HPV antibody detection by ELISA with capsid protein L1 fused to glutathione S-transferase

Sehr, Peter; Müller, Martin; Höpfl, Reinhard; Widschwendter, Andreas; Pawlita, Michael

doi:10.1016/s0166-0934(02)00134-9

Cited by 122 publications

(141 citation statements)

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“…L1 proteins were expressed in E. coli bacteria as fusion proteins with N-terminal GST and a C-terminal tag epitope as described previously. 13 Fluorescence-labeled polystyrene beads (Luminex) were derivatized with glutathione casein. 14 GST-fusion proteins were specifically bound to glutathionecoated Luminex beads directly from the lysates.…”

Section: Multiplex Serologymentioning

confidence: 99%

Prospective seroepidemiologic study on the role of Human Papillomavirus and other infections in cervical carcinogenesis: Evidence from the EPIC cohort

et al. 2014

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To evaluate prospectively the association between serological markers of selected infections, including HPV, and risk of developing cervical cancer (CC) and precancer, we performed a nested case–control study within the European Prospective Investigation into Cancer and Nutrition (EPIC) study that included 184 cases of invasive CC (ICC), 425 cases of cervical intraepithelial neoplasia (CIN) grade 3 or carcinoma in situ (CIS), and 1,218 matched control women. At enrollment participants completed lifestyle questionnaires and provided sera. Subjects were followed‐up for a median of 9 years. Immunoassays were used to detect serum antibodies to Human Herpes Virus 2 (HHV‐2), Chlamydia trachomatis (CT), Chlamydia pneumoniae, L1 proteins of mucosal and cutaneous HPV types, E6/E7 proteins of HPV16/18, as well as to four polyomaviruses. Adjusted odds ratios (OR) [and 95% confidence intervals (CI)] for CIN3/CIS and ICC risk were respectively: 1.6 (1.2–2.0) and 1.8 (1.1–2.7) for L1 seropositivity to any mucosal HPV type, 1.0 (0.4–2.4) and 7.4 (2.8–19.7) for E6 seropositivity to HPV16/18, 1.3 (0.9–1.9) and 2.3 (1.3–4.1) for CT seropositivity, and 1.4 (1.0–2.0) and 1.5 (0.9–2.6) for HHV‐2 seropositivity. The highest OR for ICC was observed for HPV16 E6 seropositivity [OR = 10.2 (3.3–31.1)]. Increasing number of sexually transmitted infections (STIs) was associated with increasing risk. Non‐STIs were not associated with CC risk. In conclusion, this large prospective study confirms the important role of HPV and a possible contribution of CT and HHV‐2 in cervical carcinogenesis. It further identifies HPV16 E6 seropositivity as the strongest marker to predict ICC well before disease development.

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Section: Multiplex Serologymentioning

confidence: 99%

Prospective seroepidemiologic study on the role of Human Papillomavirus and other infections in cervical carcinogenesis: Evidence from the EPIC cohort

et al. 2014

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“…PCR was done using hot start high fidelity DNA polymerase (Roche, Mannheim, Germany) at 94 °C for 2 min plus 30 cycles of 94 °C for 30 s, 54 °C for 1 min and 72 °C for 2 min. The PCR product was digested with EcoRI and SalI cleaving at the introduced sites, purified by gel electrophoresis and cloned into the correspondingly treated pGEX4T3tag derivative (Sehr et al, 2002). The F17/951 Elp-SU ectodomain was fused in frame between the 5′ GST domain and the 3′ SV40-tag (KPPTPPPEPET).…”

Section: Molecular Cloning and Recombinant Proteinsmentioning

confidence: 99%

“…For fusion protein expression, E. coli BL21 or E. coli BL21 Rosetta cells were transformed with pGEX-X-tag plasmids and recombinant proteins (Gag, Bet, FUV-7 ElpSU, F17/951 ElpSU) were purified as described ( [Sehr et al, 2001] and [Sehr et al, 2002]). …”

Section: Molecular Cloning and Recombinant Proteinsmentioning

confidence: 99%

“…ELISAs were performed as previously described ( [Sehr et al, 2001], [Sehr et al, 2002] and [Romen et al, 2006]). 96-well titer plates were coated with glutathione casein, pre-adsorbed with blocking buffer (0.2% (w/v) casein in PBS, 0.05% (v/v) Tween-20) and then 100 μl cleared E. coli lysates containing the GST-tag or GST-X-tag fusion proteins (0.25 μg total protein in blocking buffer) were added.…”

