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Title: Degradation of extracellular matrix by larvae of Schistosoma mansoni. II. Degradation by newly transformed and developing schistosomula

Abstract

The ability of schistosomula of Schistosoma mansoni to degrade an extracellular connective tissue matrix synthesized by rat vascular smooth muscle cells in culture was investigated. Six to 12% of the total matrix was degraded by schistosomula from the time of transformation from cercariae to adult development in vitro. Most matrix degradation occurred during the first 24 hours of incubation and was dependent on the number of schistosomula and the type of medium in which they were incubated. The use of proteinase inhibitors indicated that schistosomula activity was distinctly different from that of cercariae. Newly transformed schistosomula expressed one activity that was similar in inhibition characteristics to that of cercarial preacetabular gland secretions and another activity that was unique to schistosomula. From 1 day after transformation to adulthood, the schistosomula-derived activity was the predominant activity detected. Schistosomula degraded a smaller percentage of the total matrix than did cercariae and showed a different substrate profile. Schistosomula degraded glycoprotein components of extracellular matrix but showed little or no activity against elastin or collagen. Matrix-degrading activity was also detected in schistosomula-conditioned medium. Sedimentation of the activity and lack of permeability through filter barriers suggest that the enzyme may be initially associated with membrane andmore » then sloughed with membrane fragments. Since the schistosomula-derived activity initially overlaps with cercarial preacetabular gland proteolytic activity, the two activities may act in concert to facilitate skin penetration by newly transformed schistosomula. However, schistosomula activity probably serves some, as yet undetermined, function later in development.« less

Authors:
; ; ;
Publication Date:
Research Org.:
Univ. of California, San Francisco
OSTI Identifier:
6451738
DOE Contract Number:
AC03-76SF01012
Resource Type:
Journal Article
Resource Relation:
Journal Name: Lab. Invest.; (United States); Journal Volume: 49:2
Country of Publication:
United States
Language:
English
Subject:
59 BASIC BIOLOGICAL SCIENCES; CONNECTIVE TISSUE; PROTEOLYSIS; LARVAE; SERINE PROTEINASES; ENZYME ACTIVITY; GLUCOPROTEINS; METAMORPHOSIS; RATS; SCHISTOSOMA; SKIN; ANIMAL TISSUES; ANIMALS; BODY; CARBOHYDRATES; CHEMICAL REACTIONS; DECOMPOSITION; ENZYMES; HELMINTHS; HYDROLASES; MAMMALS; ORGANIC COMPOUNDS; ORGANS; PEPTIDE HYDROLASES; PLATYHELMINTHS; PROTEINS; RODENTS; SACCHARIDES; TISSUES; TREMATODES; VERTEBRATES; 550200* - Biochemistry

