
    Article Snippet: Up to 40 mL of whole blood was drawn from HIV-1 positive individuals on ART, PBMC were isolated by density gradient centrifugation using Histopaque 1.077 g/mol (Sigma). .. Subsequently, untouched CD4+ T cells were isolated by negative selection (CD4 T Cell Isolation Kit, Miltenyi), The individual T cell memory compartments of CD4 +T cells were separated by MACS using CD45RA microbeads (CD45RA MicroBeads human, Miltenyi), CD62L microbeads (CD62L MicroBeads human, Miltenyi), CD27 microbeads (CD27 MicroBeads human, Miltenyi) to separate T naïve (TN: CD45RA+CD62L+CD27+), T central memory (TCM: CD45RA-CD62L+CD27+), T translational memory (TTM: CD45RA-CD62L-CD27+)), and T effector memory (TEM: CD45RA-CD62L-CD27-) cells. ..

    Article Title: Tph cells are expanded in IgA vasculitis nephritis.
    Article Snippet: Ectopic lymphoid structures (ELSs) contribute to maintaining the chronic inflammation process of IgA vasculitis nephritis (IgAVN) and are associated with a more severe disease course.. PD-1hiCXCR5− CD4 T peripheral helper (Tph) cells are recognized as the main CD4 T cells that contribute to B cell immune responses and antibody production in ELSs.. However, the role of Tph cells in the pathogenesis of IgAVN is currently unknown.

    Article Title: Disruptors of sestrin-MAPK interactions rejuvenate T cells and expand TCR specificity
    Article Snippet: Peripheral blood mononuclear cells (PBMCs) were isolated by Ficoll (17-1440-03; GE Healthcare) density gradient centrifugation from blood of healthy volunteers. .. Primary human CD4 + T cells were isolated using CD4 MicroBeads (130-045-101; Miltenyi); primary human senescent CD4 + CD27 - CD28 - T cells (thereafter T sen ) were isolated using CD4 + T Cell Isolation Kit (130-096-533; Miltenyi) followed by depletion of CD27 + and CD28 + T cells using CD27 MicroBeads (130-051-601; Miltenyi) and CD28 MicroBead Kit (130-093-247; Miltenyi). .. CD3 - cells (thereafter APCs) were isolated by depleting CD3 + cells using human CD3 MicroBeads (130-050-101; Miltenyi).

    Selection:

    Article Title: HIV-1 diversity in viral reservoirs obtained from circulating T-cell subsets during early ART and beyond.
    Article Snippet: Up to 40 mL of whole blood was drawn from HIV-1 positive individuals on ART, PBMC were isolated by density gradient centrifugation using Histopaque 1.077 g/mol (Sigma). .. Subsequently, untouched CD4+ T cells were isolated by negative selection (CD4 T Cell Isolation Kit, Miltenyi), The individual T cell memory compartments of CD4 +T cells were separated by MACS using CD45RA microbeads (CD45RA MicroBeads human, Miltenyi), CD62L microbeads (CD62L MicroBeads human, Miltenyi), CD27 microbeads (CD27 MicroBeads human, Miltenyi) to separate T naïve (TN: CD45RA+CD62L+CD27+), T central memory (TCM: CD45RA-CD62L+CD27+), T translational memory (TTM: CD45RA-CD62L-CD27+)), and T effector memory (TEM: CD45RA-CD62L-CD27-) cells. ..

    Article Title: HIV-1 diversity in viral reservoirs obtained from circulating T-cell subsets during early ART and beyond
    Article Snippet: Up to 40 mL of whole blood was drawn from HIV-1 positive individuals on ART, PBMC were isolated by density gradient centrifugation using Histopaque 1.077 g/mol (Sigma). .. Subsequently, untouched CD4+ T cells were isolated by negative selection (CD4 T Cell Isolation Kit, Miltenyi), The individual T cell memory compartments of CD4 +T cells were separated by MACS using CD45RA microbeads (CD45RA MicroBeads human, Miltenyi), CD62L microbeads (CD62L MicroBeads human, Miltenyi), CD27 microbeads (CD27 MicroBeads human, Miltenyi) to separate T naïve (TN: CD45RA+CD62L+CD27+), T central memory (TCM: CD45RA-CD62L+CD27+), T translational memory (TTM: CD45RA-CD62L-CD27+)), and T effector memory (TEM: CD45RA-CD62L-CD27-) cells. ..

