High incidence of high-risk HPV in benign and malignant lesions of the larynx.
; ; et al
in International Journal of Oncology (2011), 39(1), 51-9
The aim of this study was to determine the prevalence of human papillomavirus (HPV) in patients with laryngeal benign lesions (LBLs) and laryngeal squamous cell carcinomas (LSCCs) using a sensitive E6/E7 ... [more ▼]
The aim of this study was to determine the prevalence of human papillomavirus (HPV) in patients with laryngeal benign lesions (LBLs) and laryngeal squamous cell carcinomas (LSCCs) using a sensitive E6/E7 type-specific PCR. Paraffin-embedded samples from LBL (n=39) and LSCC patients (n=67) were evaluated for the presence of HPV DNA by GP5+/GP6+ consensus PCR and E6/E7 type-specific PCR for HPV types 6, 11, 16, 18, 31, 33, 35, 39, 45, 51, 52, 53, 56, 58, 59, 66 and 68. In LSCCs, immunohistochemical staining of p16, p53 and EGFR was also assessed. The E6/E7 type-specific PCR showed that 44 out of 59 LSCC patients (i.e., 75%) had high-risk (hr) HPV types and that 27 out of 35 LBL patients (i.e., 77%) had hrHPV types. HPV-16 viral load was significantly higher in LSCC than in LBL patients (p<10-6). The presence of hrHPV DNA did not correlate with the proportion of disease-free patients. Comparable levels of p16, p53 and EGFR expression were observed in the hrHPV+ tumor group (100% p16+, 56% p53+ and 97% EGFR+) and in the HPV- or low-risk (lr) HPV+ tumor group (92% p16+, 66% p53+ and 100% EGFR+). A very high prevalence of oncogenic HPV-16 was found in a series of benign and malignant laryngeal lesions. LSCC appears to be characterized by an active hrHPV infection. In LSCCs, the hrHPV+ subgroup had a similar prognosis (in terms of risk of recurrence) as the HPV- subgroup. [less ▲]Detailed reference viewed: 17 (4 ULg)
HPV triggers NK cell cytotoxic activity and cytokine secretion
Jacobs, Nathalie ; Renoux, Virginie ; Bisig, Bettina et al
Background The immune system controls, at least partially, human papillomavirus (HPV) infection and subsequent tumor development as demonstrated by a higher tumor prevalence in immunodeficient patients ... [more ▼]
Background The immune system controls, at least partially, human papillomavirus (HPV) infection and subsequent tumor development as demonstrated by a higher tumor prevalence in immunodeficient patients. Around 90% of HPV-infected women will clear the virus within two years. However, it remains unclear which immune cells are implicated in this process and no study has been performed evaluating the direct interaction between HPV and Natural Killer (NK) cells although these cells play a key role in host resistance to virus and tumor. Methods/Results By immunochemistry, we demonstrated an NK cell infiltration in HPV+ squamous pre-neoplasic lesions. Since HPV cannot grow in vitro, virus-like particles (VLP) were used as a model for studying the NK cell response against the virus. Interestingly, NK cells displayed a higher cytotoxic activity (CD107 and chromium release assays) and cytokine production (TNF-α and IFN-γ) in the presence of HPV-VLP. Uptake of HPV-VLP by dendritic cells (DC) has been shown to induce their activation, therefore, we investigated by flow cytometry and microscopy whether the stimulation of NK cell activity is linked to VLP internalization. We observed a faster entry into these cells compared to DC. Furthermore, virus uptake by NK cells is mediated by macropinocytosis, whereas this entry is dependent on clathrin or caveolin endocytosis pathways in DC. Using NK cell lines expressing or not CD16 and blocking antibody, we demonstrated that CD16 is necessary for HPV-VLP internalization, but also for degranulation and cytokine production. Conclusion Thus, we show for the first time that NK cells interact with HPV and could participate in the immune response against HPV-induced tumors. [less ▲]Detailed reference viewed: 109 (5 ULg)
Vaccine against human papillomavirus (HPV)-associated lesions induces collaboration between natural killer and dendritic cells in vitro.
