Journal: AIDS Research and Therapy

Article Title: The dynamic changes of interferon lambdas related genes and proteins in JAK/STAT pathway in both acute and chronic HIV-1 infected patients

doi: 10.1186/s12981-017-0158-7

Figure Lengend Snippet: Nucleotide sequences of the primers used for real-time PCR

Article Snippet: Antibodies (Abs) used in this study were as follows: mouse phycoerythin (PE)-conjugated anti-human IFN-alpha/beta R2 Ab (clone MMHAR-2, R&D Systems), mouse PE-conjugated anti-human IFN-lambdas R1 Ab (clone 601106, R&D Systems), mouse PE-conjugated anti-human IFN-gamma Receptor 1 (clone GIR-208, Thermo Fisher Scientific), Mouse eFluor ® 450-conjugated anti-human STAT1 (KIKSI0803, Thermo Fisher Scientific), Rabbit FITC anti-Human STAT2 (Thermo Fisher Scientific), Mouse PE-Cyanine7-conjugated anti-human STAT3 (clone LUVNKLA, Thermo Fisher Scientific), mouse APC-conjugated anti-human STAT4 (clone 4LURPLE, Thermo Fisher Scientific), mouse FITC-conjugated anti-human STAT5 (clone SRBCZX, Thermo Fisher Scientific), mouse PerCP-eFluor ® 710-conjugated anti-human STAT6 (clone CHI2S4N, Thermo Fisher Scientific).

Techniques: Sequencing

Journal: AIDS Research and Therapy

Article Title: The dynamic changes of interferon lambdas related genes and proteins in JAK/STAT pathway in both acute and chronic HIV-1 infected patients

doi: 10.1186/s12981-017-0158-7

Figure Lengend Snippet: Correlation between the CD4 + T cells and mRNA levels of IFN-alpha receptor ( a ), IFN-gamma receptor ( c ), and IFN-lambdas receptor ( e ). Correlation between the viral loads and mRNA levels of IFN-alpha receptor ( b ), IFN-gamma receptor ( d ), and IFN-lambdas receptor ( f ). The results were performed Spearman’s rank correlation, where coefficients “r” and corresponding p values are indicated on each panel

Article Snippet: Antibodies (Abs) used in this study were as follows: mouse phycoerythin (PE)-conjugated anti-human IFN-alpha/beta R2 Ab (clone MMHAR-2, R&D Systems), mouse PE-conjugated anti-human IFN-lambdas R1 Ab (clone 601106, R&D Systems), mouse PE-conjugated anti-human IFN-gamma Receptor 1 (clone GIR-208, Thermo Fisher Scientific), Mouse eFluor ® 450-conjugated anti-human STAT1 (KIKSI0803, Thermo Fisher Scientific), Rabbit FITC anti-Human STAT2 (Thermo Fisher Scientific), Mouse PE-Cyanine7-conjugated anti-human STAT3 (clone LUVNKLA, Thermo Fisher Scientific), mouse APC-conjugated anti-human STAT4 (clone 4LURPLE, Thermo Fisher Scientific), mouse FITC-conjugated anti-human STAT5 (clone SRBCZX, Thermo Fisher Scientific), mouse PerCP-eFluor ® 710-conjugated anti-human STAT6 (clone CHI2S4N, Thermo Fisher Scientific).

Techniques:

Journal:

Article Title: Autoimmunity to type VII collagen in SKH1 mice is independent of regulatory T cells

doi: 10.1111/j.1365-2249.2006.03115.x

Figure Lengend Snippet: Effect of anti-CD25 monoclonal antibody. (a) SKH1 mice were intraperitoneally administered 500 µg purified rat anti-CD25. The mice which received the antibodies were killed at 24 h (day 1), day 3, day 5 or day 7 after the administration of the antibody. Spleen and lymph nodes were then harvested. Single-cell suspensions were stained with fluorescein isothiocyanate (FITC)-conjugated anti-mouse CD4 and phycoerythrin (PE)-labelled anti-mouse CD25 and analysed by fluorescence activated cell sorter (FACSCalibur). Results of injection of isotype rat Ig G are not shown. Day 0 indicates results of mice without anti-CD25 treatment. The numbers located above dot plots indicate the percentages of CD25+ cells in CD4+ cells. (b) To determine the expression of CD25 and forkhead boxp3 (Foxp3) in CD4+ cells after anti-CD25 antibody treatment, five groups of SKH1 mice were used. Group 1 received a single dose of 500 µg isotype IgG. Group 2 received a single dose of 500 µg anti-CD25. Group 3 received two doses of 500 µg anti-CD25, 5 days apart. Group 4 received three doses of 500 µg anti-CD25, 5 days apart. Group 5 received four doses of 500 µg anti-CD25, 5 days apart. At day 5 of the last injections, cells from the spleen and lymph node were stained by FITC-anti-CD4, PE-anti-Foxp3 and allophycocyanin (APC)-anti-CD25 and analysed by Cyan ADP flow cytometer. Data shown are results from lymph node cells in groups 1 and 2 after one anti-CD25 or IgG isotype injection. The numbers located above each dot plot in the left column indicate the percentages of CD25+ in Foxp3+ cells gated on CD4+ cells. The numbers located above each dot plot in the middle column indicate the percentages of CD25+ cells in CD4+ cells. The numbers located above each dot plot in the right column indicate the percentages of Foxp3+ cells in CD4+ cells. (c) Anti-CD25 inactivates Treg in a consistent manner: CD25+/Foxp3 (gated on CD4+), CD25+/CD4+ and Foxp3+/CD4+ cells in the lymph node were analysed after one, two, three and four cycles of anti-CD25 treatment, showing consistent inactivation of Treg, without physical depletion. Similar analyses for IgG1 isotype treatment obtained after one cycle treatment are shown on the far left for comparison. Similar results were obtained in the studies of the spleen cells (data not shown).

Article Snippet: The function of the secondary antibody was validated by using a sandwich method: the plate was coated with unlabelled monoclonal rat anti-mouse IgG1 (BD Biosciences), followed by purified standard mouse IgG1 lambda (BD Biosciences) in serial dilutions, then by biotin-labelled anti-mouse lambda chain (1,2,3) (BD Biosciences), followed by incubation of streptavidin–peroxidase polymer (Sigma) and then TMB substrate (Sigma).

Techniques: Purification, Staining, Fluorescence, Injection, Expressing, Flow Cytometry

Journal:

Article Title: Autoimmunity to type VII collagen in SKH1 mice is independent of regulatory T cells

doi: 10.1111/j.1365-2249.2006.03115.x

Figure Lengend Snippet: NC1-immunized SKH1 mice produce IgG antoantibodies specifically against recombinant NC1 protein. (a). Titratable enzyme-linked immunosorbent assay (ELISA) titres of autoantibodies. Three groups of female SKH1 mice were included. One group received rat anti-CD25 and mouse NC1 (n = 5, solid diamond), the second group received rat isotype and mouse NC1 (n = 5, solid square) and the third group received rat anti-CD25 and mouse albumin (n = 3, open circle). Mice sera were collected at the end of 5·5 months after the initial immunization and tested for the specific IgG against mouse NC1 by ELISA as described in Materials and methods. (b). Western blot analysis of NC1-specific autoantibodies. Two representative samples from each group are shown: mice treated with anti-CD25 and immunized with albumin (lanes 1 and 2); mice treated with anti-CD25 and immunized with NC1 (lanes 3 and 4); mice treated with rat IgG and immunized with NC1 (lanes 5 and 6). Each of the wells were loaded with 300 ng of recombinant mouse NC1 protein.

Article Snippet: The function of the secondary antibody was validated by using a sandwich method: the plate was coated with unlabelled monoclonal rat anti-mouse IgG1 (BD Biosciences), followed by purified standard mouse IgG1 lambda (BD Biosciences) in serial dilutions, then by biotin-labelled anti-mouse lambda chain (1,2,3) (BD Biosciences), followed by incubation of streptavidin–peroxidase polymer (Sigma) and then TMB substrate (Sigma).

Techniques: Recombinant, Enzyme-linked Immunosorbent Assay, Western Blot

Journal:

Article Title: Autoimmunity to type VII collagen in SKH1 mice is independent of regulatory T cells

doi: 10.1111/j.1365-2249.2006.03115.x

Figure Lengend Snippet: NC1-immunized SKH1 mice produce IgG autoantibodies which react specifically with skin basement membrane. Indirect immunofluorescence microscopy was used to examine the reactivity of autoreactive IgGs with the skin basement membrane zone component, as described in Materials and methods. Left and middle panels show IgG autoantibodies reacting with the skin basement membrane in mice immunized with NC1 in combinations of anti-CD25 or isotype control, respectively. The right panel shows the result of the serum from mouse treated with anti-CD25 and immunized with albumin. Images are representative for each group (original magnification ×40).

Article Snippet: The function of the secondary antibody was validated by using a sandwich method: the plate was coated with unlabelled monoclonal rat anti-mouse IgG1 (BD Biosciences), followed by purified standard mouse IgG1 lambda (BD Biosciences) in serial dilutions, then by biotin-labelled anti-mouse lambda chain (1,2,3) (BD Biosciences), followed by incubation of streptavidin–peroxidase polymer (Sigma) and then TMB substrate (Sigma).

Techniques: Immunofluorescence, Microscopy

Journal:

Article Title: Autoimmunity to type VII collagen in SKH1 mice is independent of regulatory T cells

doi: 10.1111/j.1365-2249.2006.03115.x

Figure Lengend Snippet: IgG class autoantibodies deposited to skin basement membrane in NC1-immunized mice. Direct immunofluorescence microscopy was performed to examine the deposit of IgG skin basement membrane zone, as described in Materials and methods. The left panel shows IgG deposits in mice treated with anti-CD25 and immunized with NC1, the middle panel shows IgG deposits in mice treated with rat IgG and immunized with NC1 and the right panel shows IgG deposits in mice treated with anti-CD25 and immunized with albumin. Images are representative for each group (original magnification ×40).

Article Snippet: The function of the secondary antibody was validated by using a sandwich method: the plate was coated with unlabelled monoclonal rat anti-mouse IgG1 (BD Biosciences), followed by purified standard mouse IgG1 lambda (BD Biosciences) in serial dilutions, then by biotin-labelled anti-mouse lambda chain (1,2,3) (BD Biosciences), followed by incubation of streptavidin–peroxidase polymer (Sigma) and then TMB substrate (Sigma).

Techniques: Immunofluorescence, Microscopy

Journal:

Article Title: Autoimmunity to type VII collagen in SKH1 mice is independent of regulatory T cells

doi: 10.1111/j.1365-2249.2006.03115.x

Figure Lengend Snippet: Anti-NC1 IgG subclasses and light chains [enzyme-linked immunosorbent assay (ELISA)].

Article Snippet: The function of the secondary antibody was validated by using a sandwich method: the plate was coated with unlabelled monoclonal rat anti-mouse IgG1 (BD Biosciences), followed by purified standard mouse IgG1 lambda (BD Biosciences) in serial dilutions, then by biotin-labelled anti-mouse lambda chain (1,2,3) (BD Biosciences), followed by incubation of streptavidin–peroxidase polymer (Sigma) and then TMB substrate (Sigma).

Techniques: Enzyme-linked Immunosorbent Assay