    Cell Isolation:

    Article Title: HIV-1 diversity in viral reservoirs obtained from circulating T-cell subsets during early ART and beyond.
    Article Snippet: Up to 40 mL of whole blood was drawn from HIV-1 positive individuals on ART, PBMC were isolated by density gradient centrifugation using Histopaque 1.077 g/mol (Sigma). .. Subsequently, untouched CD4+ T cells were isolated by negative selection (CD4 T Cell Isolation Kit, Miltenyi), The individual T cell memory compartments of CD4 +T cells were separated by MACS using CD45RA microbeads (CD45RA MicroBeads human, Miltenyi), CD62L microbeads (CD62L MicroBeads human, Miltenyi), CD27 microbeads (CD27 MicroBeads human, Miltenyi) to separate T naïve (TN: CD45RA+CD62L+CD27+), T central memory (TCM: CD45RA-CD62L+CD27+), T translational memory (TTM: CD45RA-CD62L-CD27+)), and T effector memory (TEM: CD45RA-CD62L-CD27-) cells. ..

    Article Title: HIV-1 diversity in viral reservoirs obtained from circulating T-cell subsets during early ART and beyond
    Article Snippet: Up to 40 mL of whole blood was drawn from HIV-1 positive individuals on ART, PBMC were isolated by density gradient centrifugation using Histopaque 1.077 g/mol (Sigma). .. Subsequently, untouched CD4+ T cells were isolated by negative selection (CD4 T Cell Isolation Kit, Miltenyi), The individual T cell memory compartments of CD4 +T cells were separated by MACS using CD45RA microbeads (CD45RA MicroBeads human, Miltenyi), CD62L microbeads (CD62L MicroBeads human, Miltenyi), CD27 microbeads (CD27 MicroBeads human, Miltenyi) to separate T naïve (TN: CD45RA+CD62L+CD27+), T central memory (TCM: CD45RA-CD62L+CD27+), T translational memory (TTM: CD45RA-CD62L-CD27+)), and T effector memory (TEM: CD45RA-CD62L-CD27-) cells. ..

    Article Title: Disruptors of sestrin-MAPK interactions rejuvenate T cells and expand TCR specificity
    Article Snippet: Peripheral blood mononuclear cells (PBMCs) were isolated by Ficoll (17-1440-03; GE Healthcare) density gradient centrifugation from blood of healthy volunteers. .. Primary human CD4 + T cells were isolated using CD4 MicroBeads (130-045-101; Miltenyi); primary human senescent CD4 + CD27 - CD28 - T cells (thereafter T sen ) were isolated using CD4 + T Cell Isolation Kit (130-096-533; Miltenyi) followed by depletion of CD27 + and CD28 + T cells using CD27 MicroBeads (130-051-601; Miltenyi) and CD28 MicroBead Kit (130-093-247; Miltenyi). .. CD3 - cells (thereafter APCs) were isolated by depleting CD3 + cells using human CD3 MicroBeads (130-050-101; Miltenyi).

    Magnetic Cell Separation:

    Article Title: HIV-1 diversity in viral reservoirs obtained from circulating T-cell subsets during early ART and beyond.
    Article Snippet: Up to 40 mL of whole blood was drawn from HIV-1 positive individuals on ART, PBMC were isolated by density gradient centrifugation using Histopaque 1.077 g/mol (Sigma). .. Subsequently, untouched CD4+ T cells were isolated by negative selection (CD4 T Cell Isolation Kit, Miltenyi), The individual T cell memory compartments of CD4 +T cells were separated by MACS using CD45RA microbeads (CD45RA MicroBeads human, Miltenyi), CD62L microbeads (CD62L MicroBeads human, Miltenyi), CD27 microbeads (CD27 MicroBeads human, Miltenyi) to separate T naïve (TN: CD45RA+CD62L+CD27+), T central memory (TCM: CD45RA-CD62L+CD27+), T translational memory (TTM: CD45RA-CD62L-CD27+)), and T effector memory (TEM: CD45RA-CD62L-CD27-) cells. ..

