Journal: bioRxiv

Article Title: Synergistic Hypoxia and Apoptosis Conditioning Unleashes Superior Mesenchymal Stem Cells Efficacy in Acute Graft-versus-Host-Disease

doi: 10.1101/2024.04.11.588248

Figure Lengend Snippet: Effect of hypoxia-preconditioned MSCs (MSCs HYP , MSCs HYP→APO , MSCs HYP+APO ) on the proliferation of aGVHD patients derived T-cell. The bar graph represents (A) the percentage of CD3 + CFSE + proliferating T-cell (n=5). (B) the ratio of CD4 + /CD8 + T cell (n=5) in the direct co-culture of MSCs and T-cell. Data shown as Mean±S.D.; Statistical analysis: Tukey’s multiple comparisons test; **≤0.01; ****≤0.0001. Abbreviations: BM: Bone marrow; WJ: Wharton’s Jelly; MSCs: Mesenchymal Stem Cells; HYP: Hypoxia-preconditioned; APO: Apoptosis .

Article Snippet: CD3 + T-cell was isolated from PBMNCs by negative selection using a Pan T cell isolation kit, human (Miltenyi Biotec, USA) as per the manufacturer’s instructions.

Techniques: Derivative Assay, Co-Culture Assay

Journal: bioRxiv

Article Title: Synergistic Hypoxia and Apoptosis Conditioning Unleashes Superior Mesenchymal Stem Cells Efficacy in Acute Graft-versus-Host-Disease

doi: 10.1101/2024.04.11.588248

Figure Lengend Snippet: Dot plots showing the proliferation of CD3 + T-cell in the direct co-culture of aGVHD patients derived T-cell and (A) Untreated (Control). (B) BM-MSCs HYP . (C) BM-MSCs HYP→APO . (D) BM-MSCs HYP+APO . (E) WJ-MSCs HYP . (F) WJ-MSCs HYP→APO . (G) WJ-MSCs HYP+APO . Abbreviations: BM: Bone marrow; WJ: Wharton’s Jelly; MSCs: Mesenchymal Stem Cells; HYP: Hypoxia-preconditioned; APO: Apoptosis .

Article Snippet: CD3 + T-cell was isolated from PBMNCs by negative selection using a Pan T cell isolation kit, human (Miltenyi Biotec, USA) as per the manufacturer’s instructions.

Techniques: Co-Culture Assay, Derivative Assay, Control

Journal: bioRxiv

Article Title: Synergistic Hypoxia and Apoptosis Conditioning Unleashes Superior Mesenchymal Stem Cells Efficacy in Acute Graft-versus-Host-Disease

doi: 10.1101/2024.04.11.588248

Figure Lengend Snippet: Effect of hypoxia-preconditioned MSCs (MSCs HYP , MSCs HYP→APO , MSCs HYP+APO ) on the induction of Tregs and polarization of helper T cell from pro-inflammatory (Th1, Th17) to anti-inflammatory (Th2) phenotype. The bar graph represents (A) the induction of CD3 + CD4 + CD25 + FOXP3 + Tregs (n=5). (B) the ratio of Th1/Th2 (n=5). (C) Th1/Th17 ratio (n=5) in the direct co-culture of MSCs and T-cell. Data shown as Mean±S.D.; Statistical analysis: Tukey’s multiple comparisons test; ***≤0.001; ****≤0.0001. Abbreviations: BM: Bone marrow; WJ: Wharton’s Jelly; MSCs: Mesenchymal stem cells; HYP: Hypoxia-preconditioned; APO: Apoptosis, Tregs: Regulatory helper T-cell, Th: Helper T cell .

Article Snippet: CD3 + T-cell was isolated from PBMNCs by negative selection using a Pan T cell isolation kit, human (Miltenyi Biotec, USA) as per the manufacturer’s instructions.

Techniques: Co-Culture Assay

Journal: bioRxiv

Article Title: Synergistic Hypoxia and Apoptosis Conditioning Unleashes Superior Mesenchymal Stem Cells Efficacy in Acute Graft-versus-Host-Disease

doi: 10.1101/2024.04.11.588248

Figure Lengend Snippet: Dot plots showing the percentage of CD3 + CD4 + CD25 + FOXP3 + in the direct co-culture of aGVHD patients derived CD3 + T-cell and (A) Untreated (Control). (B) BM-MSCs HYP . (C) BM-MSCs HYP→APO . (D) BM-MSCs HYP+APO . (E) WJ-MSCs HYP . (F) WJ-MSCs HYP→APO . (G) WJ-MSCs HYP+APO . Abbreviations: BM: Bone marrow; WJ: Wharton’s Jelly; MSCs: Mesenchymal Stem Cells; HYP: Hypoxia-preconditioned; APO: Apoptosis .

Article Snippet: CD3 + T-cell was isolated from PBMNCs by negative selection using a Pan T cell isolation kit, human (Miltenyi Biotec, USA) as per the manufacturer’s instructions.

