Review



magnetic negative selection  (Miltenyi Biotec)


Bioz Verified Symbol Miltenyi Biotec is a verified supplier
Bioz Manufacturer Symbol Miltenyi Biotec manufactures this product  
  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 96

    Structured Review

    Miltenyi Biotec magnetic negative selection
    Magnetic Negative Selection, supplied by Miltenyi Biotec, used in various techniques. Bioz Stars score: 96/100, based on 209 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/cd4+t+cell+negative+isolation+kit/Naive+CD4%2B+T+Cell+Isolation+Kit+II%2C+human/pmc13284711-33-11-21
    Average 96 stars, based on 209 article reviews
    magnetic negative selection - by Bioz Stars, 2026-10
    96/100 stars

    Images

    Related Articles

    Single Cell:

    Article Title: Lymphedema pathogenesis involves antigen-driven expansion of CD4+ T cells in skin
    Article Snippet: .. Single-cell suspensions were prepared from inguinal nodes pooled from each group and sort purified for effector TCRβ + CD4 + CD44 hi CD62L lo cells after enrichment with a CD4 + T cell negative isolation kit (cat#130-104-454; Miltenyi Biotec). .. Similarly, antigen-presenting cells (APCs) were isolated from single-cell suspensions from one spleen harvested from a sham control and irradiated after depletion of T cells using antibodies against CD90.2 (cat#14-0902-82; Invitrogen).

    Article Title: Discovery of a Selective, Novel TGF-βR1 Inhibitor GFH018 for the Treatment of Solid Tumors.
    Article Snippet: Transforming growth factor-β (TGF-β) signaling pathway plays an important role in tumorigenesis.. Inhibition of the TGF-β type I receptor kinase is considered an attractive strategy for the treatment of tumor growth and metastasis.. This paper describes the discovery of GFH018, a highly potent and selective ATP-competitive inhibitor of TGF-βR1 that entered phase II clinical trials.

    Purification:

    Article Title: Lymphedema pathogenesis involves antigen-driven expansion of CD4+ T cells in skin
    Article Snippet: .. Single-cell suspensions were prepared from inguinal nodes pooled from each group and sort purified for effector TCRβ + CD4 + CD44 hi CD62L lo cells after enrichment with a CD4 + T cell negative isolation kit (cat#130-104-454; Miltenyi Biotec). .. Similarly, antigen-presenting cells (APCs) were isolated from single-cell suspensions from one spleen harvested from a sham control and irradiated after depletion of T cells using antibodies against CD90.2 (cat#14-0902-82; Invitrogen).

    Article Title: Discovery of a Selective, Novel TGF-βR1 Inhibitor GFH018 for the Treatment of Solid Tumors.
    Article Snippet: Transforming growth factor-β (TGF-β) signaling pathway plays an important role in tumorigenesis.. Inhibition of the TGF-β type I receptor kinase is considered an attractive strategy for the treatment of tumor growth and metastasis.. This paper describes the discovery of GFH018, a highly potent and selective ATP-competitive inhibitor of TGF-βR1 that entered phase II clinical trials.

    Isolation:

    Article Title: Lymphedema pathogenesis involves antigen-driven expansion of CD4+ T cells in skin
    Article Snippet: .. Single-cell suspensions were prepared from inguinal nodes pooled from each group and sort purified for effector TCRβ + CD4 + CD44 hi CD62L lo cells after enrichment with a CD4 + T cell negative isolation kit (cat#130-104-454; Miltenyi Biotec). .. Similarly, antigen-presenting cells (APCs) were isolated from single-cell suspensions from one spleen harvested from a sham control and irradiated after depletion of T cells using antibodies against CD90.2 (cat#14-0902-82; Invitrogen).

    Article Title: Discovery of a Selective, Novel TGF-βR1 Inhibitor GFH018 for the Treatment of Solid Tumors.
    Article Snippet: Transforming growth factor-β (TGF-β) signaling pathway plays an important role in tumorigenesis.. Inhibition of the TGF-β type I receptor kinase is considered an attractive strategy for the treatment of tumor growth and metastasis.. This paper describes the discovery of GFH018, a highly potent and selective ATP-competitive inhibitor of TGF-βR1 that entered phase II clinical trials.

    Article Title: Fibrosis-memory is mediated by IL-3–producing T cells and drives progression of fibrosis
    Article Snippet: Splenic CD4+ or CD8+ T cells were isolated with CD4 or CD8 microbeads (Miltenyi). .. For isolation of splenic CD62L+CD4+ and CD62L–CD4+ T cells, the CD4+ T cells were first isolated with a CD4+ T cell negative isolation kit (Miltenyi) and subsequently separated with CD62L microbeads (Miltenyi). .. CD4+ T cells were isolated from the kidneys of RUUO+UUO donor mice in two steps: first with a CD4+ T cell negative isolation kit (Miltenyi) and then with CD4 (L3T4) microbeads (Miltenyi).

    Article Title: Fibrosis-memory is mediated by IL-3–producing T cells and drives progression of fibrosis
    Article Snippet: For isolation of splenic CD62L + CD4 + and CD62L – CD4 + T cells, the CD4 + T cells were first isolated with a CD4 + T cell negative isolation kit (Miltenyi) and subsequently separated with CD62L microbeads (Miltenyi). .. CD4 + T cells were isolated from the kidneys of RUUO+UUO donor mice in two steps: first with a CD4 + T cell negative isolation kit (Miltenyi) and then with CD4 (L3T4) microbeads (Miltenyi). ..

    Article Title: Inhibition of protein kinase R suppresses HIV replication and integration in CD4 T cells
    Article Snippet: .. Peripheral blood mononuclear cells (PBMCs) were isolated from healthy subjects (HS) using density gradient separation (Lymphoprep, SepMate50), and CD4 T cells were isolated by negative selection (CD4 T Cell Negative Isolation Kit, Miltenyi Biotec). ..

