Journal: BMC Cancer

Article Title: Exercise accelerates recruitment of CD8 + T cell to promotes anti-tumor immunity in lung cancer via epinephrine

doi: 10.1186/s12885-024-12224-7

Figure Lengend Snippet: Transcriptional targets were examined by RNA-seq in tumors after exercise. a The number of differentially expressed genes in the Ex and Ctrl mice were detected by RNA-seq. b Volcano plot of differentially expressed genes in Ex compared with Ctrl tumor tissues. Red dots represent upregulated genes, and blue dots represent downregulated genes. c Heat map of the differentially expressed genes in Ex compared with Ctrl tumor tissues. d Pathway enrichment analysis of significantly upregulated or downregulated genes. e-j RT-qPCR validates the gene expression of Cxcl10 , Cxcl12 , Cxcl14 , Ppbp , Pf4 and Ccl8 in the tumors each group ( n = 3) after tumor inoculation. k-l The expression levels of Cxcl10 and Cxcl12 were detected in the tumors in each group by ELISA ( n = 8). The results of e-l are presented as mean ± SEM. Statistical analysis was performed using two-tailed unpaired t tests. * p < 0.05, ** p < 0.01, *** p < 0.001

Article Snippet: The concentrations of Ccl5, Cxcl10 and Cxcl12 were measured in Lewis lung cancercellsculture supernatants and in the serum of mice using Ccl5 ELISA kit (Cusabio, CSB-E09256m), Cxcl10 ELISA kit (Cusabio, CSB-E08183m) and Cxcl12 (Cusabio, CSB-E04723m).

Techniques: RNA Sequencing, Quantitative RT-PCR, Gene Expression, Expressing, Enzyme-linked Immunosorbent Assay, Two Tailed Test

Journal: BMC Cancer

Article Title: Exercise accelerates recruitment of CD8 + T cell to promotes anti-tumor immunity in lung cancer via epinephrine

doi: 10.1186/s12885-024-12224-7

Figure Lengend Snippet: Exercise promotes CD8 + T cell recruitment with the assistance of Ccl5 and Cxcl10. a-d Correlation between Ccl5, Cxcl9, Cxcl10, and Cxcl11 mRNA level and CD8 + T cell using QUANTISEQ. Data were obtained from TIMER 2.0 ( http://timer.comp-genomics.org/ ). e-h RT - qPCR analyses of mRNA expression of chemokines in the tumors of Ctrl and Ex groups ( n = 3). i-j ELISA analysis of serum levels of Cxcl10 and Ccl5 in Ctrl and Ex mice ( n = 8). k-m RT-qPCR analyses of mRNA expression of cytokines and PD-L1 in the tumors of Ctrl and Ex mice ( n = 3). n Western blot analysis the expression of PD-L1 in the tumors of Ctrl and Ex mice ( n = 3). Full-length blots/gels are presented in Fig. 4n of source data. o Automatic quantification of PD-L1 IHC staining in indicated mice tumors ( n = 8). p IHC staining of PD-L1 on tumor sections from different groups ( n = 8). q Western blot analysis the expression of P53 in the tumors of Ctrl and Ex mice ( n = 3). Full-length blots/gels are presented in Fig. 4q of source data. The results of e-m and o are presented as mean ± SEM. Statistical analysis was performed using two-tailed unpaired t tests. * p < 0.05, ** p < 0.01, *** p < 0.001

Article Snippet: The concentrations of Ccl5, Cxcl10 and Cxcl12 were measured in Lewis lung cancercellsculture supernatants and in the serum of mice using Ccl5 ELISA kit (Cusabio, CSB-E09256m), Cxcl10 ELISA kit (Cusabio, CSB-E08183m) and Cxcl12 (Cusabio, CSB-E04723m).

Techniques: Quantitative RT-PCR, Expressing, Enzyme-linked Immunosorbent Assay, Western Blot, Immunohistochemistry, Two Tailed Test

Journal: BMC Cancer

Article Title: Exercise accelerates recruitment of CD8 + T cell to promotes anti-tumor immunity in lung cancer via epinephrine

doi: 10.1186/s12885-024-12224-7

Figure Lengend Snippet: Exercise inhibits tumor progression by upregulation of EPI levels. a Serum levels of EPI in Ctrl, Ex and Ex-post groups were detected by ELISA in tumor-bearing mice ( n = 8). b-d EPI mice receiving daily injections of EPI (0.5 mg/kg i.p.) ( n = 8) and TE mice with daily running for 14 days after tumor inoculation ( n = 8) compared with TC mice ( n = 8). b Representative images. c Tumor weights of indicated groups. d Tumor volumes of different groups. e IHC staining of CD8, CD4, CD3 and GZMB on tumor sections from indicated groups ( n = 8). f Automatic quantification of CD8, CD4, CD3 and GZMB IHC staining in indicated mice tumors ( n = 8). Data are presented as the mean ± SEM in each group. Statistical analysis of a was performed using two-tailed unpaired t tests. c , d and f were performed using one-way analysis of variance (ANOVA). * p < 0.05, ** p < 0.01, *** p < 0.001

Article Snippet: The concentrations of Ccl5, Cxcl10 and Cxcl12 were measured in Lewis lung cancercellsculture supernatants and in the serum of mice using Ccl5 ELISA kit (Cusabio, CSB-E09256m), Cxcl10 ELISA kit (Cusabio, CSB-E08183m) and Cxcl12 (Cusabio, CSB-E04723m).

