Countermeasure development: Immunoprophylaxis and immunotherapy of Combined Acute Radiation Syndromes. Dmitry Popov.

Advanced Medical Technology and Systems inc. Canada. intervaccine@gmail.com

Introduction: Combined Acute Radiation Syndromes ( CARS) are extremely severe type of injury . Combination of Radiation and Thermal factors are induce development of acute pathological processes: systemic inflammatory response syndrome (SIRS), toxic multiple organ injury (TMOI), toxic multiple organ dysfunction syndromes (TMOD), toxic multiple organ failure (TMOF) in irradiated mammals. High doses of Radiation and Thermal injury induce formation of groups of Toxins: A. Specific Radiation Toxins B. Specific Thermal Toxins 3.Nonspecific Histiogenic Pro-inflammatory and Inflammatory Toxins. Specific Radiation Toxins (SRT) include four major group of Toxins : Cerebrovascular Radiation Toxins (Cv RT), Cardiovascular Radiation Toxins (Cr RT),Gastrointestinal Radiation Toxins (Gi RT), Hematopoietic Radiation Toxins (Hp RT). CvRT, Cr RT, Gi RT this groups of toxins are defined as a Neurotoxins and Hp RT this group are defined as Hematotoxins. Specific Thermal Toxins (STT) isolated from burned skin (Voul S., Colker I. 1972). Group of Nonspecific Histiogenic Inflammatory Toxins (NHIT) includes high amount of tissues toxins which are peptides with medium molecular weight and this group of polypeptides can be significant factor as a part of developing of general inflammation reaction. However, NHIT toxins cant induce many specific reactions and changes which are specific for radiation. Specific Radiation Toxins can induce specific processes and reactions such as clonogenic cell death - programmed apoptotic necrosis. Although after high doses of radiation other forms of cell death such as Pyroptosis or Oncosis should be considered. We postulate that NHIT toxins similar for high doses of radiation and thermal injury. Neurotoxins and Hemato-toxins which are induced after high doses of radiation and STT toxins which formation are induced by Thermal Factor are different. Administration of STT toxins or NNHIT toxins (IV or IM) to healthy mammals induce development lymphocytosis, leukocytosis, trombocytosis and activation of blood coagulation cascade. Administration of SRT (IV or IM) to radiation nave animals induce leukopeina, thrombopenia, lymphopenia as a result of clonogenic programmed cell death, blood coagulation cascade suppression was registered. Matherials and Methods: Cow, horses, rabbits, rats, mice were used for different stages of our experiments. Animals were quarantined at laboratory conditions for three weeks prior to experimentation. Isolation of SRT were provided from central lymphatic duct of irradiated cows. Immunization of horses and rabbits for elaboration of Antiradiation Antibodies ( Specific Antiradiation Antidote SAR) were provided. Animals: cows, mice, rats were irradiated in the VSRI ( Kazan), Academy of Veterinary Medicine (Moscow), Scientific Research Institute of Radiobiology (Gomel), Scientific Research Nuclear Center (Dubna). Equipment for gammairradiation: Pyma, Panorama - Co gamma radiation source. Irradiation was performed in different doses corresponding to induction of severe forms of Acute Radiation Syndromes (ARS).Mice and rats receiving combined radiation thermal injury. Model of Thermal injury: Burns - 10% of total body surface. Third grade of burns was used as a model. Thermal Injury

was given after irradiation. Preparations of Antiradiation Vaccine contained toxoid form of Radiation Toxins was used for immunoprophylaxis. Preparations of Antiradiation Antidote IgG , contained antibodies to Radiation Toxins was used for immunotherapy. Scheme of experiments : I. Control: Group A. Animals with ARS without treatment. Group B. Animals with thermal injury without treatment.Group C.Animals with combined forms of ARS without treatment. II. Specific Immunoprophylaxis with Antiradiation Vaccine (AV). Group C. Animals with immunoprophylaxis by AV. Irradiation provided 24 days after vaccination. Group D. Animals with immunoprophylaxis by AV. Thermal injury was provided 24 days after vaccination. Group E. Animals with vaccination by AV and combined injur. Irradiation was provided 24 days after irradiation. Thermal injury was provided immediately after irradiation. III. Specific Immunotherapy with Antiradiation Antidote IgG (AA IgG). Group E. Animals with ARS and immunotherapy by AA IgG. Group F. Animals with thermal injury and immunotherapy by AA IgG. Group H. Animals with combined radiation thermal injury and treatment by AA IgG. Results: The Lethality Doses (LD) 100/30 of radiation expressed 100 % mortality rate in next 30 days after irradiation with development of different forms of ARS. The thermal injury which induced third degree burns with area of dry necrosis. Mortality rate in group with thermal injury without treatment was almost 90 % in next 30 days. Lethality rate at Combined Radiation and Thermal injury without any treatment demonstrated 100 % in next 30 days. Immunoprophylaxis with specific AV was most effective for animals with ARS and survival rate was up to 70 %. Immunotherapy with specific AA IgG although demonstrated less effectiveness and demonstrated survival rate 50 % -60% in different groups of irradiated animals. For animals with thermal injury Immunotherapy with AV and Immunoprophylaxis AA IgG were significantly uneffective and survival rate do not exceed 15 %. Results of specific immunotherapy and immunoprophylaxis provided at combined radiation thermal injury ( CRTI) demonstrated 30% of survival rate. Conclusion: Effects of Different Biological Response to specific immunoprophylaxis with AV and specific immunotherapy with AA IgG demonstrated effective radioprotection for irradiated animals with different forms of ARS. The recovery phases demonstrated shorter period of reconvalescence . Efffects of specific immunoprophylaxis with AV and immunotherapy with AA IgG provided for animals with thermal and combined injury were less effective. However Immunoprophylaxis and Immunotherapy with Specific Immunomodifiers used at combined and Thermal injury demonstrated prolonged life time after immunoprophylaxis. Demarcation zone of burns and necrotic tissues rejection were more expressed after immunotherapy. Specific Immunoprophylaxis with Thermal Injury Toxins and Specific Immunotherapy with specific anti-thermal injury antibodies ( serum of IgG preparation) could significantly improve results of therapy of thermal and combined injury.