dr3 ko mouse  (CancerTools Org)

Journal: FASEB journal : official publication of the Federation of American Societies for Experimental Biology

Article Title: The death receptor 3/TL1A pathway is essential for efficient development of antiviral CD4 + and CD8 + T-cell immunity

doi: 10.1096/fj.11-200618

Figure Lengend Snippet: Expression of DR3 on splenocytes. A–C) DR3 expression on NK and NKT (A), CD4+ T (B), and CD8+ T (C) cells. Representative flow cytometric dotplots of splenic NK1.1+ lymphocytes stained for TCRβ, and CD4+ or CD8+ T cells stained for CD44, DR3, or control Ig from DR3WT and DR3KO mice. D) Preferential expression of DR3 on naive CD44lo rather than antigen-experienced CD44hi CD4+ and CD8+ T cells. Each symbol represents a different DR3WT mouse.

Article Snippet: In other experiments, Thy1.2 + DR3 WT or DR3 KO T cells were isolated using MACS (Miltenyi Biotec) purification kits, and 4 × 10 6 cells were adoptively transferred into congenic Thy1.1 + mice before MCMV challenge.

Techniques: Expressing, Staining, Control

Journal: FASEB journal : official publication of the Federation of American Societies for Experimental Biology

Article Title: The death receptor 3/TL1A pathway is essential for efficient development of antiviral CD4 + and CD8 + T-cell immunity

doi: 10.1096/fj.11-200618

Figure Lengend Snippet: Kinetics of DR3 expression by T cells. A) Kinetics of DR3 expression by CD4+ and CD8+ T cells following stimulation through the TCR. Values represent relative mean fluorescence, calculated as the ratio of geometric mean fluorescence intensity of the sample above negative control staining, following stimulation without (○), or with anti-CD3 mAb (●) on TCRβ+ lymphocytes stained using anti-DR3 and analyzed by flow cytometry. B, C) Expression of DR3 splice variants from CD8+ T cells following TCR stimulation. RNA was isolated from naive and activated OT-1 CD8+ DR3WT T cells; DR3 transcripts were amplified, cloned, and sequenced as described in Materials and Methods. B) RT-PCR for DR3 and β-actin before and after activation. C) Splice variant analysis and frequency of sequenced clones from CD8+ T cells after activation. Arrows mark the position of primers. CRR, cysteine-rich region; TM, transmembrane; DD, death domain. Numbers and boxes indicate coding exons. Plus symbol denotes position of ATG. Asterisk denotes position of stop codon.

Article Snippet: In other experiments, Thy1.2 + DR3 WT or DR3 KO T cells were isolated using MACS (Miltenyi Biotec) purification kits, and 4 × 10 6 cells were adoptively transferred into congenic Thy1.1 + mice before MCMV challenge.

Techniques: Expressing, Fluorescence, Negative Control, Staining, Flow Cytometry, Isolation, Amplification, Clone Assay, Reverse Transcription Polymerase Chain Reaction, Activation Assay, Variant Assay

Journal: FASEB journal : official publication of the Federation of American Societies for Experimental Biology

Article Title: The death receptor 3/TL1A pathway is essential for efficient development of antiviral CD4 + and CD8 + T-cell immunity

doi: 10.1096/fj.11-200618

Figure Lengend Snippet: Regulation of T-cell responses by DR3 following viral infection. A) Time courses showing numbers of NK (i), CD4+ T (ii), and CD8+ T (iii) cells in the spleens from DR3WT and DR3KO mice following MCMV challenge. DR3WT receiving PBS (●) or MCMV (▲); DR3KO receiving PBS (○) or MCMV (). Each data point represents mean ± se from n ≥ 5 mice and is representative of 4 separate experiments. *P < 0.05 at indicated time points; Student’s t test. B) Relative expansion of CD4+ and CD8+ T cells compared to unchallenged controls following MCMV challenge at peak (d 6 for CD4+, d 7 for CD8+). Bars show means ± se from n ≥ 8 mice. P values were determined by Mann-Whitney U test. C) Time courses showing numbers of total splenocytes (i), CD4+ T cells (ii), and CD8+ T cells (iii) in the spleens from DR3WT and DR3KO mice following rVV challenge. DR3WT receiving PBS (●) or rVV (▲); DR3KO receiving PBS (○) or rVV (). Each data point represents mean ± se from n ≥ 6 mice. * P < 0.05 at indicated time points; Student’s t test. D) Relative expansion of CD4+ and CD8+ T cells compared to unchallenged controls following rVV challenge at d 8. Bars show means ± se from n ≥ 6 mice. P values were determined by Mann-Whitney U test.