Techniques: Enzyme-linked Immunosorbent Assay, Immunohistochemistry, Two Tailed Test

Journal: BMC Cancer

Article Title: Exercise accelerates recruitment of CD8 + T cell to promotes anti-tumor immunity in lung cancer via epinephrine

doi: 10.1186/s12885-024-12224-7

Figure Lengend Snippet: EPI regulates changes in chemokine levels. a-f RT-qPCR analyses the mRNA expression of chemokines and cytokines in the tumors of TC and EPI mice ( n = 3). g-h ELISA analysis serum levels of Cxcl10 and Ccl5 in TC, TE and EPI treated mice ( n = 8). i-j RT-qPCR detect the mRNA levels of Cxcl10 and Ccl5 in LLC cells after treated with different concentrations of EPI (0, 5, 10, 20 µM). k-l Cxcl10 and Ccl5 in the supernatant of LLC cells were measured by ELISA. m RT-qPCR analyses the mRNA expression of CD274 in the tumors of control TC and EPI groups ( n = 3). n Western blot analyses PD-L1 in LLC cells after EPI treatment for 24 h at different concentration (0, 5, 10, 20 µM). Full-length blots/gels are presented in Fig. 6n of source data. o IHC staining of PD-L1 on tumor sections from indicated groups ( n = 8). p Automatic quantification of PD-L1 IHC staining in indicated mice tumors ( n = 8). q Graphical abstract of this study. Exercise-induced elevation of EPI is involved in regulating Ccl5 and Cxcl10 expression, subsequently promoting CD8 + T cells recruitment, and ultimately inhibiting tumor progression. Data are presented as the mean ± SEM in each group. Statistical analysis of a-f and m were performed using two-tailed unpaired t tests. g-l and p were performed using one-way analysis of variance (ANOVA). * p < 0.05, ** p < 0.01, *** p < 0.001

Article Snippet: The concentrations of Ccl5, Cxcl10 and Cxcl12 were measured in Lewis lung cancercellsculture supernatants and in the serum of mice using Ccl5 ELISA kit (Cusabio, CSB-E09256m), Cxcl10 ELISA kit (Cusabio, CSB-E08183m) and Cxcl12 (Cusabio, CSB-E04723m).

Techniques: Quantitative RT-PCR, Expressing, Enzyme-linked Immunosorbent Assay, Control, Western Blot, Concentration Assay, Immunohistochemistry, Two Tailed Test

Journal: The Journal of Experimental Medicine

Article Title: Hematopoietic Stem Cells Are Uniquely Selective in Their Migratory Response to Chemokines

doi: 10.1084/jem.20011284

Figure Lengend Snippet: LT-HSC and ST-HSC migrate in response to SDF-1α, but are refractory to other chemokines and to G-CSF. (A) Flow cytometry contour plots showing partial enrichment of BM cells for HSC by lineage depletion (upper two panels), and gating of lineage − LT-HSC (boxed cells, lower left) and lineage lo ST-HSC (boxed cells, lower right). See the Materials and Methods section for details. (B and C) Lineage-depleted BM cells were prepared and added to inserts placed in wells containing medium alone, the listed chemokines, or G-CSF (see the Materials and Methods section for concentrations). Responding cells were harvested, stained for HSC markers, and analyzed for the presence and number of (B) LT-HSC and (C) ST-HSC by flow cytometry. The data presented are the means ±SD of two to nine independent experiments and represent the percentage of input HSC that migrated to each chemokine or to G-CSF. Numbers in parentheses indicate the number of experiments performed for each agent. CC and CXC refer to the receptor families to which the listed chemokines belong. Compared with basal migration, only migration to SDF-1α was statistically significant ( P < 0.05).

Article Snippet: The following chemokines were added to the bottom well, and cells were allowed to migrate for 2 h at 37°C: mouse JE, mouse eotaxin, human thymus- and activation-regulated chemokine, mouse regulated upon activation, normal T expressed and secreted (RANTES), mouse macrophage inflammatory protein (MIP)-1α, human MIP-3α, human I-309, mouse KC, and human SDF-1α (PeproTech); mouse MIP-1β, human MIP-3β, mouse thymus–expressed chemokine (TECK), mouse monokine induced by IFN-γ (MIG), and mouse B lymphocyte chemokine (BLC)/BCA-1 (R&D Systems); and human IL-8 (a gift from K. Matsushima, University of Tokyo, School of Medicine, Tokyo, Japan).

