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Biomedical subjects

C L Chappell

Publications and source records attributed to C L Chappell.

49 records · Page 3Linked to original sources
PubMed1989

Coenuriasis in a spectacled langur (Presbytis obscura): praziquantel treatment and the antibody response to cyst antigens.

A diagnosis of coenuriasis was made in a spectacled langur raised in captivity. Multiple cysts removed from the subcutaneous tissues and later from the abdominal cavity were identified as coenuri, typical of the genus Taenia. Post-surgical treatment of the remaining cysts with praziquantel was assessed with whole body computerized tomography (CT). CT at 6 weeks post-treatment revealed a reduction in size and increased calcification of abdominal cysts as compared to pretreatment CT. Cyst fluid antigens in ELISA assays showed a high titer (1:5, 120) IgG response in the langur serum, while no IgM response could be detected. No decrease in IgG titer was seen 6 weeks after treatment. Immunoblot analyses identified several parasite-specific antigens with apparent molecular weights of greater than 92.5 (3 bands), 88, 41, 37, and 34 kDa.

Animals↗
PubMed1988

Schistosoma mansoni: expression and role of cysteine proteinases in developing schistosomula.

The adult stage of Schistosoma mansoni utilizes host hemoglobin as a nutrient source. A proteolytic enzyme (SMw32) that has "hemoglobinase" activity is secreted into the parasite gut where it appears to be rapidly activated by glutathione released from host red blood cells. In the present study the expression of this proteinase, in developing schistosomula, has been correlated with digestive tract development and a dramatic rise in enzyme activity as early as Days 8-10 of culture. No evidence of the SMw32 proteinase was found in eggs, cercariae, or in newly transformed larvae. Further, the proteinase expressed at Days 8-10 is indistinguishable from the adult worm enzyme. In the larvae, indirect immunofluorescence with an anti-SMw32 monoclonal antibody showed that the proteinase is found throughout the developing cecum. The importance of cysteine proteinases to parasite development was also studied using a specific enzyme inhibitor, Ep-459. In cultures containing Ep-459 most (75%) of the schistosomula failed to survive the 18-day study period. Moreover, those that did survive showed a decrease in their growth (body length). These data suggest that the SMw32 proteinase is a developmentally regulated enzyme and that cysteine proteinase activity is essential in providing nutrients for the growth and survival of this parasite in its mammalian host. Thus, this proteinase may be an important target for chemotherapeutic intervention.

Animals↗
PubMed1988

The hypersensitivity response to the adult worm proteinase, SMw32, in Schistosoma mansoni infected mice.

A cysteine proteinase (SMw32) from the digestive tract of an adult Schistosoma mansoni worm has previously been purified and characterized. During the course of infection with this parasite, a strong immune response to the enzyme occurs. We now have confirmed the presence of anti-proteinase IgE and IgG in S. mansoni infected mice and have investigated the in vivo cellular response to proteinase in infected and uninfected mice. Immediate and delayed type hypersensitivities were detected in uninfected mice sensitized by multiple injections of proteinase. In S. mansoni infected mice, immediate hypersensitivity reactions were seen at 6 and 8 weeks following infection, coincident with the increase in anti-proteinase IgE antibody. Histological sections of the injection site confirmed the presence of degranulating mast cells. In contrast, delayed type hypersensitivity could not be detected at any time during the course of the infection. In the murine model of acute infection, immediate hypersensitivity to the SMw32 proteinase was predictive of infection.

Animals↗
PubMed1988

Antibody response to a purified parasite proteinase (SMw32) in Schistosoma mansoni infected mice.

Indirect immunofluorescence of Schistosoma mansoni adult worm sections has revealed that the early immunoglobulin response is directed toward the parasite digestive tract. One of the components of the worm gut is a cysteine proteinase which degrades host hemoglobin ingested by the parasite. In this report the purified proteinase (SMw32) was used in ELISA and immunoblot analyses to study the specific antibody response during the course of an acute infection. We have found high titer IgG antibody in S. mansoni infected, but not uninfected, mice. The anti-proteinase response involves IgM, IgG1, IgG2a, and IgE isotypes. Total IgM and IgG levels increased by week 3 post-infection and remained elevated throughout the study (7 weeks). Increased titers (IgM, IgG) of specific anti-proteinase were also apparent by week 3 post-infection, long before fecal eggs were detectable. Mean anti-proteinase IgG stabilized at high titer by week 5 post-infection, while IgM titers decreased to near background levels. Anti-proteinase IgE was first detectable at week 4 and reached peak titers by weeks 6 and 7. The strong antibody response to the purified SMw32 proteinase is consistent with the early reactivity of S. mansoni infected mice and humans to a 31 kDa component of the worm gut described by others.

Animals↗
PubMed1987

Purification of cysteine proteinases from adult Schistosoma mansoni.

Proteolytic activity against hemoglobin and low molecular weight synthetic substrates has been previously found in homogenates and excretion/secretion products of adult Schistosoma mansoni worms. This activity is stimulated in the presence of thiol compounds and is maximally active at acidic pH. To characterize further this proteolytic activity, lyophilized adult worms were extracted, and proteinases were isolated and purified. From extracts prepared in 0.2 M citrate buffer, pH 4.9, two proteinase species were purified to homogeneity by centrifugation, gel filtration, dialysis, and chromatofocusing chromatography. The proteinases, designated SMw32 and SMw28, have apparent molecular weights (SDS-PAGE) of 31,700 +/- 1400 and 27,800 +/- 1700, respectively. Both are thiol-dependent, acidic endopeptidases that cleave hemoglobin and a synthetic substrate, CBZ-arg-arg-AFC. A statistical comparison of amino acid compositions reveals that the proteinases are highly related.

