Journal: Infection and Immunity

Article Title: Role of T Cells and Gamma Interferon during Induction of Hypersensitivity to Lipopolysaccharide by Toxic Shock Syndrome Toxin 1 in Mice

doi: 10.1128/iai.69.3.1256-1264.2001

Figure Lengend Snippet: FIG. 4. Effects of Cs (A) or anti-IFN-g MAb (B) on LPS-induced serum TNF-a activity in BALB/c-AnNCr mice primed with TSST-1. Groups of four mice were pretreated with Cs (40 mg/kg, i.p.) or anti-IFN-g MAb (750 mg/mouse, i.p.) at 4 or 2 h, respectively, prior to injection of TSST-1. Mice were then injected i.p. with 200 mg of TSST-1 (dotted bars) per kg or PBS (clear bars). Control mice received equal volumes of PBS instead of Cs, rat IgG2a instead of anti-IFN-g MAb, or PBS instead of TSST-1. All mice were injected with LPS (400 mg/kg, i.p.) 12 h later. TNF-a activity in serum was measured 2 h postinjection of LPS. Solutions of Cs or MAb were treated with 2 mg of polymyxin B per ml to remove contaminating LPS. The percent reductions in circulating TNF-a activity caused by Cs or anti-IFN-g MAb are shown.

Article Snippet: Neutralizing rat monoclonal antibody (MAb) to mouse IFN-g, rat immunoglobulin G2a (IgG2a) isotype control MAb, and murine recombinant TNF-a were obtained from R&D Systems (Minneapolis, Minn.).

Techniques: Activity Assay, Injection, Control

Journal: Cellular immunology

Article Title: IL10 restrains autoreactive B cells in transgenic mice expressing inactive RAG1

doi: 10.1016/j.cellimm.2018.06.004

Figure Lengend Snippet: (A). Splenocytes from WT or dnRAG1 mice were cultured for 72h with or without LPS (10 μg/mL) in the absence or presence of the following: recombinant IL10 (rIL10), IL10 blocking antibody (α-IL10 Ab) or isotype control (Iso Ab), or ruxolitinib (Rux), SH-4–54, triamcinolone acetonide (TA) or vehicle (DMSO), and plasma cell subsets were analyzed by flow cytometry as in Fig. 2B. Summary data for n=3 animals/genotype are presented in bar graph format and represented as mean +/− SEM. Statistically significant differences using the Tukey correction are indicated (a, vs LPS; b, vs LPS+Iso Ab; c, vs LPS+DMSO). (B) Concentrations of IgM and IgG in supernatants from splenocyte cultures shown in panel A were measured by ELISA.

Article Snippet: Cell culture Single-cell suspensions from spleen were adjusted to 4.0×10 6 cells/mL in sterile RF10M and 2.0×10 5 cells dispensed into wells of a sterile 96-well flat bottom plate (Corning #353072) The cells were mixed with an equal volume of RF10M lacking or containing the following reagents (source and final concentration shown in parentheses): LPS (Sigma, L2880, 10 μg/mL), IL10 blocking antibody (R&D Systems, MAB417, 10 ng/mL), Rat IgG1 control for IL10 blocking antibody (R&D Systems, MAB005, 10 ng/mL in PBS), recombinant mouse IL10 (Biolegend, 575802, 100 ng/mL), ruxolitinib (Selleckchem, S1378, 10 nM), SH-4–54 (Selleckchem, S7337, 10 nM), or triamcinolone acetonide (Alfa Aesar, J6354803, 10 nM).

Techniques: Cell Culture, Recombinant, Blocking Assay, Control, Flow Cytometry, Enzyme-linked Immunosorbent Assay