Fig. 3

Zn-RS-adjuvanted gE elicited potent immune responses in young and aged mice. Young adult, 6-week-old C57BL/6 and aged, 15-month-old C57BL/6 mice received intramuscular injections on days 0 and 28 with 5 μg of gE protein with Al-001 or Zn-RS. Retro-orbital blood was collected weekly, and A binding antibody titers against gE at Week 2, B binding antibody titers against gE at Week 5, C and D dynamic changes of binding antibody titers against gE from week 1 to week 5 (6-week-old and 15-month-old C57BL/6). E Neutralizing titer of only aged individuals at Week 5. F Spleens and lymph nodes were collected 1 week flowing the final immunization and then stimulated with a gE peptide pool before being subjected to ELISpot and G to J flow cytometry, n = 6. F Quantification of IFN-γ released from cells within each well was performed via ImmunoSpot software. G to J Absolute numbers of IFN-γ⁺CD4⁺ T helper (Th1), IL-4⁺CD4⁺ T helper (Th2), IFN-γ⁺CD8⁺ T, and IL-4⁺CD8⁺ T cells were examined via flow cytometry. Statistical significance was determined via the Mann‒Whitney U test. *p < 0.05, **p < 0.01, ***p < 0.001