The Impact of T Cell Intrinsic Antigen Adaptation on Peripheral Immune Tolerance
Figure 3
Polyclonal B-Cell Activation and Autoantibody Production following the Induction of T Cell Adaptive Tolerance
(A–G) Total serum antibody levels of individual isotypes, measured by sandwich ELISA at different days after T cell transfer (solid circles). Open circles at time 0 represent serum measurements from normal mice, without T cell transfer. *Serum samples where the levels of the isotype was below detection [for two, four, three, three, and six samples each in (B), (C), (D), (E), and (F)]. Data in (A–E) are combined from two experiments between 4 and 60 d and two additional experiments at later time points, with two to five mice per experimental group. Three additional experiments between days 7 and 120 gave similar results. Data in (F) and (G) are from two separate experiments with three mice per time point. Serum IgA levels were below our limit of detection after the day 3 time point as indicated (*). Solid triangles in (G) represent measurements from solubilized fecal pellets. Note that the y-axes are scaled differently for (A), (F), and (G) to accommodate different absolute amounts of IgM, IgE, and IgA.
(H) Increased circulating immune complexes in the serum after T cell transfer, demonstrated by a C1q binding ELISA. Each data point represents an individual mouse, whose sera were all sampled on the same day after receiving T cells at various prior times (x-axis).
(I) Titers of anti-DNA antibodies in the sera of 16 individual T cell transfer recipients sampled in two separate experiments. Optical density values obtained from isotype specific ELISAs for IgM (solid circles), IgG2b (solid squares), and IgG2a (solid triangles) are shown. Empty circles (IgM), squares (IgG2b), and triangles (IgG2a) on day 0 represent sera obtained from three normal mice, assayed in the two experiments.
(J–O) Sera from mPCC,CD3ε−/− mice were assayed for anticellular antibodies of the IgM (green) and IgG (red) isotypes, by hybridizing with frozen tester sections of B10.A,Rag2−/− kidneys. Numbers in the inset denote days after T cell transfer. (O) shows an enlarged area from (N) to illustrate speckled nuclear staining. (J) was probed with serum from a normal mPCC,CD3ε−/− mouse without T cell transfer. Similar sera samples from mice 4 d (n = 3) or 7 d (n = 2) after T cell transfer did not show measurable IgG or IgM anti cellular antibodies. However, all mice tested after 28 d (n = 22) scored positive for both IgG and IgM antibodies to various extents.