    Article Title: Fibrosis-memory is mediated by IL-3–producing T cells and drives progression of fibrosis
    Article Snippet: Splenic CD4 + or CD8 + T cells were isolated with CD4 or CD8 microbeads (Miltenyi). .. For isolation of splenic CD62L + CD4 + and CD62L – CD4 + T cells, the CD4 + T cells were first isolated with a CD4 + T cell negative isolation kit (Miltenyi) and subsequently separated with CD62L microbeads (Miltenyi). .. CD4 + T cells were isolated from the kidneys of RUUO+UUO donor mice in two steps: first with a CD4 + T cell negative isolation kit (Miltenyi) and then with CD4 (L3T4) microbeads (Miltenyi).

    Article Title: Fibrosis-memory is mediated by IL-3–producing T cells and drives progression of fibrosis
    Article Snippet: For isolation of splenic CD62L+CD4+ and CD62L–CD4+ T cells, the CD4+ T cells were first isolated with a CD4+ T cell negative isolation kit (Miltenyi) and subsequently separated with CD62L microbeads (Miltenyi). .. CD4+ T cells were isolated from the kidneys of RUUO+UUO donor mice in two steps: first with a CD4+ T cell negative isolation kit (Miltenyi) and then with CD4 (L3T4) microbeads (Miltenyi). ..

    Selection:

    Article Title: Inhibition of protein kinase R suppresses HIV replication and integration in CD4 T cells
    Article Snippet: .. Peripheral blood mononuclear cells (PBMCs) were isolated from healthy subjects (HS) using density gradient separation (Lymphoprep, SepMate50), and CD4 T cells were isolated by negative selection (CD4 T Cell Negative Isolation Kit, Miltenyi Biotec). ..

    other:

    Article Title: Precursor central memory versus effector cell fate and naïve CD4 + T cell heterogeneity
    Article Snippet: T cell–depleted splenocytes were prepared using biotinylated antibodies against CD4, followed by antibiotin microbeads (Miltenyi Biotec), and irradiation at 450 rad.



    Similar Products

    96
    Miltenyi Biotec magnetic negative selection
    Magnetic Negative Selection, supplied by Miltenyi Biotec, used in various techniques. Bioz Stars score: 96/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/cd4+t+cell+negative+isolation+kit/Naive+CD4%2B+T+Cell+Isolation+Kit+II%2C+human/pmc13284711-33-11-21
    Average 96 stars, based on 1 article reviews
    magnetic negative selection - by Bioz Stars, 2026-10
    96/100 stars
      Buy from Supplier

    97
    Miltenyi Biotec cd4 negative cell isolation kit
    Longitudinal dynamics of the HIV‐1 proviral reservoir following SARS‐CoV‐2 mRNA vaccination. (a) Intact HIV‐1 reservoir, (b) total proviral HIV‐1 DNA, (c) 3′ defective and (d) 5′ defective proviral DNA (copies/million resting <t>CD4</t> + T‐cells) quantified using the Intact Proviral DNA Assay (IPDA) at baseline (pre‐vaccine, day 0, blue, n = 37), 3 months after two vaccine doses (red, n = 27) and 3 months after three doses (green, n = 24). Points represent individual values, and horizontal bars represent median values. No significant changes in reservoir size were observed following vaccination (Wilcoxon signed‐rank and Mann–Whitney U tests). Baseline versus 3 months after the second dose, p = 0.09–0.88 (Wilcoxon), 0.47–0.85 (Mann–Whitney). Baseline versus 3 months after the third dose, p = 0.19–0.41 (Wilcoxon), 0.10–0.39 (Mann–Whitney).
    Cd4 Negative Cell Isolation Kit, 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/cd4+t+cell+negative+isolation+kit/CD4%2B+T+Cell+Isolation+Kit%2C+human/pmc13340954-85-5-10
    Average 97 stars, based on 1 article reviews
    cd4 negative cell isolation kit - by Bioz Stars, 2026-10
    97/100 stars
      Buy from Supplier

    96
    Miltenyi Biotec cd4 t cell negative isolation kit
    ( A , B , and E ) Mice underwent RUUO of the right kidney from day 0–6 (first insult), were sham operated, or remained naive (Ø). Mice then underwent UUO of the left kidney from day 17 to 22 (second insult) or remained naive (Ø). ( A ) Quantification of collagen-1 (Col1), overall fibrosis (fibrotic area), and col1a1 mRNA expression in the left kidney on day 22. Two pooled experiments, with n = 4, naive; 8, received the second insult only; 17, sham; and 16, RUUO. ( B – D ) Mice underwent RUUO of the right kidney from day 0 to 6 (first insult) or remained naive (Ø). Mice then underwent UUO of the left kidney from day 11 to 16 or from day 37 to 42 (second insult). ( B ) Analysis of the left UUO kidney on day 16 or day 42. Two pooled experiments, with n = 14 naive; 10 day 16 UUO; and 8 day 37 UUO for immunofluorescence and qPCR, and 1 single experiment for Trichrome staining, with n = 8 received the second insult only; 6, day 11 UUO; and 8, day 37 UUO. ( C ) Representative images of collagen-1 immunofluorescence and Trichrome staining. ( D ) Quantification of infiltrating <t>CD4</t> + T cells, CD8 + T cells, monocytes (Monos), and neutrophils (Neutros) in the left UUO kidneys (single experiment with n = 8 received the second insult only; 6, day 11 UUO; and 8, day 37 UUO). ( E ) Quantification of CD4 + tissue-resident memory (TRM) T cells in the blood, spleen, and left UUO kidneys (single experiment with n = 6 and 4). Scale bars: 100 μm. Data are represented as mean ± SEM. One-way ANOVA with multiple comparisons. * P < 0.05; ** P < 0.01; *** P < 0.001.
    Cd4 T Cell Negative Isolation Kit, supplied by Miltenyi Biotec, used in various techniques. Bioz Stars score: 96/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/cd4+t+cell+negative+isolation+kit/CD4%2B+T+Cell+Isolation+Kit%2C+non-human+primate/pmc12987615-241-25-32
    Average 96 stars, based on 1 article reviews
    cd4 t cell negative isolation kit - by Bioz Stars, 2026-10
    96/100 stars
      Buy from Supplier