Techniques: Co-Culture Assay, Derivative Assay, Control

Journal: Heliyon

Article Title: Binding domain on CD22 molecules contributing to the biological activity of T cell-engaging bispecific antibodies

doi: 10.1016/j.heliyon.2023.e17960

Figure Lengend Snippet: CD3 + T cells proliferation and CD69 expression frequency on CD8 + and CD4 + T cells. (A) FACS histograms of CD3 + T cells proliferation for G5/44 BsAb (up line) and M971 BsAb (next line) on 2, 4 and 6 days. Green signal was the proliferated CD3 + T cells with the lower fluorescence intensity contributed by CD22-TCBs. (B) Column diagram of CD3 + T cells proliferation mediated by all six CD22-TCBs. (C) Expression frequency of CD69 on CD8 + and CD4 + T cells after 20 h incubation of huPBMCs with presence or absence of Raji cells or 100 ng/mL CD22-TCBs. Data were shown as mean ± SD (n = 3). (For interpretation of the references to colour in this figure legend, the reader is referred to the Web version of this article.)

Article Snippet: Next, the cell mixture was harvested on day 2, 4 and 6 with labeling of mouse anti-human CD3 APC-labeled mAb (Sino Biological).

Techniques: Expressing, Fluorescence, Incubation

Journal: eLife

Article Title: Cytotoxic T-cells mediate exercise-induced reductions in tumor growth

doi: 10.7554/eLife.59996

Figure Lengend Snippet: ( A ) Wild type (WT) and MMTV-PyMT (PyMT) mice on the FVB background were allowed to exercise voluntarily (in running wheels, runners) or left non-exercised (locked running wheels, non-runners) between 4 and 12 weeks of age. ( B ) Running distance (km/day) in WT and PyMT mice. *p<0.05, two-tailed unpaired t test. ( C ) Tumor volume measured twice weekly for PyMT running and non-running mice. Mean and SEM (n = 10–11). ( D ) Survival of PyMT mice, runners, and non-runners. Survival curve ns = not significant, Log-rank (Mantel-Cox) test, (n = 10–11). ( E ) Tumor initiation as age (day) of first indication of a palpable tumor in running (Runners) and non-running (Non-runners) PyMT mice. n = 11, ns = not significant, two-tailed t test. ( F ) Tumor stage from histological scoring (1-4) in mammary glands of running and non-running PyMT mice at 12 (n = 6–10) and 8 (n = 4) weeks of age, ns = not significant, two-tailed t test. ( G ) Individual body weight for WT and PyMT running and non-running mice. n = 7–10, ns = not significant, one-way ANOVA with Tukey’s multiple comparison test. ( H ) Immunohistological characterization of PyMT tumors from non-running and running mice using CD3, F4/80, PCNA, Podocalyxin (PODXL), and Granzyme B (GZMB) antibodies, respectively. n = 8–15, ns = not significant, *p<0.05, Two-tailed unpaired t test. ( I ) Flow-cytometry-based frequency of macrophages within I3TC tumor. Ns = not significant, two-tailed t-test. ( J ) Flow-cytometry-based frequency of neutrofiles within I3TC tumor. Ns = not significant, two-tailed t-test.

Article Snippet: The cells were characterized using Flow cytometry described below or plated at 1 × 10 6 (24-well plate) or 5 × 10 5 (48-well plate) and activated with Dynabeads Mouse T-Activator CD3/CD28 (Thermo Fisher, 11456D ) in a 1:1 beat to T-cell ratio and cultured in 2 ml (24WP) or 1 ml (48WP) RPMI 1640 (Thermo Fisher, 21875 ) supplemented with 10% Fetal Bovine Serum, (Thermo Fisher, 10270 – 106 ), 50 µM 2-mercaptoethanol (Thermo Fisher, 21985023 ), 10 U/ml penicillin-streptomycin (Thermo Fisher, 15140122 ), and 10 U/ml recombinant human IL-2 (Sigma, 11011456001 ), and incubated at 37 °C for 3 days in a humidified CO 2 incubator.

Techniques: Two Tailed Test, Flow Cytometry

Journal: eLife

Article Title: Cytotoxic T-cells mediate exercise-induced reductions in tumor growth

doi: 10.7554/eLife.59996

Figure Lengend Snippet:

Article Snippet: The cells were characterized using Flow cytometry described below or plated at 1 × 10 6 (24-well plate) or 5 × 10 5 (48-well plate) and activated with Dynabeads Mouse T-Activator CD3/CD28 (Thermo Fisher, 11456D ) in a 1:1 beat to T-cell ratio and cultured in 2 ml (24WP) or 1 ml (48WP) RPMI 1640 (Thermo Fisher, 21875 ) supplemented with 10% Fetal Bovine Serum, (Thermo Fisher, 10270 – 106 ), 50 µM 2-mercaptoethanol (Thermo Fisher, 21985023 ), 10 U/ml penicillin-streptomycin (Thermo Fisher, 15140122 ), and 10 U/ml recombinant human IL-2 (Sigma, 11011456001 ), and incubated at 37 °C for 3 days in a humidified CO 2 incubator.

Techniques: Transgenic Assay, Marker, Derivative Assay, Staining, Software, Plasmid Preparation, Avidin-Biotin Assay