Detection of human papillomavirus oncoprotein E7 in liquid-based cytology.
The selection and characterization of a set of mouse mAbs against high-risk human papillomavirus (HPV) E7 oncoprotein and the development of protocols for immunocytochemistry (ICC) are described here. A large number of antibodies raised towards HPV16 and 18 E7 were tested for high-risk specificity by ELISA using a panel of HPV E7 proteins. Antibodies detecting low-risk E7 were discarded, resulting in 38 high-risk HPV E7-specific antibodies. The corresponding epitopes were mapped using overlapping HPV E7 fragments displayed on phage particles. Functionality in ICC against formalin-fixed cervical cancer cell lines was demonstrated for ten mAbs; their high-risk specificity was confirmed by Western blot analysis and ICC on transiently transformed cells expressing high- or low-risk HPV E7. These mAbs were specific for one or several of the high-risk strains HPV16, 18, 31, 35 and 45. Specific E7 staining of liquid-based cytology (LBC) samples was demonstrated for seven mAbs and optimized protocols were established. The E716-41 and E718-79 mAbs demonstrated particularly strong and specific staining of cells stored in LBC fluid for at least 6 months. It is proposed that the high-risk HPV E7 staining protocols established in this study may have the potential to be included in a complementary test for the detection and identification of malignantly transformed cells, in for example atypical squamous cells of undetermined significance samples.
Institute of Biomedicine, Department of Microbiology and Immunology, Gothenburg University, SE-405 30 Gothenburg, Sweden.|Fujirebio Diagnostics AB, Elof Lindälvs gata 13, SE-414 55 Gothenburg, Sweden.
Maria
M
Lidqvist
Fujirebio Diagnostics AB, Elof Lindälvs gata 13, SE-414 55 Gothenburg, Sweden.
Olle
O
Nilsson
Institute of Biomedicine, Department of Microbiology and Immunology, Gothenburg University, SE-405 30 Gothenburg, Sweden.
Jan
J
Holmgren
Department of Gynaecology and Obstetrics, Jena University Hospital, Friedrich-Schiller-University, Jena, 07743 Jena, Germany.
Sebastian
S
Hölters
Fujirebio Diagnostics AB, Elof Lindälvs gata 13, SE-414 55 Gothenburg, Sweden.
Eva
E
Röijer
Department of Gynaecology and Obstetrics, Jena University Hospital, Friedrich-Schiller-University, Jena, 07743 Jena, Germany.
Matthias
M
Dürst
Fujirebio Diagnostics AB, Elof Lindälvs gata 13, SE-414 55 Gothenburg, Sweden.
Christian
C
Fermér
D000818:Animals; D000911:Antibodies, Monoclonal; D000914:Antibodies, Viral; D015153:Blotting, Western; D055088:Early Detection of Cancer / Q000379:methods; D018604:Epitope Mapping; D005260:Female; D006801:Humans; D007150:Immunohistochemistry / Q000379:methods*; D051379:Mice; D008807:Mice, Inbred BALB C; D050725:Papillomavirus E7 Proteins / Q000032:analysis*; D030361:Papillomavirus Infections / Q000175:diagnosis*; D012680:Sensitivity and Specificity
D000911:Antibodies, Monoclonal; D000914:Antibodies, Viral; D050725:Papillomavirus E7 Proteins