human lymphocyte separation kit  (Beijing Solarbio Science)

Journal: Advanced Science

Article Title: Hyperbaric Oxygen Boosts PD‐1 Antibody Delivery and T Cell Infiltration for Augmented Immune Responses Against Solid Tumors

doi: 10.1002/advs.202100233

Figure Lengend Snippet: HBO enhances T cell infiltration into tumor parenchyma. A) Experimental scheme of HBO enabling T cell infiltration in H22 orthotopic tumor‐bearing BALB/c mice. Proportion of infiltrated CD45 + lymphocyte B), CD3 + T cells C), CD8 + T cells D), and CD4 + T cells E) in tumors ( n = 10). F) Immunofluorescence staining of CD4 + T cell and CD8 + T cell images. The scale bar is 50 µm. Quantification of CD4 + T cells G) and CD8 + T cells H) in tumor tissue sections ( n = 20). I) Experimental procedures of HBO enhancing T cell infiltration in H22 subcutaneous tumor‐bearing BALB/c nu/nu mice. J,K) Infiltrated CD4 + T cells and CD8 + T cells in tumors were detected by flow cytometry ( n = 6) after adoptive lymphocytes transfer. M) MASSON staining images, immunofluorescence staining images of T cells (CD4 + and CD8 + ) of tumor tissue. L) Quantification of collagen in MASSON staining of tumor tissue sections ( n = 20). N) Average infiltrated T cells in tumor tissue ( n = 20). O) The infiltration depth of T cells from the tumor edge ( n = 20). Error bars indicate SEM. Statistical significance was calculated by t ‐test. P ‐values: *, P < 0.05; **, P < 0.01; ***, P < 0.001.

Article Snippet: The lymphocytes were extracted by a mouse lymphocyte extraction kit (Cedarlane: CL5010).

Techniques: Immunofluorescence, Staining, Flow Cytometry

Journal: Advanced Science

Article Title: Hyperbaric Oxygen Boosts PD‐1 Antibody Delivery and T Cell Infiltration for Augmented Immune Responses Against Solid Tumors

doi: 10.1002/advs.202100233

Figure Lengend Snippet: HBO synergizes with PD‐1 Ab to inhibit the relapse of the H22 tumor. A) Experimental scheme of HBO helping PD‐1 Ab trigger a long‐term immunological memory effect. B) Tumor growth curves and D) tumor weight of the first tumors ( n = 8). C) Tumor formation curves and E) tumor weight of the rechallenged tumors ( n = 8). Proportions of effector memory T cells gated on lymphocyte F) and gated on CD8 + T cells G) from spleen analyzed by flow cytometry at day 51 before mice were rechallenged with the secondary tumors ( n = 6). IFN‐ γ H) and granzyme B I) expression of CD8 + T cells in lymph node from mice isolated 10 days after mice were rechallenged with the secondary tumors (at day 61) ( n = 8). Cytokine of J) IL‐2 and K) IFN‐ γ in serum from mice isolated 10 days after mice were rechallenged with the secondary tumors (at day 61) ( n = 4). Error bars indicate SEM. Statistical significance was calculated by t ‐test. P ‐values: *, P < 0.05; **, P < 0.01; ***, P < 0.001.

Article Snippet: The lymphocytes were extracted by a mouse lymphocyte extraction kit (Cedarlane: CL5010).

Techniques: Flow Cytometry, Expressing, Isolation