Section: Gst Capture Elisa and Cut-off Definitionmentioning

confidence: 99%

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Pattern of seroreactivity against feline foamy virus proteins in domestic cats from Germany

Bleiholder

Mühle

Hechler

et al. 2011

Veterinary Immunology and Immunopathology

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“…OVA protein (grade V) was purchased from Sigma (Taufkirchen, Germany). E7 his(6)-tagged protein produced in Escherichia coli 40 was kindly provided by Dr. P. Sehr (Deutsches Krebsforschungszentrum, Heidelberg, Germany). Proteins were dissolved in PBS/DMSO.…”

Section: Peptides Proteins and Adjuvantsmentioning

confidence: 99%

Transcutaneous immunization in mice: Induction of T-helper and cytotoxic T lymphocyte responses and protection against human papillomavirus-induced tumors

2006

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Previous reports have shown that transcutaneous immunization (TCI) with proteins or peptides in combination with adjuvants efficiently induces specific cellular and humoral immune responses. However, depending on the kind of skin pretreatment, induction of cellular immune responses was restricted to generation of either specific cytotoxic T lymphocytes (CTLs) or T-helper (Th) cells. In this study, we induced antigen-specific CTL responses together with the appropriate Th responses by TCI of C57BL/6 mice. We applied ovalbumin protein or an ovalbuminderived fusion peptide containing a CTL and Th epitope together with a combination of cholera toxin (CT) and CpG oligodeoxynucleotide (CpG) onto cold wax-depilated and hydrated bare skin. TCI with the ovalbumin fusion peptide induced more robust CTL and Th responses than that with ovalbumin protein. The fusion peptide in combination with the nontoxic CT derivative CTA1-D2D1 and CpG induced an antigen-specific CTL response, albeit less efficiently than in combination with complete CT. Further, we compared the potency of HPV-16 E7 oncoprotein-derived peptides containing single (CTL) or multiple (CTL 1 Th 1 B cell) epitopes to induce effective CTL responses. Strong E7-specific CTL responses were detected only after TCI with the E7 multiepitope peptide. This peptide was also shown to protect mice against tumor growth after challenge with HPV-16 E7-positive tumor cells. TCI with E7 protein and CT/CpG led to formation of an E7-specific humoral immune response. ' 2005 Wiley-Liss, Inc.Key words: transcutaneous immunization; human papillomavirus type 16 E7 (HPV-16 E7); CTA1-D2D1; cytotoxic T lymphocyte; T helper cell; ovalbumin; tumor protection TCI is a novel strategy in modern vaccinology. By this application, blood-borne transmission of diseases through the reuse of needles, which is a common practice in remote parts of the world, can be circumvented and better compliance of vaccinees obtained. Apart from its benefits toward vaccine safety, TCI targets LCs, a dense population of highly accessible APCs within the skin. 1 An important feature of vaccination against viral infections and virus-based tumors is the induction of CTLs. 2-4 Upon activation by APCs, predominantly DCs, CTLs are able to selectively kill antigen-expressing cells. 5 For clearance of certain viral infections, the aid of Th cells for CTL activation is necessary. 6 The bias to one of the 2 Th subsets (Th1 > Th2) is a prerequisite for specific CTL induction. 7 Generation of specific CTLs against proteinaceous or peptidebased vaccines in combination with adjuvants after TS of the skin is possible. 8-10 TS leads to removal of the outermost horny layer of the skin, resulting in enhanced permeability and maturation and activation of LCs via inflammatory cytokines secreted from keratinocytes. 11,12 The barrier function of the SC can also be bypassed by hydration of the skin, which leads to swelling of the keratinocytes and pooling of fluid into intercellular spaces. This kind of pretreatment of the skin leads to ...

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HPV antibody detection by ELISA with capsid protein L1 fused to glutathione S-transferase

Cited by 122 publications

References 23 publications

Prospective seroepidemiologic study on the role of Human Papillomavirus and other infections in cervical carcinogenesis: Evidence from the EPIC cohort

Prospective seroepidemiologic study on the role of Human Papillomavirus and other infections in cervical carcinogenesis: Evidence from the EPIC cohort

Pattern of seroreactivity against feline foamy virus proteins in domestic cats from Germany

Transcutaneous immunization in mice: Induction of T-helper and cytotoxic T lymphocyte responses and protection against human papillomavirus-induced tumors

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