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  • CANCER  (2)
  • antibodies  (2)
  • 1
    Keywords: CANCER ; Germany ; human ; CLASSIFICATION ; QUANTIFICATION ; DISEASE ; DISEASES ; PROTEIN ; PROTEINS ; COMPLEX ; RESPONSES ; COMPLEXES ; INFECTION ; RISK-FACTORS ; ANTIGEN ; FLOW ; papillomavirus ; ASSOCIATION ; antibodies ; antibody ; virus ; IN-SITU ; ASSAY ; GLUTATHIONE ; NUMBER ; REPRODUCIBILITY ; CERVICAL-CANCER ; FUSION ; FUSION PROTEINS ; human papillomavirus ; HPV ; E6 ; case-control studies ; LINKED-IMMUNOSORBENT-ASSAY ; glutathione-S-transferase ; FUSION PROTEIN ; E6 ONCOPROTEIN ; SERUM ; CHEMISTRY ; ELISA ; case-control study ; PROGRAM ; RE ; case control studies ; multiplex ; MICROSPHERE IMMUNOASSAY
    Abstract: Background: More than 100 different human papillo-maviruses (HPVs) can cause proliferative diseases, many of which are malignant, such as cervical cancer. HPV serology is complex because infection and disease lead to distinct type-specific antibody responses. Using bead-based technology, we have developed an assay platform that allows the simultaneous detection of antibodies against up to 100 in situ affinity-purified recombinant HPV proteins. Methods: Twenty-seven HPV proteins were expressed as glutathione S-transferase fusion proteins and affinity-purified in one step by incubation of glutathione-displaying beads in bacterial lysate. Spectrally distinct bead sets, each carrying one particular antigen, were mixed, incubated with serum, and differentiated in a flow cytometer-like analyzer (xMAP; Luminex Corp). Antibodies bound to the antigens were detected via fluorescent secondary reagents. We studied 756 sera from 2 case-control studies of cervical cancer. Results: Glutathione S-transferase fusion proteins bound with high affinity to glutathione-displaying beads (K-d = 6.9 X 10(-9) mol/L). The dynamic range of multiplex serology covered 1.5 orders of magnitude, and antibodies were detected at serum dilutions 〉 1:1 000 000. Imprecision (median CV) was 〈= 5.4%, and assay reproducibility was high (R-2 = 0.97). Results on clinical samples showed high concordance with ELISA (kappa = 0.846), but multiplex serology exhibited increased detection of weak antibody responses. Antibodies to the E6 oncoproteins of the rare HPV types 52 and 58 were associated with cervical cancer (P 〈 0.001). Conclusion: Multiplex serology enables antibody analyses of large numbers of sera against up to 100 antigens in parallel and has the potential to replace ELISA technology. (c) 2005 American Association for Clinical Chemistry
    Type of Publication: Journal article published
    PubMed ID: 16099939
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  • 2
    Keywords: CANCER ; EXPRESSION ; CLINICAL-TRIAL ; Germany ; human ; INFORMATION ; PROTEIN ; PROTEINS ; PATIENT ; RESPONSES ; INFECTION ; T cell ; T-CELLS ; E7 ; IMMUNE-RESPONSES ; antibodies ; antibody ; PARTICLES ; TRIAL ; NEOPLASIA ; HUMANS ; WOMEN ; cervical cancer ; cervical intraepithelial neoplasia ; CERVICAL-CANCER ; EFFICACY ; FUSION ; LYMPHOCYTES ; VIRUS-LIKE PARTICLES ; HPV ; VACCINE ; SAFETY ; immune response ; IMMUNE-RESPONSE ; IMMUNOTHERAPY ; intraepithelial neoplasia ; T-LYMPHOCYTES ; vaccination ; HUMAN-PAPILLOMAVIRUS TYPE-16 ; L1 ; T lymphocyte ; DOUBLE-BLIND ; PREVALENCE ; T lymphocytes ; glutathione-S-transferase ; YOUNG-WOMEN ; IMMUNIZATION ; ONCOLOGY ; PERSISTENT ; RECOMBINANT ; END ; GRADE ; RECIPIENTS ; development ; EVENTS ; PERSISTENCE ; USA ; HUMAN PAPILLOMAVIRUSES ; IMPROVEMENT ; immune responses ; clinical trial ; CIN
    Abstract: Persistent infection with human papillomaviruses (HPV) is a prerequisite for the development of cervical cancer. Vaccination with virus-like particles (VLP) has demonstrated efficacy in prophylaxis but lacks therapeutic potential. HPV16 L1E7 chimeric virus-like particles (CVLP) consist of a carboxy-terminally truncated HPV16L1 protein fused to the amino-terminal part of the HPV16 E7 protein and self-assemble by recombinant expression of the fusion protein. The CVLP are able to induce L1- and E7-specific cytotoxic T lymphocytes. We have performed a first clinical trial to gain information about the safety and to generate preliminary data on the therapeutic potential of the CVLP in humans. A randomized, double blind, placebo-controlled clinical trial has been conducted in 39 HPV16 mono-infected high grade cervical intraepithelial neoplasia (CIN) patients (CIN 2/3). Two doses (75 mu g or 250 mu g) of CVLP were applied. The duration of the study was 24 weeks with 2 optional visits after another 12 and 24 weeks. The vaccine showed a very good safety profile with only minor adverse events attributable to the immunization. Antibodies with high titers against HPV16 L1 and low titers against HPV16 E7 as well as cellular immune responses against both proteins were induced. Responses were equivalent for both vaccine concentrations. A trend for histological improvement to CIN I or normal was seen in 39% of the patients receiving the vaccine and only 25% of the placebo recipients. Fifty-six percent of the responders were also HPV16 DNA-negative by the end of the study. Therefore, we demonstrated evidence for safety and a nonsignificant trend for the clinical efficacy of the HPV16 L1E7 CVLP vaccine. (c) 2007 Wiley-Liss, Inc
    Type of Publication: Journal article published
    PubMed ID: 17721997
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