Abstract
BackgroundThe diagnostic value of CSF tau for Creutzfeldt-Jakob disease (CJD) has been widely evaluated, showing a markedly disease-relative manner. However, the profiles of tau isoforms in CSF of CJD patients remain unknown. Here, we prepared the exon-specific antibodies against the peptides encoded by exon-2, exon-3 and exon-10 of human tau protein and evaluated the reactive profiles of tau in CSF samples from the patients with probable CJD.Methodology/Principal FindingsSequences encoding exon-2, exon-3 and exon-10 of human tau protein were cloned into a prokaryotic expression vector pGEX-2T. Using recombinant fusion proteins GST-E2, GST-E3 and GST-E10, three tau exon-specific antibodies were elicited. Reliable specificities of the prepared antibodies were obtained after a serial of purification processes, not only in recognizing the tau peptides encoded by exon-2, -3 and -10, but also in distinguishing six recombinant tau isoforms by Western blot and ELISA. Three predominant tau-specific bands were observed in CSF samples with the exon-specific and the commercial tau antibodies, respectively, showing different reactive profiles between the groups of probable CJD and non-CJD. A 65 KD band was detected only in the CSF samples from probable CJD patients, especially with the antibodies against exon-2 (Anti-tE2) and exon-10 (Anit-tE10). The appearances of 65 KD band in CSF correlated well with positive 14-3-3 in CSF and typical abnormality in EEG. Such band was not observed in the CSF samples of six tested genetic CJD patients.Conclusions/SignificanceThree exon-specific polyclonal antibodies were successfully prepared. Based on these antibodies, different CSF tau profiles in Western blots were observed between the groups of probable CJD and non-CJD. A disease-specific tau band emerged in the CSF samples from probable sporadic CJD, which may supply a new biomarker for screening sporadic CJD.
Highlights
Creutzfeldt-Jakob disease (CJD) is a rapidly progressive and fatal disorder of the central nervous system believed to be caused by prion [1]
The profiles of tau isoforms in cerebrospinal fluid (CSF) of CJD patients remain unknown, possibly due to lacking of tau isoform- or exon-specific antibodies
Our study provide the reliable human tau exon-specific polyclonal antibodies and the 65 KD tau-specific band may be used as a new biomarker for screening sporadic CJD
Summary
Creutzfeldt-Jakob disease (CJD) is a rapidly progressive and fatal disorder of the central nervous system believed to be caused by prion [1]. A definite diagnosis can only be made by neuropathological examination and demonstration of the pathological isoform of the prion protein (PrPSc) in central nervous tissues, either at biopsy or autopsy [3]. The diagnostic value of CSF tau for Creutzfeldt-Jakob disease (CJD) has been widely evaluated, showing a markedly disease-relative manner. The profiles of tau isoforms in CSF of CJD patients remain unknown. We prepared the exon-specific antibodies against the peptides encoded by exon-2, exon-3 and exon-10 of human tau protein and evaluated the reactive profiles of tau in CSF samples from the patients with probable CJD
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