    97
    Miltenyi Biotec naïve cd4 t cell negative isolation kit
    ( A , B , and E ) Mice underwent RUUO of the right kidney from day 0–6 (first insult), were sham operated, or remained naive (Ø). Mice then underwent UUO of the left kidney from day 17 to 22 (second insult) or remained naive (Ø). ( A ) Quantification of collagen-1 (Col1), overall fibrosis (fibrotic area), and col1a1 mRNA expression in the left kidney on day 22. Two pooled experiments, with n = 4, naive; 8, received the second insult only; 17, sham; and 16, RUUO. ( B – D ) Mice underwent RUUO of the right kidney from day 0 to 6 (first insult) or remained naive (Ø). Mice then underwent UUO of the left kidney from day 11 to 16 or from day 37 to 42 (second insult). ( B ) Analysis of the left UUO kidney on day 16 or day 42. Two pooled experiments, with n = 14 naive; 10 day 16 UUO; and 8 day 37 UUO for immunofluorescence and qPCR, and 1 single experiment for Trichrome staining, with n = 8 received the second insult only; 6, day 11 UUO; and 8, day 37 UUO. ( C ) Representative images of collagen-1 immunofluorescence and Trichrome staining. ( D ) Quantification of infiltrating <t>CD4</t> + T cells, CD8 + T cells, monocytes (Monos), and neutrophils (Neutros) in the left UUO kidneys (single experiment with n = 8 received the second insult only; 6, day 11 UUO; and 8, day 37 UUO). ( E ) Quantification of CD4 + tissue-resident memory (TRM) T cells in the blood, spleen, and left UUO kidneys (single experiment with n = 6 and 4). Scale bars: 100 μm. Data are represented as mean ± SEM. One-way ANOVA with multiple comparisons. * P < 0.05; ** P < 0.01; *** P < 0.001.
    Naïve Cd4 T Cell Negative Isolation Kit, 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/cd4+t+cell+negative+isolation+kit/CD4%2B+T+Cell+Isolation+Kit%2C+mouse/pm41830088-161-35-44
    Average 97 stars, based on 1 article reviews
    naïve cd4 t cell negative isolation kit - by Bioz Stars, 2026-10
    97/100 stars
      Buy from Supplier

    97
    Miltenyi Biotec cd4 negative isolation kit
    ( A , B , and E ) Mice underwent RUUO of the right kidney from day 0–6 (first insult), were sham operated, or remained naive (Ø). Mice then underwent UUO of the left kidney from day 17 to 22 (second insult) or remained naive (Ø). ( A ) Quantification of collagen-1 (Col1), overall fibrosis (fibrotic area), and col1a1 mRNA expression in the left kidney on day 22. Two pooled experiments, with n = 4, naive; 8, received the second insult only; 17, sham; and 16, RUUO. ( B – D ) Mice underwent RUUO of the right kidney from day 0 to 6 (first insult) or remained naive (Ø). Mice then underwent UUO of the left kidney from day 11 to 16 or from day 37 to 42 (second insult). ( B ) Analysis of the left UUO kidney on day 16 or day 42. Two pooled experiments, with n = 14 naive; 10 day 16 UUO; and 8 day 37 UUO for immunofluorescence and qPCR, and 1 single experiment for Trichrome staining, with n = 8 received the second insult only; 6, day 11 UUO; and 8, day 37 UUO. ( C ) Representative images of collagen-1 immunofluorescence and Trichrome staining. ( D ) Quantification of infiltrating <t>CD4</t> + T cells, CD8 + T cells, monocytes (Monos), and neutrophils (Neutros) in the left UUO kidneys (single experiment with n = 8 received the second insult only; 6, day 11 UUO; and 8, day 37 UUO). ( E ) Quantification of CD4 + tissue-resident memory (TRM) T cells in the blood, spleen, and left UUO kidneys (single experiment with n = 6 and 4). Scale bars: 100 μm. Data are represented as mean ± SEM. One-way ANOVA with multiple comparisons. * P < 0.05; ** P < 0.01; *** P < 0.001.
    Cd4 Negative Isolation Kit, 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/cd4+t+cell+negative+isolation+kit/CD4%2B+T+Cell+Isolation+Kit%2C+human/us12491159-640-10-14
    Average 97 stars, based on 1 article reviews
    cd4 negative isolation kit - by Bioz Stars, 2026-10
    97/100 stars
      Buy from Supplier

    Image Search Results


    Longitudinal dynamics of the HIV‐1 proviral reservoir following SARS‐CoV‐2 mRNA vaccination. (a) Intact HIV‐1 reservoir, (b) total proviral HIV‐1 DNA, (c) 3′ defective and (d) 5′ defective proviral DNA (copies/million resting CD4 + T‐cells) quantified using the Intact Proviral DNA Assay (IPDA) at baseline (pre‐vaccine, day 0, blue, n = 37), 3 months after two vaccine doses (red, n = 27) and 3 months after three doses (green, n = 24). Points represent individual values, and horizontal bars represent median values. No significant changes in reservoir size were observed following vaccination (Wilcoxon signed‐rank and Mann–Whitney U tests). Baseline versus 3 months after the second dose, p = 0.09–0.88 (Wilcoxon), 0.47–0.85 (Mann–Whitney). Baseline versus 3 months after the third dose, p = 0.19–0.41 (Wilcoxon), 0.10–0.39 (Mann–Whitney).

    Journal: HIV Medicine

    Article Title: HIV ‐1 viral load and reservoir size remain stable following SARS ‐ CoV ‐2 mRNA vaccination in people with HIV

    doi: 10.1111/hiv.70235

    Figure Lengend Snippet: Longitudinal dynamics of the HIV‐1 proviral reservoir following SARS‐CoV‐2 mRNA vaccination. (a) Intact HIV‐1 reservoir, (b) total proviral HIV‐1 DNA, (c) 3′ defective and (d) 5′ defective proviral DNA (copies/million resting CD4 + T‐cells) quantified using the Intact Proviral DNA Assay (IPDA) at baseline (pre‐vaccine, day 0, blue, n = 37), 3 months after two vaccine doses (red, n = 27) and 3 months after three doses (green, n = 24). Points represent individual values, and horizontal bars represent median values. No significant changes in reservoir size were observed following vaccination (Wilcoxon signed‐rank and Mann–Whitney U tests). Baseline versus 3 months after the second dose, p = 0.09–0.88 (Wilcoxon), 0.47–0.85 (Mann–Whitney). Baseline versus 3 months after the third dose, p = 0.19–0.41 (Wilcoxon), 0.10–0.39 (Mann–Whitney).

