Journal:

Article Title: Compound heterozygosity for Pten and SHIP augments T-dependent humoral immune responses and cytokine production by CD4 + T cells

doi: 10.1111/j.1365-2567.2004.01901.x

Figure Lengend Snippet: Pten+/– SHIP+/– (phosphatase with tensin homology/SH2-containing inositol phosphatase) mice show an increased humoral response to the T-dependent antigen, nitrophenylacetyl (NP)-keyhole limpet haemocyanin (NP-KLH). Serum antibodies against NP-bovine serum albumin (NP-BSA) were assessed from six or seven mice of each genotype that had been challenged with NP-Ficoll or NP-KLH. Anti-NP titres [immunoglobulin M (IgM) and pan immunoglobulin G (IgG)] were determined by enzyme-linked immunosorbent assay (ELISA) on serially diluted serum samples. Bars indicate the average titre, and triangles represent individual mice. No differences were observed in either the IgM or IgG response to NP-Ficoll (a, b). However, statistically significant differences in IgM and IgG responses to NP-KLH were detected (c, d). *P < 0·05 compared with all genotypes, ***P < 0·005 compared with wild type and SHIP+/–, but not significant over Pten+/–. **P < 0·05 compared with wild type and SHIP+/–. WT, wild type; SH, SHIP+/–; PH, Pten+/–; DH, Pten+/– SHIP+/–.

Article Snippet: Enzyme-linked immunosorbent assays (ELISAs) for serum immunoglobulins, NP-specific antibody production and cytokines ELISA quantification of serum immunoglobulin levels was performed, as previously described, 19 using capture and biotinylated detection antibodies for immunoglobulin G1 (IgG1), immunoglobulin G2b (IgG2b) and immunoglobulin M (IgM) (BD Pharmingen).

Techniques: Enzyme-linked Immunosorbent Assay

Journal:

Article Title: Compound heterozygosity for Pten and SHIP augments T-dependent humoral immune responses and cytokine production by CD4 + T cells

doi: 10.1111/j.1365-2567.2004.01901.x

Figure Lengend Snippet: Increased levels of serum immunoglobulin G1 (IgG1) in Pten+/– SHIP+/– (phosphatase with tensin homology/SH2-containing inositol phosphatase) mice. (a) Spontaneous increases in circulating serum IgG1 levels were detected in the sera of 8-week-old Pten+/– SHIP+/– mice, as compared to the other genotypes, and this pattern remained steady and elevated over time. (b) Serum levels of circulating immunoglobulin M (IgM) were not significantly increased over all genotypes at the 8-week point, but steadily increased, becoming significant at later time-points. (c) Levels of circulating immunoglobulin G2b (IgG2b) did not achieve statistical significance compared to all genotypes until the 24-week time-point. Values were representative of sera from at least nine animals per genotype, assessed in duplicate. Error bars represent the standard error of the mean (SEM) of the duplicate readings. *P < 0·05 compared with all genotypes.

Article Snippet: Enzyme-linked immunosorbent assays (ELISAs) for serum immunoglobulins, NP-specific antibody production and cytokines ELISA quantification of serum immunoglobulin levels was performed, as previously described, 19 using capture and biotinylated detection antibodies for immunoglobulin G1 (IgG1), immunoglobulin G2b (IgG2b) and immunoglobulin M (IgM) (BD Pharmingen).

Techniques:

Journal:

Article Title: Co-ordinate expression of the pre-T-cell receptor complex and a novel immature thymocyte-specific antigen, IMT-1, during thymocyte development

doi: 10.1046/j.1365-2567.1999.00656.x

Figure Lengend Snippet: Cell-surface expression of IMT-1, TCR-β or CD3 during fetal thymocyte development. Thymocytes were prepared from fetuses at days 13·5–18·5 of gestation, or from newborn or adult (4–5-week-old) ICR mice, and stained with anti-IMT-1, anti-TCR-β, or anti-CD3 antibodies, followed by anti-CD8 and anti-CD4 antibodies. Stained cells were analysed as described in the legend to Figure 1. On the left, dot-plot profiles of CD4 versus CD8 expression on thymocytes are shown together with the percentage of cells in each quadrant. On the right, histograms of IMT-1, TCR-β and CD3 expression on total thymocytes are shown, together with percentages of IMT-1+, TCRβ+ and CD3+ cells (mean±SD). Dotted lines show the staining profile with rat IgM (a negative control of anti-IMT-1 antibody) or hamster IgG (a negative control of anti-TCR-β and anti-CD3 antibodies). Representative data of three to four independent experiments is shown.

Article Snippet: Fluorescein isothiocyanate (FITC)-conjugated anti-CD8 antibody, phycoerythrin (PE)-conjugated anti-CD4 antibody, FITC-conjugated anti-CD69 antibody, FITC-conjugated anti-CD25 antibody, anti-NK1.1 antibody, biotinylated anti-CD8 antibody and biotinylated rat immunoglobulin M (IgM) antibody were purchased from Pharmingen (San Diego, CA).

Techniques: Expressing, Staining, Negative Control

Journal:

Article Title: Co-ordinate expression of the pre-T-cell receptor complex and a novel immature thymocyte-specific antigen, IMT-1, during thymocyte development

doi: 10.1046/j.1365-2567.1999.00656.x

Figure Lengend Snippet: Phenotypic change of RAG2−/− thymocytes by stimulation with anti-CD3 antibody. (A) Flow cytometric analysis of anti-CD3-stimulated thymocytes in RAG-2−/− mice. RAG-2−/− mice (4–6 weeks of age) were given a single intraperitoneal injection of 145-2C11 anti-CD3 antibody and thymocytes, prepared on the indicated days after treatment, were stained with anti-CD8, anti-CD4 and anti-IMT-1 antibody or anti-CD69 antibody, or with anti-CD25 antibody, as described in the Materials and methods. Stained cells were analysed as described in the legend to Fig. 1. On the left, dot-plot profiles of CD4 versus CD8 expression on thymocytes are shown together with the percentage of cells in each quadrant. On the right, histograms of IMT-1, CD69 and CD25 expression on total thymocytes are presented together with percentages (mean±SD) of IMT-1+, CD69+ and CD25+ cells. Dotted lines show the staining profile with rat IgM (a negative control of anti-IMT-1 antibody) or PBS (a negative control of anti-CD69). A representative dot-plot and a histogram profile of three independent experiments are shown. (B) Expression of IMT-1 on double negative (DN) and double positive (DP) subpopulations of anti-CD3-stimulated RAG2−/− thymocytes on day 5. On the left, a dot-plot profile of CD4 versus CD8 expression is shown. Gates for DN thymocytes (a) and DP thymocytes (b) are indicated. On the right, histograms of IMT-1 expression on DN thymocytes (a) and DP thymocytes (b) are shown along with percentages (mean±SD) of IMT-1+ cells.

Article Snippet: Fluorescein isothiocyanate (FITC)-conjugated anti-CD8 antibody, phycoerythrin (PE)-conjugated anti-CD4 antibody, FITC-conjugated anti-CD69 antibody, FITC-conjugated anti-CD25 antibody, anti-NK1.1 antibody, biotinylated anti-CD8 antibody and biotinylated rat immunoglobulin M (IgM) antibody were purchased from Pharmingen (San Diego, CA).

Techniques: Injection, Staining, Expressing, Negative Control