rosetteseptm human cd4+ t cell enrichment cocktail  (STEMCELL Technologies Inc)

Journal: Viruses

Article Title: Expanding Insights: Harnessing Expansion Microscopy for Super-Resolution Analysis of HIV-1–Cell Interactions

doi: 10.3390/v16101610

Figure Lengend Snippet: U-ExM allows expansion of various cell types and cellular structures. ( A ) TZM-bl cells, resting and activated CD4+ T-cells, and MDM fixed with PFA/GA, expanded and stained for chromatin (Hoechst, cyan), membranes (BODIPY, green), and Nup153 (magenta). Scale bar, 10 µm. ( B ) Cropped region of the nucleus of an MDM fixed with PFA/GA, expanded and stained for chromatin (Hoechst, cyan), membranes (BODIPY, green), and Nup153 (magenta) showing double membrane layers of the nuclear envelope in membrane staining. Scale bar, 2 µm. ( C ) Cropped region of the nucleus of an MDM fixed with PFA/GA, expanded and stained for chromatin (Hoechst, cyan), membranes (BODIPY, not shown in merge), and Nup153 (magenta), showing heterochromatin-free channels beneath the NPCs in the nucleus. Scale bar, 2 µm; arrowheads mark the position of NPC. ( D ) A TZM-bl cell fixed with PFA/GA, expanded and stained for chromatin (Hoechst, cyan), protein density (NHS-ester, green), SRRM2 (yellow), and SON (magenta), showcasing different intranuclear compartments. Scale bar, 10 µm. ( E ) Enlargements from ( D ) highlighting nuclear speckles and nucleolus. Scale bar, 5 µm. All scale bars in expanded samples reflect post-expansion sizes.

Article Snippet: Negative CD4+ T-cell isolation was performed using the RosetteSepTM Human CD4+ T-Cell Enrichment Cocktail (STEMCELL Technologies Inc., Vancouver, BC, Canada) following the manufacturer’s instructions.

Techniques: Staining, Membrane

Journal: Clinical & Translational Immunology

Article Title: Engineered CAR‐T cells targeting the non‐functional P2X purinoceptor 7 (P2X7) receptor as a novel treatment for ovarian cancer

doi: 10.1002/cti2.1512

Figure Lengend Snippet: Effect of nfP2X7‐CAR‐T cells in patient‐derived explant assays. Representative images of cleaved caspase 3 immunostaining in patient‐derived ovarian cancer explants (patient 5) following treatment with PBS (a) , carboplatin (CBP, 100 μm (b) , un‐transduced (UT) T cells, (CD4:CD8 1:1) (c) , nfP2X7‐CAR‐T (CD4:CD8 1:1) (d) for 48 h. (e) Cleaved caspase 3 quantitation. Data are expressed as % of PBS or UT CD3 controls ( n = 10, patients 4–13), The bar graphs show the median values. *, significantly increased compared to PBS control and UT T cell treatment, Wilcoxon signed‐rank test ( P < 0.05). CD3 immunostaining in explant tissue that responded to nfP2X7‐CAR‐T treatment (patient 11); treatment with un‐transduced (UT, CD4:CD8 1:1 cells) (f) and nfP2X7‐CAR‐T (CD4:CD8 1:1) (g) for 48 h. CD3 quantitation using quPath analysis in explant tissues that responded to nfP2X7‐CAR‐T treatment (h) , n = 7). Data are expressed as % of UT CD3 control. * P = 0.046, Wilcoxon signed‐rank test. CD3 immunostaining in ovarian cancer explant tissue that did not respond to treatment with nfP2X7‐CAR‐T (patient 10) treatment with UT (CD4:CD8 1:1 cells) (i) and nfP2X7‐CAR‐T (CD4:CD8 1:1) (j) for 48 h. CD3 quantitation using quPath analysis in explant tissues that did not respond to nfP2X7‐CAR‐T treatment (k) , n = 3). Data are expressed as % of UT CD3 control. (l) Correlation between CD3 positivity (cells mm −2 ) and response to nfP2X7‐CAR‐T treatment measured by cleaved caspase 3 positivity (% of UT control), Spearman correlation test, r = 0.783, P = 0.0172. (a–d) scale bar = 50 μm, (f–j) scale bar = 100 μm. (f–j) pink asterisks indicate CD3 positive cells.

Article Snippet: T cells were isolated from whole blood using RosetteSepTM Human CD4, CD8 or CD3 T‐cell enrichment cocktail (StemCell Technologies, Tullamarine, VIC, AUS) from donors following the manufacturer's protocol.

Techniques: Derivative Assay, Immunostaining, Quantitation Assay, Control

Journal: Clinical & Translational Immunology

Article Title: Engineered CAR‐T cells targeting the non‐functional P2X purinoceptor 7 (P2X7) receptor as a novel treatment for ovarian cancer

doi: 10.1002/cti2.1512

Figure Lengend Snippet: Characterisation of OVCAR3 xenograft tumours. Representative images and quantitation of CD3 (a) , CD4 (b) , CD8 (c) and PD1 (d) immunostaining in OVCAR3 xenograft tumours from mice treated with UT CD3 T cells or nfP2X7‐CAR‐T cells. OVCAR3 tumours were collected between 106 and 139 days post T‐cell treatment. All images are the same magnification. Scale bar = 100 μm. Data are the % positive CD3, CD4, CD8, or PD1 cells in tumour area. The bar represents the median value of 5 or 6 tumours/group. * P < 0.05, ** P < 0.001. Mann–Whitney U ‐test.

Article Snippet: T cells were isolated from whole blood using RosetteSepTM Human CD4, CD8 or CD3 T‐cell enrichment cocktail (StemCell Technologies, Tullamarine, VIC, AUS) from donors following the manufacturer's protocol.

Techniques: Quantitation Assay, Immunostaining, MANN-WHITNEY