Journal: Frontiers in Virology

Article Title: Complex Interactions Between Human Immunodeficiency Virus Type-1, Sex, and Osteopontin Influence Viral Replication and Leukocyte Proportions in Tissues

doi: 10.3389/fviro.2021.690360

Figure Lengend Snippet: FIGURE 1 | Experimental design and correlation of viral loads with engraftment levels. (A) Illustrates the experimental design for the generation of hCD34+ humanized mice, HIV infection, groups implemented and the tissues harvested for the study. The time point for the tissue harvest reflects chronic infection. Representative flow cytometric dot plot of mouse blood-stained for human CD45-PerCP and mouse CD45-FITC used to identify successfully engrafted animals. The 2 × 2 design for female and male mice is indicated. (B) Correlation analyses between measured variables %hCD45+ cells (at time of engraftment) and viral loads (at study endpoint) for each mouse in the four HIV-infected groups. The Spearman coefficients (rs) indicated, while positive, did not reach significance (P < 0.05) in any of the groups. F-HIV-Opn+ n = 5, F-HIV-Opn– n = 9, M-HIV-Opn+ n = 8 and M-HIV-Opn– n = 5.

Article Snippet: Tissues were evaluated at the study endpoint to validate and quantify human leukocytes by seven-color flow cytometry with a cocktail containing the following antibodies: human pan-CD45-Viogreen or PerCP (Miltenyi Biotec), 130-110-638, ThermoFisher (MHCD4531), CD3-PE (ThermoFisher, MHCD0304), CD4-Vioblue (Miltenyi Biotec, 130-113-219), CD8-APC-Vio770 (Miltenyi Biotec, 130- 113-155), CD14-APC (BioLegend, 301808), CD19-FITC or PerCP (eBiosciences, 11-0199-42; BioLegend, 302228) and mouse CD45-FITC (eBiosciences, 11-0451-82).

Techniques: Infection, Staining

Journal: Journal of immunology (Baltimore, Md. : 1950)

Article Title: CD45 tyrosine phosphatase controls common gamma-chain cytokine-mediated STAT and extracellular signal-related kinase phosphorylation in activated human lymphoblasts: inhibition of proliferation without induction of apoptosis.

doi: 10.4049/jimmunol.166.10.6034

Figure Lengend Snippet: FIGURE 1. CD45 mAbs inhibit proliferation in quiescent PBMCs. PB- MCs were isolated as described in Materials and Methods. One 3 105

Article Snippet: Hybridoma cells producing CD3-mAb OKT-3 (IgG2a, 1 mg/ml), panCD45 mAb NIH45-2 (IgG1, 10 mg/ml), or pan-CD45 mAb GAP8.3 (IgG2a, 10 mg/ml) were obtained from American Type Culture Collection (Manassas, VA).

Techniques: Isolation

Journal: Journal of immunology (Baltimore, Md. : 1950)

Article Title: CD45 tyrosine phosphatase controls common gamma-chain cytokine-mediated STAT and extracellular signal-related kinase phosphorylation in activated human lymphoblasts: inhibition of proliferation without induction of apoptosis.

doi: 10.4049/jimmunol.166.10.6034

Figure Lengend Snippet: FIGURE 4. Influence of CD45 signals on mRNA quantities in lympho- blasts. Lymphoblasts were generated as described. After stimulation under the various conditions as specified, cells were lysed, RNA isolated, mRNA reverse transcribed, and amplified as described in Materials and Methods. The results shown are representative of three different experiments after PCR amplification for 28 cycles.

Article Snippet: Hybridoma cells producing CD3-mAb OKT-3 (IgG2a, 1 mg/ml), panCD45 mAb NIH45-2 (IgG1, 10 mg/ml), or pan-CD45 mAb GAP8.3 (IgG2a, 10 mg/ml) were obtained from American Type Culture Collection (Manassas, VA).

Techniques: Generated, Isolation, Reverse Transcription

Journal: Journal of immunology (Baltimore, Md. : 1950)

Article Title: CD45 tyrosine phosphatase controls common gamma-chain cytokine-mediated STAT and extracellular signal-related kinase phosphorylation in activated human lymphoblasts: inhibition of proliferation without induction of apoptosis.

doi: 10.4049/jimmunol.166.10.6034

Figure Lengend Snippet: FIGURE 7. Influence of CD45 signals on IL-5 secretion by lympho- blasts. Lymphoblasts were generated as described. Sixteen to 20 h after stimulation under the various conditions as specified, supernatants were collected and concentrations of IL-5 were determined as described in Ma- terials and Methods. Data are mean values 6 SD from three different experiments.

