Isolation of cDNA clones for differentially expressed genes of the human parasite Schistosoma mansoni.

Davis, Alan H.; Blanton, Ronald E.; Rottman, Fritz M.; Maurer, Russell

doi:10.1073/pnas.83.15.5534

Cited by 14 publications

(12 citation statements)

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“…We previously reported the isolation of a 313-bp adult cDNA clone, clone 10-3, from adult S. mansoni (10). Initial Northern blot hybridization of adult and cercarial mRNAs with the 10-3 cDNA used as a probe indicated the presence of two adult transcripts and a single cercarial transcript.…”

Section: Resultsmentioning

confidence: 99%

“…The data are consistent with the fact that the largest band on Northern blots (950 bases) represents at least two transcripts which contain five tandem repeats. One of these transcripts contained a 27-base insertion 5 10 ,ug of genomic DNA digested with the respective restriction endonuclease, fractionated on 0.75% agarose gels, transferred to a Gene Screen nylon membrane, cross-linked to the membrane with UV light, and hybridized with radiolabeled probe. (A) End-labeled oligonucleotide G was used as the hybridization probe.…”

Section: Discussionmentioning

confidence: 99%

“…We have previously described the isolation of an adult S. mansoni cDNA clone, designated , that encodes an antigen that is recognized by sera from infected humans (10). Expression of transcripts homologous to 10-3 is developmentally regulated, and different-sized transcripts are expressed in the adult and cercarial stages.…”

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confidence: 99%

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Tandemly repeated exons encode 81-base repeats in multiple, developmentally regulated Schistosoma mansoni transcripts.

Davis¹,

Davis²,

Carroll³

et al. 1988

Mol. Cell. Biol.

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The adult Schistosoma mansoni cDNA clone 10-3 encodes an antigen that is recognized by sera from infected humans. We characterized multiple developmentally regulated transcripts homologous to the 10-3 cDNA and portions of the complex genomic loci encoding those transcripts. Transcripts of approximately 950, 870, and 780 nucleotides were expressed in adults, whereas only the 780-nucleotide transcript was observed in the larval stage. These transcripts were highly similar, containing variable numbers of identical direct tandem repeats of 81 bases. Although the sequence of the repeating elements and sequences 3' to them were identical in ali the transcripts, sequences 5' of the repeating elements exhibited variations, including a 27-base insertion, alternative start sites for transcription, and alternate 5' exon usage. These transcripts appeared to be derived in part by the developmentally controlled alternative splicing of small exons and the use of alternative transcription initiation sites from the one or two complex loci of at least 40 kilobase pairs. Each 81-base repeat in the transcripts was encoded by three dispersed 27-base-pair exons. These 27-base-pair exons were contained within highly conserved, reiterated 3-kilobase-pair genomic tandem arrays.

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Section: Resultsmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

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Tandemly repeated exons encode 81-base repeats in multiple, developmentally regulated Schistosoma mansoni transcripts.

Davis¹,

Davis²,

Carroll³

et al. 1988

Mol. Cell. Biol.

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“…This antigen named Sm44 has a specific antigenic expression since only serum related to the chronic phase is able to recognize it although in terms of RNA expression the Sm44 antigen of S. mansoni is also present in other evolutionary phases of parasitism but is excluded by the immune system perhaps by some block due to host molecule incorporation or different distribution during the development of the parasite in vertebrate hosts, similar to other antigens described previously (Aronstein & Strand 1983, Harn et al 1985, Davies et al 1986, Stein & David 1986, Bobek et al 1986, Grossman et al 1990, Menrath et al 1995.…”

Section: Discussionmentioning

confidence: 80%

Cloning and Characterization of a Schistosoma mansoni cDNA Clone with a Specific Antigenic Expression during Development in a Vertebrate Host

Coelho-Castelo¹,

Rodrigues²

1997

Mem. Inst. Oswaldo Cruz

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“…Several S. mansoni proteins have recently been molecularly cloned and expressed in Escherichia coli; however, none of these appears to be localized on the surface of early developmental stages (2)(3)(4)(5).…”

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confidence: 99%

Cloning of a cDNA encoding a surface antigen of Schistosoma mansoni schistosomula recognized by sera of vaccinated mice.

Dalton¹,

Tom²,

Strand³

1987

Proc. Natl. Acad. Sci. U.S.A.

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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. 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 constructed in Xgtll. 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.High levels of resistance to Schistosoma mansoni challenge infection can be induced in mice, rats, and baboons by vaccination with radiation-attenuated cercariae. At least in mice and rats, this resistance can be passively transferred to naive recipients using serum or purified immunoglobulins, indicating the importance of humoral immune responses in this resistance. The most consistent transfer of resistance is achieved when immune sera are administered while the schistosomula are in residence in the lungs. All such sera, including those of vaccinated mice, rats, and rabbits and of infected rats, contain antibodies that bind to the surface of lung-stage and newly transformed schistosomula. In contrast, sera from mice chronically infected with S. mansoni do not consistently transfer resistance, and antibodies in these sera do not bind to the surface of lung-stage schistosomula (reviewed in ref. 1). It is therefore likely that resistance following vaccination with radiation-attenuated cercariae is mediated by antibody binding to epitopes exposed on the surface of newly transformed and lung-stage schistosomula.Several S. mansoni proteins have recently been molecularly cloned and expressed in Escherichia coli; however, none of these appears to be localized on the surface of early developmental stages (2-5).Our approach to identifying potential immunoprophylactic antigens was to prepare monoclonal antibodies (mAb) from spleens of vaccinated mice and to screen such antibodies for their ability to immunoprecipitate surface-labeled schistosomular polypeptides. By this method we obtained a mAb, designated 654B2/1, that reacted with a single 18-kDa surface-labeled polypeptide. We purified this antigen and prepared a polyclonal antiserum (anti-654) against it. Immunofluorescence studies using anti-654 showed that some epitopes on the antigen are exposed not only on the surface of newly transformed schistosomula but also on the surface of lung-stage schistos...

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Isolation of cDNA clones for differentially expressed genes of the human parasite Schistosoma mansoni.

Cited by 14 publications

References 38 publications

Tandemly repeated exons encode 81-base repeats in multiple, developmentally regulated Schistosoma mansoni transcripts.

Tandemly repeated exons encode 81-base repeats in multiple, developmentally regulated Schistosoma mansoni transcripts.

Cloning and Characterization of a Schistosoma mansoni cDNA Clone with a Specific Antigenic Expression during Development in a Vertebrate Host

Cloning of a cDNA encoding a surface antigen of Schistosoma mansoni schistosomula recognized by sera of vaccinated mice.

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