    Article Snippet: For the first step, the CD4 negative cell isolation kit (Miltenyi Biotec, Cat# 130–096‐533) was used according to the manufacturer's protocol.

    Techniques: MANN-WHITNEY

    SARS‐CoV‐2 mRNA vaccine‐induced immune response in relation to HIV‐1 RNA and fold change of intact HIV‐1 reservoir. (a) SARS‐CoV‐2‐Spike antibody titre (U/mL) in people with HIV stratified by HIV‐1 RNA <20 (LLOQ) (blue) and ≥ 20 copies/mL (red), 2 weeks after second vaccine dose ( n = 36), 6 months after second dose ( n = 36) and 3 months after third dose ( n = 18) of SARS‐CoV‐2 mRNA vaccine (Mann–Whitney U test, p = 0.07, 0.4, 0.29). (b) Correlation between fold change of intact HIV‐1 DNA (IPDA) from baseline (day 0) to 3 months after second dose (blue) or 3 months after third dose (red) and vaccine‐induced SARS‐CoV‐2‐Spike antibody titres (Spearman's correlation). (c) Correlation between fold change of intact HIV‐1 DNA from baseline to 3 months after second dose and the proportion of Spike‐specific CD4 + T‐cells (Spearman's correlation).

    Journal: HIV Medicine

    Article Title: HIV ‐1 viral load and reservoir size remain stable following SARS ‐ CoV ‐2 mRNA vaccination in people with HIV

    doi: 10.1111/hiv.70235

    Figure Lengend Snippet: SARS‐CoV‐2 mRNA vaccine‐induced immune response in relation to HIV‐1 RNA and fold change of intact HIV‐1 reservoir. (a) SARS‐CoV‐2‐Spike antibody titre (U/mL) in people with HIV stratified by HIV‐1 RNA <20 (LLOQ) (blue) and ≥ 20 copies/mL (red), 2 weeks after second vaccine dose ( n = 36), 6 months after second dose ( n = 36) and 3 months after third dose ( n = 18) of SARS‐CoV‐2 mRNA vaccine (Mann–Whitney U test, p = 0.07, 0.4, 0.29). (b) Correlation between fold change of intact HIV‐1 DNA (IPDA) from baseline (day 0) to 3 months after second dose (blue) or 3 months after third dose (red) and vaccine‐induced SARS‐CoV‐2‐Spike antibody titres (Spearman's correlation). (c) Correlation between fold change of intact HIV‐1 DNA from baseline to 3 months after second dose and the proportion of Spike‐specific CD4 + T‐cells (Spearman's correlation).

    Article Snippet: For the first step, the CD4 negative cell isolation kit (Miltenyi Biotec, Cat# 130–096‐533) was used according to the manufacturer's protocol.

    Techniques: MANN-WHITNEY

    Immunological profiles and intact proviral HIV‐1 reservoir at baseline and post mRNA vaccination. (a) Intact HIV‐1 reservoir (copies/million resting CD4 + T‐cells) dynamics in INR (CD4 < 350 cells/mm 3 , n = 15) and IR (CD≥350 cells/mm 3 , n = 22) at baseline and after two and three doses of vaccine. A larger intact reservoir was observed in INR at baseline (Mann–Whitney U test, p = 0.02). No significant within‐group changes or between‐group differences of absolute values and fold change were observed following vaccination (Wilcoxon signed‐rank and Mann–Whitney U tests, all p ≥ 0.25). (b–d) Correlation between fold change in intact HIV‐1 DNA from baseline (day 0) to 3 months after the second (blue) and third vaccine dose (red) and (b) CD4 + T‐cell count (cells/mm 3 ), (c) CD4 + /CD8 + ratio and (d) nadir CD4 + T‐cell count (cells/mm 3 ). No significant correlations were observed (Spearman's correlation).

    Journal: HIV Medicine

    Article Title: HIV ‐1 viral load and reservoir size remain stable following SARS ‐ CoV ‐2 mRNA vaccination in people with HIV

    doi: 10.1111/hiv.70235

    Figure Lengend Snippet: Immunological profiles and intact proviral HIV‐1 reservoir at baseline and post mRNA vaccination. (a) Intact HIV‐1 reservoir (copies/million resting CD4 + T‐cells) dynamics in INR (CD4 < 350 cells/mm 3 , n = 15) and IR (CD≥350 cells/mm 3 , n = 22) at baseline and after two and three doses of vaccine. A larger intact reservoir was observed in INR at baseline (Mann–Whitney U test, p = 0.02). No significant within‐group changes or between‐group differences of absolute values and fold change were observed following vaccination (Wilcoxon signed‐rank and Mann–Whitney U tests, all p ≥ 0.25). (b–d) Correlation between fold change in intact HIV‐1 DNA from baseline (day 0) to 3 months after the second (blue) and third vaccine dose (red) and (b) CD4 + T‐cell count (cells/mm 3 ), (c) CD4 + /CD8 + ratio and (d) nadir CD4 + T‐cell count (cells/mm 3 ). No significant correlations were observed (Spearman's correlation).

    Article Snippet: For the first step, the CD4 negative cell isolation kit (Miltenyi Biotec, Cat# 130–096‐533) was used according to the manufacturer's protocol.