Techniques: Flow Cytometry, Staining, Migration

Journal: The Journal of Experimental Medicine

Article Title: Hematopoietic Stem Cells Are Uniquely Selective in Their Migratory Response to Chemokines

doi: 10.1084/jem.20011284

Figure Lengend Snippet: PCR Primers Used for Analysis of Chemokine Receptor mRNA Expression by HSC. DHFR, Dihydrofolate Reductase

Article Snippet: The following chemokines were added to the bottom well, and cells were allowed to migrate for 2 h at 37°C: mouse JE, mouse eotaxin, human thymus- and activation-regulated chemokine, mouse regulated upon activation, normal T expressed and secreted (RANTES), mouse macrophage inflammatory protein (MIP)-1α, human MIP-3α, human I-309, mouse KC, and human SDF-1α (PeproTech); mouse MIP-1β, human MIP-3β, mouse thymus–expressed chemokine (TECK), mouse monokine induced by IFN-γ (MIG), and mouse B lymphocyte chemokine (BLC)/BCA-1 (R&D Systems); and human IL-8 (a gift from K. Matsushima, University of Tokyo, School of Medicine, Tokyo, Japan).

Techniques: Expressing, Sequencing

Journal: The Journal of Experimental Medicine

Article Title: Hematopoietic Stem Cells Are Uniquely Selective in Their Migratory Response to Chemokines

doi: 10.1084/jem.20011284

Figure Lengend Snippet: Chemokines Used and Their Known Receptors. Standardized Chemokine Names Are Given in Parentheses

Article Snippet: The following chemokines were added to the bottom well, and cells were allowed to migrate for 2 h at 37°C: mouse JE, mouse eotaxin, human thymus- and activation-regulated chemokine, mouse regulated upon activation, normal T expressed and secreted (RANTES), mouse macrophage inflammatory protein (MIP)-1α, human MIP-3α, human I-309, mouse KC, and human SDF-1α (PeproTech); mouse MIP-1β, human MIP-3β, mouse thymus–expressed chemokine (TECK), mouse monokine induced by IFN-γ (MIG), and mouse B lymphocyte chemokine (BLC)/BCA-1 (R&D Systems); and human IL-8 (a gift from K. Matsushima, University of Tokyo, School of Medicine, Tokyo, Japan).

Techniques:

Journal: The Journal of Experimental Medicine

Article Title: Hematopoietic Stem Cells Are Uniquely Selective in Their Migratory Response to Chemokines

doi: 10.1084/jem.20011284

Figure Lengend Snippet: LT-HSC and ST-HSC contain mRNA for CCR3, CCR9, and CXCR4. (A) RT-PCR analysis of mRNA for chemokine receptors of combined LT-HSC and ST-HSC (Thy-1.1 lo Sca-1 + Lin −/lo c-Kit + cells). RT-PCR was performed on RNA isolated from the equivalent of 1,000 double-sorted HSC, or from 1,000 unfractionated WBM cells. Representative data are shown (see for data summary). (B) Additional RT-PCR was performed on mRNA isolated from the equivalent of 1,000 LT-HSC or 1,000 ST-HSC for the receptors found to be positive in the first screen. Both LT-HSC and ST-HSC contained mRNA for CXCR4, CCR3, and CCR9. See the Materials and Methods section for RT-PCR protocol. DHFR, dihydrofolate reductase; WBM, whole bone marrow.

Article Snippet: The following chemokines were added to the bottom well, and cells were allowed to migrate for 2 h at 37°C: mouse JE, mouse eotaxin, human thymus- and activation-regulated chemokine, mouse regulated upon activation, normal T expressed and secreted (RANTES), mouse macrophage inflammatory protein (MIP)-1α, human MIP-3α, human I-309, mouse KC, and human SDF-1α (PeproTech); mouse MIP-1β, human MIP-3β, mouse thymus–expressed chemokine (TECK), mouse monokine induced by IFN-γ (MIG), and mouse B lymphocyte chemokine (BLC)/BCA-1 (R&D Systems); and human IL-8 (a gift from K. Matsushima, University of Tokyo, School of Medicine, Tokyo, Japan).

Techniques: Reverse Transcription Polymerase Chain Reaction, Isolation

Journal: The Journal of Experimental Medicine

Article Title: Hematopoietic Stem Cells Are Uniquely Selective in Their Migratory Response to Chemokines

doi: 10.1084/jem.20011284

Figure Lengend Snippet: Summary of RT-PCR Analysis of Chemokine Receptor mRNA Expression by HSC

Article Snippet: The following chemokines were added to the bottom well, and cells were allowed to migrate for 2 h at 37°C: mouse JE, mouse eotaxin, human thymus- and activation-regulated chemokine, mouse regulated upon activation, normal T expressed and secreted (RANTES), mouse macrophage inflammatory protein (MIP)-1α, human MIP-3α, human I-309, mouse KC, and human SDF-1α (PeproTech); mouse MIP-1β, human MIP-3β, mouse thymus–expressed chemokine (TECK), mouse monokine induced by IFN-γ (MIG), and mouse B lymphocyte chemokine (BLC)/BCA-1 (R&D Systems); and human IL-8 (a gift from K. Matsushima, University of Tokyo, School of Medicine, Tokyo, Japan).

Techniques: Expressing