anti sell  (Santa Cruz Biotechnology)

Journal: Frontiers in Immunology

Article Title: Ex vivo- generated lymphoid progenitors encompass both T cell and innate lymphoid cell fates

doi: 10.3389/fimmu.2025.1617707

Figure Lengend Snippet: Both CD161 + and CD161 - ProTcell subsets retain T and NK cell potential as well as thymus seeding ability. (A–E, G, H) ProTcells were produced from CB CD34 + cells in 7 days as indicated in Figure 1A and sorted by FACS regarding their CD161 expression for further NK or T cell differentiation and functional in vivo experiment. To ensure the consistency, we used the same cell lot and the same sorted cell populations to perform the experiments shown in this figure, including NK cell differentiation, T cell differentiation, and the in vivo engraftment assay. (A) Experimental scheme for CD7 + CD161 - (red frame) and CD7 + CD161 + (green frame) cell subset sorting before NK or T cell differentiation. Differentiating cell phenotype was assessed by flow cytometry. (B, C) CD7 + CD161 +/- lymphoid progenitors were cultured for 14 days in the presence of IL15 without any feeder cell for NK differentiation. Bar plots represent the mean ± SD of (B) the proportion and (C) number of CD56 + NK-primed cells in the culture. (D, E) CD7 + CD161 +/- lymphoid progenitors were co-cultured for 4 weeks with OP9-DLL1 to advance T cell differentiation. Line plots depicting the mean ± SD of the (D) proportion and (E) number of CD3 + developing T cell throughout the culture. *p<0.05 values are for multiple paired Student’s t-test. (F) ProTcells were produced from mPB and CB CD34 + cells and their protein expression (CyTOF) was assessed in a single cell fashion, and restricted to CD7 + progenitors, as described in Figures 1A, B . CyTOF data was visualized by UMAP dimension reduction technic. UMAP projections illustrate the protein expression of several adhesion/homing molecules CCR9, CCR7, CXCR4, CD44, CD62L, SELPLG, ITGA4 and ITGB4. The black line separates the CD161-enriched from the CD161-devoid cells. (G) CD7 + CD161 - or CD7 + CD161 + cell subset has been injected separately into NSG neonates (<4-day-old). T cell reconstitution was analyzed by flow cytometry in the thymus at 6-week-post-transplantation. Bar plot representing the human chimerism (human CD45 + cells/human + murine CD45 + cells) and (H) the proportion of mature CD3 + TCRαβ + T cells in the thymus of mice injected either CD161 + or CD161 - ProTcells. ns, non significant for (B, C) Wilcoxon signed-rank test, (D) multiple paired Student's t-test, (G, H) Mann-Whitney U test.

Article Snippet: 153Eu , CD62L (SELL) , Fluidigm , DREG-56 , 3153004C.

Techniques: Produced, Expressing, Cell Differentiation, Functional Assay, In Vivo, Flow Cytometry, Cell Culture, Injection, Transplantation Assay, MANN-WHITNEY

Journal: International Journal of Biological Sciences

Article Title: Lung Cancer Cell-intrinsic Asparagine Synthetase Potentiates Anti-Tumor Immunity via Modulating Immunogenicity and Facilitating Immune Remodeling in Metastatic Tumor-draining Lymph Nodes

