Louisiana State University LSU Digital Commons LSU Historical Dissertations and Theses Graduate School 1969 Immunological Studies on the Genus Spirometra (Cestoda: Pseudophyllidea). Harry L. Henson Jr Louisiana State University and Agricultural & Mechanical College Follow this and additional works at: https://digitalcommons.lsu.edu/gradschool_disstheses Recommended Citation Henson, Harry L. Jr, "Immunological Studies on the Genus Spirometra (Cestoda: Pseudophyllidea)." (1969). LSU Historical Dissertations and Theses. 1662. https://digitalcommons.lsu.edu/gradschool_disstheses/1662 This Dissertation is brought to you for free and open access by the Graduate School at LSU Digital Commons. It has been accepted for inclusion in LSU Historical Dissertations and Theses by an authorized administrator of LSU Digital Commons. For more information, please contact [email protected]. This dissertation has been microfilmed exactly as received 70-9062 HENSON, Jr., Harry L., 1931- IMMUNOLOGICAL STUDIES ON THE GENUS SPIROMETRA (CESTODA.'PSEUDOPHYLLIDEA). The Louisiana State University and Agricultural and Mechanical College, Ph.D., 1969 Zoology University Microfilms, Inc., Ann Arbor, Michigan IMMUNOLOGICAL STUDIES ON THE GENUS SPIROMETRA (CESTODA: PSEUDOPHYLLIDEA) A Dissertation Submitted to the Graduate Faculty of the Louisiana State University and Agricultural and Mechanical College in partial fulfillment of the requirements for the degree of Doctor of Philosophy in The Department of Zoology and Physiology by Harry L. Henson, Jr. B.S., Southeastern Louisiana College, 1956 M.Ed., Louisiana State University, 1960 M.S., Louisiana State University, 1966 August, 1969 ACKNOWLEDGMENT The author wishes to express his sincere gratitude to Dr. Kenneth C. Corkum for his interest, guidance and suggestions during the work leading to and preparation of this manuscript. A special thanks is extended to his wife, Gerry, and sons for their understanding, patience and encouragement throughout the course of study. TABLE OF CONTENTS Page ACKNOWLEDGMENT.......................................... ii LIST OF TABLES................................... iv LIST OF FIGURES........................................ vi LIST OF PLATES..................... vii ABSTRACT................... viii INTRODUCTION............................... 1 Acquired Immunity ................................ 5 In Vitro Action of Immune Serum on the Life Stages of Helminths........................... 16 Changes in Serum Proteins........................ 19 Antigens.......................................... 22 Diagnostic Tests for Helminth Infections......... 26 MATERIALS AND METHODS.................................. 31 RESULTS................................................ 39 Electrophoresis.............................. 41 Passive Immunization. ...................... 45 Gel Diffusion ........ .................... 47 Intradermal Test. ............ 48 EXPLANATION OF PLATES........................ 74 DISCUSSION AND CONCLUSIONS.............................. 77 Electrophoretic Studies .......................... 78 Gel Diffusion 83 Passive Immunization.......................... 86 Intradermal Tests ........ ................ 87 SUMMARY................................................ 88 LITERATURE CITED........................................ 90 VITA......................... 114 iii LIST OF TABLES Table Page I. Absolute values for protein fractions in sera from control mice (gms/100 ml)................ 49 II. Electrophoretic analysis of sera from control. mice (relative concentrations in percent). 50 III. Electrophoretic analysis of sera from mice infected with the plerocercoids of Spirometra mansoni (relative concentrations in per cent) .... 51 IV. Absolute values for protein fractions in sera from mice infected with plerocercoids of Spirometra mansoni (gms/100 m l ) ..................... 52 V. Electrophoretic analysis of sera from mice infected with the plerocercoids of Spirometra urichi (relative concentrations in per cent)..... 53 VI. Absolute values for protein fractions in sera from mice infected with plerocercoids of Spirometra urichi (gms/100 ml) . 54 VII. Electrophoretic analysis of sera from mice infected with the plerocercoids of Spirometra mansonoides "A" (relative concentrations in per c e n t ) ................................ 55 VIII. Absolute values for protein fractions in sera from mice infected with plerocercoids of Spirometra mansonoides "A" (gms/100 ml) ....... 56 IX. Electrophoretic analysis of sera from mice infected with the plerocercoids of Spirometra mansonoides "B" (relative concentrations in per c ent).......................................... 57 X. Absolute values for protein fractions in sera from mice infected with plerocercoids of Spirometra mansonoides "B" (gms/100 m l ) ............ 