Fig. 5

LA-ISA immunization induces responses of DC, CD8⁺ T, and NK cells in HLA-A2.1 transgenic mice. a HLA-A2.1 transgenic mice were immunized subcutaneously for three times with the same volume of YL-ISA, LA-ISA, ISA, or control. The dose of LA or YL was 1 mg/20 g body weight. After final immunization, splenocytes or peripheral blood cells were harvested and analysed by flow cytometry. b DC phenotype of peripheral blood cells (n = 3). c The frequency of CD8⁺ T cells in splenocytes (n = 3). d The expression levels of perforin and granzymes-B in CD8⁺ T cells. e The frequency of NK cells in splenocytes (n = 3). f The expression levels of perforin and granzymes-B in NK cells (n = 3). g No significant difference could be detected in body weight of HLA-A2.1 transgenic mice immunized subcutaneously with YL-ISA, LA-ISA, ISA, or control (n = 6). h No significant difference was found in liver function (ALT, AST) and kidney function (CRE, UA) of HLA-A2.1 transgenic mice immunized subcutaneously with YL-ISA, LA-ISA, ISA, or control (n = 6). i No significant difference was identified in pathological examination of vital organs of HLA-A2.1 transgenic mice immunized subcutaneously with YL-ISA, LA-ISA, ISA, or control. Leukocytes (Wright Giemsa staining, scale bars, 10 μm). Liver, kidney, heart, lung, stomach, intestine (H&E staining, scale bars, 50 μm). j The stability of YL-FITC-ISA or LA-FITC-ISA in mice was detected periodically by an in-vivo imaging system. Control, HLA-A2.1 transgenic mice were treated with normal saline. ISA, HLA-A2.1 transgenic mice were treated with ISA. YL-ISA, HLA-A2.1 transgenic mice were treated with YL-ISA. LA-ISA, HLA-A2.1 transgenic mice were treated with LA-ISA. b–f P values were obtained with independent-samples t-test; error bars denote standard deviation (SD). Compared with control, ^*P < 0.05, ^**P < 0.01, ^***P < 0.001. Compared with ISA, ^#P < 0.05, ^##P < 0.01, ^###P < 0.001. Compared with YL-ISA, ^&P < 0.05, ^&&P < 0.01