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Immunologic Properties of Anti-Rabbit Globulin Sera

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Immunologic Properties of Anti-Rabbit Globulin Sera This dissertation has been 63-2547 microfilmed exactly as received ROGUL, Marvin, 1932- IMMUNOLOGIC PROPERTIES OF ANTI-RABBIT GLjOBUUN CHICKEN SERA. The Ohio State University, Ph.D., 1962 Bacteriology University Microfilms, Inc., Ann Arbor, Michigan IMMUNOLOGIC PROPERTIES OP ANT I-RABBIT GLOBULIN CHICKEN SERA DISSERTATION Presented in Partial Fulfillment of the Requirements for the Degree Doctor of Philosophy in the Graduate School of The Ohio State University By MARVIN ROGUL, A .B ., H .S, •a*#***#** Tho Ohio State University 1962 Approved by L d v iser Department of Microbiology ACOOWLEDGMEHTS The author wishes to express sincere appreciation to Dr. II. C. Dodd for his help and thoughtfulness throughout this investigation. The author also wishes to thank Drs, If. J. Bigley, C. Cohen, H, S. Rheins, and Mrs. V. B. Geyer for their aid. Gratitude is also expressed to Ilessrs. P. H, Aldenderfer, 0. Pogue, C. F. Thayer, J. S. Widder, R. J. Zahransky, and Hiss Joan Shillis for their assistance. CONTENTS Page INTRODUCTION....................................................................................................... 1 LITERATURE R E V IE W ......................................................................................... 3 MATERIALS AND M ETH O D S.................................................................................... 18 RESULTS .......................................................................................................................31 DISCUSSION..................................................................................................................83 SUMMARY.......................................................................................................................93 BIBLIOGRAPHY.............................................................................................................95 AUTOBIOGRAPHY............................................ 100 ill TABLES Table Page 1 Electrophoretic Analysis of Rabbit Serum Preparations Used in Immunizing Chickens ............................ 32 2 Titration of Normal and Imnune Chicken Sera with Isosensitized Group A Rabbit Erythrocytes in 0.85 Per Cent Sodium C hloride ................................................................. 35 3 Normal and Immune Chicken Sera Adsorbed with Rabbit Red Cells Titrated with Isosensitized Group A Rabbit Brythrocytos in O.85 P®r Cent Sodium C h l o r i d e ....................................................................................................... 36 4 Titration of Normal and Immune Chicken Sera with Isosensitized Group A Rabbit Erythrocytes in 8.0 Per Cent Sodium C hloride ................................................................. 37 5 Normal and Immune Chicken Sera Adsorbed with Rabbit Red Cells Titrated with Isosensitized Group A Rabbit Erythrocytes in 8,0 Per Cent Sodium C hloride .................................................................................... 39 6 Titration of Normal and Immune Chicken Sera with Sensitized Sheep Erythrocytes in 0.85 Per Cent Sodium C hloride .................................................................................... 40 7 Titration of Normal and Immune Chicken Sera with Normal Sheep Erythrocytes in O.85 Per Cent Sodium C h l o r i d e ....................................................................................................... 41 8 Normal and Immune Chicken Sera Adsorbed with Rabbit Erythrocytes Titrated with Sensitized Sheep Erythrocytes in O.85 Per Cent Sodium Chloride . 43 9 Titration of Normal and Immune Chicken Sera with Sensitized Sheep Erythrocytes in 8.0 Per Cent Sodium C hloride ........................................................................ ....... 44 10 Normal and Immune Chicken Sera Adsorbed with Rabbit Erythrocytes T itrated with Sensitized Sheep Erythrocytes in 8.0 Per Cent Sodium Chloride. 45 iv TABLES— (Continued) Page 11 T itration of Normal and Imnune Chicken Sera with. Rabbit Serum Cohn Fraction II (Gamma Globulin) Coupled to Human E r y th ro c y te s................................................ 48 12 T itration of Normal and Immune Chicken Sera with Rabbit Serum Cohn Fraction III (Beta Globulin) Coupled to Human E r y th ro c y te s . ....................................... 49 13 T itration of Normal and Immune Chicken Sera with Rabbit Serum Cohn Fraction IV (Alpha Globulin) C oupled to Human E ry th ro c y te s . ...................................... 