doi: 10.7150/ijbs.114791

Figure Lengend Snippet: ASNS shapes the immune landscapes in metastatic TdLN and primary tumor site. Figure A-C . LN metastasis model was conducted on C57BL/6 mice with LLC-ASNS WT (n=6), ASNS C2A overexpression cells(n=6) and control group(n=6), primary tumor and popliteal lymph nodes were isolated at the end of the experiment, and primary tumor volume(B) and TdLN volume/tumor volume(C) was measured and analyzed. 5D-F . (D) Representative FACS profiles of CD8+T cells are shown. The percentage(E) and number(F) of CD8+ subset in TIL cells isolated from primary tumor is shown. 5G-I . (G) Representative FACS profiles of the co-expression pattern of CD44 and CD62L, or the co-expression pattern of TCF-1and TOX in CD8+ T cells are shown. The number(H) and percentage(I) of CD44+, CD44+CD62L+, CD44+CD62L-, Tsl and TTSM subset in CD8+T cells isolated from TdLN is shown. 5J-L . (J) Representative FACS profiles of the co-expression pattern of CD44 and CD62L, or the co-expression pattern of TCF-1and TOX in CD8+ T cells are shown. The number(K) and percentage(L) of CD44+, CD44+CD62L+, CD44+CD62L-, Tsl and TTSM subset in CD8+T cells isolated from primary tumor is shown.

Article Snippet: For mIHC, the mouse tissue slides were sequentially labeled with monoclonal antibodies against CD44 (Bosterbio, A00052), CD62L (Bosterbio, PB9389), CD8 (Bosterbio, A02236-1), Ovabumin (Abcam, ab181688), and TCF-1 (Cell Signaling Technology, 2203T).

Techniques: Over Expression, Control, Isolation, Expressing

Journal: International Journal of Biological Sciences

Article Title: Lung Cancer Cell-intrinsic Asparagine Synthetase Potentiates Anti-Tumor Immunity via Modulating Immunogenicity and Facilitating Immune Remodeling in Metastatic Tumor-draining Lymph Nodes

doi: 10.7150/ijbs.114791

Figure Lengend Snippet: ASNS-high-expression metastases generated lymphocyte niches enriched with activated T cells, memory T cells, Tsl and TTSM. Figure A-C . Representative immunofluorescence staining images of metastatic TdLNs from LN metastasis model. 6D . The number of CD8+ T cells in the metastasis locations within TdLNs (ASNS WT , n=6, ASNS C2A , n=5, and EV, n=4). 6E-F . The number(E) and percentage(F) of CD44+CD8+T cells among all CD8 T cells in the metastasis locations within TdLNs (ASNS WT , n=6, ASNS C2A , n=5, and EV, n=4). 6G-H . The number(G) and percentage(H) of Tsl cells among all CD8 T cells in the metastasis locations within TdLNs (ASNS WT , n=6, ASNS C2A , n=5, and EV, n=4). 6I-J . The number(I) and percentage(J) of TTSM cells among all CD8 T cells in the metastasis locations within TdLNs (ASNS WT , n=5, ASNS C2A , n=4, and EV, n=3). 6K . Quantitative estimates of the distance from ova+ to CD8+CD44+CD62L+TCF+(TTSM) (ASNS WT , n=6, ASNS C2A , n=5, and EV, n=4). 6L-M . Representative immunofluorescence staining images of metastatic TdLNs from NSCLC patients. 6N-O . The number(C) and percentage(D) of CD8+ T cells in the metastasis locations within TdLNs(ASNS high group, n=7, and ASNS low group, n=6). 6P-Q . The number (E) and percentage(F) of CD45RO+CD8+ T cells in CD8+T cells in the metastasis locations within TdLNs(ASNS high group, n=7, and ASNS low group, n=6). 6R-S . The number (G) and percentage(H) of Tsl cells in CD8+T cells in the metastasis locations within TdLNs(ASNS high group, n=7, and ASNS low group, n=6). 6T-U . The number (I) and percentage(J) of TTSM cells in CD8+T cells in the metastasis locations within TdLNs(ASNS high group, n=7, and ASNS low group, n=6).

Article Snippet: For mIHC, the mouse tissue slides were sequentially labeled with monoclonal antibodies against CD44 (Bosterbio, A00052), CD62L (Bosterbio, PB9389), CD8 (Bosterbio, A02236-1), Ovabumin (Abcam, ab181688), and TCF-1 (Cell Signaling Technology, 2203T).

Techniques: Expressing, Generated, Immunofluorescence, Staining