Linked Life Data

19329278

Source:http://linkedlifedata.com/resource/pubmed/id/19329278

Statements in which the resource exists as a subject.
PredicateObject
rdf:type
lifeskim:mentions
pubmed:issue
1
pubmed:dateCreated
2009-6-1
pubmed:abstractText
In primary adrenocortical failure (Addison's disease) caused by autoimmunity, autoantibodies to the steroidogenic cytochrome P450 enzyme 21-hydroxylase (21OH) are detected in the majority of patients. It is currently uncertain whether the autoantibodies themselves participate in the pathogenesis, or if they merely reflect an on-going T cell mediated response. The identification of T cells reactive with 21OH, if any, has been hampered by the lack of a high-quality antigen. In the current study recombinant human 21OH has been expressed in Spodoptera frugiperda insect cells using a baculovirus expression system. Recombinant enzymatically active 21OH was purified to apparent homogeneity by immobilized metal ion affinity chromatography. The purified enzyme was highly immunogenic in immunized SJL/J mice, and immune responses to 21OH-derived peptides assayed as T cell proliferation and interferon gamma production could be invoked after priming with the recombinant protein. Furthermore, purified 21OH was recognized by sera from patients with autoimmune Addison's disease, and it could block the binding of radiolabeled in vitro translated 21OH in a sensitive fluid-phase radioimmunoassay. We conclude that the recombinant preparation of 21OH presented here is of sufficient purity and quality to be used for studies of cellular and humoral immunity in autoimmune Addison's disease.
pubmed:language
eng
pubmed:journal
pubmed:citationSubset
IM
pubmed:chemical
pubmed:status
MEDLINE
pubmed:month
Aug
pubmed:issn
1095-9157
pubmed:author
pubmed:issnType
Electronic
pubmed:volume
33
pubmed:owner
NLM
pubmed:authorsComplete
Y
pubmed:pagination
58-67
pubmed:meshHeading
pubmed-meshheading:19329278-Addison Disease, pubmed-meshheading:19329278-Animals, pubmed-meshheading:19329278-Autoantibodies, pubmed-meshheading:19329278-Autoantigens, pubmed-meshheading:19329278-Baculoviridae, pubmed-meshheading:19329278-Cell Line, pubmed-meshheading:19329278-Cell Proliferation, pubmed-meshheading:19329278-Chromatography, Affinity, pubmed-meshheading:19329278-Cytochrome P-450 Enzyme System, pubmed-meshheading:19329278-Humans, pubmed-meshheading:19329278-Immune Sera, pubmed-meshheading:19329278-Immunization, pubmed-meshheading:19329278-Interferon-gamma, pubmed-meshheading:19329278-Lymphocyte Activation, pubmed-meshheading:19329278-Mice, pubmed-meshheading:19329278-Peptides, pubmed-meshheading:19329278-Spodoptera, pubmed-meshheading:19329278-Steroid 21-Hydroxylase, pubmed-meshheading:19329278-T-Lymphocytes, pubmed-meshheading:19329278-Transgenes
pubmed:year
2009
pubmed:articleTitle
The purification and application of biologically active recombinant steroid cytochrome P450 21-hydroxylase: the major autoantigen in autoimmune Addison's disease.
pubmed:affiliation
Department of Medicine, Section for Endocrinology, Institute of Medicine, Haukeland University Hospital, Bergen, Norway. eirik.bratland@med.uib.no
pubmed:publicationType
Journal Article, Research Support, Non-U.S. Gov't