Journal: Frontiers in Physiology

Article Title: Integrative Transcriptomic and microRNAomic Profiling Reveals Immune Mechanism for the Resilience to Soybean Meal Stress in Fish Gut and Liver

doi: 10.3389/fphys.2018.01154

Figure Lengend Snippet: Immunohistochemistry analysis for the main lymphocytes and key cytokines involved in SBMIE. (A) IHC results for IgM, CD4, IL17 and IL10 in the gut and liver in all time points in 40SBM group as well as 0 days in FM group (negative control) and 3 week in 70SBM group (positive control). The green signals represent the examined protein, while the blue signals (stained by Hoechst 33342) represent the nuclei. The white arrows points out typical immunohistochemistry signals. Scale bar: 30 μm. (B) The quantification of IHC signals in both the gut (left) and liver (right). In the gut, the significant ( p < 0.01) increase of signals compared to the sample at 0 days in LP or in IEL was indicated by stars or hashes. In the liver, for each protein, the signals were calculated for significant changes, indicated by lowercase letters “a, b, c, d, e”. For each protein, the same label means no difference. For comparing T cell and B cell signals, CD4 and IgM signals were calculated for p -value at each time point, the significant ( p < 0.01) more CD4 was indicated by the star, whereas the significant result for IgM ( p < 0.1) was indicated by hashes.

Article Snippet: Commercial antibodies [anti-zebrafish CD4 monoclonal antibody (mAb), Genetex] or the antibodies developed by our group, including mouse anti-grass carp IgM (GenBank: DQ417927.1 ) mAb, rabbit anti-grass carp IL17 (NCBI Accession: KC978892.1 ) polyclonal antibody (pAb), and rabbit anti-grass carp IL10 (NCBI Accession: JQ768312.1 ) pAb were used in the study.

Techniques: Immunohistochemistry, Negative Control, Positive Control, Staining

Journal: Journal of investigative medicine : the official publication of the American Federation for Clinical Research

Article Title: Nitric Oxide Induces Apoptosis in Spleen Lymphocytes from MRL/ lpr Mice

doi: 10.1136/jim-52-01-29

Figure Lengend Snippet: Oral Administration of the NOS Inhibitor NMMA Inhibited Apoptosis of Spleen Lymphocytes from MRL/ lpr but not BALB/cJ Control Mice

Article Snippet: To resuspend pelleted cells, 0.2 μg of phycoerythrin (PE)-conjugated anti-CD4, -CD8, or -immunoglobulin M (IgM) antibodies (Promega, Madison, WI) in 50 μL of 1% BSA in PBS was added to wells.

Techniques: Control, Marker

Journal: Journal of investigative medicine : the official publication of the American Federation for Clinical Research

Article Title: Nitric Oxide Induces Apoptosis in Spleen Lymphocytes from MRL/ lpr Mice

doi: 10.1136/jim-52-01-29

Figure Lengend Snippet: Apoptosis of MRL/lpr spleen lymphocyte subsets with exogenous nitric oxide in vitro. MRL/lpr spleen lymphocytes were isolated and cultured overnight in varying concentrations of Deta NONOate. Cells were harvested and analyzed for apoptosis using flow cytometry with annexin V–fluorescein isothiocyanate and phycoerythrin cell surface marker (CD4+, CD8+, and IgM+) staining. A, The results were normalized to the percentage of apoptotic cells relative to control wells (not exposed to Deta NONOate) ± standard error and were calculated from the average of five experiments. Apoptosis of subsets was reported as follows: CD4+ cells as closed circles, CD8+ cells as open inverted triangles, IgM+ cells as closed squares, and CD4−CD8−IgM− cells as open diamonds. Deta NONOate induced significant apoptosis among CD4+, CD8+, and CD4−CD8−IgM− cells (asterisk signifies p < .05). B, The results of a representative single experiment reported as the percentage of apoptotic intact cells in each subset. Subset analysis was not performed for BALB/cJ spleen lymphocytes.

Article Snippet: To resuspend pelleted cells, 0.2 μg of phycoerythrin (PE)-conjugated anti-CD4, -CD8, or -immunoglobulin M (IgM) antibodies (Promega, Madison, WI) in 50 μL of 1% BSA in PBS was added to wells.