Article Snippet: In other experiments, Thy1.2 + DR3 WT or DR3 KO T cells were isolated using MACS (Miltenyi Biotec) purification kits, and 4 × 10 6 cells were adoptively transferred into congenic Thy1.1 + mice before MCMV challenge.

Techniques: Infection, MANN-WHITNEY

Journal: FASEB journal : official publication of the Federation of American Societies for Experimental Biology

Article Title: The death receptor 3/TL1A pathway is essential for efficient development of antiviral CD4 + and CD8 + T-cell immunity

doi: 10.1096/fj.11-200618

Figure Lengend Snippet: Antigen-specific responses following MCMV infection in the absence of DR3. A) IFN response of T cells in DR3WT and DR3KO mice following MCMV challenge. Representative flow cytometric dotplots of T cells showing CD8 and intracellular IFNγ staining in spleens from DR3WT and DR3KO mice, 7 d after MCMV challenge, to the M45 peptide. B) Intracellular staining for TNF-α and circulation of CD107a on splenic CD8+ T cells in response to M45 and m139 peptides following MCMV challenge. Values are means ± se of n ≥ 6 mice. C, D) Numbers of IFNγ-producing CD4+ T cells specific for the indicated peptide (left panel) and CD8+ T cells to IE3, M38, M45 and m139 in DR3WT (▲) and DR3KO () mice (right panel) either 7 d (C) or 32 d (D) following MCMV challenge. Bars indicate means. Each symbol represents a mouse. One of 3 experiments is shown. P values were determined by Student’s t test.

Article Snippet: In other experiments, Thy1.2 + DR3 WT or DR3 KO T cells were isolated using MACS (Miltenyi Biotec) purification kits, and 4 × 10 6 cells were adoptively transferred into congenic Thy1.1 + mice before MCMV challenge.

Techniques: Infection, Staining

Journal: FASEB journal : official publication of the Federation of American Societies for Experimental Biology

Article Title: The death receptor 3/TL1A pathway is essential for efficient development of antiviral CD4 + and CD8 + T-cell immunity

doi: 10.1096/fj.11-200618

Figure Lengend Snippet: Weight loss and viral titers in the absence of DR3. A) Weight loss in DR3WT or DR3KO mice on a C57Bl/6 background following infection with MCMV. No significant difference observed using 2-way ANOVA. B, C) MCMV (B) and rVV (C) titers (PFU/g tissue) in the indicated tissues of C57Bl/6 DR3WT or DR3KO mice at d 7 following MCMV or rVV challenge, respectively. Bars represent median and interquartile range of n ≥ 6 mice. D) Weight loss in DR3WT or DR3KO mice on a DBA-1 background following infection with a low dose (1×104 PFU/mouse) of MCMV. P value determined by 2-way ANOVA. E) MCMV titers (PFU/g tissue) in the salivary glands of DBA DR3WT or DR3KO mice at d 7 following low-dose challenge. Bars represent median and interquartile range of n ≥ 6 mice. P values were determined by Mann-Whitney U test. One of 2 experiments is shown. F) Weight loss (left panel) and survival curves (right panel) of DR3WT or DR3KO mice on a DBA-1 background following infection with a high dose (4×104 PFU/mouse) of MCMV. P value determined by Fisher’s exact test. One of 2 experiments is shown.

Article Snippet: In other experiments, Thy1.2 + DR3 WT or DR3 KO T cells were isolated using MACS (Miltenyi Biotec) purification kits, and 4 × 10 6 cells were adoptively transferred into congenic Thy1.1 + mice before MCMV challenge.

