anti cd8α rat igg2b monoclonal antibody 2 43  (Bio X Cell)

Journal: Advanced Science

Article Title: CD8 T Cell‐Derived Exosomal miR‐186‐5p Elicits Renal Inflammation via Activating Tubular TLR7/8 Signal Axis

doi: 10.1002/advs.202301492

Figure Lengend Snippet: External source miR‐186‐5p initiates renal tubular inflammation and tissue injury in the ADR mouse model. A,B) ADR treatment elevated miR‐186‐5p level in A) mouse plasma particularly B) circulating CD8 T cells. C) Depletion of CD8 T cell with anti‐CD8 antibody in ADR mouse decreased miR‐186‐5p level in plasma. D,E) ADR treatment increased the level of D) miR‐186‐5p but not E) pre‐miR‐186‐5p in the mouse kidney. F) Top: Schematic of adenovirus‐based miR‐186‐5p sponge treatment. Bottom: miR‐186‐5p sponge abolished the increase of miR‐186‐5p level in mouse plasma (left) and kidney (right) by ADR treatment. G) Adenovirus‐based miR‐186‐5p sponge attenuated renal tubular injury in ADR mice. H) Flow cytometry analysis of infiltration of macrophages (F4/80 + CD11b + ) and T cells (CD45 + CD3 + ) in ADR mice with or without miR‐186‐5p sponge treatment. I) Tissue staining of macrophages and CD8 T cells in mouse kidney with or without ADR plus miR‐186‐5p sponge treatment. Scale bars, 50 µm. G,I) There were five mice per group; 6–8 fields were analyzed for each mouse. A—H) Data were analyzed by unpaired two‐sided Student's t‐ test. *, p < 0.05 and **, p < 0.01. ns, no significance.

Article Snippet: To deplete mouse CD8 T cells, mice were injected with 100 µg per mouse anti‐CD8 mAb (rat IgG2b mAb, clone 2.43; BioXCell, BE0061) or isotype control mAb (rat IgG2b mAb, clone LTF‐2; BioXCell, BE0090) via tail vein 1 day prior to ADR treatment.

Techniques: Flow Cytometry, Staining

Journal: Advanced Science

Article Title: CD8 T Cell‐Derived Exosomal miR‐186‐5p Elicits Renal Inflammation via Activating Tubular TLR7/8 Signal Axis

doi: 10.1002/advs.202301492

Figure Lengend Snippet: CD8 T cell depletion alleviates renal inflammation and tissue injury in the ADR mouse model. A) Depletion of CD8 T cells by CD8 antibody attenuated renal tubular injury in ADR mice. B) Flow cytometry analysis of kidney infiltration of macrophages (F4/80 + CD11b + ) and T cells (CD45 + CD3 + ) in ADR mice with isotype or anti‐CD8 Ab treatment. C,D) Tissue staining of mouse kidney infiltration of C) macrophages and D) CD8 T cells with or without ADR plus isotype or anti‐CD8 Ab treatment. Scale bars, 50 µm. A,C,D) There were five mice per group; 6–8 fields were analyzed for each mouse. A—D) Data were analyzed by unpaired two‐sided Student's t‐ test. **, p < 0.01.

Article Snippet: To deplete mouse CD8 T cells, mice were injected with 100 µg per mouse anti‐CD8 mAb (rat IgG2b mAb, clone 2.43; BioXCell, BE0061) or isotype control mAb (rat IgG2b mAb, clone LTF‐2; BioXCell, BE0090) via tail vein 1 day prior to ADR treatment.

Techniques: Flow Cytometry, Staining

Journal: Advanced Science

Article Title: CD8 T Cell‐Derived Exosomal miR‐186‐5p Elicits Renal Inflammation via Activating Tubular TLR7/8 Signal Axis

doi: 10.1002/advs.202301492

Figure Lengend Snippet: Circulating CD8 T‐EVs from FSGS patients contain higher level of miR‐186‐5p compared to that from healthy donors. A) Level of miR‐186‐5p in various peripheral leukocytes isolated from 5 FSGS patients and healthy donors. B) Distribution of plasma miR‐186‐5p from FSGS patients in EVs and EV‐free fraction. C) Level of T‐EVs in peripheral blood from FSGS patients and healthy donors (HC). D) miR‐186‐5p level in the T‐EVs from FSGS patients and healthy donors. E) miR‐186‐5p level in exosomes secreted from T cells that were isolated from FSGS patients or healthy donors. A—D) There were five patients or healthy donors per group and three independent tests from each group. In panel (E), n = 5. B—E) Data were analyzed by unpaired two‐sided Student's t‐ test. **, p < 0.01.

Article Snippet: To deplete mouse CD8 T cells, mice were injected with 100 µg per mouse anti‐CD8 mAb (rat IgG2b mAb, clone 2.43; BioXCell, BE0061) or isotype control mAb (rat IgG2b mAb, clone LTF‐2; BioXCell, BE0090) via tail vein 1 day prior to ADR treatment.

Techniques: Isolation

Journal: Advanced Science

Article Title: CD8 T Cell‐Derived Exosomal miR‐186‐5p Elicits Renal Inflammation via Activating Tubular TLR7/8 Signal Axis

doi: 10.1002/advs.202301492

Figure Lengend Snippet: Transfer of miR‐186‐5p from circulating CD8 T cells to renal tubulointerstitium via T‐EVs. A) Schematic of T‐EVs/miR‐186‐5p tracing experiment. B) Distribution of DiI ‐labeled T‐EVs in mouse glomeruli and renal tubulointerstitium. C) ADR treatment promoted the accumulation of DiI‐labeled T‐EVs in mouse renal tubulointerstitium. D) Distribution of miR‐186‐5p in mouse glomeruli and renal tubulointerstitium following tail vein injection of miR‐186‐5p‐Cy5. E) qRT‐PCR detection of miR‐186‐5p in isolated mouse glomerulus and renal tubules 0.5 h after injection of miR‐186‐5p‐Cy5. Scale bars, 50 µm. B—E) There were five mice per group; B—D) 6–7 fields were analyzed for each mouse. Data were analyzed by unpaired two‐sided Student's t‐ test (B–E). **, p < 0.01.

