International Journal of Molecular Zoology 2024, Vol.14, No.2, 72-83 http://animalscipublisher.com/index.php/ijmz 76 Figure 1 The immunogenicity of GGTA1−/−β2M−/−CIITA−/− triple gene knockout (GBC-3KO) pigs in vitro and in vivo (Adopted from Fu et al., 2020) Image caption: A, Human peripheral blood mononuclear cells (hPBMCs) from healthy volunteers’ blood were labeled with carboxyfluorescein succinimidyl ester (CFSE) and cocultured with irradiated splenocytes (2:1 ratio) from the wild-type (WT) or GBC-3KO fetal pigs for 6 d. Representative data of CFSE density dilution in CD4 and CD8 populations are shown. B, Comparison between WT and GBC-3KO piglet with respect to the CFSE density dilution in CD4 and CD8 populations (n[GBC-3KO] = 6; n[WT] = 3). C, Percentage of CD25 cells in the CD4 and CD8 population of the lymphocytes stimulated by GBC-3KO or WT splenocytes. D, Comparison of the percentage of CD25-positive cells in the CD4 and CD8 population that were stimulated by irradiated splenocytes from the WT or GBC-3KO fetal pigs. E, hPBMCs from healthy volunteers’ blood were cocultured with the irradiated splenocytes (2:1 ratio) from the GBC-3KO or WT fetal pigs for 5 d. Bromodeoxyuridine (BrdU) detection was made by ELISA. The experiment was performed 3×: 3 different GBC-3KO samples and 1 WT sample. F, The absorbance values of 3 experimental results were converted into cell proliferation rate (cell proliferation rate % = experimental well/control well × 100). Each sample has 4 replicates. G, Pig skin grafts (12 × 10 mm) from WT or GBC-3KO pigs were transplanted into C57B6/L mice. Transplant rejection was monitored after day 5. Photographs of the transplanted skin were taken daily. Representative graphs of days 5, 8, 11, and 14 posttransplanted skins are shown. H, Comparison of the survival time of the skin from WT pigs (median survival time [MST], 13.5 d) and from GBC-3KO pigs (MST, 16 d) in C57BL/6 mice. (n[NCG] = 2; n[GBC-TKO] = 15; n[WT] = 10). Plot graphs represent the mean ± SD. Statistical analysis was done using t test or ANOVA with Tukey post hoc test. *P < 0.05, **P < 0.01, ****P < 0.001. HCD8, anti-human CD8 antibody; KO, GBC-KO pig cells with hPBMCs; NCG, the mice that lacking T, B, and NK cells; R, hPBMCs; WT, WT pig cells with hPBMCs (Adopted from Fu et al., 2020)
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