Skip to main content
. 2015 Jul 21;6:7725. doi: 10.1038/ncomms8725

Figure 4. Maturation of neonatal CD4 T lymphocytes following mucosal challenge.

Figure 4

(a) Lymphocyte maturation status in the adult lymphopenic host after the induction of Rag in InduRag mice. FACS analysis (left panel) and gMFI of CD44 (right panel) on CD4+TCRβ+ lymphocytes obtained from the SI of 11- and 56-day-old wt (wt d11 and wt d56) and adult InduRag mice 11 days after Tamoxifen administration (InduRag d56+TAM11). (n=3–4, representative of two experiments, mean; one-way ANOVA, Bonferroni's post test, ***P<0.001; NS, not significant). (b) Ex vivo activation of CD4+TCRβ+ lymphocytes from PP of 11- and 56-day-old mice after 3 days of culture in presence of anti-CD3/CD28 beads (n=4, one experiment). (c) H&E staining and (d) percentage of infiltrating CD4+TCRβ+ lymphocytes of CD45+ in the colonic tissue of adult Rag2 recipients 5–6 weeks after transfer of 1–5 × 104 SI CD4+ T cells from 11-day-old or adult Foxp3-GFP donor mice after TReg depletion (T-cell transfer colitis model). Magnification, × 40 (bar graph 100 μm). (n=3–4, representative of two experiments, mean±s.d.; unpaired Student's t-test, ***P<0.001). (e) Percentage of CD44hi cells among CD4+TCRβ+ lymphocytes (using the 3% of CD4 T cells in the non-infected control mice with the highest CD44 expression as a reference gate) in the neonate SI after infection with rotavirus (infected at d4 post parturition, pp and analysed at 8 d.p.i.), S. Typhimurium (infected at d4 pp and analysed at 4 d.p.i.) and Giardia lamblia (infected d4 pp and analysed at 8 d.p.i.). (One litter (n=6–11) from the infected group and four age-matched non-infected controls were analysed per experiment; representative of two experiments, mean±s.d.; unpaired Student's t-test, *P<0.05, ***P<0.001; NS, not significant). (f) Quantification of rotavirus antigen by ELISA in colon homogenate of wt and TCRα−/− neonates at 8 d.p.i. infected at 4d pp (n=12–13 from two experiments, mean; Mann–Whitney U-test, ***P<0.001). (g) FACS plots (left panel) and percentages (right panel) of CD44hiCD62L− of CD4+TCRβ+ lymphocytes in PPs. 11-day-old DO11.10 neonates were gavaged daily with 10 mg OVA or PBS starting at d3 pp (n=5, representative of two experiments, mean±s.d.; unpaired Student's t-test, ***P<0.001).