VII. IDENTIFICATION OF ATYPICAL MYCOBACTERIA
335
tioning, column chromatography and preparative thin-layer chromatography. Two serologically active fractions from serotype 8 differ in their
content of carbohydrate components, and have a similar distribution of
simple fatty acids (principally palmitic, stearic, oleic (minor), tuberculostearic, and tetracosanoic) and of amino-acids, principally alanine and
threonine. T h e serologically active component(s) of M . sxulgai and M.
Kansasii appear to be simpler in structure, with little or no peptide content.
Both contain only a small, but similar amount of a nitrogenous, as yet
unidentified, substance. This may be a basic amino-acid, or an aminoalcohol.
V. T H E PROTOPLASMIC ANTIGENS
I n addition to their type-specific surface antigens mycobacteria have
numerous protoplasmic antigens. They are present in culture filtrates and
in bacterial extracts obtained by sonic disintegration. At least 17 different
antigens have been detected in the filtrates of M . tuberculosis. The specificities of the protoplasmic antigens from various mycobacterial species have
been investigated using the Ouchterlony immunodiffusion technique.
These studies have shown that some of the antigens are common to many
species, others are common to only some of them and still others are species
specific (Lind, 1965).
Stanford and Muse (1969) used the Ouchterlony technique to determine
whether the strains of the M . avium-Battey group belonged to a single or
different species. The M . avium antiserum tested against the homologous
extract gave 12 precipitin lines. Tested with extracts of other strains the sera
gave either 12 precipitin lines (serotype A) or ten precipitin lines (serotype
B). Virulence tests in chickens and rabbits indicated that the strains giving
12 precipitin lines (serotype A) were virulent for chickens and rabbits and
therefore were M. avium. The strains giving ten precipitin lines (serotype
B) were either avirulent for chicken and rabbits or virulent for the chicken
only. These latter strains therefore were either Battey strains or attenuated
M. avium strains. Thus, this test enables us to differentiate fully virulent
strains from attenuated M . avium or M. intracellulare strains.
The most remarkable property of the protoplasmic antigens is their
ability to elicit delayed hypersensitivity reactions of the tuberculin type.
Stottmeyer et al. (1969) reported that the tuberculin activity and specificity of purified protein derivatives and protoplasmic extracts of various
species of mycobacteria paralleled their antigenic activity and specificity
in immunodiffusion tests.
Considerable work has been done, using immunoelectrophoresis
(Castelnuovo and Morellini, 1965), ion-exchange chromatography (Kniker,
335
tioning, column chromatography and preparative thin-layer chromatography. Two serologically active fractions from serotype 8 differ in their
content of carbohydrate components, and have a similar distribution of
simple fatty acids (principally palmitic, stearic, oleic (minor), tuberculostearic, and tetracosanoic) and of amino-acids, principally alanine and
threonine. T h e serologically active component(s) of M . sxulgai and M.
Kansasii appear to be simpler in structure, with little or no peptide content.
Both contain only a small, but similar amount of a nitrogenous, as yet
unidentified, substance. This may be a basic amino-acid, or an aminoalcohol.
V. T H E PROTOPLASMIC ANTIGENS
I n addition to their type-specific surface antigens mycobacteria have
numerous protoplasmic antigens. They are present in culture filtrates and
in bacterial extracts obtained by sonic disintegration. At least 17 different
antigens have been detected in the filtrates of M . tuberculosis. The specificities of the protoplasmic antigens from various mycobacterial species have
been investigated using the Ouchterlony immunodiffusion technique.
These studies have shown that some of the antigens are common to many
species, others are common to only some of them and still others are species
specific (Lind, 1965).
Stanford and Muse (1969) used the Ouchterlony technique to determine
whether the strains of the M . avium-Battey group belonged to a single or
different species. The M . avium antiserum tested against the homologous
extract gave 12 precipitin lines. Tested with extracts of other strains the sera
gave either 12 precipitin lines (serotype A) or ten precipitin lines (serotype
B). Virulence tests in chickens and rabbits indicated that the strains giving
12 precipitin lines (serotype A) were virulent for chickens and rabbits and
therefore were M. avium. The strains giving ten precipitin lines (serotype
B) were either avirulent for chicken and rabbits or virulent for the chicken
only. These latter strains therefore were either Battey strains or attenuated
M. avium strains. Thus, this test enables us to differentiate fully virulent
strains from attenuated M . avium or M. intracellulare strains.
The most remarkable property of the protoplasmic antigens is their
ability to elicit delayed hypersensitivity reactions of the tuberculin type.
Stottmeyer et al. (1969) reported that the tuberculin activity and specificity of purified protein derivatives and protoplasmic extracts of various
species of mycobacteria paralleled their antigenic activity and specificity
in immunodiffusion tests.
Considerable work has been done, using immunoelectrophoresis
(Castelnuovo and Morellini, 1965), ion-exchange chromatography (Kniker,