    Techniques: MANN-WHITNEY, Cell Characterization

    ( A , B , and E ) Mice underwent RUUO of the right kidney from day 0–6 (first insult), were sham operated, or remained naive (Ø). Mice then underwent UUO of the left kidney from day 17 to 22 (second insult) or remained naive (Ø). ( A ) Quantification of collagen-1 (Col1), overall fibrosis (fibrotic area), and col1a1 mRNA expression in the left kidney on day 22. Two pooled experiments, with n = 4, naive; 8, received the second insult only; 17, sham; and 16, RUUO. ( B – D ) Mice underwent RUUO of the right kidney from day 0 to 6 (first insult) or remained naive (Ø). Mice then underwent UUO of the left kidney from day 11 to 16 or from day 37 to 42 (second insult). ( B ) Analysis of the left UUO kidney on day 16 or day 42. Two pooled experiments, with n = 14 naive; 10 day 16 UUO; and 8 day 37 UUO for immunofluorescence and qPCR, and 1 single experiment for Trichrome staining, with n = 8 received the second insult only; 6, day 11 UUO; and 8, day 37 UUO. ( C ) Representative images of collagen-1 immunofluorescence and Trichrome staining. ( D ) Quantification of infiltrating CD4 + T cells, CD8 + T cells, monocytes (Monos), and neutrophils (Neutros) in the left UUO kidneys (single experiment with n = 8 received the second insult only; 6, day 11 UUO; and 8, day 37 UUO). ( E ) Quantification of CD4 + tissue-resident memory (TRM) T cells in the blood, spleen, and left UUO kidneys (single experiment with n = 6 and 4). Scale bars: 100 μm. Data are represented as mean ± SEM. One-way ANOVA with multiple comparisons. * P < 0.05; ** P < 0.01; *** P < 0.001.

    Journal: The Journal of Clinical Investigation

    Article Title: Fibrosis-memory is mediated by IL-3–producing T cells and drives progression of fibrosis

    doi: 10.1172/JCI192095

    Figure Lengend Snippet: ( A , B , and E ) Mice underwent RUUO of the right kidney from day 0–6 (first insult), were sham operated, or remained naive (Ø). Mice then underwent UUO of the left kidney from day 17 to 22 (second insult) or remained naive (Ø). ( A ) Quantification of collagen-1 (Col1), overall fibrosis (fibrotic area), and col1a1 mRNA expression in the left kidney on day 22. Two pooled experiments, with n = 4, naive; 8, received the second insult only; 17, sham; and 16, RUUO. ( B – D ) Mice underwent RUUO of the right kidney from day 0 to 6 (first insult) or remained naive (Ø). Mice then underwent UUO of the left kidney from day 11 to 16 or from day 37 to 42 (second insult). ( B ) Analysis of the left UUO kidney on day 16 or day 42. Two pooled experiments, with n = 14 naive; 10 day 16 UUO; and 8 day 37 UUO for immunofluorescence and qPCR, and 1 single experiment for Trichrome staining, with n = 8 received the second insult only; 6, day 11 UUO; and 8, day 37 UUO. ( C ) Representative images of collagen-1 immunofluorescence and Trichrome staining. ( D ) Quantification of infiltrating CD4 + T cells, CD8 + T cells, monocytes (Monos), and neutrophils (Neutros) in the left UUO kidneys (single experiment with n = 8 received the second insult only; 6, day 11 UUO; and 8, day 37 UUO). ( E ) Quantification of CD4 + tissue-resident memory (TRM) T cells in the blood, spleen, and left UUO kidneys (single experiment with n = 6 and 4). Scale bars: 100 μm. Data are represented as mean ± SEM. One-way ANOVA with multiple comparisons. * P < 0.05; ** P < 0.01; *** P < 0.001.

    Article Snippet: For isolation of splenic CD62L + CD4 + and CD62L – CD4 + T cells, the CD4 + T cells were first isolated with a CD4 + T cell negative isolation kit (Miltenyi) and subsequently separated with CD62L microbeads (Miltenyi).

    Techniques: Expressing, Immunofluorescence, Staining

    Mice underwent RUUO of the right kidney from day 0 to 6 (first insult), followed by UUO of the left kidney from day 16 to 22 (second insult). On days 13 and 14, mice were injected with a depleting CD4 antibody (α-CD4) or an isotype control antibody (control). ( A ) Quantification of collagen-1 (Col1), overall fibrosis (fibrotic area), and col1a1 mRNA expression in the left UUO kidney on day 22. Four pooled experiments, with n = 18/group. Controls are identical to those in A. ( B ) Representative images of collagen-1 immunofluorescence and Trichrome staining. ( C ) Quantification of CD4 + T cells, CD8 + T cells, B cells (CD19 + ), monocytes (Monos), and neutrophils (Neutros) in the spleen and left UUO kidneys by flow cytometry (single experiment with n = 6–7/group). Scale bars: 100 μm. Data are represented as mean ± SEM. Unpaired 2-sided t test. * P < 0.05; ** P < 0.01; *** P < 0.001.

    Journal: The Journal of Clinical Investigation

    Article Title: Fibrosis-memory is mediated by IL-3–producing T cells and drives progression of fibrosis

    doi: 10.1172/JCI192095

    Figure Lengend Snippet: Mice underwent RUUO of the right kidney from day 0 to 6 (first insult), followed by UUO of the left kidney from day 16 to 22 (second insult). On days 13 and 14, mice were injected with a depleting CD4 antibody (α-CD4) or an isotype control antibody (control). ( A ) Quantification of collagen-1 (Col1), overall fibrosis (fibrotic area), and col1a1 mRNA expression in the left UUO kidney on day 22. Four pooled experiments, with n = 18/group. Controls are identical to those in A. ( B ) Representative images of collagen-1 immunofluorescence and Trichrome staining. ( C ) Quantification of CD4 + T cells, CD8 + T cells, B cells (CD19 + ), monocytes (Monos), and neutrophils (Neutros) in the spleen and left UUO kidneys by flow cytometry (single experiment with n = 6–7/group). Scale bars: 100 μm. Data are represented as mean ± SEM. Unpaired 2-sided t test. * P < 0.05; ** P < 0.01; *** P < 0.001.

