Immunopathology of Insulin

A. Introduction.- B. Brief Review of the Immunology of Insulin.- I. Antigenicity of Insulin.- II. Humoral Antibodies Against Insulin and Their Demonstration.- 1. Immunological Reactions and Serologic Methods.- 2. Neutralization of the Biological Effect of Insulin..- 3. Binding of 131lodine-labelled Insulin.- 4. The Types of Insulin Antibodies.- III. Cell Mediated Immunity Against Insulin.- 1. Some General Aspects of Cell Mediated Immunity.- 2. Previous Immunocytological Studies.- IV. The Clinical Symptoms of the Immune Response to Insulin.- 1. Insulin Allergy.- a) The Clinical Picture.- b) Frequency.- c) Etiology and Pathogenesis.- 2. Insulin Resistance.- a) Clinical Picture.- b) Frequency.- c) Methods for Measurement of Insulin-neutralizing Antibodies.- d) Relationships between Maximum Insulin-binding Capacity of the Serum and the Daily Requirement of Insulin.- V. Proinsulin and Newer Concepts in Insulin Immunology.- C. Investigations of Insulin Allergy in Diabetes.- I. Investigations of Delayed Insulin Allergy.- 1. Introductory Remarks.- 2. Cytological Investigations.- a) Immunofluorescence.- ?) The Different Techniques of Immunofluorescence.- ?) Principles of Labelling with Fluorochromes.- ?) The Biological Characteristics of FITC Insulin.- ?) The Immunological Characteristics of FITC Insulin.- ?) The Conjugation of Insulin with FITC.- ?)The Conjugation of Insulin Antibodies with FITC.- ?) The Isolation of Blood Leucocytes.- ?) Separation of Lymphocytes and Granulocytes.- ?) The Application of Immunofluorescence to Blood Cells.- ?) The Specificity Tests for Immunofluorescence.- ?) The Evaluation of the Cytologic Preparations.- b) Immune Adherence.- ?) Introductory Remarks.- ?) Technique.- ?) The Specificity Tests for Immune Adherence.- ?) The Evaluation of the Cytologic Preparations.- c) Lymphocyte Transformation.- ?) Introductory Remarks.- ?) Technique.- ?) Evaluation of the Cell Preparations.- d) The Cytology of the Peripheric Blood Cells.- 3. The Intracutaneous Skin Test.- 4. Histological and Immunofluorescent Studies in Skin Biopsies.- 5. Investigations of Humoral Insulin Antibodies.- a) Insulin-Neutralizing Antibodies.- ?) Measurement of the Maximal Insulin-binding Capacity.- ?) Passive Hemagglutination.- b) Skin-sensitizing Antibodies (Reagin).- ?) Remarks.- ?) The "Allergic Serum Transfer Test" (AST Test).- c) The Investigation of Cytophilic Antibodies.- 6. Results.- a) Case Reports.- b) Summary of the Findings.- ?) Clinical Picture.- ?) Antigen-binding by Sensitized White Blood Cells.- ?) Behaviour of Serum Antibodies.- ?) Cytophilic Antibodies.- ?) Histological Findings of Skin Biopsies..- ?) Findings with Lymphocyte Transformation.- 7. Discussion.- 8. Summary and Recommended Therapeutic Measures.- II. Investigations of Immediate Allergy to Insulin.- III. Coexistence of Insulin Allergy and Insulin Resistance.- D. Investigations of the Delayed Immune Reaction and Formation of Antibody to Insulin in Experimental Animals.- I. Investigations of the Delayed Immune Reaction to Insulin in Guinea Pigs.- 1. Material and Methods.- a) Animals, Mode of Sensitization, Skin Test.- b) Cytological Examination of Peripheral White Blood Cells.- c) Transfer Test.- d) Migration Inhibition Test.- e) Studies of Humoral Antibodies to Insulin.- 2. Results.- a) Cytological Investigations of the White Cells.- b) Transfer Experiments.- c) Migration Inhibition Test.- d) Humoral Antibodies.- 3. Discussion.- 4. Summary.- II. Investigations of the Production of Humoral Antibodies.- 1. Material and Methods.- a) Mode of Immunization.- b) Detection of Antibody-forming Cells in the Peripheral Blood.- ?) Isolation of Blood Cells.- ?) Plaque Technique (According to Cunningham).- c) Binding of 131Iodine-labelled Insulin.- d) Passive Hemagglutination.- e) Immunohistological Techniques.- f) Histological Techniques.- 2. Results.- a) Antibody-forming Cells in Blood.- b) Humoral Antibodies.- ?) Binding of 131lodine-labelled Insulin.- ?) Passive Hemagglutination.- ?) Binding of Insulin in Pancreatic Tissue..- ?) The Effect of Labelling with FITC on the Immunological Properties of Insulin and Insulin Antibodies.- c) Immunofluorescence Studies of Various Organs.- ?) Local Granulomatous Tissue.- ?) Regional Lymph Nodes.- ?) Spleen.- ?) Bone Marrow.- ?) Liver.- ?)Kidney.- ?) Pancreas.- d) Histological Studies.- 3. Discussion.- a) Blood Cells Producing Antibodies to Insulin.- b) Characteristics of the Humoral Insulin Antibodies in Guinea Pigs.- c) The Localization of Antigen and Antibody Formation.- d) The Kidney in Long-term Immunization with Insulin.- 4. Summary.- E. Appendix: Studies of Insulin-immunized Sheep.- I. Statement of Purpose.- II. Results.- III. Discussion.- IV. Summary.- F. General Summary.- References.