Journal: Clinical and Experimental Immunology

Article Title: Cytotoxic T lymphocyte antigen 4-immunoglobulin G is a potent adjuvant for experimental allergen immunotherapy

doi: 10.1111/cei.12041

Figure Lengend Snippet: Experimental layout: In experiment 1, to evaluate that whether cytotoxic T lymphocyte antigen 4-immunoglobulin (CTLA-4–Ig) can enhance the suppressive effects of allergen-specific immunotherapy (SIT), it was combined with ovalbumin (OVA)-SIT (100 μg/mouse) in BALB/c mice. In experiment 2, CTLA-4–Ig is combined with OVA)-SIT in indoleamine 2,3 dioxygenase (IDO–/–) mice to examine whether the effect of CTLA-4–Ig is mediated by IDO. In experiment 3, to analyse the effects of CTLA-4–Ig on regulatory T cells the frequency of CD4+CD25+ forkhead box protein 3 (FoxP3)+ regulatory T cells (Treg) cells and Th2 cells were analysed 24 h after the last SIT injection and 24 h after inhalation challenges.

Article Snippet: Flow cytometry and antibodies Peridinin chlorophyll (Per-CP)-anti-CD4 (BD Pharmingen), fluorescein isothiocyanate (FITC)-anti-T1ST2 (also known as IL-33Ra) (MD-Biosciences, Zurich, Switzerland), phycoerythrin (PE)-anti-forkhead box protein 3 (FoxP3) and eFluor450-anti-CD25 (eBioscience, San Jose, CA, USA) were used for fluorescence activated cell sorting (FACS).

Techniques: Injection

Journal: Clinical and Experimental Immunology

Article Title: Cytotoxic T lymphocyte antigen 4-immunoglobulin G is a potent adjuvant for experimental allergen immunotherapy

doi: 10.1111/cei.12041

Figure Lengend Snippet: The effects of co-administration of cytotoxic T lymphocyte antigen 4-immunoglobulin (CTLA-4–Ig) with allergen-specific immunotherapy–ovalbumin (OVA-SIT) on the frequency of CD4+CD25+ forkhead box protein 3 (FoxP3)+ regulatory T cells (Treg) cells and T helper type 2 (Th2) cells in the blood. (a) Frequency of CD4+CD25+FoxP3+Treg cells in the blood 24 h after the last SIT injection; (b) frequency of CD4+CD25+FoxP3+Treg cells in the blood 24 h after the last inhalation challenge; (c) frequency of CD4+T1ST2+ [also known as interleukin (IL)-33Ra] T cells in the blood 24 h after the last SIT injection.

Article Snippet: Flow cytometry and antibodies Peridinin chlorophyll (Per-CP)-anti-CD4 (BD Pharmingen), fluorescein isothiocyanate (FITC)-anti-T1ST2 (also known as IL-33Ra) (MD-Biosciences, Zurich, Switzerland), phycoerythrin (PE)-anti-forkhead box protein 3 (FoxP3) and eFluor450-anti-CD25 (eBioscience, San Jose, CA, USA) were used for fluorescence activated cell sorting (FACS).

Techniques: Injection

Journal: Cell Death & Disease

Article Title: HIV integrase inhibitor, Elvitegravir, impairs RAG functions and inhibits V(D)J recombination

doi: 10.1038/cddis.2017.237

Figure Lengend Snippet: Evaluation of effect of Elvitegravir on B cells progression in mice by FACS analysis. ( A ) Schematic representation of steps involved during the in vivo experiment. Balb/c mice (vehicle control and Elvitegravir-treated) were fed with Elvitegravir (8 days; 30 mg/kg). Mice were sacrificed and bone marrow cells were collected, stained with CD45, CD25 surface markers and FACS analysed. ( B ) Representative FACS dot plots of CD45 + CD25 + cells from vehicle control and Elvitegravir-treated mice are shown. Two mice each from independent batches of vehicle control (a,b batch I, c,d batch II) and Elvitegravir-treated groups (e,f batch I, g,h batch II) are presented. ( C ) Table showing percentage of CD45 + CD25 + cells obtained following flow cytometric analysis from control ( n =11) and Elvitegravir-treated ( n =16) mice from all three batches. Mice, from the Elvitegarvir treated group, that are affected ( n =11) and unaffected ( n =5) by Elvitegravir are shown. ( D ) Histogram showing CD45 + CD25 + B cells. Double-positive B cells detected following FACS analysis from bone marrow cells of vehicle control (white) are depicted in comparison to Elvitegravir-treated group which is divided into affected (black) and unaffected (grey). While vehicle control mice possessed average 78.03% of CD45 + CD25 + B cells in bone marrow it was reduced to average 62.5% following Elvitegravir treatment. ~30% mice were insensitive to treatment. P value <0.001

Article Snippet: Anti-CD3-FITC (Cat No. 555274), anti-CD8-APC-Cy7 (Cat No. 557654), anti-CD19-APC (Cat No. 550992), anti-CD25-PECy7 (Cat No. 552880) and anti-CD45-APC (Cat No. 559864) were from BD Biosciences.

Techniques: In Vivo, Staining