RESUMEN
Injection of acid-fast bacilli, obtained from spleens of human leprosy patients, into normal rabbits did not lead to the development of an antiserum reacting with the serum of leprosy patients in such a way as to indicate the presence of specify leprosy bacillus antigen in leprous serum.
Asunto(s)
Mycobacterium leprae/aislamiento & purificación , Mycobacterium leprae/patogenicidad , Pruebas Cutáneas/métodosRESUMEN
The Seibert techniques for the preparation of specific proteins of the tubercle bacillus were applied to spleens rich in acid-fast bacilli, from leprosy patients, in the hope that there could be obtained specific proteins of the microorganism of leprosy which could be used in serological or skin tests for the diagnosis of leprosy. Preparations were made from three such spleens. In the case of the first one the organ was ground, the minced tissue was extracted with water, and the aqueous extract was concentrated by ultrafiltration. A corresponding preparation was made from a normal spleen. The concentrated extract from the leprosy spleen reacted with 14 of 31 leprosy sera in the precipitin test, but in none of 14 sera from healthy persons or 2 from Kahn-positive patients in Philadelphia. The preparation from normal spleen did not react with any of the sera. The preparation from the leprous spleen, which appeared to act as an antigen with the 14 leprosy sera, did not react with rabbit antiserum to proteins from various strains of supposed M. leprae. In the case of the second spleen, what was believed to be a superior method of grinding, extraction and protein purification was used. Extraction was carried out with faintly alkaline buffered phosphate solution, more thorough grinding was employed, and the trichloracetic acid method was used to precipitate the proteins . The refined proteins thus obtained were used as antigens in the precipitin test with the sera of 32 leprous patients, but in no case to demonstrate the presence of protein other than that derived from the spleen used, for after rabbit antiserum immune to the leprosy spleen protein was saturated with a corresponding protein caused no further precipitate. It was hoped, however, that a protein comparable to tuberculin protein, reactive in the skin test, might te present. A comittee in Manila, the report of which appears in the preceding article. No evidence for the presence of an antigen reacting specifically in the skin of lepers was obtained. Preparations were made from a third leprous spleen by methods believed to combine any advantages of the methods employed in making preparations from the first two.
Asunto(s)
Lepromina , Lepromina/aislamiento & purificación , Lepra/inmunologíaRESUMEN
Theantigenic relationships of sixteen strains of mycobacteria isolated from leprosy, and of this group of organisms and eleven other strains of acid-fast bacteria, have been determined by means of the precipition reaction, using purified specific proteins and sera of rabbits immunized against those proteins. The organisms employed all grew readily on Long´s synthetic medium, which was used for all cultures; obviously, strains not growing on the artificial medium could not be included. With but few exceptions the antisera tested gave the highest precipitin titers with their homologous antigens. Thirteen of the sixteen lleprosy strains gave strong precipitin reactions with one another (text-figure 1). In view of this it is considered justifiable to place them in one large, antigenically related group and may therefore be considered as antigenically unrelated. The one remaining strains, Levy-Kedrowsky, while it failed to give definite cross reactions with all members of the large lepra group, did react with two of them, the Ota-Sato and Karlinski strains, and therefore has an antigenic relationship to the group. All of the acid-fast bacteria studied, including the bacilli from leprosy, various types of tubercle bacilli, and acid-fast saprophytes, tend to fall into certain groups (text-figure 2). A relationship between the leprosy strains and other acid-fast bacteria is indicated by their common cross reactions. Striking relationships are noted in the case of the strains of Daines, Karlinski and Duval (nonchromogenic), either directly or through linkage with M. avium, M.tuberculosis, hominis, M. marinum and with M. phlei. It is interesting to note that the avian tubercle bacillus studied has some antigenic relationship to Duval´s nonchromogenic organism, while it has no direct relationship with the large leprosy group represented by Daines' strain. The bovine tubercle bacillus strain studied apparently bears no antigenic relationship to any of the other acid-fast organisms employed except the human tubercle bacillus. Chromogenesis apparently bears no relationship to the antigenic composition of these micro-organisms, in so far as that is related to their protein elements.