Langers, Inge ; Reschner, Anca ; Renoux, Virginie et al
Poster (2010, December)
Cervical cancer, the second most frequent gynaecological malignancy in the world, is caused by infection with high-risk human papillomaviruses (HPV). HPV16 and/or 18 are detected in more than 70% of these ... [more ▼]
Cervical cancer, the second most frequent gynaecological malignancy in the world, is caused by infection with high-risk human papillomaviruses (HPV). HPV16 and/or 18 are detected in more than 70% of these tumours. Prophylactic HPV-L1 virus like particle (VLP) vaccines are highly efficient to protect against HPV16 and HPV18 infection, but not against established infection. In this context, we study the effect of HPV-VLP on natural killer cells (NK) and on the crosstalk between NK and Dendritic Cells (DC). In order to know if HPV-VLP are able to enter in NK cells, we used fluorescent HPV-VLP with flow cytometry and confocal microscopy. HPV-VLP were internalised more rapidly in NK cells than in DC. They were already detected inside NK cells after 10 min of contact at 37°C. We also observed in CD107 assays, that HPV-VLP induce degranulation of NK cytotoxic granules. Previous works have shown that HPV-VLP were able to activate DC. We confirmed these results and observed an increase of CD69 cell surface expression and IFN-γ production by NK cells in the presence of DC activated by VLP. Interestingly, NK cells seemed to further activate DC in the presence of VLP as shown by an up-regulation of HLA-DR and CD86 on DC. Moreover, NK cells in the presence of HPV-VLP induced the production of IL12p70, but not the immunosuppressive cytokine IL10. Our results suggest that NK cells could play a role in the activation of DC induced by HPV-VLP during the vaccination against cervical cancer. Supported by the Belgian FNRS-Télévie [less ▲]Detailed reference viewed: 53 (4 ULg)
POLYPLEXES TARGETING E6: OPTIMIZATION OF THE FORMULATIONS IN TERMS OF TRANSFECTION, TOXICITY AND EFFICIENCY
Piel, Géraldine ; Collard, Laurence ; Evrard, Brigitte et al
Poster (2010, October)Detailed reference viewed: 29 (10 ULg)
Implication of plasmacytoid dendritic cells in the malignant transformation of cervical epithelial metaplasia
Demoulin, Stéphanie ; Herfs, Michael ; Somja, Joan et al
Poster (2010, September)
The cervical transformation zone is a dynamic area of a few millimeters in which a glandular epithelium has been replaced by a squamous epithelium through a metaplastic process. Interestingly, a ... [more ▼]
The cervical transformation zone is a dynamic area of a few millimeters in which a glandular epithelium has been replaced by a squamous epithelium through a metaplastic process. Interestingly, a substantial majority (87%) of cervical (pre)cancerous lesions develops within this peculiar microenvironment. Our previous studies reported that intrinsic immune features altered in the metaplastic epithelium could contribute to cancer development by preventing efficient antitumor/antiviral immune response. Plasmacytoid dendritic cells (pDC) are key effectors in host innate immunity and orchestrate adaptive immune responses. Recently, infiltration by these subtypes of dendritic cells has been shown in different cancers. However their implication in antitumor response is largely debated. The present study was performed to determine the implication of pDC in the cervical “metaplasia-dysplasia-cancer” sequence. We demonstrated that the density of pDC increases in the epithelium of metaplastic and (pre)cancerous cervical tissues as well as in underlying stroma as compared with normal exocervical epithelium. This could be partially explained by the increased expression of chemerin, their chemotactic peptide, observed in those areas. We developed a method to efficiently generate pDC cells exhibiting morphological and immunohistochemical features of blood pDC from a limited number of CD34+ cord blood progenitors. Using these in vitro generated pDC, we demonstrated that medium conditioned by transformed keratinocytes modified the activation status of pDC, by inducing a decreased expression of costimulatory molecules such as CD86 and HLA-DR. Moreover, malignant keratinocytes diminished the ability of pDC to produce IFNα in response to an oligonucleotide containing CpG motifs, a defined microbial stimulus for pDC. These results suggest that pDC could be educated within the metaplastic and/or (pre)cancerous microenvironment to acquire a tolerogenic phenotype that could promote carcinogenesis. In agreement with those results, we observed that both metaplastic areas and (pre)cancerous lesions of the cervix are infiltrated by T regulatory cells. [less ▲]Detailed reference viewed: 38 (7 ULg)
Systematic chromosomal aberrations found in murine bone marrow-derived mesenchymal stem cells.
