Fig. 4.

AFS98 treatment does not affect T cell division in lymph nodes but prevents T cell entry into islets of Langerhans. NOD mice were injected with AFS98 antibody at a dose of 0.5 mg at 2 wk of age and 2.0 mg at 4 wk of age. Two TCR transgenic T cells, the CD4⁺ BDC2.5 and the CD8⁺ NY8.3, were isolated from lymph nodes and spleens of their respective mice. T cells were then labeled with CFSE and transferred into 6-wk-old NOD mice that had either been left untreated or treated with AFS98. (A and B) Seven days after T cell transfer, the pancreatic and inguinal lymph nodes were isolated and analyzed by flow cytometry. (A) Dilution of CFSE in either inguinal (Upper) or pancreatic (Lower) lymph nodes for an individual mouse per treatment is shown. Cells were gated on forward and side-scatter, CD45, CD3, and either CD4 (BDC2.5) or CD8 (NY.8.3). (B) Summary of division index and proliferation index for individual mice examined as in A. Results show three or four individual mice per group. (C) Ten days after TCR transgenic T cell transfer, islets of Langerhans were isolated and examined for entry of either the BDC2.5 or NY8.3 T cells by flow cytometry. The Left two images are gated on forward and side-scatter, CD45, and CD3. The Right images are gated on forward and side-scatter. BDC T cells were identified using a clonotypic antibody to its T cell receptor. NY8.3 T cells were identified by a CD45.2 congenic label. Numbers indicate the percent of cells in each selection as a function of CD45⁺ cells. T cell islet entry results are representative of two to three independent experiments with two to four individual mice per group.