Article Snippet: To deplete mouse CD8 T cells, mice were injected with 100 µg per mouse anti‐CD8 mAb (rat IgG2b mAb, clone 2.43; BioXCell, BE0061) or isotype control mAb (rat IgG2b mAb, clone LTF‐2; BioXCell, BE0090) via tail vein 1 day prior to ADR treatment.

Techniques: Labeling, Injection, Quantitative RT-PCR, Isolation

Journal: Advanced Science

Article Title: CD8 T Cell‐Derived Exosomal miR‐186‐5p Elicits Renal Inflammation via Activating Tubular TLR7/8 Signal Axis

doi: 10.1002/advs.202301492

Figure Lengend Snippet: Injection of miR‐186‐5p initiates mouse renal inflammation via specific activating renal tubular TLR7 signal. A) kidney miR‐186‐5p level before and after miR‐186‐5p or miR‐186‐5p‐Mut injection. B) Mouse renal tubule damage quantitated by PAS. C) Proteinuria and creatinine level in mice on day 7 post‐injection of miR‐186‐5p or miR‐186‐5p‐Mut. D) Flow cytometry analysis of infiltration of macrophages (F4/80 + CD11b + ) and T cells (CD45 + CD3 + ) in mice after miR‐186‐5p or miR‐186‐5p‐Mut injection. E) Tissue staining of macrophage and CD8 T cell infiltration in mouse kidney with miR‐186‐5p or miR‐186‐5p‐Mut treatment. F) Immunohistochemical detection (left) and analysis (right) of protein level of TLR7 and NF‐ κ B‐p65 in mouse kidney tissues after miR‐186‐5p or miR‐186‐5p‐Mut treatment. G) Western blot analysis of nuclear and total NF‐ κ B‐p65 level in HK2 cells transfected with miR‐186‐5p or miR‐186‐5p‐Mut. Scale bars, 50 µm. A,C,D) There were five mice per group and three tests for each mouse; B,E,F) 6–8 fields were analyzed for each mouse. A—D,F) Data were analyzed by unpaired two‐sided Student's t ‐test. *, p < 0.05 and **, p < 0.01. ns, no significance.

Article Snippet: To deplete mouse CD8 T cells, mice were injected with 100 µg per mouse anti‐CD8 mAb (rat IgG2b mAb, clone 2.43; BioXCell, BE0061) or isotype control mAb (rat IgG2b mAb, clone LTF‐2; BioXCell, BE0090) via tail vein 1 day prior to ADR treatment.

Techniques: Injection, Flow Cytometry, Staining, Immunohistochemistry, Western Blot, Transfection

Journal: Advanced Science

Article Title: CD8 T Cell‐Derived Exosomal miR‐186‐5p Elicits Renal Inflammation via Activating Tubular TLR7/8 Signal Axis

doi: 10.1002/advs.202301492

Figure Lengend Snippet: TLR7‐deficient mice resist renal inflammation and tissue injury induced by miR‐186‐5p or ADR administration. A) Immunofluorescence labeling (left) and western blot analysis (right) of TLR7 in mouse kidney tissues. B,D) Renal tubular injury induced by B) miR‐186‐5p or D) ADR treatment in WT and Tlr7 −/− mice. C,E) Flow cytometry analysis of renal infiltration of macrophages (F4/80 + CD11b + ) and T cells (CD45 + CD3 + ) induced by C) miR‐186‐5p or E) ADR treatment in WT and Tlr7 −/− mice. F,G) Tissue staining of F) macrophage and G) CD8 T cell infiltration in WT and Tlr7 −/− mouse kidney with or without miR‐186‐5p treatment. H,I) Tissue staining of H) macrophage and I) CD8 T cell infiltration in WT and Tlr7 −/− kidney with or without ADR treatment. Scale bars, 50 µm. B–I) There were five mice per group; B,D,F—I) 6–8 fields were analyzed for each mouse. B—I) Data were analyzed by unpaired two‐sided Student's t ‐test. **, p < 0.01. ns, no significance.

Article Snippet: To deplete mouse CD8 T cells, mice were injected with 100 µg per mouse anti‐CD8 mAb (rat IgG2b mAb, clone 2.43; BioXCell, BE0061) or isotype control mAb (rat IgG2b mAb, clone LTF‐2; BioXCell, BE0090) via tail vein 1 day prior to ADR treatment.

Techniques: Immunofluorescence, Labeling, Western Blot, Flow Cytometry, Staining

Journal: Advanced Science

Article Title: CD8 T Cell‐Derived Exosomal miR‐186‐5p Elicits Renal Inflammation via Activating Tubular TLR7/8 Signal Axis

doi: 10.1002/advs.202301492

Figure Lengend Snippet: Schematic depiction of CD8 T cell exosomal miR‐186‐5p as a circulating pathogenic factor in causing renal inflammation and tissue injury via activating tubular TLR7/8 signal.

Article Snippet: To deplete mouse CD8 T cells, mice were injected with 100 µg per mouse anti‐CD8 mAb (rat IgG2b mAb, clone 2.43; BioXCell, BE0061) or isotype control mAb (rat IgG2b mAb, clone LTF‐2; BioXCell, BE0090) via tail vein 1 day prior to ADR treatment.

Techniques: