Journal: Clinical and Vaccine Immunology

Article Title: Association of Reduced Tumor Necrosis Factor Alpha, Gamma Interferon, and Interleukin-1β (IL-1β) but Increased IL-10 Expression with Improved Chest Radiography in Patients with Pulmonary Tuberculosis

doi: 10.1128/cvi.00381-09

Figure Lengend Snippet: FIG. 1. Schematic timeline of therapeutic regimen and clinical as- sessment. The workup for TB included sputum smear, chest radio- graphs, and plasma cytokine level determination.

Article Snippet: A mixture of antibodies for the detection of nine biotinylated anti-human cytokine antibodies (R&D Systems, Inc.) were diluted with PBSBT at 40 g/ml (1:200) and added to the cytokine arrays, which were kept at room temperature for 30 min.

Techniques: Clinical Proteomics

Journal:

Article Title: Anti-IL-9 vaccination prevents worm expulsion and blood eosinophilia in Trichuris muris -infected mice

doi:

Figure Lengend Snippet: Induction of IL-9 specific autoantibodies. (A) Vaccination against IL-9. Groups of four C57BL/6 mice were injected subcutaneously with IL-9-OVA complexes in CFA (IL-9-OVA), with noncomplexed IL-9 and OVA (IL-9 + OVA) or with CFA alone (CFA). Boosts were carried out in IFA after 2 and 4 wk. Serially diluted sera, collected 2 wk after the last immunization, were tested for IL-9 inhibition in a TS1 cell proliferation assay. Cell growth was evaluated by measuring hexosaminidase activity. Means ± SD are indicated. (B) Specificity of anti-IL-9 activity. Pools of 1/320 diluted sera from immunized (IL-9-OVA) or control (CFA) mice were tested for growth inhibition of TS1 cells, in the presence of either natural mIL-9, baculovirus-derived mIL-9 (rec.), or mIL-4. Cell proliferation was measured after 3 days of culture. Results are given as means ± SD.

Article Snippet: IL-9-OVA complexes were detected in column fractions by ELISA by using 2C12, a hamster monoclonal anti-IL-9 antibody produced in our laboratory for capture, and a rabbit anti-OVA antiserum followed by peroxidase-conjugated anti-rabbit antibody (Santa Cruz Biotechnology) for detection of complexes.

Techniques: Injection, Inhibition, Proliferation Assay, Activity Assay, Derivative Assay

Journal:

Article Title: Anti-IL-9 vaccination prevents worm expulsion and blood eosinophilia in Trichuris muris -infected mice

doi:

Figure Lengend Snippet: Production of anti-IL-9 autoantibodies in different mouse strains. Eight-week-old C57BL/6, FVB, BALB/c, NOD, DBA/2, or NZW mice were injected with IL-9-OVA complexes (four mice) or adjuvant only (four mice), as described in Materials and Methods. Serially diluted sera, collected 2 wk after the last immunization, were tested in a TS1 cell proliferation assay. Inhibition titers, given for each mouse serum, correspond to serum dilutions inhibiting IL-9-induced cell proliferation by 50%.

Article Snippet: IL-9-OVA complexes were detected in column fractions by ELISA by using 2C12, a hamster monoclonal anti-IL-9 antibody produced in our laboratory for capture, and a rabbit anti-OVA antiserum followed by peroxidase-conjugated anti-rabbit antibody (Santa Cruz Biotechnology) for detection of complexes.

Techniques: Injection, Proliferation Assay, Inhibition

Journal:

Article Title: Anti-IL-9 vaccination prevents worm expulsion and blood eosinophilia in Trichuris muris -infected mice

doi:

Figure Lengend Snippet: Persistence of the anti-IL-9 response in immunized mice. Three 8-wk-old C57BL/6 mice were injected s.c. with IL-9-OVA complexes in CFA. Mice were boosted with IFA after 2 and 4 wk. Serum was collected 2, 4, 6, 8, 12, 26, and 56 wk after priming and tested for anti-IL-9 activity in a TS1 cell proliferation assay. Results are given as the inhibition titer for individual mouse sera.

Article Snippet: IL-9-OVA complexes were detected in column fractions by ELISA by using 2C12, a hamster monoclonal anti-IL-9 antibody produced in our laboratory for capture, and a rabbit anti-OVA antiserum followed by peroxidase-conjugated anti-rabbit antibody (Santa Cruz Biotechnology) for detection of complexes.

Techniques: Injection, Activity Assay, Proliferation Assay, Inhibition

Journal:

Article Title: Anti-IL-9 vaccination prevents worm expulsion and blood eosinophilia in Trichuris muris -infected mice

doi:

Figure Lengend Snippet: Inhibition of IL-9-induced mast-cell activation and eosinophilia in immunized mice. Seven-week-old C57BL/6 mice subjected to the IL-9-OVA immunization protocol or immunized with noncoupled IL-9 and OVA or left untreated (five mice per group) were injected i.p. with 107 IL-9-producing TS1.G6 cells, 5 mo after the last boost. Five age-matched mice served as negative controls. All mice were tested 25 days later for MMCP-1 serum levels by ELISA (A), and blood leukocytes were isolated for eosinophil counting (B). Eosinophil percentages were determined by enumerating 20,000 cells per slide. Means ± SEM are indicated.

Article Snippet: IL-9-OVA complexes were detected in column fractions by ELISA by using 2C12, a hamster monoclonal anti-IL-9 antibody produced in our laboratory for capture, and a rabbit anti-OVA antiserum followed by peroxidase-conjugated anti-rabbit antibody (Santa Cruz Biotechnology) for detection of complexes.

Techniques: Inhibition, Activation Assay, Injection, Enzyme-linked Immunosorbent Assay, Isolation

Journal:

Article Title: Anti-IL-9 vaccination prevents worm expulsion and blood eosinophilia in Trichuris muris -infected mice

doi:

Figure Lengend Snippet: Altered response to T. muris in IL-9-OVA-vaccinated mice. (A) Failure of IL-9-OVA-immunized C57BL/6 mice to expel T. muris. C57BL/6 mice immunized with IL-9-OVA complexes or vehicle (eight mice per group) were infected by oral gavage of 200 T. muris eggs. Worms were counted in ceca on day 34 after infection, and results are given as worm burden in each individual. P value between groups is 0.001. On day 13, the worm burden, assessed to verify infectivity, was similar in all animals. (B) Inhibition of T. muris-induced blood eosinophilia in IL-9-vaccinated mice. Blood eosinophils were counted 23 days after infection from infected control or IL-9-vaccinated C57BL/6 mice or uninfected naive animals. Eosinophil percentages were determined by enumerating 500 cells per slide. Results are presented as the mean percentage of blood eosinophils ± SEM (four mice per group). P value is 0.029 for the difference between OVA and IL-9-OVA groups.

Article Snippet: IL-9-OVA complexes were detected in column fractions by ELISA by using 2C12, a hamster monoclonal anti-IL-9 antibody produced in our laboratory for capture, and a rabbit anti-OVA antiserum followed by peroxidase-conjugated anti-rabbit antibody (Santa Cruz Biotechnology) for detection of complexes.

Techniques: Infection, Inhibition