Journal: Science Advances

Article Title: CD38 reduces mitochondrial fitness and cytotoxic T cell response against viral infection in lupus patients by suppressing mitophagy

doi: 10.1126/sciadv.abo4271

Figure Lengend Snippet: ( A ) Percentage of depolarized mitochondria in control, CD38-overexpressing CD8 + T cells, or CD38-overexpressing cells treated with SRT1720, or one of the CD38 inhibitors (78c or MK-0159). ( B to D ) Percentage of positive cells expressing CD107a (B), granzyme B (C), and IFN-γ (D) in control, CD38-overexpressing CD8 + T cells, or CD38-overexpressing cells treated with SRT1720, or one of the CD38 inhibitors (78c or MK-0159). ( E ) MFI of LC3B staining in control or CD38-overexpressed CD8 + T cells. ( F ) Counts of LC3B-positive dots per cell of control or CD38-overexpressed CD8 + T cells. ( G ) MFI of PINK1 in control, CD38-overexpressing CD8 + T cells, or CD38-overexpressing cells treated with SRT1720. ( H ) Percentage of Parkin positive of the TOMM20-positive area in control, CD38-overexpressing CD8 + T cells, or CD38-overexpressing cells treated with SRT1720. ( I ) MFI of PINK1 in CD38 hi CD8 + and CD38 lo CD8 + T cells from lupus patient peripheral blood. ( J ) Percentage of Parkin positive of the TOMM20-positive area in CD38 hi CD8 + and CD38 lo CD8 + T cells from lupus patient peripheral blood. Data are means ± SD; statistical analysis by two-tailed t test (A to H and J), paired t test (I), ns = P > 0.05, * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001.

Article Snippet: For flow cytometry, CD8 + T cells from human peripheral blood were stained with the following antibodies or reagents, listed with clones, vendor, and catalog numbers: anti-human CD4 (clone RPA-T8, 300521), anti-human CD8a (clone SK1, BioLegend, 344718), anti-human CD8a (clone HIT8a, BioLegend, 300908), anti-human CD38 (clone HIT2, BioLegend, 303516 and 303510), allophycocyanin (APC) mouse immunoglobulin G1 (IgG1), κ isotype control (BD, 555751), anti–green fluorescent protein (GFP) antibody (Rockland, 600-102-215), anti–IFN-γ (clone 4S.B3, BioLegend, 502509), anti–granzyme B (clone GB11, BD, 560212), anti-CD107a (clone H4A, BioLegend, 328608), PINK1 antibody (Rockland, 600-401-GU5), anti-Rab4 antibody (EPR3043, Abcam, ab109009), goat anti-rabbit IgG secondary antibody, and APC (Invitrogen, A-10931).

Techniques: Control, Expressing, Staining, Two Tailed Test

Journal: Science Advances

Article Title: CD38 reduces mitochondrial fitness and cytotoxic T cell response against viral infection in lupus patients by suppressing mitophagy

doi: 10.1126/sciadv.abo4271

Figure Lengend Snippet: ( A and B ) Percentage of positive cells expressing CD107a (A) and granzyme B (B) in gp33 + CD8 + T cells 8 days after LCMV Armstrong infection from diseased BXD2 treated with control water or MK-0159. ( C ) Percentage of depolarized mitochondria in gp33 + CD8 + T cells 8 days after LCMV Armstrong infection from diseased BXD2 treated with control water or MK-0159. ( D ) LCMV viral load 8 days after LCMV Armstrong infection using qPCR of glycoprotein gene normalized by tissue ACTB in diseased BXD2 treated with control water or MK-0159. ( E to G ) Representative liver histology 8 days after LCMV Armstrong infection from diseased BXD2 treated with control water (E) or MK-0159 (F) and their respective hepatitis activity index (G). ( H and I ) Percentage of positive cells expressing CD107a (H) and granzyme B (I) in gp33 + CD8 + T cells 8 days after LCMV Armstrong infection from diseased BXD2 treated with control NP or NP-MK-0159. ( J to M ) Representative liver histology 8 days after LCMV Armstrong infection from diseased BXD2 treated with NP-control with the presentation of necrosis (J) or portal and periportal inflammation (K) or treated with NP-MK-0159 (L) and their respective hepatitis activity index (M). ( N ) LCMV viral load 8 days after LCMV Armstrong infection using qPCR of glycoprotein gene normalized by tissue ACTB in diseased BXD2 treated with NP-control or NP-MK-0159. ( O ) Percentage of depolarized mitochondria in gp33 + CD8 + T cells 8 days after LCMV Armstrong infection from diseased BXD2 treated with NP-control or NP-MK-0159. ( P and Q ) MFI of PINK1 (P) and LysoSensor Green (Q) in gp33 + CD8 + T cells 8 days after LCMV Armstrong infection from diseased BXD2 treated with NP-control or NP-MK-0159. Data are means ± SD; statistical analysis by two-tailed t test. * P < 0.05, ** P < 0.01.

Article Snippet: For flow cytometry, CD8 + T cells from human peripheral blood were stained with the following antibodies or reagents, listed with clones, vendor, and catalog numbers: anti-human CD4 (clone RPA-T8, 300521), anti-human CD8a (clone SK1, BioLegend, 344718), anti-human CD8a (clone HIT8a, BioLegend, 300908), anti-human CD38 (clone HIT2, BioLegend, 303516 and 303510), allophycocyanin (APC) mouse immunoglobulin G1 (IgG1), κ isotype control (BD, 555751), anti–green fluorescent protein (GFP) antibody (Rockland, 600-102-215), anti–IFN-γ (clone 4S.B3, BioLegend, 502509), anti–granzyme B (clone GB11, BD, 560212), anti-CD107a (clone H4A, BioLegend, 328608), PINK1 antibody (Rockland, 600-401-GU5), anti-Rab4 antibody (EPR3043, Abcam, ab109009), goat anti-rabbit IgG secondary antibody, and APC (Invitrogen, A-10931).

Techniques: Expressing, Infection, Control, Activity Assay, Two Tailed Test