Techniques: Infection, MANN-WHITNEY

Journal: FASEB journal : official publication of the Federation of American Societies for Experimental Biology

Article Title: The death receptor 3/TL1A pathway is essential for efficient development of antiviral CD4 + and CD8 + T-cell immunity

doi: 10.1096/fj.11-200618

Figure Lengend Snippet: T-cell apoptosis following MCMV infection in the absence of DR3. DR3WT or DR3KO mice were challenged with MCMV, and T-cell apoptosis was measured using mAbs to CD4, CD8, and TCR and staining with annexin V and 7AAD at d 7. A) Representative flow cytometric dotplots for CD8+ T cells. B) Summary for CD4+ and CD8+ T cells following MCMV challenge. Analysis for different groups was carried out using the plots shown in A: live, annexin Vlo7AAD−; apopototic, annexin Vhi7AADlo; dead, annexin Vhi7AADhi. Data represent means ± se for n ≥ 5 mice. One experiment of 3 is shown. No significant differences were observed.

Article Snippet: In other experiments, Thy1.2 + DR3 WT or DR3 KO T cells were isolated using MACS (Miltenyi Biotec) purification kits, and 4 × 10 6 cells were adoptively transferred into congenic Thy1.1 + mice before MCMV challenge.

Techniques: Infection, Staining

Journal: FASEB journal : official publication of the Federation of American Societies for Experimental Biology

Article Title: The death receptor 3/TL1A pathway is essential for efficient development of antiviral CD4 + and CD8 + T-cell immunity

doi: 10.1096/fj.11-200618

Figure Lengend Snippet: In vitro T cell proliferation in the absence of DR3. A) Effect of TL1A on purified naive CD8+ T cells. CD8+ T cells were purified from spleens and treated as shown. Cells were counted at the indicated time points. Data are means ± se from n ≥ 5 mice. B) Proliferation of CD8+ T cells stimulated with anti-CD3 and TL1A. DR3WT (left panel) or DR3KO T cells (right panel) were loaded with CFSE and stimulated as described in Materials and Methods. Plots are histograms demonstrating loss of CFSE signal after 4 d stimulation. C) Density plots showing staining of CD8+ T cells for CD25, CD44, and CD69, 4 d after stimulation with anti-CD3 and TL1A.

Article Snippet: In other experiments, Thy1.2 + DR3 WT or DR3 KO T cells were isolated using MACS (Miltenyi Biotec) purification kits, and 4 × 10 6 cells were adoptively transferred into congenic Thy1.1 + mice before MCMV challenge.

Techniques: In Vitro, Purification, Staining

Journal: FASEB journal : official publication of the Federation of American Societies for Experimental Biology

Article Title: The death receptor 3/TL1A pathway is essential for efficient development of antiviral CD4 + and CD8 + T-cell immunity

doi: 10.1096/fj.11-200618

Figure Lengend Snippet: T-cell proliferation following MCMV infection in the absence of DR3. DR3WT or DR3KO mice were challenged with MCMV, and T-cell proliferation was measured using mAbs to CD4, CD8, TCR, and Ki-67 at d 7. A) Representative flow cytometric dotplots of Ki-67 expression in unchallenged CD8+ T cells (i) and in CD8+ T cells following MCMV challenge (ii). B) Numbers of CD4+ T cells and CD8+ T cells expressing Ki-67 from uninfected DR3WT (●) and DR3KO (○) or MCMV-challenged DR3WT (■) and DR3KO (□) mice. Data represent means ± se of n ≥ 5 mice. One of 3 experiments is shown. *P < 0.05 at indicated time points; Student’s t test. C) Correlation between increasing numbers of CD8+ T cells, expression of Ki-67, and DR3 genotype. Increase in CD8+ T-cell numbers was calculated by subtracting the average number of splenic CD8+ T cells recovered from uninfected animals from the number at 7 d after MCMV infection. DR3WT (●) and DR3KO (○) mice are shown. Line represents best-fit curve; dotted lines indicate 95% confidence limits. D) Numbers of CD8+, Ki-67+, and IFNγ+ T cells responding to M38, M45, and m139 peptide challenge in DR3WT (▲) and DR3KO () mice 7 d following MCMV challenge. Bars indicate means. Each symbol represents a mouse. One experiment is shown. P values were determined by Student’s t test.

Article Snippet: In other experiments, Thy1.2 + DR3 WT or DR3 KO T cells were isolated using MACS (Miltenyi Biotec) purification kits, and 4 × 10 6 cells were adoptively transferred into congenic Thy1.1 + mice before MCMV challenge.

Techniques: Infection, Expressing