title: Neutral protease activity in lymphocytes of Lewis rats with acute experimental allergic encephalomyelitis.
abstract: Lymphocytes from popliteal and inguinal lymph nodes of Lewis rats with acute EAE as a result of injection of lyophilized guinea pig myelin in Freund's complete adjuvant exerted strong proteolytic activity at neutral pH toward myelin basic protein. After injection of myelin the level of proteolytic activity remained about the same as that in lymphocytes from Freund's adjuvant-injected controls until about day 10 after injection, just before the onset of paralytic symptoms; then the proteolytic activity increased to approximately double its former level. Myelin basic protein was hydrolyzed by whole lymphocytes, but more activity was unmasked by homogenization. Similar results were also obtained using lymphocytes from thymus of EAE and control animals. Lymphocytes with high levels of proteolytic activity were not absorbed by glass wool, did not stain with neutral red, nor did they phagocytose antibody-coated sheep red blood cells. Thymus and lymph node lymphocytes cleaved myelin basic protein to three major peptides and a fourth minor peptide, while spleen lymphocytes hydrolyzed basic protein at only one point resulting in two peptides whose molecular weights added up to that of myelin basic protein. The protease activity was inhibited by 5 X 10(-3) M p-chloromercuribenzoate and by phenylmethyl sulfonyl fluoride, TPCK, and soybean trypsin inhibitor, therefore the enzymatic activity probably depends on a serine residue and a sulfhydryl group. The bulk of the enzymatic activity is mostly membrane bound with the highest specific activity and total activity contained in a lysosomal-mitochondrial fraction. In view of the infiltration of lymphocytes into the brain substance in acute EAE, it is suggested that these cells may contribute to the destruction of myelin which is usually attributed to the monocyte or macrophage.
mesh_terms: D000818:Animals; D001402:B-Lymphocytes / Q000201:enzymology*; D002462:Cell Membrane / Q000201:enzymology; D004591:Electrophoresis, Polyacrylamide Gel; D004681:Encephalomyelitis, Autoimmune, Experimental / Q000201:enzymology*; D004791:Enzyme Inhibitors; D005260:Female; D005620:Freund's Adjuvant; D008247:Lysosomes / Q000201:enzymology; D008297:Male; D004676:Myelin Basic Protein / Q000378:metabolism; D010447:Peptide Hydrolases / Q000378:metabolism*; D011480:Protease Inhibitors; D051381:Rats; D011917:Rats, Inbred Lew; D012967:Sodium Dodecyl Sulfate; D013601:T-Lymphocytes / Q000201:enzymology
chemical_list: D004791:Enzyme Inhibitors; D004676:Myelin Basic Protein; D011480:Protease Inhibitors; D012967:Sodium Dodecyl Sulfate; D005620:Freund's Adjuvant; D010447:Peptide Hydrolases