Rabbits injected intravenously with extracellular products ("toxins") of group A streptococci develop myocardial, muscular, and hepatic lesions. When such animals are then challenged with fluorochrome-labeled group A streptococci or with titanium oxide particles the labeled bacteria or particles localize within phagocytic cells in the tissue lesions caused by the toxins. Similarly, labeled streptococci or titanium oxide particles will also localize within phagocytic cells in skin lesions of guinea pigs that develop delayed hypersensitivity to tuberculin or to bovine gamma globulin. It is proposed that a combined mechanism of injury and localization of bacteria in damaged tissues may be responsible for poststreptococcal sequelae or other chronic inflammatory diseases.
Studies on the host parasite relationship in experimental streptococcal infections have been greatly aided by fluorescent antibody techniques (1). By such methods it is possible to follow the localization and persistence of group A streptococcal antigen in cells and tissues of laboratory animals (2-6). To detect such antigens by fluorescent techniques, frozen sections are often employed and the antisera to streptococcal antigens must be repeatedly absorbed with organ powders to remove nonspecific staining.
The present communication describes a simple method for applying a label of fluorescein isothiocyanate (FITC) to living group A streptococci, streptococcal cell-wall fragments,mucopeptides, and streptococcal Lforms. Such labeled streptococci and products were found to persist for long periods of time in inflammatory sites in the muscle and heart of mice and rabbits.
Intraartikuläre Injektion von Streptolysin-S-freien extrazellulären Produkten der Streptokokken Gruppe A verursacht eine zunächst akute, in der Folge aber subkutane Synovitis. Die Veränderungen gleichen denjenigen nach Injektion von Streptolollensonikaten, so dass angenommen wird, ausser Streptolysin S bedingen auch andere streptokokkale Faktoren eine Arthritis.
Extracellular antigens have been isolated from Streptococcus mitis, Streptococcus
salivarius and group A streptococcal cultures grown in a synthetic medium.
Analysis of the antigens was performed by immunoelectrophoretic and double
diffusion techniques using rabbit immune sera. S. mitis cultures produced 10
antigens, S. salivarius six antigens and group A streptococcus 12 antigens, when
tested with their corresponding antisera. S. mitis and S. salivarius antigens had
only one common antigen when tested with antisera to both antigen pools. No
cross reaction was found between the exo-antigens of group A and viridans
streptococci. While the extracellular antigen pool of group A streptococci contained
ribonuclease, deoxyribonuclease, hyaluronidase, diphosphopyridine-nucleotidase,
streptokinase and streptolysin 0, that of S. mitis contained only ribonuclease
and that of S. salivarius contained hyaluronidase and collagenase.
Each of the three streptococcal antigen pools contained a hemosensitizing
factor which sensitized mammalian cells to passive immune kill. Sonicates produced
from the mitis, salivarius and group A streptococcus contained six antigens,
most of which cross reacted with each other. S. mitis sonicates were separated
into six fractions by ion exchange chromatography on ECTEOLA cellulose, and
into three major fractions following gel-filtration on Sephadex 0-200 columns.
Rabbits injected i.v. with sonicates derived from S. mitis developed cardiac
and hepatic lesions which, in some cases, were accompanied by a steep rise in
serum glutamic oxalacetic transaminase, sorbitol dehydrogenase and total lipids.
The relationship of tissue damage to enzyme rise is discussed in relation to the
possible early diagnosis of tissue damage following streptococcal infection.
Rabbits injected with streptococcal extracellular protein (SEP) developed degenerative and infiltrative lesions in the heart and liver, with coagulation necrosis and multinucleated giant cells in the latter organ. The majority of these rabbits also showed elevated levels of glutamic-oxalacetic transaminase, or of sorbitol dehydrogenase, and all showed elevated serum lipids. These biochemical indications of cell injury could be found within 2 to 4 hours after a first injection of SEP. No such biochemical or pathologic effects were found following an injection of heated SEP or a commercially available streptokinase-streptodornase preparation from group C streptococcal culture. In a preliminary fractionation of SEP all the activity was found in a fraction not adsorbed to DEAE cellulose at pH 7.4 (0.05 M P04). The active fraction contained at least five antigens and four of the known streptococcal enzymes. Injection of extracts of sonically disrupted streptococci produced similar biochemical and pathologic changes. There was some cross-reacting material between the sonicates and anti-SEP serum.