Amino Acids↗
PubMed1987

Glutathione activation of a cysteine proteinase from Schistosoma mansoni.

A cysteine proteinase isolated from Schistosoma mansoni adults requires reduction by thiols for activation. The proteinase is located in the parasite digestive tract where it degrades hemoglobin released from host red blood cells. Reduced glutathione (GSH) has been shown to be effective in activation. Total glutathione concentration and the GSH/oxidized glutathione (GSSG) ratio were measured in whole blood lysate (3.2 mM, 284), serum (24 microM, 9.8) and material collected from the parasite digestive tract (4.2 mM, 137). The ratio of GSH/GSSG at which the enzyme displays half-maximal activity (Kox) is 1.0. Proteinase activation as a function of glutathione concentration and time was determined. The first-order reaction yielded a half-time of activation of 13 min at 5 mM. The second-order rate constant was 12.7 M-1 X min-1. The function of the proteinase and its possible regulation by glutathione activation are discussed.

Animals↗
PubMed1987

Schistosoma mansoni: anti-SMw32 proteinase response in vaccinated and challenged baboons.

Antibodies to a cysteinyl proteinase of the trematode Schistosoma mansoni have been detected in serum from infected mice and humans. We have evaluated antiproteinase responses in infected baboons and in baboons vaccinated with irradiated, cryopreserved schistosomules prior to challenge. Prechallenge sera and normal, uninfected control sera were nonreactive by ELISA and immunoblots. Serum antibodies were first detectable by ELISA at two months post-challenge in both challenged (C) and vaccinated-challenged (V-C) baboons (serum dilution 1:200). By four months post-challenge, ELISA absorbance values for subgroup C baboons were significantly higher than for V-C counterparts. The immunoblot technique provided a more sensitive means of detecting antibody early in the infection. One month post-challenge, 7 of 12 C and V-C sera (diluted 1:100) contained measurable anti-proteinase antibody. By month two, 12 of 12 were immunoblot-positive. Baboons vaccinated but not challenged (subgroup V) remained negative. The presence of the anti-proteinase antibody appears to be a sensitive and early marker for infection by S. mansoni.

Animals↗
PubMed1986

Schistosoma mansoni: proteinase activity of "hemoglobinase" from the digestive tract of adult worms.

A method of collecting samples from the Schistosoma mansoni digestive tract was used to study proteinase activity. Activity against hemoglobin and a low molecular weight synthetic substrate, carbobenzoxy-arginyl-arginyl-7-amino-4-trifluoromethylcoumarin, was demonstrated in the soluble fraction of material regurgitated by S. mansoni adults and was dependent on the addition of a thiol compound, cysteine, to the assays. Using sodium dodecyl sulfate-polyacrylamide gel electrophoresis and gel filtration chromatography (AcA54), two proteins with estimated mol wt of 32,500 and 28,500 were found in the regurgitant and were associated with proteinase activity against both hemoglobin and the synthetic substrate. Homogenates of intact worms showed greater specific activity (synthetic substrate) in the females. Further, in bisected worms proteinase activity paralleled protein content, suggesting that, once secreted into the lumen, proteinase activity was distributed throughout the worm digestive tract.

Animals↗
PubMed1981

Properties of the acid thiol proteinase from Schistosoma mansoni adults.

Seven beta-naphthylamine-linked peptides were tested as substrates for a previously described thiol proteinase of adult Schistosoma mansoni. The enzyme was not active on carbobenoxy-arginyl-arginyl-4-methoxy-2-naphthylamide and carbobenzoxy-alanyl-arginyl-arginyl-4-methoxy-2-naphthylamide. Enzyme activity was maximal at acidic pH (4.9-5.5) with similar optima for both macromolecular and peptide substrates. Activity of partially purified enzyme preparations against carbobenzoxy-arginyl-arginyl-4-methoxy-2-naphthylamide was stimulated more than 10-fold by thiols. The properties of this proteinase differ from those of proteolytic enzymes from the cercariae ad eggs of S. mansoni.

Animals↗
PubMed1979

Pili of Neisseria meningitidis: effect of media on maintenance of piliation, characteristics of Pili, and colonial morphology.

In contrast to information in the literature which indicates that meningococci rapidly lose pili upon cultivation in vitro, we found that piliation of meningococci could be maintained in vitro for 15 or more passages. Pili were present on all eight isolates tested, whether from asymptomatic carriers or from subjects with meningococcal disease. Complete loss of piliation occurred in the same two strains on two of the three media tested. On one medium (Thayer-Martin medium with supplement B), there was partial or complete loss of pili by all strains. The optimal medium for maintaining pili was chocolate agar with 1% IsoVitaleX; 95% or more of the microorganisms of six of the eight strains tested were piliated after 15 passages in vitro, and more than 60% of the microorganisms of the other two strains were piliated. Meningococci passed on this medium generally maintained their initial density of piliation (3 to 34 pili per diplococcus). The ability to predictably cultivate piliated meningococci in vitro and to select piliated and nonpiliated clones of the same strain should allow investigation of the biochemical and immunological properties of meningococcal pili as well as their possible role in the pathogenicity of Neisseria meningitidis.

Culture Media↗