EXPLANATION OF DESIGNATION OF THERMAL ANTIBODY [58R1-756]


  "It was postulated that the formation of both natural and


artificial antibody might be due to oxidation of antigen. 


Accordingly, suspensions of streptococci in NaCl solution were


subjected to the oxidative action of prolonged heat (96 hours)


without hydrogen peroxide and for but three hours on adding 1.5%


hydrogen peroxide to the autoclave.  As this was done specific


agglutinins and other antibodies developed which had respective


curative action on subcutaneous injection in therapeutic dosage in


respective persons ill."  58R1


  " ... The material in the supernatant of NaCl solution


suspensions of streptococci and other bacteria, after the


application of prolonged heat in the autoclave and the application


of heat for a far shorter period on the addition of 1.5% of the


oxidizing agent, hydrogen peroxide, are designated as 'antibody'


because (1) the supernatant of respective solutions agglutinated


specifically the organisms from which they were prepared, (2) they


precipitated specifically the respective dissolved antigens and (3)


had curative action in the treatment of diseases in which they were


causative, in a manner similar to convalescent serum and the serum


of horses hyperimmunized with the streptococcus."






CANCER, LEUKEMIA; AGGLUTINATION TITERS FOR STREPTOCOCCI [58R1-759]


  "In keeping with the concept generally held that leukemia is


'cancer' of the blood are the fairly comparable crosswise


agglutinative titers for the respective streptococci isolated in


leukemia (69 and 63%) and carcinoma (50 and 81% respectively." 58R1






DIVERSE SPECIFICITIES OF STREPTOCOCCI ACQUIRED IN HOSTS [58R1-760]


"Evidence ... indicates that the diverse specifities of


nonhemolytic streptococci ... is acquired in the respective hosts,


rather than being due to streptococci from diverse extraneous


sources, such as from milk and other foods or from air or water


supplies." 58R1






ISOLATION AND PRODUCTION OF STREPTOCOCCI   [58R1]


  The following particularly detailed account of Dr. Rosenow's


methodology is to be found in his last published article:


  "Serial dilution cultures at steps of 10-1, 10-6, and 10-10 were


made of nasopharyngeal swabbings, with cotton-wrapped aluminum wire


swabs, of the nasopharynx of persons having diverse chronic


disease.  The material thus obtained was inoculated into tall (8-10


cm.) columns of 0.2 percent dextrose broth adjusted to pH 7.0, to


which pieces of fresh calf or beef brain were added, comprising


approximately one part of brain substance to six or seven parts of


broth, and autoclaved at 17 pounds' pressure for 20 minutes.


  "Cultures were made in this freshly prepared medium directly or


soon after sterilization, but after prolonged storage the test


tubes containing the tall columns of medium were heated in a


boiling water bath for 15 minutes to remove absorbed oxygen."


  "The nasopharyngeal swabbings were obtained without touching the


tongue. The material on the swabs was washed off in 2 ml. of NaCl


solution, and then a tube of dextrose-brain broth was inoculated


with the swab, representing an estimated dilution of the material


swabbed of 10-2.  Two serial transfers were then made, with a


nicrome wire, the length of the column of dextroxe brain-broth


sterilized in a Bunsen flame at each step, representing dilutions


of inocula of 10-6 and 10-10 respectively.  Growths after incubation


at 35 degrees C. for 18 to 24 hours, at a dilution of 10-2,


consisted of predominating numbers of gram-staining, short-chained


streptococci and, usually, of moderate numbers of gram-staining


micococci and sometimes also gram-negative, gas-producing bacilli. 


Growth at the dilution of 10-6 usually consisted of a pure culture


of short-chained, alpha-type streptococci.  Growths usually did not


occur at dilutions of 10-10, but if positive such growth consisted


of a pure culture of streptococci.


  One milliliter of pure cultures from such end points of growth in


dextrose-brain broth were inoculated into [previously warmed, per


54R1] 200 ml. or gallon lots of 0.2 percent dextrose broth, and the


streptococci, thus grown at 35 degrees C. for 18 hours in the


smaller lots, were harvested in a cup-type centrifuge.  Pure


cultures of streptococci in the gallon lots of 0.2 percent dextrose


broth were harvested in the revolving bowl of the Sharples super-


centrifuge.  The sedimented streptococci from the cup-type


centrifuge were in turn suspended in dense suspension of an


estimated 200 billion streptocci per milliliter, and those from the


larger lots from the bowl of the super centrifuge at 1000 billion


per milliliter of two parts of chemically pure glycerol and one


part of saturated NaCl solution and stored in the dark in the


refrigerator at 10 degrees C.  Some of the streptococci in this


menstruum remained viable for months.  All remained gram-positive


and antigenically specific for many months on storage at 10 degrees


C.  Such storage made it readily possible to maintain serologic and


other specific properties of streptococci as isolated in diverse


diseases.[58R1]






SIMILAR ORGANISMS, DIFFERENT DISEASES  [58R1]


  --"The streptococci isolated in dextrose-brain broth from


nasopharyngeal swabbings of persons having diverse diseases were


much alike in size, chain formation and staining reactions.  All


were gram-positive and none produced zones of clear hemolysis on


blood-agar, but instead, small colonies surrounded by a narrow


green or indifferent zone were formed.  The freshly isolated


strains from nasopharyngeal swabbings of persons who had diverse



acute or chronic disease on intravenous injection into mice and


rabbits, localized and produced lesions "electively" in the organ


or organs corresponding to those involved in patients from whom the


streptococci were isolated."[58R1]






PREPARATION OF "THERMAL" ANTIBODY  58R1


  "...  Thermal antibody solutions ... were prepared by autoclaving


the respective specific and control NaCl solution suspensions


containing 20 billion streptococci per milliter for 96 hours, and


diluting the supernatant of such autoclaved suspensions with an


equal volume of NaCl solution and adding 0.2 percent phenol as a


preservative."[58R1]