    Article Snippet: For isolation of splenic CD62L + CD4 + and CD62L – CD4 + T cells, the CD4 + T cells were first isolated with a CD4 + T cell negative isolation kit (Miltenyi) and subsequently separated with CD62L microbeads (Miltenyi).

    Techniques: Injection, Control, Expressing, Immunofluorescence, Staining, Flow Cytometry

    ( A and B ) CD4 + and CD8 + T cells were purified on day 0 from the spleens of naive donor mice (no OP) or donor mice after RUUO of the right kidney from day –18 to –12 and UUO of the same kidney from day –7 to 0 (RUUO+UUO). Where indicated, RPMI-1640 medium (Ø) or 5 × 10 6 CD4 + T cells or CD8 + T cells were adoptively transferred into recipients on day 0 and UUO was performed on all recipients on day 0. ( A ) Quantification of collagen-1 (Col1), overall fibrosis (fibrotic area), and col1a1 mRNA expression in the recipient’s UUO kidneys on day 7. Two pooled experiments with n = 11, no cell transfer; 13, CD4 + T cells from naïve mice; 13, CD8 + T cells from naïve mice; 14, CD4 + T cells from mice after RUUO+UUO; and 13, CD8 + T cells from mice after RUUO+UUO. ( B ) Representative images of collagen-1 immunofluorescence and Trichrome staining. ( C and D ) CD4 + T cells were purified on day 0 from the spleens or fibrotic kidneys of donor mice, with renal fibrosis induced as described above (RUUO+UUO). Where indicated, medium or 130,000 CD4 + T cells were adoptively transferred into recipients on day 0 and UUO was performed on all recipients on day 0. ( C ) Analysis of the recipient’s UUO kidneys on day 7. Two pooled experiments with n = 14, no cell transfer; 8, CD4 + T cells from the spleen; and 12, CD4 + T cells from the UUO kidney. ( D ) Representative images of collagen-1 immunofluorescence and Trichrome staining. Scale bars: 100 μm. Data are represented as mean ± SEM. One-way ANOVA with multiple comparisons ( A and C ). * P < 0.05; ** P < 0.01; *** P < 0.001.

    Journal: The Journal of Clinical Investigation

    Article Title: Fibrosis-memory is mediated by IL-3–producing T cells and drives progression of fibrosis

    doi: 10.1172/JCI192095

    Figure Lengend Snippet: ( A and B ) CD4 + and CD8 + T cells were purified on day 0 from the spleens of naive donor mice (no OP) or donor mice after RUUO of the right kidney from day –18 to –12 and UUO of the same kidney from day –7 to 0 (RUUO+UUO). Where indicated, RPMI-1640 medium (Ø) or 5 × 10 6 CD4 + T cells or CD8 + T cells were adoptively transferred into recipients on day 0 and UUO was performed on all recipients on day 0. ( A ) Quantification of collagen-1 (Col1), overall fibrosis (fibrotic area), and col1a1 mRNA expression in the recipient’s UUO kidneys on day 7. Two pooled experiments with n = 11, no cell transfer; 13, CD4 + T cells from naïve mice; 13, CD8 + T cells from naïve mice; 14, CD4 + T cells from mice after RUUO+UUO; and 13, CD8 + T cells from mice after RUUO+UUO. ( B ) Representative images of collagen-1 immunofluorescence and Trichrome staining. ( C and D ) CD4 + T cells were purified on day 0 from the spleens or fibrotic kidneys of donor mice, with renal fibrosis induced as described above (RUUO+UUO). Where indicated, medium or 130,000 CD4 + T cells were adoptively transferred into recipients on day 0 and UUO was performed on all recipients on day 0. ( C ) Analysis of the recipient’s UUO kidneys on day 7. Two pooled experiments with n = 14, no cell transfer; 8, CD4 + T cells from the spleen; and 12, CD4 + T cells from the UUO kidney. ( D ) Representative images of collagen-1 immunofluorescence and Trichrome staining. Scale bars: 100 μm. Data are represented as mean ± SEM. One-way ANOVA with multiple comparisons ( A and C ). * P < 0.05; ** P < 0.01; *** P < 0.001.

    Article Snippet: For isolation of splenic CD62L + CD4 + and CD62L – CD4 + T cells, the CD4 + T cells were first isolated with a CD4 + T cell negative isolation kit (Miltenyi) and subsequently separated with CD62L microbeads (Miltenyi).

    Techniques: Purification, Expressing, Immunofluorescence, Staining

    ( A and B ) RUUO of the right kidney (day 0–6), followed by UUO of the left kidney (day 17–22). ( A ) Clonal diversity of UUO kidney and splenic CD4 + T cells on day 22. ( B ) TCR repertoire overlaps between UUO kidney and splenic CD4 + T cells. UMI counts represent individual TCR-β mRNAs. ( C – I ) RUUO of the right kidney (day 0–6) and UUO of the same kidney (day 11–18) (RUUO+UUO). Analysis on day 18. Naive mice (no OP) served as controls. ( C ) CD62L + and CD62L – CD4 + T cells in the spleens of naive mice ( n = 8), and in the spleens and fibrotic kidneys of RUUO+UUO mice ( n = 6). ( D ) CD4 + T cells expressing IL-3, IL-2, GM-CSF, IFN-γ ( n = 8 each), and IL-17 ( n = 5) in the spleens and fibrotic kidneys of RUUO+UUO mice. ( E ) IL-3 mRNA expression of CD4 + T cells from the spleens of naive and RUUO+UUO mice ( n = 8) and the fibrotic kidney of RUUO+UUO mice ( n = 3). ( F ) IL-3 mRNA expression of CD62L + CD4 + T cells ( n = 12) and CD62L – CD4 + T cells ( n = 6) from the spleens of naive and RUUO+UUO mice. ( G ) IL-3 in the supernatant of anti-CD3–activated CD4 + T cells, CD62L + CD4 + , and CD62L – CD4 + T cells from the spleens of RUUO+UUO mice ( n = 3 per group). ( H ) IL-3 in the supernatant of anti-CD3–activated PBMCs from the blood of RUUO+UUO mice ( n = 6) or naive mice ( n = 7). ( I ) CFSE-labeled CD4 + T cells from the spleens of RUUO+UUO mice were injected into recipients 3 days after UUO operation (5 × 10 6 cells/mouse, n = 5). Two days later, CFSE + T cells were quantified in relation to endogenous CFSE – CD4 + T cells in various organs. Data are represented as mean ± SEM. Unpaired 2-sided t test ( A , D , F , and H ) and 1-way ANOVA with multiple comparisons ( C , E , G , and I ). * P < 0.05; ** P < 0.01; *** P < 0.001.

