Journal: The Journal of Experimental Medicine

Article Title: Omega-1, a glycoprotein secreted by Schistosoma mansoni eggs, drives Th2 responses

doi: 10.1084/jem.20082460

Figure Lengend Snippet: Immunological and biochemical characterization of ESP from S. mansoni eggs. (A) Monocyte-derived DCs pulsed for 48 h with the different antigen preparations in the absence (top) or presence (bottom) of 100 ng/ml LPS as a maturation factor were co-cultured with allogeneic naive CD4 + T cells for 12 d in the presence of staphylococcal enterotoxin B and IL-2. Intracellular cytokine production was assayed by FACS 6 h after the stimulation of primed T cells with PMA and ionomycin. The frequencies of each population are indicated as percentages in the plot. One representative result from three independent experiments is shown. (B) 5 µg/cm ESP was separated under nonreducing conditions by SDS-PAGE and silver stained. (C) The presence of omega-1 and IPSE/alpha-1 was confirmed on Western blots by staining with specific anti–IPSE/alpha-1 and anti–omega-1 monoclonal antibodies.

Article Snippet: To determine T cell polarization, 5 × 10 3 48-h-pulsed DCs were co-cultured with 2 × 10 4 naive T cells that were purified using a human CD4 + /CD45RO − column kit (R&D Systems) in the presence of 100 pg/ml staphylococcal enterotoxin B (Sigma-Aldrich) in 96-well flat-bottom plates (Costar).

Techniques: Derivative Assay, Cell Culture, SDS Page, Staining, Western Blot, Bioprocessing

Journal: The Journal of Experimental Medicine

Article Title: Omega-1, a glycoprotein secreted by Schistosoma mansoni eggs, drives Th2 responses

doi: 10.1084/jem.20082460

Figure Lengend Snippet: Omega-1 is sufficient to drive Th2 polarization in vivo, independently of IL-4R signaling. (A) 4get/KN2 IL-4 dual reporter mice were injected s.c. with 20 µg SEA, 2 µg omega-1, or 2 µg IPSE/alpha-1 into the footpad. After 7 d, the frequency of GFP + and huCD2 + within the CD4 + CD44 high effector T cell population was determined by flow cytometry in the draining popliteal lymph nodes. (B) 4get/KN2 mice were injected with recombinant proteins and analyzed as in A. (C) IL-4Rα −/− 4get/KN2 mice were treated and analyzed as in A. In all panels, representative plots are shown with the frequencies of each population indicated as percentages. Graphs depict the combined data of three to four individual mice per group and show one of at least two independent experiments. Error bars represent the mean ± SD. * P < 0.05; **, P < 0.01; ***, P < 0.001 for values significantly different from the PBS control (two-sided paired Student's t test).

Article Snippet: To determine T cell polarization, 5 × 10 3 48-h-pulsed DCs were co-cultured with 2 × 10 4 naive T cells that were purified using a human CD4 + /CD45RO − column kit (R&D Systems) in the presence of 100 pg/ml staphylococcal enterotoxin B (Sigma-Aldrich) in 96-well flat-bottom plates (Costar).

Techniques: In Vivo, Injection, Flow Cytometry, Recombinant, Control

Journal: Transplantation Direct

Article Title: Alemtuzumab as Antirejection Therapy

doi: 10.1097/TXD.0000000000000595

Figure Lengend Snippet: T cell repopulation after alemtuzumab therapy. A, Typical example of the gating strategy; CD4+ and CD8+ naive (CCR7+CD45RO−), central memory (CCR7+CD45RO+), effector memory (CCR7−CD45RO+) and EMRA (CCR7−CD45RO−) T cells. B, Absolute number of the total (left), CD4+ (middle) and CD8+ (right) T cell population before and at months 1, 3, 6, and 12 after alemtuzumab treatment. C, Percentage CD4+ (left) and CD8+ (right) naive, total memory, central memory, effector memory and EMRA T cells before and at months 1, 3, 6, 12 after alemtuzumab therapy. D, Typical example of the gating strategy of CD4+CD25+CD127− regulatory T cells. E, The percentage of CD4+CD25+CD127− Treg cells before and after alemtuzumab therapy. Data are shown as median ± IQR, * P < 0.05, ** P < 0.01; significant differences comparing pre-transplant values to post transplant values the Wilcoxon Signed-Rank test. Due to low cell numbers not all samples could be analysed reliable and are therefore not depicted. IQR, interquartile range.

Article Snippet: Regulatory T cell samples were stained with CD3-AmCyan, CD4-APC-Cy7 (BD Bioscience), CD127-PE-Cy7 (BD Bioscience), CD45RO-PerCP-Cy5 (BD Bioscience), and CD31-PE.

Techniques:

Journal: Transplantation Direct

Article Title: Alemtuzumab as Antirejection Therapy

doi: 10.1097/TXD.0000000000000595

Figure Lengend Snippet: Thymopoiesis and homeostatic proliferation. A, Typical example of the gating strategy: expression of CD31 within CD4+ and CD8+ naive (CCR7+CD45RO−) T cells. B, percentage of RTE; naive T cells expressing CD31, before and after alemtuzumab therapy, before and after alemtuzumab therapy. C, Typical example of the gating strategy of CD4+ and CD8+ Ki-67+ T cells. D, the percentage Ki-67 expressing T cells within the total CD4+, CD4+ naïve, CD4+ total memory and CD4+ central memory and effector memory T cells before and after alemtuzumab therapy. E, the percentage Ki-67 expressing T cells within the total CD8+, CD8+ naive, CD8+ total memory, and CD8+ effector memory and EMRA T cells before and after alemtuzumab therapy. F, The percentage of Ki-67+ Treg cell. Because of low number of cells, we could not determine the percentage RTE and Ki-67+ cells for all patients. Data are shown as median, * P < 0.05; significant differences comparing pretransplant values to posttransplant values using the Wilcoxon Signed-Rank test. RTE, recent thymic emigrants.

Article Snippet: Regulatory T cell samples were stained with CD3-AmCyan, CD4-APC-Cy7 (BD Bioscience), CD127-PE-Cy7 (BD Bioscience), CD45RO-PerCP-Cy5 (BD Bioscience), and CD31-PE.

Techniques: Expressing