58 XI. Absolute values for protein fractions in sera from sham injected rabbits (gms/100 ml) ............ 59 XII. Electrophoretic analysis of sera from sham injected rabbits (concentrations in per cent) .... 60 iv LIST OF TABLES (continued) Table Page XIII. Absolute values for protein fractions in sera from rabbits immunized against Spirometra mansoni (gms/100 m l ) .......... 61 XIV. Electrophoretic analysis of sera from rabbits immunized against Spirometra mansoni (concentration in per cent)........................ 62 XV. Absolute values for protein fractions in sera from rabbits immunized against Spirometra urichi (gms/100 ml)................................ 63 XVI. Electrophoretic analysis of sera from rabbits immunized against Spirometra urichi (concentrations in per c e n t ) ...................... 64 XVII. Absolute values for protein fractions in sera from rabbits immunized against Spirometra mansonoides "A” (gms/100 m l ) ............ 65 XVIII. Electrophoretic analysis of sera from rabbits immunized against Spirometra mansonoides "A” (concentrations in per c e n t ) ...................... 66 XIX. Absolute values for protein fractions in sera from rabbits immunized against Spirometra mansonoides "B" (gms/100 m l ) ................ 67 XX. Electrophoretic analysis of sera from rabbits immunized against Spirometra mansonoides "B" (concentrations in per cent) ........... 68 v LIST OF FIGURES Figure Page 1 Pattern of changes In serum proteins In control mice.............................. 69 2 Pattern of changes In serum proteins In mice experimentally infected with the plerocercoids of Spirometra ma n s o n i.......................... 70 3 Pattern of changes in serum proteins in mice experimentally infected with the plerocercoids of Spirometra urichi................ 71 4 Pattern of changes in serum proteins in mice experimentally infected with the plerocercoids of Spirometra mansonoides "A" .................. 72 5 Pattern of changes in serum proteins in mice experimentally infected with the plerocercoids of Spirometra mansonoides "B" .......... 73 6 Average of changes in serum protein fractions in mice experimentally infected with the plerocercoids of Spirometra mansoni, j>. urichi and S. mansonoides "A" and "B".......... 89 vi LIST OF ELATES Plate Page I. Double gel diffusion of sera from mice experimentally infected with the plerocercoids of Spirometra.................... 75 II. Double gel diffusion of sera from rabbits passively immunized against the plerocercoids of Spirometra.................................... 76 vii ABSTRACT Total serum proteins were determined, electrophoresis and double gel diffusion were performed on the sera from animals infected with the plerocercoids or passively immunized with larval antigen from Spirometra mansoni, jS. urichi and £S. mansonoides "A" and "B". Serum samples from mice were taken at three, 12 and 24 hours post infection and at four day intervals thereafter for a period of 12 weeks. Sera from these groups analyzed by electrophoresis exhibited decreased serum albumin concentrations and generally increases in the alpha, beta and gamma globulins. Rabbits passively immunized against the plerocercoids of these species exhibited little change in the total proteins after immuniza­ tion. Changes in the protein fractions were expressed as slight decreases in albumin and a corresponding increase in the gamma globulins. Precipitin bands were demonstrable by double gel diffusion in sera from all groups of mice infected with the plerocercoids of the three species of worms. Sera from rabbits passively immunized against the three species formed a maximum of three precipitin bands in agar gel six weeks post immunization. Intradermal tests in mice passively immunized against Spirometra mansoni and S. urichi gave widely variable results. Reactions obtained with injection of larval antigens indicate the presence of delayed hypersensitivity. viii INTRODUCTION Faust, et al. (1929) divided the genus Diphyllobothrium Cobbold, 1858, into two subgenera, Diphyllobothrium and Spirometra, based on uterine configuration and the shape of the eggs. Diphyllobothrium was to include those species with the outer uterine coils forming a. rosette and eggs with rounded ends. The subgenus Spirometra was to include those species with the outer uterine coils spiralled and eggs with pointed ends. Mueller (1937) elevated Spirometra to generic rank and further suggested the establishment of two other genera, subsequently named Dibothriocephalus and Cordicephalus by Wardle, et al. (1947). The cirrus, vagina and uterus open separately in Spirometra, whereas, the other two genera have common cirro-vaginal atria and separate pores. As further evidence for separation, Mueller (1937) noted