    Journal: The Journal of Clinical Investigation

    Article Title: Fibrosis-memory is mediated by IL-3–producing T cells and drives progression of fibrosis

    doi: 10.1172/JCI192095

    Figure Lengend Snippet: ( A and B ) RUUO of the right kidney (day 0–6), followed by UUO of the left kidney (day 17–22). ( A ) Clonal diversity of UUO kidney and splenic CD4 + T cells on day 22. ( B ) TCR repertoire overlaps between UUO kidney and splenic CD4 + T cells. UMI counts represent individual TCR-β mRNAs. ( C – I ) RUUO of the right kidney (day 0–6) and UUO of the same kidney (day 11–18) (RUUO+UUO). Analysis on day 18. Naive mice (no OP) served as controls. ( C ) CD62L + and CD62L – CD4 + T cells in the spleens of naive mice ( n = 8), and in the spleens and fibrotic kidneys of RUUO+UUO mice ( n = 6). ( D ) CD4 + T cells expressing IL-3, IL-2, GM-CSF, IFN-γ ( n = 8 each), and IL-17 ( n = 5) in the spleens and fibrotic kidneys of RUUO+UUO mice. ( E ) IL-3 mRNA expression of CD4 + T cells from the spleens of naive and RUUO+UUO mice ( n = 8) and the fibrotic kidney of RUUO+UUO mice ( n = 3). ( F ) IL-3 mRNA expression of CD62L + CD4 + T cells ( n = 12) and CD62L – CD4 + T cells ( n = 6) from the spleens of naive and RUUO+UUO mice. ( G ) IL-3 in the supernatant of anti-CD3–activated CD4 + T cells, CD62L + CD4 + , and CD62L – CD4 + T cells from the spleens of RUUO+UUO mice ( n = 3 per group). ( H ) IL-3 in the supernatant of anti-CD3–activated PBMCs from the blood of RUUO+UUO mice ( n = 6) or naive mice ( n = 7). ( I ) CFSE-labeled CD4 + T cells from the spleens of RUUO+UUO mice were injected into recipients 3 days after UUO operation (5 × 10 6 cells/mouse, n = 5). Two days later, CFSE + T cells were quantified in relation to endogenous CFSE – CD4 + T cells in various organs. Data are represented as mean ± SEM. Unpaired 2-sided t test ( A , D , F , and H ) and 1-way ANOVA with multiple comparisons ( C , E , G , and I ). * P < 0.05; ** P < 0.01; *** P < 0.001.

    Article Snippet: For isolation of splenic CD62L + CD4 + and CD62L – CD4 + T cells, the CD4 + T cells were first isolated with a CD4 + T cell negative isolation kit (Miltenyi) and subsequently separated with CD62L microbeads (Miltenyi).

    Techniques: Expressing, Labeling, Injection

    CD62L + CD4 + T cells (CD62L + ) and CD62L – CD4 + T cells (CD62L – ) were purified from the spleens of donor mice with renal fibrosis induced by RUUO of the right kidney from day –18 to –12 and UUO of the same kidney from day –7 to 0 (RUUO+UUO). Medium (Ø) or 4 × 10 6 purified cells were injected into recipients on day 0 and UUO was performed in all recipients on day 0. ( A ) Quantification of collagen-1 (Col1), overall fibrosis (fibrotic area), and col1a1 mRNA expression in the recipient’s UUO kidneys on day 7. Three pooled experiments for histology with n = 22, no cell transfer; 21, transfer of CD62L + ; 21, CD62L – , and 2 pooled experiments for qPCR with n = 14, no cell transfer; 15, transfer of CD62L + ; 14, transfer of CD62L – . ( B ) Representative images of collagen-1 immunofluorescence and Trichrome staining. ( C ) Quantification of IL-3 in the supernatant of shredded recipient’s UUO kidneys ( n = 6/group). ( D ) CD4 + T cells, monocytes (Monos), and neutrophils (Neutros) as well as percentage of CD62L + and CD62L – CD4 + T cells in the recipient’s UUO kidneys on day 7 ( n = 7/group). Scale bars: 100 μm. Data are represented as mean ± SEM. One-way ANOVA with multiple comparisons. * P < 0.05; ** P < 0.01; *** P < 0.001.

    Journal: The Journal of Clinical Investigation

    Article Title: Fibrosis-memory is mediated by IL-3–producing T cells and drives progression of fibrosis

    doi: 10.1172/JCI192095

    Figure Lengend Snippet: CD62L + CD4 + T cells (CD62L + ) and CD62L – CD4 + T cells (CD62L – ) were purified from the spleens of donor mice with renal fibrosis induced by RUUO of the right kidney from day –18 to –12 and UUO of the same kidney from day –7 to 0 (RUUO+UUO). Medium (Ø) or 4 × 10 6 purified cells were injected into recipients on day 0 and UUO was performed in all recipients on day 0. ( A ) Quantification of collagen-1 (Col1), overall fibrosis (fibrotic area), and col1a1 mRNA expression in the recipient’s UUO kidneys on day 7. Three pooled experiments for histology with n = 22, no cell transfer; 21, transfer of CD62L + ; 21, CD62L – , and 2 pooled experiments for qPCR with n = 14, no cell transfer; 15, transfer of CD62L + ; 14, transfer of CD62L – . ( B ) Representative images of collagen-1 immunofluorescence and Trichrome staining. ( C ) Quantification of IL-3 in the supernatant of shredded recipient’s UUO kidneys ( n = 6/group). ( D ) CD4 + T cells, monocytes (Monos), and neutrophils (Neutros) as well as percentage of CD62L + and CD62L – CD4 + T cells in the recipient’s UUO kidneys on day 7 ( n = 7/group). Scale bars: 100 μm. Data are represented as mean ± SEM. One-way ANOVA with multiple comparisons. * P < 0.05; ** P < 0.01; *** P < 0.001.