    Article Title: HIV-1 diversity in viral reservoirs obtained from circulating T-cell subsets during early ART and beyond
    Article Snippet: Up to 40 mL of whole blood was drawn from HIV-1 positive individuals on ART, PBMC were isolated by density gradient centrifugation using Histopaque 1.077 g/mol (Sigma). .. Subsequently, untouched CD4+ T cells were isolated by negative selection (CD4 T Cell Isolation Kit, Miltenyi), The individual T cell memory compartments of CD4 +T cells were separated by MACS using CD45RA microbeads (CD45RA MicroBeads human, Miltenyi), CD62L microbeads (CD62L MicroBeads human, Miltenyi), CD27 microbeads (CD27 MicroBeads human, Miltenyi) to separate T naïve (TN: CD45RA+CD62L+CD27+), T central memory (TCM: CD45RA-CD62L+CD27+), T translational memory (TTM: CD45RA-CD62L-CD27+)), and T effector memory (TEM: CD45RA-CD62L-CD27-) cells. ..

    Transmission Electron Microscopy:

    Article Title: HIV-1 diversity in viral reservoirs obtained from circulating T-cell subsets during early ART and beyond.
    Article Snippet: Up to 40 mL of whole blood was drawn from HIV-1 positive individuals on ART, PBMC were isolated by density gradient centrifugation using Histopaque 1.077 g/mol (Sigma). .. Subsequently, untouched CD4+ T cells were isolated by negative selection (CD4 T Cell Isolation Kit, Miltenyi), The individual T cell memory compartments of CD4 +T cells were separated by MACS using CD45RA microbeads (CD45RA MicroBeads human, Miltenyi), CD62L microbeads (CD62L MicroBeads human, Miltenyi), CD27 microbeads (CD27 MicroBeads human, Miltenyi) to separate T naïve (TN: CD45RA+CD62L+CD27+), T central memory (TCM: CD45RA-CD62L+CD27+), T translational memory (TTM: CD45RA-CD62L-CD27+)), and T effector memory (TEM: CD45RA-CD62L-CD27-) cells. ..

    Article Title: HIV-1 diversity in viral reservoirs obtained from circulating T-cell subsets during early ART and beyond
    Article Snippet: Up to 40 mL of whole blood was drawn from HIV-1 positive individuals on ART, PBMC were isolated by density gradient centrifugation using Histopaque 1.077 g/mol (Sigma). .. Subsequently, untouched CD4+ T cells were isolated by negative selection (CD4 T Cell Isolation Kit, Miltenyi), The individual T cell memory compartments of CD4 +T cells were separated by MACS using CD45RA microbeads (CD45RA MicroBeads human, Miltenyi), CD62L microbeads (CD62L MicroBeads human, Miltenyi), CD27 microbeads (CD27 MicroBeads human, Miltenyi) to separate T naïve (TN: CD45RA+CD62L+CD27+), T central memory (TCM: CD45RA-CD62L+CD27+), T translational memory (TTM: CD45RA-CD62L-CD27+)), and T effector memory (TEM: CD45RA-CD62L-CD27-) cells. ..

    Labeling:

    Article Title: Tph cells are expanded in IgA vasculitis nephritis.
    Article Snippet: Ectopic lymphoid structures (ELSs) contribute to maintaining the chronic inflammation process of IgA vasculitis nephritis (IgAVN) and are associated with a more severe disease course.. PD-1hiCXCR5− CD4 T peripheral helper (Tph) cells are recognized as the main CD4 T cells that contribute to B cell immune responses and antibody production in ELSs.. However, the role of Tph cells in the pathogenesis of IgAVN is currently unknown.