Techniques: In Vitro, Isolation, Cell Culture, Flow Cytometry, Marker, Staining, Control

Journal: PLoS ONE

Article Title: Anti-CD154 mAb and Rapamycin Induce T Regulatory Cell Mediated Tolerance in Rat-to-Mouse Islet Transplantation

doi: 10.1371/journal.pone.0010352

Figure Lengend Snippet: Percentages of CD25 + Foxp3 + Treg gated in CD4 + T cell in the peripheral blood. Representative example of the gating in CD4 + T cells and the isotype controls used for CD25 (IgG1: pC61, IgG2b: 3C7) and Foxp3 (IgG2a) are shown in the upper panel ( A , B , C ). Representative analysis of CD25 + Foxp3 + Treg in Group 1, i.e. islet transplantation without further therapy, at day 20 post-transplantation ( D ) in naive mice ( E ) and in Group 7, i.e early anti-CD25 mAb at day 20 post-transplantation ( F ).

Article Snippet: Briefly serial frozen sections were stained for characterization of infiltrating cells with anti CD4 mAb and anti-CD8 mAb and anti mac-1 (Morphosys AbD Düsseldorf, Germany).

Techniques: Transplantation Assay

Journal: PLoS ONE

Article Title: Anti-CD154 mAb and Rapamycin Induce T Regulatory Cell Mediated Tolerance in Rat-to-Mouse Islet Transplantation

doi: 10.1371/journal.pone.0010352

Figure Lengend Snippet: Cryostat sections were stained by anti-mouse CD4 ( A – D ), CD8 ( B – D ) and macrophage antibodies ( C – E ) in Group 1 ( A – B – C ) and Group 4 at 200 days post transplantation. In Group 1 of rejecting mice, immunohistology for cellular immune responses to concordant islet xenografts at time of rejection has detected mixed cellular infiltrates with presence of CD4+ ( A ), CD8+ ( B ), and macrophages ( C ). In Group 4 of tolerant mice, immunohistology for cellular immune responses to concordant islet xenografts of tolerant mice at 200 days post-transplantation detected only minimal cellular infiltrate with absence of CD4+ ( D ), CD8+ ( E ), and macrophages ( F ). (Magnification in A – C (200x), Magnification in D – F (100x)).

Article Snippet: Briefly serial frozen sections were stained for characterization of infiltrating cells with anti CD4 mAb and anti-CD8 mAb and anti mac-1 (Morphosys AbD Düsseldorf, Germany).

Techniques: Staining, Transplantation Assay

Journal: PLoS ONE

Article Title: Anti-CD154 mAb and Rapamycin Induce T Regulatory Cell Mediated Tolerance in Rat-to-Mouse Islet Transplantation

doi: 10.1371/journal.pone.0010352

Figure Lengend Snippet: Treg were isolated form the spleen of Group 1 (rejecting mice) at day 10 post transplantation before rejection occurrence and of Group 4 (tolerant mice) at day 20 after tolerance induction. Three mice per Group in 3 separate experiments showed similar mean fluorescence intensity ( A ) and purity ( B ) of Foxp3 + cells (gated on CD4 + CD25 + ). Treg of Group 1 and 4 were used at different ratio (1∶1, 1∶2, 1∶4, and 1∶8) in co-culture with syngeneic responders (CD4 + CD25 + ) and stimulators and anti CD3e mAb ( C ). Treg xenospecificity was assed by co-culture of naive mice splenocytes with donor Sprague Dawley or third party Lewis stimulators plus minus rejector or tolerant Treg. Percentage of suppression are shown ( D ). Foxp3, TGFβ-1 and IL-10 mRNA expression were measured by real time PCR. Results were calculated on basis of relative mRNA expression compared to naive mice (relative expression = 1, E ).

Article Snippet: Briefly serial frozen sections were stained for characterization of infiltrating cells with anti CD4 mAb and anti-CD8 mAb and anti mac-1 (Morphosys AbD Düsseldorf, Germany).

Techniques: Isolation, Transplantation Assay, Fluorescence, Co-Culture Assay, Expressing, Real-time Polymerase Chain Reaction