Please use this identifier to cite or link to this item: https://ahro.austin.org.au/austinjspui/handle/1/13039
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dc.contributor.authorApostolidis, V Aen
dc.contributor.authorBrowne, K Aen
dc.contributor.authorSmyth, Mark Jen
dc.contributor.authorTrapani, Joseph Aen
dc.date.accessioned2015-05-16T02:48:53Z
dc.date.available2015-05-16T02:48:53Z
dc.date.issued1995-08-01en
dc.identifier.citationMolecular Immunology; 32(12): 909-17en
dc.identifier.govdoc7565817en
dc.identifier.otherPUBMEDen
dc.identifier.urihttp://ahro.austin.org.au/austinjspui/handle/1/13039en
dc.description.abstractGranzyme B (also termed fragmentin 2) is a prototypic member of a subfamily of serine proteases expressed in the cytoplasmic granules of cytotoxic T lymphocytes and natural killer cells, and has been implicated in the destruction of targeted cells. Studies on the role of all granzymes in the cytolytic response would be greatly facilitated by the availability of specific anti-granzyme antisera. Three synthetic peptides corresponding to amino acid residues 1-17, 92-109 and 139-157 of human granzyme B were predicted to be immunogenic in the mouse, based on their hydrophilicity, accessibility to solvent, polymorphism with respect to mouse granzyme B and by comparison with X-ray crystallographic models of the rat mast cell protease II. Each peptide was conjugated to keyhole limpet hemocyanin and used to produce monoclonal antibodies in BALB/c mice. The monoclonal antibodies produced generally exhibited strong and specific reactivity with the respective immunizing peptide. However, only those antibodies detecting the peptide corresponding to residues 139-157 were able to detect native or denatured granzyme B, in direct binding studies with purified granzyme B or by immunoblotting. As an alternative approach for antiserum production, mice were immunized with whole, proteolytically active granzyme B isolated by immuno-affinity purification from NK tumour cell lysates, using one of the monoclonal antibodies generated. Despite the overall structural similarities between the various human granzymes, these mouse antisera surprisingly reacted only with granzyme B. Indeed, the reactivity of these polyclonal antisera was specifically abrogated by preincubation with the peptide corresponding to amino acid residues 139-157. This peptide stretch therefore represents an immunodominant portion of the granzyme B molecule in the mouse. Given the analogous structures of serine protease families expressed in leukocytes, these findings have implications for the production of monospecific antisera to granzymes and related proteases.en
dc.language.isoenen
dc.subject.otherAmino Acid Sequenceen
dc.subject.otherAnimalsen
dc.subject.otherAntibodies, Monoclonalen
dc.subject.otherGranzymesen
dc.subject.otherHumansen
dc.subject.otherImmunodominant Epitopes.chemistry.geneticsen
dc.subject.otherKiller Cells, Natural.immunologyen
dc.subject.otherMiceen
dc.subject.otherMolecular Sequence Dataen
dc.subject.otherPeptide Fragments.chemistry.genetics.immunologyen
dc.subject.otherProtein Denaturationen
dc.subject.otherRatsen
dc.subject.otherSerine Endopeptidases.chemistry.genetics.immunologyen
dc.subject.otherT-Lymphocytes, Cytotoxic.immunologyen
dc.titleThe peptide loop consisting of amino acids 139-157 of human granzyme B (fragmentin 2) contains an immunodominant epitope recognized by the mouse.en
dc.typeJournal Articleen
dc.identifier.journaltitleMolecular immunologyen
dc.identifier.affiliationCellular Cytotoxicity Laboratory, Austin Research Institute, Austin Hospital, Heidelberg, Victoria, Australiaen
dc.description.pages909-17en
dc.relation.urlhttps://pubmed.ncbi.nlm.nih.gov/7565817en
dc.type.austinJournal Articleen
item.languageiso639-1en-
item.openairecristypehttp://purl.org/coar/resource_type/c_18cf-
item.openairetypeJournal Article-
item.grantfulltextnone-
item.cerifentitytypePublications-
item.fulltextNo Fulltext-
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