    FACS:

    Article Title: Tph cells are expanded in IgA vasculitis nephritis.
    Article Snippet: Ectopic lymphoid structures (ELSs) contribute to maintaining the chronic inflammation process of IgA vasculitis nephritis (IgAVN) and are associated with a more severe disease course.. PD-1hiCXCR5− CD4 T peripheral helper (Tph) cells are recognized as the main CD4 T cells that contribute to B cell immune responses and antibody production in ELSs.. However, the role of Tph cells in the pathogenesis of IgAVN is currently unknown.

    Flow Cytometry:

    Article Title: Tph cells are expanded in IgA vasculitis nephritis.
    Article Snippet: Ectopic lymphoid structures (ELSs) contribute to maintaining the chronic inflammation process of IgA vasculitis nephritis (IgAVN) and are associated with a more severe disease course.. PD-1hiCXCR5− CD4 T peripheral helper (Tph) cells are recognized as the main CD4 T cells that contribute to B cell immune responses and antibody production in ELSs.. However, the role of Tph cells in the pathogenesis of IgAVN is currently unknown.



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    (A) Senescence-associated β-galactosidase expression in DOS-treated (DOS-juvenated) or untreated T sen . Cells were purified and cultured for one week in the presence of anti-CD3 (0.5 μg/mL) and rh-IL-2 (5 ng/mL), then stained to detect β-galactosidase activity. Representative image on the inverted phase-contrast microscope (left) and relative quantification (right, n = 8 donors). (B) Population doublings of human T sen (transduced with irrelevant scramble) and sestrin null CD4 + T sen (transduced by triple lentiviral depletion of sestrins) and cultured as in (A) (left). Donor-matched <t>CD27</t> + CD28 + CD4 + T cells (herafter, T erl ) were cultured in parallel but activated with anti-CD3 and anti-CD28 ( n = 5 donors). Cells were cultured over two weeks with restimulation every 7 days. DOS-driven population doublings (right) were calculated as delta between DOS treated and DOS untreated T cells, with or without depletion of sestrins. (C) DOS-juvenation of human CD4 + T cells. Terminally differentiated effector memory CD45RA - CD28 - CD27 - CD4 + T cells (hereafter, T EM ) were purified and cultured over 20 days, as in (A). At day 1 (18 hours), 7, and 21 cell phenotypes were assessed by flow cytometry. Quantifications of rejuvenated stem like (CD28 + CD45RA + CCR7 + CD95 + CD62L + TCF1 + ) among human CD4 + T cells are shown (left; n = 5 donors). Decay of T EM and CD28 - CD27 - CD45RA + CD4 + T cells (hereafter, T EMRA ) undergoing rejuvenation is shown (right). (D) Adoptive transfer of DOS-juvenated T cells, experimental design. Donor T cells were derived from twenty-month-old mice 15 days after Fluad vaccination with or without DOS treatment (0.1 mg/Kg throughout), labeled with Cell Trace Violet (CTV) dye or congenic CD45.1 tracking, then transferred into young naïve CD45.2 recipients (3 months). In parallel, young mice (3 months) were used as young donor control. Recipient animals were rested for 28 days, then analysed for donor T cell persistence and maintenance of stem phenotype after transfer. (E) Maintenance of donor DOS-juvenated CD45.1 CD4 + T cells, their aged-matched controls, and that of young donor T cells, 28 days after transfer (day 43) in recipient mouse lymph nodes. Representative flow cytometry plots and poled data ( n = 5 mice per group) are shown. (F) Assessment of mouse T cell memory programs in stem like transferred T cells (among CD45.1 CD44 - CD62L + CD95 + CD4 + T cells) and terminally differentiated cells (TE, among CD45.1 CD44 - CD62L - CD4 + T cells) following adoptive transfer as in (D) ( n = 5 mice). (G) IL7R gene expression and (H) CD95 and Sca-1 mean fluorescent intensity (MFI; throughout) in stem cells derived from CD4 + CD45.1 + transferred stem T cells among lymph nodes of recipient CD45.2 mice, 28 days after transfer ( n = 5 mice). (I) Representative plots of CD45.1 transferred cells in quiescent state before and after transfer assessed by cycle related intra-nuclear Ki67 staining. Representative of n = 5 mice per group. (J) G1 (Ki67 + ) to G0 (Ki67 - ) transition in stem like CD45.1 CD4 + T cells before and after adoptive transfer as indicated ( n = 5 mice per group). (K) Assessment of T cell longevity following DOS-juvenation. Cells from lymph nodes were stained using the Annexin-PI Apoptosis detection Kit 28 days after transfer. Representative FACS plot (left) and quantification of dead CD45.1 + CD4 + transferred T cells (right). (L) DOS-juvenated T cell maintenance, in vivo model. In (A and B, right) two tailed paired T test was used. In (B, left-C, E-H and J-K) one-way Anova with Bonferroni post-correction for multiple comparisons was used, *p<0,05, **P<0,01; ***P<0,001; ****P<0,0001. Error bars indicate SEM.
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    Image Search Results