Citation Formats

Keene, W.E., Jeong, K.H., McKerrow, J.H., and Werb, Z. Degradation of extracellular matrix by larvae of Schistosoma mansoni. II. Degradation by newly transformed and developing schistosomula. United States: N. p., 1983. Web.
Keene, W.E., Jeong, K.H., McKerrow, J.H., & Werb, Z. Degradation of extracellular matrix by larvae of Schistosoma mansoni. II. Degradation by newly transformed and developing schistosomula. United States.
Keene, W.E., Jeong, K.H., McKerrow, J.H., and Werb, Z. 1983. "Degradation of extracellular matrix by larvae of Schistosoma mansoni. II. Degradation by newly transformed and developing schistosomula". United States. doi:.
@article{osti_6451738,
title = {Degradation of extracellular matrix by larvae of Schistosoma mansoni. II. Degradation by newly transformed and developing schistosomula},
author = {Keene, W.E. and Jeong, K.H. and McKerrow, J.H. and Werb, Z.},
abstractNote = {The ability of schistosomula of Schistosoma mansoni to degrade an extracellular connective tissue matrix synthesized by rat vascular smooth muscle cells in culture was investigated. Six to 12% of the total matrix was degraded by schistosomula from the time of transformation from cercariae to adult development in vitro. Most matrix degradation occurred during the first 24 hours of incubation and was dependent on the number of schistosomula and the type of medium in which they were incubated. The use of proteinase inhibitors indicated that schistosomula activity was distinctly different from that of cercariae. Newly transformed schistosomula expressed one activity that was similar in inhibition characteristics to that of cercarial preacetabular gland secretions and another activity that was unique to schistosomula. From 1 day after transformation to adulthood, the schistosomula-derived activity was the predominant activity detected. Schistosomula degraded a smaller percentage of the total matrix than did cercariae and showed a different substrate profile. Schistosomula degraded glycoprotein components of extracellular matrix but showed little or no activity against elastin or collagen. Matrix-degrading activity was also detected in schistosomula-conditioned medium. Sedimentation of the activity and lack of permeability through filter barriers suggest that the enzyme may be initially associated with membrane and then sloughed with membrane fragments. Since the schistosomula-derived activity initially overlaps with cercarial preacetabular gland proteolytic activity, the two activities may act in concert to facilitate skin penetration by newly transformed schistosomula. However, schistosomula activity probably serves some, as yet undetermined, function later in development.},
doi = {},
journal = {Lab. Invest.; (United States)},
number = ,
volume = 49:2,
place = {United States},
year = 1983,
month = 1
}
  • The ability of cercariae of Schistosoma mansoni to degrade a model extracellular connective tissue matrix produced by rat vascular smooth muscle cells in culture was investigated. In this model, connective tissue macromolecules are present in the interactive framework that characterizes their structure in vivo. Cercariae were stimulated to degrade the matrix by skin lipid or linoleic acid. At the maximally stimulating concentration of linoleic acid (25 ..mu..g/cm/sup 2/), 68% of the total matrix was degraded, including 57% of the glycoprotein, 79% of the elastin, and 8% of the collagen. Degradation of matrix was inhibited by ..cap alpha../sub 1/-proteinase inhibitor andmore » soybean trypsin inhibitor. Ethylenediaminetetraacetic acid inhibited degradation by unstimulated but not linoleic acid-stimulated cercariae. Preacetabular gland secretions collected from cercariae also degraded the matrix with an activity 86% of that of live cercariae. Preacetabular gland proteolytic activity was also inhibited by ..cap alpha../sub 1/-proteinase inhibitor, soybean trypsin inhibitor, and ethylenediaminetetraacetic acid. The similar characteristics of matrix degradation by both live cercariae and cercarial preacetabular gland secretions support the idea that a proteinase secreted from cercarial preacetabular glands facilitates invasion of skin and connective tissue by these larvae. Degradation of elastin and glycoprotein constituentes of extracellular matrix is probably essential for skin penetration.« less
  • Surface components of mechanically transformed schistosomula of Schistosoma mansoni were labeled by lactoperoxidase-catalyzed iodination. After solubilization with Triton X-100, antigens were identified by immunoprecipitation. Serum from chronically infected Swiss mice reproducibly precipitated seven major polypeptides with approximate molecular weights (X 10/sup 3/) of 94, 68, 45, 40 to 32, 22, and 16. The antigens of molecular weights (X 10/sup 3/) of 94, 40 to 32, 22, and 16 were shown to be exposed on the parasite surface by interaction of the antibodies with intact labeled schistosomula. Sera from several strains of infected inbred mice precipitated the same polypeptides. The antibodiesmore » produced during chronic infection were found to be stimulated by adult