Detection of T-cell receptor beta chain mRNA in frozen and paraffin-embedded biopsy tissue using digoxigenin-labelled oligonucleotide probes in situ.
In situ hybridization techniques using a cocktail of digoxigenin-labelled T-cell receptor (TcR) constant (C) region beta oligonucleotide probes were used to detect TcR beta mRNA in frozen and paraffin-embedded tissue sections. The specificity of the C beta cocktail was confirmed by Northern blot analysis. The TcR C beta cocktail successfully hybridized to T cells in frozen and paraffin-embedded tissue obtained from patients with inflammatory arthropathies, B- and T-cell non-Hodgkin's lymphoma (NHL), and reactive tonsillitis, and showed staining patterns comparable to those obtained by conventional immunohistological detection of T cells. This is the first report of in situ studies using labelled TcR C beta oligonucleotide probes and may indicate the feasibility of investigating clonal T-cell populations using digoxigenin-labelled clonospecific probes in clinical samples in situ.
Hodges
E
E
Tenovus Research Laboratory, Southampton University Hospitals, Hants, U.K.
Howell
W M
WM
Tyacke
S R
SR
Wong
R
R
Cawley
M I
MI
Smith
J L
JL
D001706:Biopsy; D015152:Blotting, Northern; D004076:Digoxigenin; D005629:Frozen Sections; D006801:Humans; D017403:In Situ Hybridization; D015345:Oligonucleotide Probes; D016612:Paraffin Embedding; D012333:RNA, Messenger / Q000032:analysis*; D016693:Receptors, Antigen, T-Cell, alpha-beta / Q000032:analysis* / Q000235:genetics
D015345:Oligonucleotide Probes; D012333:RNA, Messenger; D016693:Receptors, Antigen, T-Cell, alpha-beta; D004076:Digoxigenin