Cryptosporidium: Isolate variation and humoral responses to sporozoite antigens.

Persistent Link:
http://hdl.handle.net/10150/184391
Title:
Cryptosporidium: Isolate variation and humoral responses to sporozoite antigens.
Author:
Mead, Jan Renee.
Issue Date:
1988
Publisher:
The University of Arizona.
Rights:
Copyright © is held by the author. Digital access to this material is made possible by the University Libraries, University of Arizona. Further transmission, reproduction or presentation (such as public display or performance) of protected items is prohibited except with permission of the author.
Abstract:
The humoral response of humans, calves and horses to Cryptosporidium sporozoite antigens was evaluated using a western blot technique. Sera from calves, humans and horses were obtained at various times following the detection of infection. Sera were reacted with detergent-solubilized, sporozoite antigens form sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE). The number of antigens recognized by immune sera from humans and animals increased with time post infection (P.I.). A 20 kDa antigen appeared to be a major sporozoite surface determinant since it was labelled via membrane protein biotinylation and recognized by mouse monoclonal antibodies using indirect immunofluorescence and western blotting. Detectable recognition of the 20 kDa band occurred in 3 week post infection (P.I.) sera from all species tested. Sera reactivity to the 20 kDa band diminished significantly within 5 months P.I. when infected humans had no further recurrence of cryptosporidial diarrhea. In contrast, 12 month P.I. sera from an individual constantly exposed to oocysts under working conditions was as strongly reactive as the 3 week convalescent sera. Therefore, reactivity to the 20 kDa antigen appeared to be a good indicator of exposure to Cryptosporidium. Anti-sporozoite indirect immunofluorescent titers decrease in reactivity from convalescent to post convalescent sera which correlated with western blot results. Chromosomal DNA of five Cryptosporidium parvum isolates and one Cryptosporidium baileyi isolate were compared by field inversion gel electrophoresis (FIGE). FIGE analyses of parasite DNA prepared from purified sporozoites versus intact oocysts showed no observable differences. Chromosomal DNA migration patterns of the five Cryptosporidium parvum isolates were indistinguishable. Distinct differences in chromosomal DNA were evident between the Cryptosporidium baileyi and Cryptosporidium parvum isolates, yet the overall pattern was similar. Five C. parvum isolates were also compared using two dimensional electrophoretic analyses. Silver stained patterns of sporozoite proteins showed a shift in a 106 kDa protein in three of the isolates. One isolate (Mexico) showed a complete absence of this protein (106 kDa) and the presence of an additional 40 kDa protein not found in any other isolate.
Type:
text; Dissertation-Reproduction (electronic)
Keywords:
Cryptosporidium.; Parasite antigens.; Apicomplexa.
Degree Name:
Ph.D.
Degree Level:
doctoral
Degree Program:
Microbiology and Immunology; Graduate College
Degree Grantor:
University of Arizona
Advisor:
Sterling, Charles R.

Full metadata record

DC FieldValue Language
dc.language.isoenen_US
dc.titleCryptosporidium: Isolate variation and humoral responses to sporozoite antigens.en_US
dc.creatorMead, Jan Renee.en_US
dc.contributor.authorMead, Jan Renee.en_US
dc.date.issued1988en_US
dc.publisherThe University of Arizona.en_US
dc.rightsCopyright © is held by the author. Digital access to this material is made possible by the University Libraries, University of Arizona. Further transmission, reproduction or presentation (such as public display or performance) of protected items is prohibited except with permission of the author.en_US
dc.description.abstractThe humoral response of humans, calves and horses to Cryptosporidium sporozoite antigens was evaluated using a western blot technique. Sera from calves, humans and horses were obtained at various times following the detection of infection. Sera were reacted with detergent-solubilized, sporozoite antigens form sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE). The number of antigens recognized by immune sera from humans and animals increased with time post infection (P.I.). A 20 kDa antigen appeared to be a major sporozoite surface determinant since it was labelled via membrane protein biotinylation and recognized by mouse monoclonal antibodies using indirect immunofluorescence and western blotting. Detectable recognition of the 20 kDa band occurred in 3 week post infection (P.I.) sera from all species tested. Sera reactivity to the 20 kDa band diminished significantly within 5 months P.I. when infected humans had no further recurrence of cryptosporidial diarrhea. In contrast, 12 month P.I. sera from an individual constantly exposed to oocysts under working conditions was as strongly reactive as the 3 week convalescent sera. Therefore, reactivity to the 20 kDa antigen appeared to be a good indicator of exposure to Cryptosporidium. Anti-sporozoite indirect immunofluorescent titers decrease in reactivity from convalescent to post convalescent sera which correlated with western blot results. Chromosomal DNA of five Cryptosporidium parvum isolates and one Cryptosporidium baileyi isolate were compared by field inversion gel electrophoresis (FIGE). FIGE analyses of parasite DNA prepared from purified sporozoites versus intact oocysts showed no observable differences. Chromosomal DNA migration patterns of the five Cryptosporidium parvum isolates were indistinguishable. Distinct differences in chromosomal DNA were evident between the Cryptosporidium baileyi and Cryptosporidium parvum isolates, yet the overall pattern was similar. Five C. parvum isolates were also compared using two dimensional electrophoretic analyses. Silver stained patterns of sporozoite proteins showed a shift in a 106 kDa protein in three of the isolates. One isolate (Mexico) showed a complete absence of this protein (106 kDa) and the presence of an additional 40 kDa protein not found in any other isolate.en_US
dc.typetexten_US
dc.typeDissertation-Reproduction (electronic)en_US
dc.subjectCryptosporidium.en_US
dc.subjectParasite antigens.en_US
dc.subjectApicomplexa.en_US
thesis.degree.namePh.D.en_US
thesis.degree.leveldoctoralen_US
thesis.degree.disciplineMicrobiology and Immunologyen_US
thesis.degree.disciplineGraduate Collegeen_US
thesis.degree.grantorUniversity of Arizonaen_US
dc.contributor.advisorSterling, Charles R.en_US
dc.contributor.committeememberFriedman, Richard L.en_US
dc.contributor.committeememberJoens, Lynn A.en_US
dc.contributor.committeememberSammons, David W.en_US
dc.contributor.committeememberSinclair, Norval A.en_US
dc.identifier.proquest8814260en_US
dc.identifier.oclc701245573en_US
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