    Article Snippet: For isolation of splenic CD62L + CD4 + and CD62L – CD4 + T cells, the CD4 + T cells were first isolated with a CD4 + T cell negative isolation kit (Miltenyi) and subsequently separated with CD62L microbeads (Miltenyi).

    Techniques: Purification, Injection, Expressing, Immunofluorescence, Staining

    ( A ) RUUO of the right kidney (day 0–6, first insult) followed by UUO of the left kidney (day 16–22, second insult). Treatment with anti–IL-3 ( n = 20) or isotype control antibody ( n = 18; 4 pooled experiments, same controls as in ) from day 15–21. Analysis of the left UUO kidneys on day 22. ( B ) WT or IL-3–KO mice underwent RUUO of the right kidney (day 0–6, first insult) or remained naive (Ø). All mice then underwent UUO of the left kidney (day 16–22). Analysis of the left UUO kidneys on day 22. One experiment with n = 7, WT RUUO+UUO; 7, IL-3-KO RUUO+UUO; 6, WT UUO; and 7, IL-3-KO UUO. ( C ) Splenic CD62L – CD4 + or renal CD4 + T cells were purified from the spleens or fibrotic kidneys of RUUO+UUO donor mice (RUUO from day –18 to –12 and UUO of the same kidney from day –7 to 0). Medium, 4 × 10 6 splenic CD62L – CD4 + or 67,000 renal CD4 + T cells were injected into recipients on day 0. UUO in all recipients on day 0. Treatment with anti–IL-3 or an isotype control antibody (Ø) from day 0–6. Analysis of the recipient’s UUO kidneys on day 7. Single experiment with n = 8, no cell transfer; 5, no cell transfer and anti–IL-3; 7, transfer of CD62L - from spleen; 7, transfer of CD62L - from spleen and anti–IL-3; 3, transfer of CD4 + from UUO kidney; and 3, transfer of CD4 + from UUO kidney and anti–IL-3. ( D ) Splenic CD62L – CD4 + or renal CD4 + T cells were purified from the spleens or fibrotic kidneys of WT or IL-3–deficient (IL-3 KO) RUUO+UUO mice, as in C . Medium (Ø), 4 × 10 6 splenic CD62L – CD4 + or 116,000 renal CD4 + T cells were injected on day 0. UUO in all recipients on day 0. Analysis of the recipient’s UUO kidneys on day 7. Single experiment with n = 7, no cell transfer; 8, CD62L + from spleen of WT; 7, CD62L + from spleen of IL-3 KO; 5, CD62L + from UUO-kidney of WT; and 8, CD62L + from UUO-kidney of IL-3 KO. Data are represented as mean ± SEM. Unpaired 2-sided t test. * P < 0.05; ** P < 0.01; *** P < 0.001.

    Journal: The Journal of Clinical Investigation

    Article Title: Fibrosis-memory is mediated by IL-3–producing T cells and drives progression of fibrosis

    doi: 10.1172/JCI192095

    Figure Lengend Snippet: ( A ) RUUO of the right kidney (day 0–6, first insult) followed by UUO of the left kidney (day 16–22, second insult). Treatment with anti–IL-3 ( n = 20) or isotype control antibody ( n = 18; 4 pooled experiments, same controls as in ) from day 15–21. Analysis of the left UUO kidneys on day 22. ( B ) WT or IL-3–KO mice underwent RUUO of the right kidney (day 0–6, first insult) or remained naive (Ø). All mice then underwent UUO of the left kidney (day 16–22). Analysis of the left UUO kidneys on day 22. One experiment with n = 7, WT RUUO+UUO; 7, IL-3-KO RUUO+UUO; 6, WT UUO; and 7, IL-3-KO UUO. ( C ) Splenic CD62L – CD4 + or renal CD4 + T cells were purified from the spleens or fibrotic kidneys of RUUO+UUO donor mice (RUUO from day –18 to –12 and UUO of the same kidney from day –7 to 0). Medium, 4 × 10 6 splenic CD62L – CD4 + or 67,000 renal CD4 + T cells were injected into recipients on day 0. UUO in all recipients on day 0. Treatment with anti–IL-3 or an isotype control antibody (Ø) from day 0–6. Analysis of the recipient’s UUO kidneys on day 7. Single experiment with n = 8, no cell transfer; 5, no cell transfer and anti–IL-3; 7, transfer of CD62L - from spleen; 7, transfer of CD62L - from spleen and anti–IL-3; 3, transfer of CD4 + from UUO kidney; and 3, transfer of CD4 + from UUO kidney and anti–IL-3. ( D ) Splenic CD62L – CD4 + or renal CD4 + T cells were purified from the spleens or fibrotic kidneys of WT or IL-3–deficient (IL-3 KO) RUUO+UUO mice, as in C . Medium (Ø), 4 × 10 6 splenic CD62L – CD4 + or 116,000 renal CD4 + T cells were injected on day 0. UUO in all recipients on day 0. Analysis of the recipient’s UUO kidneys on day 7. Single experiment with n = 7, no cell transfer; 8, CD62L + from spleen of WT; 7, CD62L + from spleen of IL-3 KO; 5, CD62L + from UUO-kidney of WT; and 8, CD62L + from UUO-kidney of IL-3 KO. Data are represented as mean ± SEM. Unpaired 2-sided t test. * P < 0.05; ** P < 0.01; *** P < 0.001.

    Article Snippet: For isolation of splenic CD62L + CD4 + and CD62L – CD4 + T cells, the CD4 + T cells were first isolated with a CD4 + T cell negative isolation kit (Miltenyi) and subsequently separated with CD62L microbeads (Miltenyi).

    Techniques: Control, Purification, Injection