Figure 3.

Enrichment of antigen-specific high-affinity antibody-producing cells in artTLO upon transplantation into immunodeficient SCID mice. ArtTLO were first formed in eight NP-OVA preimmunized Balb/c mice as described in Figure 2. Four artTLO (two artTLO in each kidney) in one mouse were constructed. Then, artTLO were individually extirpated and collected from Balb/c mice and re-transplanted into renal subcapsular space of eight SCID mice. Four artTLO were re-transplanted into each SCID mouse. Two weeks after re-transplantation, four of artTLO-carrying SCID mice were intravenously immunized with 10 μg NP-OVA. The other four SCID mice were not immunized. One week after, artTLO and recipient spleen were extirpated individually from SCID mice and single cell suspensions were prepared. Numbers of high affinity as well as total IgG class anti-NP antibody-forming cells (AFCs) in individual artTLO and in spleens of recipient SCID mice were counted by an immunospot analyzer. Numbers of AFCs were calculated from 16 individual artTLO for each group.