IgE immune complex challenge induces similar lung MCp levels in CD23^−/− as in wild type.

<p>(A–B) A representative experiment showing the quantification of MCp from wild type (WT) or CD23^−/− mice sensitized with OVA/alum and challenged with IgE-anti-TNP/OVA-TNP. The error bars shown are SEM. (C) A summary of all the four experiments performed expressed as a stimulation index (S.I.) where the numbers represent the mean number of MCp/lung or MCp/10⁶MNC per experiment of sensitized WT mice challenged with IgE-anti-TNP/OVA-TNP divided by the mean number obtained from CD23^−/− mice treated in parallel. The mean of the S.I. from all experiments is given in bold text at the bottom. The representative experiment shown in (A–B) is indicated in bold italics. There was no statistical difference (ns) in the tested parameters using a two-way ANOVA from a comparison of all individual mice from each group from the four experiments.</p

IgE immune complex-induced enhancement of lung MCp is dependent on an FcRγ chain associated receptor.

<p>(A–B) A representative experiment showing the quantification of MCp from or FcRγ^−/− sensitized with OVA/alum and challenged with either IgE-anti-TNP/OVA-TNP or OVA-TNP alone (C) A summary of all the three experiments performed expressed as a stimulation index (S.I.) where the numbers represent the mean number of MCp/lung or MCp/10⁶MNC of sensitized FcRγ^−/− mice challenged with IgE-anti-TNP/OVA-TNP divided by the mean number obtained in FcRγ^−/− mice challenged with OVA-TNP alone. The mean of the S.I. from all experiments is given in bold text at the bottom. The representative experiment shown in (A–B) is indicated in bold italics. There was no statistical difference (ns) in the tested parameters using a two-way ANOVA from a comparison of all individual mice from each group from the three experiments.</p

Challenge with IgE immune complex enhance lung MCp numbers compared to antigen alone.

<p>(A–C) A representative experiment showing the quantification of MCp and MNC from wild type (WT) mice sensitized with OVA/alum and challenged with OVA-TNP, IgE-anti-TNP/OVA-TNP or IgE-anti-TNP alone. The error bars shown are SEM. (D) A summary of all nine experiments performed expressed as a stimulation index (S.I.) where the numbers represent the mean number of MCp/lung, MNC/lung or MCp/10⁶ MNC per experiment of sensitized mice challenged with IgE-anti-TNP/OVA-TNP divided by the mean number obtained from sensitized mice challenged with OVA-TNP alone for each experiment. (E) A summary of all the experiments where B-cells, T-cells and dendritic cells (DC) were analyzed with flow cytometry. In some experiments flow cytometry was not done (nd). The mean of the S.I. from all experiments is given in bold text at the bottom. The experiment shown in (A–C) is indicated in bold italics. The p-values shown are derived from two-way ANOVA from a comparison of all individual mice in each group from the nine experiments.</p

FcRγ-chain deficient mice have a reduced number of lung MCp after IgE immune complex challenge.

<p>(A–B) A representative experiment showing the quantification of MCp from wild type (WT) or FcRγ^−/− sensitized with OVA/alum and challenged with IgE-anti-TNP/OVA-TNP (C) A summary of all the four experiments performed expressed as a stimulation index (S.I.) where the numbers represent the mean number of MCp/lung or MCp/10⁶ MNC of sensitized WT mice challenged with IgE-anti-TNP/OVA-TNP divided by the mean number obtained in FcRγ^−/− mice treated in parallel. The mean of the S.I. from all experiments is given in bold text at the bottom. The representative experiment shown in (A–B) is indicated in bold italics. The p-values shown are derived using a two-way ANOVA from a comparison of all individual mice from each group from the four experiments.</p

Intranasal challenge with OVA-TNP does not enhance the lung MCp number over basal levels.

<p>(A–C) One experiment showing the quantification of MCp and MNC from wild type (WT) mice sensitized with OVA/alum and challenged with either OVA-TNP alone, OVA aerosol or left unchallenged. The error bars shown are SEM. (D) A comparison of the OVA-TNP challenged and the unchallenged group from the two experiments performed expressed as a stimulation index (S.I.) where the numbers represent the mean number of MCp/lung, MNC/lung or MCp/10⁶ MNC per experiment of sensitized mice challenged with OVA-TNP divided by the mean number obtained from sensitized mice left unchallenged. The mean of the S.I. from the experiments is given in bold text at the bottom. The experiment shown in (A–C) is indicated in bold italics. There was no statistical difference (ns) between the OVA-TNP challenged and the unchallenged mice in the tested parameters using a two-way ANOVA from a comparison of all individual mice from each group from the two experiments.</p

01_A_S3A_069

Κωδικός τεκμηρίου: 01_A_S3A_069-019. Είδος τεκμηρίου: SLIDES 10Χ12,5 Χ 1 ΚΑΡΕ - ΕΓΧΡΩΜΑ. Ανήκει σε: 01_A_S3A_069 - ΚΟΥΤΙ "ΔΙΑΦΑΝΕΙΕΣ ΜΕΓ.

CR1/2 on FDCs are required for a robust IgG anti-SRBC response to SRBC.

<p>BALB/c and <i>Cr2^−/−</i> mice were irradiated and reconstituted with either BALB/c or <i>Cr2^−/−</i> bone marrow. Six weeks after reconstitution, mice (n = 6/group) were immunized i.v. with 5×10⁶, 5×10⁷, or 5×10⁸ SRBC. All mice were bled at indicated time points. Sera were diluted 1∶125 (A) or 1∶625 (B and C) and screened for IgG anti-SRBC in ELISA. P-values represent comparisons between the responses in recipients with the same background; ns = p>0.05; * = p<0.05; ** = p<0.01; *** = p<0.001. Representative of two (A) or one (B, C) experiments.</p

CR1/2 on B cells and FDCs is required for optimal antibody responses to IgM-SRBC complexes.

<p>BALB/c and <i>Cr2^−/−</i> mice were irradiated and reconstituted with either BALB/c or <i>Cr2^−/−</i> bone marrow. Six weeks after transplantation, mice were immunized with 5×10⁵ (A–D) or 5×10⁶ (E–H) SRBC alone (open squares) or together with IgM anti-SRBC with a hemagglutination titer of 1∶32 (filled squares) or with IgM anti-SRBC alone (open triangles) (n = 6/group). All mice were bled at indicated time points. Sera were diluted 1∶25 (A–D) or 1∶625 (E–H) and screened for IgG anti-SRBC. Two statistical comparisons were made, both using Student's <i>t</i>-test. First, comparisons between the responses in mice immunized with SRBC alone versus IgM and SRBC (to determine whether IgM enhanced antibody responses significantly; filled versus open symbols), where ns = p>0.05; * = p<0.05; ** = p<0.01; *** = p<0.001. Second, comparisons between the responses between various chimeras immunized with IgM-SRBC (to determine whether CR1/2⁺ B cells contributed significantly to the antibody response to IgM-SRBC in mice with CR1/2⁺ FDCs (A vs B; E vs F) and CR1/2⁻ FDCs (C vs D; G vs H)), where ns = p>0.05; ° = p<0.05; °° = p<0.01; °°° = p<0.001. For graphic clarity, non-significant differences are not indicated. Representative of one (A–D) and two (E–H) experiments.</p

IgE-mediated enhancement of T cell proliferation <i>in vivo</i> is dependent on CD11c⁺ cells.

<p>CD11c-DTR mice, and in B) also wild type littermates, were treated with 100 ng diphtheria toxin or left untreated. The same day, the mice were transferred i.v. with 2.3−3.0×10⁶ CD4⁺ DO11.10 spleen cells. Twenty-four hours after diphtheria toxin treatment, mice were immunized i.v. with 20 µg OVA-TNP with or without 50 µg IgE anti-TNP. Three days later, spleens were analyzed for CD4⁺ KJ1-26⁺ T cells by flow cytometry (A, B) or for KJ1-26⁺ cells in confocal microscopy (C, D). A) Representative dot-plot of flow cytometry data; values represent % CD4⁺KJ1-26⁺ cells of gated lymphocytes. B) Mean values ± SEM of % CD4⁺KJ1-26⁺ of CD4⁺ cells analyzed in flow cytometry (3 mice/group) C) Mean values ± SEM of KJ1-26⁺ cells in non-consecutive sections of T cell zones analyzed by confocal microscopy (3 mice/group; 3–12 T cell zones/mouse). D) Visualization of representative T cell zones from each group: KJ1-26⁺ cells (red) and B220⁺ cells (blue). B) is representative of 4 experiments and C) of 1 experiment.</p

Diphtheria toxin depletes CD11c-DTR mice of CD11c⁺ cells.

<p>CD11c-DTR mice were treated with 100 ng diphtheria toxin or left untreated. Twenty-four hours later total spleen cells were analyzed in flow cytometry and dendritic cells were defined as CD11c⁺, MHC-II^hi cells. The numbers show % of live cells in a representative experiment with 97% depletion of dendritic cells.</p