    Fig. 2 Single-cell secreting proteomics-defined auto-clustering of tumor-infiltrating SELL + and TNFRSF7 + NK cells in TNBC patients. A. Single-cell se creting proteomics-defined UMAP plot visualizing secreting tumor infiltrating SELL + and TNFRSF7 + NK cell clusters. B. Single-cell secreting proteomics- defined UMAP plot visualizing cells grouped by specimen (SELL + sorted and TNFRSF7 + sorted cells). C. Bar plot showing cell proportions of specimen group in each cluster. D. Dot plot showing single-cell cytokine levels and cytokine-secreting cell proportions in each cluster. E. UMAP plot visualizing expression of featured cytokines in each cluster

    Journal: Breast cancer research : BCR

    Article Title: Phenotypes and cytokines of NK cells in triple-negative breast cancer resistant to checkpoint blockade immunotherapy.

    doi: 10.1186/s13058-025-02003-y

    Figure Lengend Snippet: Fig. 2 Single-cell secreting proteomics-defined auto-clustering of tumor-infiltrating SELL + and TNFRSF7 + NK cells in TNBC patients. A. Single-cell se creting proteomics-defined UMAP plot visualizing secreting tumor infiltrating SELL + and TNFRSF7 + NK cell clusters. B. Single-cell secreting proteomics- defined UMAP plot visualizing cells grouped by specimen (SELL + sorted and TNFRSF7 + sorted cells). C. Bar plot showing cell proportions of specimen group in each cluster. D. Dot plot showing single-cell cytokine levels and cytokine-secreting cell proportions in each cluster. E. UMAP plot visualizing expression of featured cytokines in each cluster

    Article Snippet: Cell subclusters of interest (SELL + NK cells and TNFRSF7 + NK cells in single-cell suspensions of tumor tissue) were isolated with SELL microbeads (#130-091-758, Miltenyi Biotec) as well as TNFRSF7 + microbeads (#130-051-601, Miltenyi Biotec) as positive selection and resuspended in complete RPMI media.

    Techniques: Expressing

    (A) Senescence-associated β-galactosidase expression in DOS-treated (DOS-juvenated) or untreated T sen . Cells were purified and cultured for one week in the presence of anti-CD3 (0.5 μg/mL) and rh-IL-2 (5 ng/mL), then stained to detect β-galactosidase activity. Representative image on the inverted phase-contrast microscope (left) and relative quantification (right, n = 8 donors). (B) Population doublings of human T sen (transduced with irrelevant scramble) and sestrin null CD4 + T sen (transduced by triple lentiviral depletion of sestrins) and cultured as in (A) (left). Donor-matched CD27 + CD28 + CD4 + T cells (herafter, T erl ) were cultured in parallel but activated with anti-CD3 and anti-CD28 ( n = 5 donors). Cells were cultured over two weeks with restimulation every 7 days. DOS-driven population doublings (right) were calculated as delta between DOS treated and DOS untreated T cells, with or without depletion of sestrins. (C) DOS-juvenation of human CD4 + T cells. Terminally differentiated effector memory CD45RA - CD28 - CD27 - CD4 + T cells (hereafter, T EM ) were purified and cultured over 20 days, as in (A). At day 1 (18 hours), 7, and 21 cell phenotypes were assessed by flow cytometry. Quantifications of rejuvenated stem like (CD28 + CD45RA + CCR7 + CD95 + CD62L + TCF1 + ) among human CD4 + T cells are shown (left; n = 5 donors). Decay of T EM and CD28 - CD27 - CD45RA + CD4 + T cells (hereafter, T EMRA ) undergoing rejuvenation is shown (right). (D) Adoptive transfer of DOS-juvenated T cells, experimental design. Donor T cells were derived from twenty-month-old mice 15 days after Fluad vaccination with or without DOS treatment (0.1 mg/Kg throughout), labeled with Cell Trace Violet (CTV) dye or congenic CD45.1 tracking, then transferred into young naïve CD45.2 recipients (3 months). In parallel, young mice (3 months) were used as young donor control. Recipient animals were rested for 28 days, then analysed for donor T cell persistence and maintenance of stem phenotype after transfer. (E) Maintenance of donor DOS-juvenated CD45.1 CD4 + T cells, their aged-matched controls, and that of young donor T cells, 28 days after transfer (day 43) in recipient mouse lymph nodes. Representative flow cytometry plots and poled data ( n = 5 mice per group) are shown. (F) Assessment of mouse T cell memory programs in stem like transferred T cells (among CD45.1 CD44 - CD62L + CD95 + CD4 + T cells) and terminally differentiated cells (TE, among CD45.1 CD44 - CD62L - CD4 + T cells) following adoptive transfer as in (D) ( n = 5 mice). (G) IL7R gene expression and (H) CD95 and Sca-1 mean fluorescent intensity (MFI; throughout) in stem cells derived from CD4 + CD45.1 + transferred stem T cells among lymph nodes of recipient CD45.2 mice, 28 days after transfer ( n = 5 mice). (I) Representative plots of CD45.1 transferred cells in quiescent state before and after transfer assessed by cycle related intra-nuclear Ki67 staining. Representative of n = 5 mice per group. (J) G1 (Ki67 + ) to G0 (Ki67 - ) transition in stem like CD45.1 CD4 + T cells before and after adoptive transfer as indicated ( n = 5 mice per group). (K) Assessment of T cell longevity following DOS-juvenation. Cells from lymph nodes were stained using the Annexin-PI Apoptosis detection Kit 28 days after transfer. Representative FACS plot (left) and quantification of dead CD45.1 + CD4 + transferred T cells (right). (L) DOS-juvenated T cell maintenance, in vivo model. In (A and B, right) two tailed paired T test was used. In (B, left-C, E-H and J-K) one-way Anova with Bonferroni post-correction for multiple comparisons was used, *p<0,05, **P<0,01; ***P<0,001; ****P<0,0001. Error bars indicate SEM.

    Journal: bioRxiv

    Article Title: Disruptors of sestrin-MAPK interactions rejuvenate T cells and expand TCR specificity

    doi: 10.1101/2024.05.17.594698

    Figure Lengend Snippet: (A) Senescence-associated β-galactosidase expression in DOS-treated (DOS-juvenated) or untreated T sen . Cells were purified and cultured for one week in the presence of anti-CD3 (0.5 μg/mL) and rh-IL-2 (5 ng/mL), then stained to detect β-galactosidase activity. Representative image on the inverted phase-contrast microscope (left) and relative quantification (right, n = 8 donors). (B) Population doublings of human T sen (transduced with irrelevant scramble) and sestrin null CD4 + T sen (transduced by triple lentiviral depletion of sestrins) and cultured as in (A) (left). Donor-matched CD27 + CD28 + CD4 + T cells (herafter, T erl ) were cultured in parallel but activated with anti-CD3 and anti-CD28 ( n = 5 donors). Cells were cultured over two weeks with restimulation every 7 days. DOS-driven population doublings (right) were calculated as delta between DOS treated and DOS untreated T cells, with or without depletion of sestrins. (C) DOS-juvenation of human CD4 + T cells. Terminally differentiated effector memory CD45RA - CD28 - CD27 - CD4 + T cells (hereafter, T EM ) were purified and cultured over 20 days, as in (A). At day 1 (18 hours), 7, and 21 cell phenotypes were assessed by flow cytometry. Quantifications of rejuvenated stem like (CD28 + CD45RA + CCR7 + CD95 + CD62L + TCF1 + ) among human CD4 + T cells are shown (left; n = 5 donors). Decay of T EM and CD28 - CD27 - CD45RA + CD4 + T cells (hereafter, T EMRA ) undergoing rejuvenation is shown (right). (D) Adoptive transfer of DOS-juvenated T cells, experimental design. Donor T cells were derived from twenty-month-old mice 15 days after Fluad vaccination with or without DOS treatment (0.1 mg/Kg throughout), labeled with Cell Trace Violet (CTV) dye or congenic CD45.1 tracking, then transferred into young naïve CD45.2 recipients (3 months). In parallel, young mice (3 months) were used as young donor control. Recipient animals were rested for 28 days, then analysed for donor T cell persistence and maintenance of stem phenotype after transfer. (E) Maintenance of donor DOS-juvenated CD45.1 CD4 + T cells, their aged-matched controls, and that of young donor T cells, 28 days after transfer (day 43) in recipient mouse lymph nodes. Representative flow cytometry plots and poled data ( n = 5 mice per group) are shown. (F) Assessment of mouse T cell memory programs in stem like transferred T cells (among CD45.1 CD44 - CD62L + CD95 + CD4 + T cells) and terminally differentiated cells (TE, among CD45.1 CD44 - CD62L - CD4 + T cells) following adoptive transfer as in (D) ( n = 5 mice). (G) IL7R gene expression and (H) CD95 and Sca-1 mean fluorescent intensity (MFI; throughout) in stem cells derived from CD4 + CD45.1 + transferred stem T cells among lymph nodes of recipient CD45.2 mice, 28 days after transfer ( n = 5 mice). (I) Representative plots of CD45.1 transferred cells in quiescent state before and after transfer assessed by cycle related intra-nuclear Ki67 staining. Representative of n = 5 mice per group. (J) G1 (Ki67 + ) to G0 (Ki67 - ) transition in stem like CD45.1 CD4 + T cells before and after adoptive transfer as indicated ( n = 5 mice per group). (K) Assessment of T cell longevity following DOS-juvenation. Cells from lymph nodes were stained using the Annexin-PI Apoptosis detection Kit 28 days after transfer. Representative FACS plot (left) and quantification of dead CD45.1 + CD4 + transferred T cells (right). (L) DOS-juvenated T cell maintenance, in vivo model. In (A and B, right) two tailed paired T test was used. In (B, left-C, E-H and J-K) one-way Anova with Bonferroni post-correction for multiple comparisons was used, *p<0,05, **P<0,01; ***P<0,001; ****P<0,0001. Error bars indicate SEM.

    Article Snippet: Primary human CD4 + T cells were isolated using CD4 MicroBeads (130-045-101; Miltenyi); primary human senescent CD4 + CD27 - CD28 - T cells (thereafter T sen ) were isolated using CD4 + T Cell Isolation Kit (130-096-533; Miltenyi) followed by depletion of CD27 + and CD28 + T cells using CD27 MicroBeads (130-051-601; Miltenyi) and CD28 MicroBead Kit (130-093-247; Miltenyi).

    Techniques: Expressing, Purification, Cell Culture, Staining, Activity Assay, Microscopy, Quantitative Proteomics, Transduction, Flow Cytometry, Adoptive Transfer Assay, Derivative Assay, Labeling, Control, Gene Expression, In Vivo, Two Tailed Test