1. Adaptive immune responses and NK activity.
Adaptive immune responses and NK activity. (A and B) CFSE-labeled splenocytes were stimulated with anti-CD3 and anti-CD28 mAbs or unstimulated. Three days later, cells were stained with allophycocyanin-conjugated anti-CD3 and allophycocyanin–Cy7-conjugated anti-CD4 mAbs. (A) Proliferation (CFSE dilution) of CD4+ T (CD3+CD4+) cells and CD8+ T (CD3+CD4−) cells was examined by flow cytometry. Representative flow cytometry profiles of cells with (outlined area) or without (shaded area) stimulation from three independent experiments using eight total WT and eight total PYNOD-KO mice are shown. The vertical dashed lines are arbitrary lines that are provided as an aid for comparison of CFSE dilution levels between the upper and lower profiles. (B) Concentration of IL-4 or IFN-γ in the culture supernatants was measured by ELISA. The average of two independent experiments using six total WT and six total PYNOD-KO mice are shown. (C) WT and PYNOD-KO mice were sensitized with TNCB painted on the shaved abdomen. Seven days later, mice were challenged with TNCB and control solvent painted on the skin of the right and left ears, respectively. Ear swelling 24 h after the challenge is shown. Cumulative data from two independent experiments using nine total WT and eight total PYNOD-KO mice are shown. (D) WT and PYNOD-KO mice were immunized with OVA in CFA. Ten days later, mice were challenged with OVA and control solvent injected into the left and right hind footpads, respectively. Footpad swelling 24 h after the challenge is shown. Cumulative data from two independent experiments using nine total WT and eight total PYNOD-KO mice are shown. (E) FBL-3 tumor cells (5 × 106 cells) were intradermally injected into the dorsal skin of WT and PYNOD-KO mice (each n = 9). Tumor size was measured every 3 d. Consistent results were obtained in three other similar experiments. (F) WT and PYNOD-KO mice (each n = 8) were immunized by OVA with Imject Alum on day 0 and day 10. On day 15, the concentrations of OVA-specific IgM and IgG and of total IgE in sera were determined by ELISA. Consistent results were obtained in four other similar experiments. (G) Naive splenocytes from WT and PYNOD-KO mice (each n = 3) were independently cocultured with PKH67-labeled Yac-1 cells at the indicated effector/target ratios. Four hours later, cells were stained with PI, and the proportion of PI+ dead cells in the Yac-1 cells was determined by flow cytometry. Consistent results were obtained in two other similar experiments. (B–G) No significant differences were observed between WT and PYNOD-KO cells. ****p < n.s., not significant.
Shinsuke Nakajima et al. ImmunoHorizons 2018;2:
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