Alterations in cysteine proteinase content of rat lung associated with development of Pneumocystis carinii infection.
The rate of hydrolysis of three cysteine-type proteinase substrates, N-benzyloxycarbonyl-Arg-Arg-4-methyl-7-coumarylamide (AMC) (cathepsin B), Arg-AMC (cathepsin H), and N-benzyloxycarbonyl-Phe-Arg-AMC (cathepsin L), were determined in rat lung throughout the time course of the induction of Pneumocystis carinii infection by immunosuppression. Cathepsin B-like and cathepsin L-like activities fell below control values initially, but from week 8 of the immunosuppressive treatment significant increases above the control were noted. Cathepsin H-like activity was greater than control levels from week 3, and by week 12 it was 7,600% of the mean control value. When compared with the relative degree of infection, as assessed from the number of cysts present in lung impression smears, cathepsin B-like and cathepsin L-like activities were significantly increased only at heavy parasite burdens while cathepsin H-like activity displayed a close correlation with parasite number (r = 0.884; P less than 0.001). Activity was detected in lysates of purified P. carinii with all three substrates. Treatment of heavily infected animals with co-trimoxazole cleared the lungs of P. carinii, and this was accompanied by a marked reduction in proteinase activity, in particular, cathepsin H-like activity, which fell from 108- to 3-fold the mean control value following drug treatment. Analysis of cathepsin H isozyme patterns by fluorography following isoelectric focusing revealed differences between treated and control lung samples. In the immunosuppressed group, there was a time-dependent increase in the intensity of some of the bands observed in the controls and an appearance of several novel bands which corresponded to bands observed in lysates of P. carinii. It is likely, therefore, that the increased proteinase activity observed in the treated group is due, at least in part, to isozymes from P. carinii; consequently, cathepsin H-like activity might be of use diagnostically in the identification of P. carinii infection and in the estimation of parasite burden.
Hayes
D J
DJ
Biochemical Sciences, Wellcome Research Laboratories, Beckenham, Kent, United Kingdom.
Stubberfield
C R
CR
McBride
J D
JD
Wilson
D L
DL
D000818:Animals; D001835:Body Weight; D002401:Cathepsin B / Q000032:analysis*; D056655:Cathepsin H; D056668:Cathepsin L; D002403:Cathepsins / Q000032:analysis*; D003546:Cysteine Endopeptidases; D010450:Endopeptidases; D007525:Isoelectric Focusing; D007930:Leucine / Q000031:analogs & derivatives / Q000494:pharmacology; D008168:Lung / Q000201:enzymology*; D008297:Male; D011010:Pneumocystis / Q000201:enzymology; D011020:Pneumonia, Pneumocystis / Q000201:enzymology*; D051381:Rats; D011919:Rats, Inbred Strains; D013997:Time Factors; D015662:Trimethoprim, Sulfamethoxazole Drug Combination / Q000494:pharmacology
D015662:Trimethoprim, Sulfamethoxazole Drug Combination; D002403:Cathepsins; D010450:Endopeptidases; D003546:Cysteine Endopeptidases; D002401:Cathepsin B; D056668:Cathepsin L; C534280:Ctsl protein, rat; D056655:Cathepsin H; C534128:Ctsh protein, rat; D007930:Leucine; C024974:E 64