worms since sera from 6-week-infected animals precipitated none of the surface antigens, and the pattern produced by precipitation with antibodies from a mouse infected with male worms only was indistinguishable from the pattern obtained with sera from mice with bisexual infections. Antibodies from mice immunized with highly irradiated cercariae reproducibly precipitated major polypeptides of approximately (X 10/sup 3/) 94, 68, 45, 32, 22, 19, and 15 daltons. The antigens of (X 10/sup 3/) 94, 43, 32, 22, and 15 daltons were shown to be exposed on the parasite surface by interaction of the antibodies with intact labeled schistosomula. The 15 X 10(3)-dalton surface protein was recognized by sera from vaccinated, but not chronically infected, mice, suggesting that it represents a stage-specific immunogen present on schistosomula but not on adult worms. Sera from two inbred strains of mice which develop different degrees of immunity recognized the same antigens.« less
  • The in vitro effect of the lethal antibody on schistosomula in sera of rhesus monkeys immunized with highly x-irradiated cercariae of Schistosoma mansoni or S. japonicum was studied. In all 6 experimental monkeys, 4 unchallenged and 2 challenged, the effects of lethal antibody on schistosomula were demonstrated. The sera of the challenged monkeys had no stronger lethal effect than those of the unchallenged monkeys. This shows that the lethal antibody can be produced by the antigenic stimulation of schistosomula alone. The mortality rates of schistosomula in immune sera were already high at day 1, increased to a certain extent frommore » day 1 to day 4, but showed no significant further increase in days 5 and 6. Two kinds of immunological reactions were observed: perischistosomular precipitate (PSP) and perischistosomular envelope (PSE). Schistosomula surrounded with PSP were usually dying or dead and those enclosed in PSE were usually alive and motile. Thus PSP may be related with the lethal antibody and PSE with a kind of enhancing antibody. Schistosomula with PSP showed a positive fluorescent reaction when stained with fluorescein-labeled rabbit anti-rhesus IgG. Scanning electron micrographs of schistosomula with PSP showed a highly degenerated tegument. These facts indicate that the antibody in PSP contains a fraction of IgG which acts on the tegument of schistosomula.« less
  • Spleen cells of mice vaccinated with radiation-attenuated Schistosoma mansoni cercariae were used to produce monoclonal antibodies directed against newly transformed schistosomular surface antigens. One of these monoclonal antibodies recognized a polypeptide of 18 kDa. Binding was measured by radioimmunoassay. This glycoprotein was purified by monoclonal antibody immunoaffinity chromatography and a polyclonal antiserum was prepared against it. Immunofluorescence assays showed that the polyclonal antiserum bound to the surface of newly transformed schistosomula and lung-stage organisms but not to the surface of liver-stage and adult worms. Using this polyclonal antiserum we isolated recombinant clones from an adult worm cDNA expression library constructedmore » in lambdagt11. Clone 654.2 contained an insert of 0.52 kilobase and hybridized to a 1.2-kilobase mRNA species from adult worms. Most importantly, clone 654.2 produced a fusion protein of 125 kDa that was reactive with sera of vaccinated mice that are capable of transferring resistance. This result encourages future vaccination trials with the fusion protein.« less
  • Experiments were performed to compare the migration and survival of 75Se-labeled schistosomes, introduced by percutaneous cercarial exposure or by intravascular administration of 7-day-old lung stage schistosomula, in control and irradiated cercaria-immunized mice. Schistosomula were intravascularly introduced into the lungs, systemic organs and liver by injection via the femoral vein (FV), left ventricle (LV), and superior mesenteric vein (SMV), respectively. The fate of challenge larvae was examined by autoradiography of host tissues and by recovery of adult worms. It was found that both normal and immune elimination were site-dependent. In control mice 45%-60% of cercarial penetrants and lung schistosomula injected intomore » the FV and LV were recoverable as adult worms, while a significantly greater number (70%-85%) were recoverable when lung schistosomula were injected into the SMV. In immunized mice, parasites introduced as either cercariae or FV-injected schistosomula were both highly sensitive to immune elimination. LV-injected schistosomula were also sensitive but to a slightly lesser degree. In contrast, schistosomula placed directly in the liver by SMV injection were totally insensitive to immune elimination. It was concluded that elimination of schistosomula in irradiated cercaria-immunized mice occurs in the lungs and/or in the systemic organs, but not in the liver. Also, it was concluded that immune elimination is not a rapid process, since more than 7 days were required after intravascular challenge for the development of demonstrable differences between control and immunized mice.« less