50 14 T itration of Normal and Immune Chicken Sera with Rabbit Serum Cohn Fraction IV-1 (Alpha Globulin) C oupled to Human E ry th ro c y te s............................................... 51 15 T itration of Normal and Immune Chicken Sera with Rabbit Serum Cohn Fraction IV-4 (Alpha Globulin) C oupled to Human E r y t h r o c y t................................................ e s 52 16 T itration of Normal and Immune Chicken Sera with Rabbit Serum Cohn Fraction V (Albumin) Coupled to Human E r y th ro c y te s ....................................................................... 53 17 T itration of Normal and Immune Chicken Sera with Chicken Egg Albumin Coupled to Human Erythrocytes . 54 18 Composite of Titrations of Normal and Immune Chicken Sera with Rabbit Serum Cohn Fractions C oupled to Human E ry th ro c y te s................................................ 55 19 Electrophoretic Analysis of Commercially Prepared Rabbit Serum Cohn Fractions .................................................... 57 20 The Hemagglutination of Sensitized Erythrocytes by Anti-Na„+Al Chicken Sera After Adsorption with Cohn Rabbit Fraction II (Gamma Globulin) ................................. 66 21 The Hemagglutination of Sensitized Erythrocytes by Anti-NRS Chicken Sera A fter Adsorption with Cohn Rabbit Fraction II (Gamma Globulin) .................................. 67 22 The Hemagglutination of Sensitized Erythrocytes by Anti-NH.+F Chicken Sera After Adsorption with Cohn Rabbit Fraction II (Gamma Globulin) ................................. 68 v TABLES— (Continued) Page 23 The Hemagglutination of Sensitized Erythrocytes hy Anti-HRS+F Chicken Sera After Adsorption with Cohn Rabbit Fraction II (Gamma Globulin) ....................... 69 24 The Hemagglutination of Sensitized Erythrocytes by Anti-Na«+Al Chicken Sera After Adsorption with Cohn Rabbit Fraction III (Beta Globulin) ................... • 70 25 The Hemagglutination of Sensitized Erythrocytes by Anti-HRS Chicken Sera After Adsorption with Cohn Rabbit Fraction III (Beta Globulin) ....................... 71 26 The Hemagglutination of Sensitized Erythrocytes by Anti-NH.+F Chicken Sera After Adsorption with Cohn Rabbit Fraction III (Beta Globulin) ....................... 72 27 The Hemagglutination of Sensitized Erythrocytes by Anti-HRS+F Chicken Sera After Adsorption with Cohn R ab b it F r a c tio n I I I (B eta G lo b u lin ) ........................ 73 28 The Hemagglutination of Sensitized Erythrocytes by Anti-Nap+Al Chicken Sera After Adsorption with Cohn Rabbit Fraction IV (Alpha Globulin) ....................... 74 29 The Hemagglutination of Sensitized Erythrocytes by Anti-HRS Chicken Sera After Adsorption with Cohn Rabbit Fraction IV (Alpha Globulin) ....................... 75 30 The Hemagglutination of Sensitized Erythrocytes by Anti-HH.+F Chicken Sera After Adsorption with Cohn Rabbit Fraction IV (Alpha Globulin) ....................... 76 31 The Hemagglutination of Sensitized Erythrocytes by Anti-HRS+F Chicken Sera After Adsorption with Cohn Rabbit Fraction IV (Alpha Globulin) ....................... 77 32 The Hemagglutination of Sensitized Erythrocytes by Anti-Na-+Al Chicken Sera After Adsorption with Cohn Rabbit Fraction V (Albumin) .......................................... 79 33 The Hemagglutination of Sensitized Erythrocytes by Anti-HRS Chicken Sera After Adsorption with Cohn Rabbit Fraction V (Albumin) .......................................... 80 34 The Hemagglutination of Sensitised Erythrocytes by Anti-HH.+F Chicken Sera After Adsorption with Cohn Rabbit Fraction V (Albumin) .......................................... 81 vi TABLES— (Continued) T a b le P age 35 The Hemagglutination of Sensitized Erythrocytes hy Anti-HRS+F Chicken Sera A fter Adsorption with Cohn Rabbit Fraction V (Album in) ............................................... 82 vii FIGURES Figure Page 1 Electrophoretic Patterns Representative of Rabbit Serum Components Used to Immunize C hicks............................... 33 2 Electrophoretic Patterns of Rabbit Serum Cohn Fractions at Different Dilutions .................................................. 58 3 Total nitrogen Precipitated From 0*5 ml of Chicken Antisera by Adding Various Amounts of Rabbit Serum Cohn Fraction I I ...................................................................................... 60 4 Total Nitrogen Precipitated From 0*5 ml Chicken Antisera by Adding Various Amounts of Rabbit Serum Cohn Fraction I I ................................................................................. 6 l 5 Total Nitrogen Precipitated From 0*5 nil of Chicken Antisera by Adding Various Amounts of Rabbit Serum Cohn Fraction I I I ............................................................................ 62 6 Total Nitrogen Precipitated From 0*5 nil of Chicken Antisera by Adding Various Amounts of Rabbit Serum Cohn Fraction IV ................................................................................. 63 7 Total Nitrogen Precipitated From 0*5 ml of Chicken Antisera by Adding Various Amounts of Rabbit Serum Cohn Fraction V ...............................................................................
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