The application of immunodiffusion test to the estimation of hydrolysis of mycobacteria proteins by proteolytic enzymes of mice peritoneal cells.
The possibility of applying the immunodiffusion (ID) test in agarose gel to detect the degradation of the somatic proteins of mycobacteria BCG-Poland by proteolytic enzymes of non activated and activated mice peritoneal cells was investigated. Degradation of those proteins was determined by disappearance of some precipitin lines resulting from the hydrolysis. The immunodiffusion test was found to be useful to show the degradation of protein of mycobacteria by cathepsins of mice peritoneal cells. Disappearance of 4 precipitin lines in ID test occurred after 3 h incubation of mycobacteria proteins with extracts of non activated mice peritoneal cells at pH 3, which gives evidence of degradation of some mycobacteria antigens. The extention of the time of incubation up to 24 h gave the same results. At the incubation at pH 4.5 no disappearance of precipitin lines was observed. It was found that cathepsins no specifically activated mice peritoneal cells degradate mycobacteria proteins at pH 3 and pH 4.5. Degradation occurred more rapidly at pH 3 achieving maximal effect after 6 h. The degree of degradation at pH 4.5 increased during the whole period of examination, i.e. up to 24 h of incubation time after which a disappearance of 5 precipitin lines was found. No degradation was found after incubation at pH 6 and pH 8.
D000135:Acid Phosphatase / Q000378:metabolism*; D000818:Animals; D001202:Ascitic Fluid / Q000201:enzymology*; D001426:Bacterial Proteins; D006868:Hydrolysis; D005779:Immunodiffusion; D008264:Macrophages / Q000201:enzymology*; D051379:Mice; D009163:Mycobacterium bovis; D010447:Peptide Hydrolases / Q000378:metabolism*; D005719:gamma-Globulins
D001426:Bacterial Proteins; D005719:gamma-Globulins; D000135:Acid Phosphatase; D010447:Peptide Hydrolases