Objectives and Specific Aims CHAPTER 2: LITERATURE REVIEW CHAPTER 3: MATERIALS AND METHODS 15 Stx2 and rStxB2 Isolation of Porcine Granulocytes 16 Flow Cytometric Evaluation of Stx2 and rStxB2 Binding and Gb 3 /CD77 Expression by Porcine Granulocytes Stx2 Binding to Porcine Granulocytes at 4°C 17 Flow Cytometric Analysis Fluorescence Microscopic Evaluation of Stx2 Binding Flow Cytometric rStxB2 Binding Assays 20 Flow Cytometric Gb 3 /CD77 Receptor Assay Stx2 and rStxB2 Binding to Porcine Granulocytes Under Conditions Required for Functional Studies 21 Internalization of Stx2 and rStxB2 Effect of Stx2 on Granulocyte Apoptosis Effect of Stx2 on Granulocyte Oxidative Burst Statistical Analysis iii CHAPTER 4: RESULTS Characterization of Granulocyte Preparations 26 Stx2 Binding to Porcine Granulocytes at 4°C 26 Stx2 and rStxB2 Binding and Gb 3 /CD77 Expression at 4°C 27 Stx2 Binding Under Conditions Required for Functional Studies rStxB2 Binding Under Conditions Required for Functional Studies Internalization of Stx2 and rStxB2 Effect of Stx2 on Apoptosis in Porcine Granulocytes Effect of Stx2 on the Oxidative Burst of Porcine Granulocytes Figures CHAPTER 5: DISCUSSION 52