Article Snippet: Hybridoma cells producing CD3-mAb OKT-3 (IgG2a, 1 mg/ml), panCD45 mAb NIH45-2 (IgG1, 10 mg/ml), or pan-CD45 mAb GAP8.3 (IgG2a, 10 mg/ml) were obtained from American Type Culture Collection (Manassas, VA).

Techniques: Generated

Journal: Journal of immunology (Baltimore, Md. : 1950)

Article Title: CD45 tyrosine phosphatase controls common gamma-chain cytokine-mediated STAT and extracellular signal-related kinase phosphorylation in activated human lymphoblasts: inhibition of proliferation without induction of apoptosis.

doi: 10.4049/jimmunol.166.10.6034

Figure Lengend Snippet: FIGURE 5. Influence of CD45 signals on IFN-g secretion by lympho- blasts. Lymphoblasts were generated as described. Sixteen to 20 h after stimulation under the various conditions as specified, supernatants were collected and concentrations of IFN-g were determined as described in Materials and Methods. Data are mean values 6 SD from three different experiments.

Article Snippet: Hybridoma cells producing CD3-mAb OKT-3 (IgG2a, 1 mg/ml), panCD45 mAb NIH45-2 (IgG1, 10 mg/ml), or pan-CD45 mAb GAP8.3 (IgG2a, 10 mg/ml) were obtained from American Type Culture Collection (Manassas, VA).

Techniques: Generated

Journal: Journal of immunology (Baltimore, Md. : 1950)

Article Title: CD45 tyrosine phosphatase controls common gamma-chain cytokine-mediated STAT and extracellular signal-related kinase phosphorylation in activated human lymphoblasts: inhibition of proliferation without induction of apoptosis.

doi: 10.4049/jimmunol.166.10.6034

Figure Lengend Snippet: FIGURE 6. Influence of CD45 signals on TNF-a secretion by lympho- blasts. Lymphoblasts were generated as described. Sixteen to 20 h after stimulation under the various conditions as specified, supernatants were collected and concentrations of TNF-a were determined as described in Materials and Methods. Data are mean values 6 SD from three different experiments.

Article Snippet: Hybridoma cells producing CD3-mAb OKT-3 (IgG2a, 1 mg/ml), panCD45 mAb NIH45-2 (IgG1, 10 mg/ml), or pan-CD45 mAb GAP8.3 (IgG2a, 10 mg/ml) were obtained from American Type Culture Collection (Manassas, VA).

Techniques: Generated

Journal: Journal of Biological Chemistry

Article Title: Peptide-Major Histocompatibility Complex Dimensions Control Proximal Kinase-Phosphatase Balance during T Cell Activation

doi: 10.1074/jbc.m109.039966

Figure Lengend Snippet: FIGURE 4. Small pMHC dimensions allow efficient segregation from CD45 following TCR engagement. A, experiments were performed as in Fig. 3. The data were acquired as 8-bit 1024 1024 pixel images. B, back- ground subtracted images acquired in the CD3 and CD45 channels were analyzed for colocalization by deter- mining Manders colocalization coefficients using ImageJ. The mean proportion of CD3 colocalized with CD45 expressed as a percentage (Manders M1 coefficient) was calculated from 20 interfaces chosen at random for each coated surface. Manders M2 coefficient (proportion of CD45 colocalized with CD3) is 100% for all. The horizontal black bars represent the means. Statistical significance was determined by analysis of variance with correction for multiple comparisons. *, p 0.05; ***, p 0.001. The scale bar represents 4 m.

Article Snippet: The sampleswere stained forCD3 , Zap70, and CD45 by serial incubation of each antibody followedwhere appropriate by a species-specific and fluorophoreconjugated secondary antibody in the order mouse anti-CD3 (UCHT1; AbD Serotec), anti-mouse Alexa 488 (Molecular Probes), rabbit anti-Zap70 (99F2; Cell Signaling Technology), anti-rabbit IgG Alexa 546 (Molecular Probes), and anti-human pan CD45 antibody directly conjugated with Alexa 647 (F1089-4; AbD Serotec).

Techniques: