Use of Pressure Cell to Prepare Cell Walls from Mycobacteria.

Ribi, Edgar; Perrine, Theodore D.; List, Robert; Brown, William E.; Goode, Granville

doi:10.3181/00379727-100-24730

Cited by 46 publications

(17 citation statements)

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“…Because of the complex nature of this organism, many fractionation schemes have been devised. Intact cells and culture filtrates have been subjected to chemical fractionation (36)(37)(38)(39), and some components have been isolated by a variety of methods including enzyme digestion, electrophoresis, gel filtration, and chromatography (40)(41)(42)(43)(44)(45)(46)(47)(48). As a result, many fractions have been isolated and partially defined with respect to physical structure and biological properties, but none has been applied to development of a primary test to detect and measure humoral antibodies.…”

Section: Discussionmentioning

confidence: 99%

Binding Between Components of the Tubercle Bacillus and Humoral Antibodies

Minden

Farr

1969

The Journal of Experimental Medicine

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Studies were undertaken to find a substance or substances for use in primary binding types of tests to detect humoral antibodies in rabbits and monkeys exposed to the tubercle bacillus that would distinguish between strains of mycobacteria. The antigen employed was a component of the 5159 strain of Mycobacterium tuberculosis that was obtained following sonification, ultracentrifugation, electrophoresis, and elution from a preparative polyacrylamide column. When the antigen was labeled with 131I, specific binding was observed in sera from immunized rabbits and BCG-protected rhesus monkeys by the radio-gel electrophoresis and ammonium sulfate tests. This component was partially characterized, and its major antigenic determinants were associated with anionic mucopolysaccharides. Electrophoretically at pH 8.3 it migrated anodally to albumin, its molecular weight was between 9,000 and 12,000, and it was soluble in 50% saturated ammonium sulfate. Binding to antibody was destroyed after treatment with Pronase, but not after DNase or RNase. Inhibition of the reaction, as measured by the ammonium sulfate test, between the 131I-labeled component and antisera from rabbits that had been immunized with sonicated 5159 organisms, was studied. These experiments demonstrated a capacity to define subtle similarities and differences among different mycobacteria and mycobacterial components. Some microorganisms not clearly related to mycobacteria also partially inhibited this reaction, suggesting that they shared antigenic groups with the component derived from 5159 organisms. The studies described suggested the advisability of using direct primary tests and purified components of mycobacteria to differentiate further the antigenic groups between individual pathogenic mycobacteria and between pathogenic and nonpathogenic organisms.

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Section: Discussionmentioning

confidence: 99%

Binding Between Components of the Tubercle Bacillus and Humoral Antibodies

Minden

Farr

1969

The Journal of Experimental Medicine

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“…The induction of a positive experimental Mitsuda reaction to lepromin by a variety of mycobacteria [12] ; the ability of lepromin to induce hypersensitivity to BCG and its fractions, particularly the cell wall ; the gross and histological appearances of these responses ; and the doubtful cross reactivity with tuberculin, suggest that the experimental Mitsuda reaction is like accelerated tubercle formation in tuberculosis; i.e., a response to whole mycobacteria, as had been proposed by O lm os C a s t r o [24], and to mycobacterial cell walls, as has been previously described [19,20,21,32,33].…”

Section: Ms Ms Ms Ms Ms Ms Ms Ms Ms Ms Ms Ms Ms Ms Ms Msmentioning

confidence: 68%

Cross Reactivities of Lepromin

Goihman-Yahr¹,

Raffel²,

Ferraresi³

1969

Int Arch Allergy Immunol

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“…The disintegration of mycelia cells was carried out by Sorval-Ribi Refrigator Cell Fractionator (Model RF-1) at a pressure of 30,000 psi (9,10). For electron microscopic examinations, drops of suspension of cell wall preparations were mounted on grids, dried down in the air, and the specimens were shadowed with carbon vapor for 20 sec.…”

Section: Methodsmentioning

confidence: 99%