Figure 5

Moderate SMFs enhance CTLs cytotoxicity. (A and B) The cytotoxicity of CTLs generated from OT-I mice was assessed by LDH release with EL4 cells pulsing OVA (A, n = 4) or without pulsing OVA (B, n = 3). (C) Cytokine/granule production of CTLs generated from OT-I mice. Cell samples were stimulated with anti-CD3 and anti-CD28 antibodies in the presence of 0.3 T magnets, and control cells were treated without magnets. Cell samples with no stimulation were used to show the baseline of cytokine secretion. (D and E) Percentage (D) and MFI (E) statistics for the expression of GzmB, IFNγ and TNFα of CTLs generated from OT-I mice (n = 4). Cell samples were treated with 0.3 T magnets, and samples treated without magnets were used as controls. (F) In vivo cytotoxicity assay as indicated by the CFSE^low and CFSE^high populations analyzed by flow cytometry. SMF-treated and control CTLs (3 × 10⁶) were intravenously injected into recipient mice, followed by injection of OVA_257–264-pulsed (CFSE^low) and nonpulsed (CFSE^high) splenocytes (5 × 10⁶) to measure in vivo cytotoxicity. (G) The killing percentage as revealed by the in vivo cytotoxicity assay (n = 4). (H) The negative control of in vivo cytotoxicity assay as indicated by the CFSE^low and CFSE^high populations analyzed by flow cytometry. OVA_257–264-pulsed (CFSE^low) and nonpulsed (CFSE^high) splenocytes without getting CTLs were injected into recipient mice to measure in vivo cytotoxicity. The cytotoxicity was calculated as [1 − (% CFSE^low)/(% CFSE^high)] × 100%. Data were analyzed by Student’s t-test; NS, no significance, *p < 0.05; **p < 0.01. Error bars indicate the s.e.m.