Abstract |
The U1-70K protein is specifically bound to stemloop I of the U1 small nuclear RNA contained in the U1 small nuclear ribonucleoprotein complex ( U1 snRNP), which is involved in the splicing of pre-mRNA. All components of the U1 snRNP complex, including the U1-70K protein, are important autoantigens in patients with systemic lupus erythematosus (SLE) and mixed connective tissue disease ( MCTD). Here we describe for the first time the selection and characterization of recombinant human anti-U1-70K single chain autoantibody fragments (anti-hU1-70K scFv) from autoimmune patient-derived phage display antibody libraries. All scFv specifically recognize parts of the hU1-70K protein and its apoptotic 40-kDa cleavage product. In Western blotting assays a number of scFv preferentially recognize the 40-kDa apoptotic cleavage fragment of the U1-70K protein, suggesting a possible involvement of this apoptotic cleavage product in the autoimmune response of patients. The germline gene usage of these recombinant autoantibodies was also determined. Using several U1-70K deletion and point mutants of both human (h) and Drosophila melanogaster (Dm) origin, it was established that the U1-70K epitope that is recognized by the anti-hU1-70K scFv is located within the RNA binding domain.
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Authors | W G Degen, M Pieffers, E Welin-Henriksson, F H van den Hoogen, W J van Venrooij, J M Raats |
Journal | European journal of immunology
(Eur J Immunol)
Vol. 30
Issue 10
Pg. 3029-38
(Oct 2000)
ISSN: 0014-2980 [Print] Germany |
PMID | 11069087
(Publication Type: Journal Article, Research Support, Non-U.S. Gov't)
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Chemical References |
- Antibodies, Monoclonal
- Autoantibodies
- Autoantigens
- Complementarity Determining Regions
- DNA, Complementary
- Immunoglobulin Fragments
- Immunoglobulin Variable Region
- Peptide Fragments
- Recombinant Fusion Proteins
- Ribonucleoprotein, U1 Small Nuclear
- SNRNP70 protein, human
- immunoglobulin Fv
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Topics |
- Amino Acid Sequence
- Antibodies, Monoclonal
(immunology)
- Apoptosis
- Autoantibodies
(chemistry, genetics, immunology)
- Autoantigens
(immunology)
- Autoimmune Diseases
(immunology)
- Complementarity Determining Regions
- DNA, Complementary
(genetics)
- Genes, Immunoglobulin
- Humans
- Immunoglobulin Fragments
(immunology)
- Immunoglobulin Variable Region
(genetics)
- Jurkat Cells
- Lupus Erythematosus, Systemic
(immunology)
- Molecular Sequence Data
- Peptide Fragments
(pharmacology)
- Recombinant Fusion Proteins
(immunology)
- Ribonucleoprotein, U1 Small Nuclear
(immunology)
- Sequence Alignment
- Sequence Homology, Amino Acid
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