Journal: Frontiers in Microbiology

Article Title: Immune cell early activation, apoptotic kinetic, and T-cell functional impairment in domestic pigs after ASFV CADC_HN09 strain infection

doi: 10.3389/fmicb.2024.1328177

Figure Lengend Snippet: Virulent ASFV infection led to predominant reduction of B cells, CD4 T cells, monocytes, and DCs in periphery blood. At the indicated time points, PBMCs were prepared and stained for distinct leukocyte subsets and were analyzed by FCM. The absolute amount of each leukocyte subset per milliliter of blood was calculated based on the percentage of the specific cell type in live PBMCs. The numbers of monocyte (A) , cDC1 (B) , cDC2 (C) , neutrophil (D) , lymphocyte (E) , B cell (F) , γδ T cell (G) , CD3 T cell (H) , CD4 T cell (I) , CD4 + CD8 + T cell (J) , CD8 T cell (K) , and NK cell (L) in 1 mL blood. Data shown are mean ± SD. * p < 0.05. ** p < 0.01.

Article Snippet: Lung samples were predigested with collagenase IV and DNase I (Sigma, Germany) for 30 min at 37°C in a water bath and then ground, followed by isolation with a Porcine Lymphocyte Isolation Kit (Tianjin Haoyang Co. Ltd., China).

Techniques: Infection, Staining

Journal: Frontiers in Microbiology

Article Title: Immune cell early activation, apoptotic kinetic, and T-cell functional impairment in domestic pigs after ASFV CADC_HN09 strain infection

doi: 10.3389/fmicb.2024.1328177

Figure Lengend Snippet: Early activation of different lymphocyte subsets after ASFV infection in blood and organs. Single-cell suspensions of peripheral blood, mLN, lung, and spleen were prepared at 3 dpi and stained for lymphocyte subsets and were analyzed by FCM. The percentages of CD69 + lymphocyte subsets in PBMC (A) , mLN (B) , lung (C) , and spleen (D) were compared between ASFV-uninfected and ASFV-infected groups. Data shown are mean ± SD. * p < 0.05.

Article Snippet: Lung samples were predigested with collagenase IV and DNase I (Sigma, Germany) for 30 min at 37°C in a water bath and then ground, followed by isolation with a Porcine Lymphocyte Isolation Kit (Tianjin Haoyang Co. Ltd., China).

Techniques: Activation Assay, Infection, Staining

Journal: Frontiers in Microbiology

Article Title: Immune cell early activation, apoptotic kinetic, and T-cell functional impairment in domestic pigs after ASFV CADC_HN09 strain infection

doi: 10.3389/fmicb.2024.1328177

Figure Lengend Snippet: ASFV infection induced cytokine-secreting T cells and NK cells. PBMC of both ASFV-infected and ASFV-uninfected piglets were separated at 5 dpi and stained for lymphocyte subsets, followed by intracellular cytokine staining for TNF-α, IFN-γ, and IL-2, and then, the cells were analyzed by FCM. Representative pseudocolor plots (left panel: the upper one, uninfected; the lower one, infected) and increased frequencies of IFN-γ + , IFN-γ + TNF-α + or IL-2 + CD4 T (A) , CD8 T (B) , CD4 + CD8 + T (C) , NK (D) and γδ T cells (E,F) (right panel). Data shown are mean ± SD. * p < 0.05.

Article Snippet: Lung samples were predigested with collagenase IV and DNase I (Sigma, Germany) for 30 min at 37°C in a water bath and then ground, followed by isolation with a Porcine Lymphocyte Isolation Kit (Tianjin Haoyang Co. Ltd., China).

Techniques: Infection, Staining

Journal: Frontiers in Microbiology

Article Title: Immune cell early activation, apoptotic kinetic, and T-cell functional impairment in domestic pigs after ASFV CADC_HN09 strain infection

doi: 10.3389/fmicb.2024.1328177

Figure Lengend Snippet: ASFV infection impaired the capability of T cells to produce cytokines. At the indicated time points, PBMC of ASFV-infected and uninfected piglets were separated and stimulated with mitogen and were stained for lymphocyte subsets, followed by an intracellular cytokine staining step and cells were determined by FCM. Representative pseudocolor plots (A) and the differential expression of cytokines in CD4 + CD8 + T cell (B) . Representative pseudocolor plots (left panels) and the differential expression of cytokines in CD4 + CD8 + T cell (C) , CD4 T cell (D) and CD8 T cell (E) (right panels). Each point represents data from a single pig while bars represent the mean of each group. Data shown are mean ± SD. ns, no statistical significance. * p < 0.05; ** p < 0.01.

Article Snippet: Lung samples were predigested with collagenase IV and DNase I (Sigma, Germany) for 30 min at 37°C in a water bath and then ground, followed by isolation with a Porcine Lymphocyte Isolation Kit (Tianjin Haoyang Co. Ltd., China).

Techniques: Infection, Staining, Quantitative Proteomics