Reactivity pattern of 92 monoclonal antibodies with 15 human papillomavirus types.
Most anti-human papillomavirus (HPV) capsid antibody assays are based on virus-like particles (VLP). We evaluated glutathione S-transferase (GST)-L1 fusion proteins as ELISA antigens for determining type specificity and cross-reactivity of 92 VLP-specific monoclonal antibodies (mAb) generated against nine mucosal alpha papillomavirus types of species 7, 9 and 10. The antibody panel included 25 new mAb, and 24 previously published mAb are further characterized. We determined the cross-reactivity patterns with 15 different HPV types representing 6 species (alpha1, 2, 4, 7, 9 and 10) and neutralization and cross-neutralization properties with HPV types 6, 11, 16, 18 and 45. Eighty-nine (97 %) of the antibodies including 34, 71 and 14 recognizing neutralizing, conformational and linear epitopes, respectively, reacted with the GST-L1 protein of the HPV type used as immunogen, with log titres ranging from 2.0 to 7.3. Of these 89 antibodies, 52 % were monotypic, 20 % showed intra-species and 28 % inter-species cross-reactivity. Log neutralization titres to the immunogen HPV ranged from 1.7 to 5.6. A single cross-neutralizing mAb (H6.L12) was found. ELISA titres were always higher than neutralization titres. All neutralizing epitopes were conformational and mostly type-specific. Our data show that bacterially expressed, affinity-purified GST-L1 fusion proteins display a broad variety of epitopes and thus are well suited for detection of HPV antibodies. Cross-reactivity is associated with linear as well as conformational epitopes. Distantly related mucosal and skin alpha papillomaviruses share some conformational epitopes and the phylogenetic L1-based species definition may not define a serological unit since no species-specific epitope was found.
Rizk
Raeda Z
RZ
Department of Genome Modifications and Cancer, Infection and Cancer Program, German Cancer Research Center (DKFZ), Im Neuenheimer Feld 242, 69120 Heidelberg, Germany.
Christensen
Neil D
ND
Departments of Pathology and Microbiology and Immunology, The Jake Gittlen Cancer Research Foundation, College of Medicine, Pennsylvania State University, Hershey, PA 17033, USA.
Michael
Kristina M
KM
Department of Genome Modifications and Cancer, Infection and Cancer Program, German Cancer Research Center (DKFZ), Im Neuenheimer Feld 242, 69120 Heidelberg, Germany.
Müller
Martin
M
Department of Genome Modifications and Cancer, Infection and Cancer Program, German Cancer Research Center (DKFZ), Im Neuenheimer Feld 242, 69120 Heidelberg, Germany.
Sehr
Peter
P
Department of Genome Modifications and Cancer, Infection and Cancer Program, German Cancer Research Center (DKFZ), Im Neuenheimer Feld 242, 69120 Heidelberg, Germany.
Waterboer
Tim
T
Department of Genome Modifications and Cancer, Infection and Cancer Program, German Cancer Research Center (DKFZ), Im Neuenheimer Feld 242, 69120 Heidelberg, Germany.
Pawlita
Michael
M
Department of Genome Modifications and Cancer, Infection and Cancer Program, German Cancer Research Center (DKFZ), Im Neuenheimer Feld 242, 69120 Heidelberg, Germany.
D052685:Alphapapillomavirus / Q000145:classification / Q000235:genetics* / Q000276:immunology*; D000911:Antibodies, Monoclonal; D000914:Antibodies, Viral / Q000276:immunology; D000918:Antibody Specificity; D036022:Capsid Proteins / Q000276:immunology; D003429:Cross Reactions; D000939:Epitopes / Q000032:analysis; D052162:Human papillomavirus 16 / Q000235:genetics / Q000276:immunology; D006801:Humans; D009500:Neutralization Tests; D009856:Oncogene Proteins, Viral / Q000276:immunology
D000911:Antibodies, Monoclonal; D000914:Antibodies, Viral; D036022:Capsid Proteins; D000939:Epitopes; D009856:Oncogene Proteins, Viral; C063317:L1 protein, Human papillomavirus type 16