Efficient Acquisition of Fully Human Antibody Genes against Self-Proteins by Sorting Single B Cells Stimulated with Vaccines Based on Nitrated T Helper Cell Epitopes

Figure 3

Cotransplantation of autologous DCs allowed HIS mice to produce specific human IgG against various antigens. NCG mice were engrafted with PBMCs on day 0 and then immunized with KLH-loaded DCs on day 7 and KLH-mixed alum adjuvant on day 21 or with KLH-mixed alum adjuvant on days 7 and 21. Data were expressed as (). (a) Splenocytes were harvested from vaccinated mice on day 28, and the percentages of human DCs (gated as hCD11c⁺HLA-DR⁺) were measured by FACS. compared with non-DCs. (b) (A, B) Immunofluorescent examination of hCD11c⁺ DCs (green) and hCD3⁺ T cells (red) in the spleen sections from HIS mice immunized with KLH-loaded DCs. Original magnification: ×5. (C, D) Immunofluorescent colocalization of autologous hCD11c⁺ DCs and hCD3⁺ T cells. Original magnification: ×5 (C) and ×10 (D). (c, d) The sera of vaccinated mice were collected on day 21, and the production of KLH-specific human antibodies was detected at a 1 : 800 titer by ELISA assay. n.s.: not significant. compared with PBS; ^#, ^## compared with non-DCs. (e, f) Sera were collected on the 14th day after immunization with OVA-loaded DCs, and the production of OVA-specific human antibodies was detected by ELISA assay. , compared with PBS.

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