Josse, Claire ; ; Niessen, Neville-Andrew et al
in Stem Cells & Development (2010), 19(8), 1167-1173
Mesenchymal stem cells (MSCs) are studied as a cellular source for the treatment of various diseases. In this work, we isolated and cultivated murine bone marrow-derived MSCs. After a first observation of ... [more ▼]
Mesenchymal stem cells (MSCs) are studied as a cellular source for the treatment of various diseases. In this work, we isolated and cultivated murine bone marrow-derived MSCs. After a first observation of a solid tumour in a mouse injected with these cells, we systematically explored their chromosomal stability. We observed in all the cytogenetically analysed cases gross chromosomal alterations every time the MSCs went through the senescence crisis while the lymphocytes from the same animals showed a normal chromosome count. This observation was confirmed in different mouse strains, with different culture protocols, and even in short-term cultures after an hematopoietic cell negative immunodepletion performed in order to accelerate the isolation procedure. Therefore, we conclude that murine MSCs display high chromosomal instability, can generate tumours, and that care must be taken before using them for the evaluation of MSC therapeutic potential. [less ▲]Detailed reference viewed: 59 (17 ULg)
Validation of ThermoFisher's Papspin for human papillomavirus detection in cervicovaginal specimens using PCR with GP5+/GP6+ primers and the Hybrid Capture II assay.
; Delvenne, Philippe ; et al
in Clinical Microbiology & Infection (2010), 16(6), 671-5Detailed reference viewed: 34 (4 ULg)
Cross talk between dendritic and natural killer cells in the presence of vaccine agent against cervical cancer
Langers, Inge ; Renoux, Virginie ; Reschner, Anca et al
in Belgian Journal of Medical Oncology [=BJMO] (2010, January 30), 4Detailed reference viewed: 62 (24 ULg)
Immune suppression in head and neck cancers: a review.
; Demoulin, Stéphanie ; Hubert, Pascale et al
in Clinical & Developmental Immunology (2010), 2010
Head and neck squamous cell carcinomas (HNSCCs) are the sixth most common cancer in the world. Despite significant advances in the treatment modalities involving surgery, radiotherapy, and concomitant ... [more ▼]
Head and neck squamous cell carcinomas (HNSCCs) are the sixth most common cancer in the world. Despite significant advances in the treatment modalities involving surgery, radiotherapy, and concomitant chemoradiotherapy, the 5-year survival rate remained below 50% for the past 30 years. The worse prognosis of these cancers must certainly be link to the fact that HNSCCs strongly influence the host immune system. We present a critical review of our understanding of the HNSCC escape to the antitumor immune response such as a downregulation of HLA class I and/or components of APM. Antitumor responses of HNSCC patients are compromised in the presence of functional defects or apoptosis of T-cells, both circulating and tumor-infiltrating. Langerhans cells are increased in the first steps of the carcinogenesis but decreased in invasive carcinomas. The accumulation of macrophages in the peritumoral areas seems to play a protumoral role by secreting VEGF and stimulating the neoangiogenesis. [less ▲]Detailed reference viewed: 26 (12 ULg)
Mechanisms of cell entry by human papillomaviruses: an overview.
; ; Renoux, Virginie et al
in Virology Journal (2010), 7
ABSTRACT: As the primary etiological agents of cervical cancer, human papillomaviruses (HPVs) must deliver their genetic material into the nucleus of the target cell. The viral capsid has evolved to ... [more ▼]
ABSTRACT: As the primary etiological agents of cervical cancer, human papillomaviruses (HPVs) must deliver their genetic material into the nucleus of the target cell. The viral capsid has evolved to fulfil various roles that are critical to establish viral infection. The particle interacts with the cell surface via interaction of the major capsid protein, L1, with heparan sulfate proteoglycans. Moreover, accumulating evidence suggests the involvement of a secondary receptor and a possible role for the minor capsid protein, L2, in cell surface interactions.The entry of HPV in vitro is initiated by binding to a cell surface receptor in contrast to the in vivo situation where the basement membrane has recently been identified as the primary site of virus binding. Binding of HPV triggers conformational changes, which affect both capsid proteins L1 and L2, and such changes are a prerequisite for interaction with the elusive uptake receptor. Most HPV types that have been examined, appear to enter the cell via a clathrin-dependent endocytic mechanism, although many data are inconclusive and inconsistent. Furthermore, the productive entry of HPV is a process that occurs slowly and asynchronously and it is characterised by an unusually extended residence on the cell surface.Despite the significant advances and the emergence of a general picture of the infectious HPV entry pathway, many details remain to be clarified. The impressive technological progress in HPV virion analysis achieved over the past decade, in addition to the improvements in general methodologies for studying viral infections, provide reasons to be optimistic about further advancement of this field.This mini review is intended to provide a concise overview of the literature in HPV virion/host cell interactions and the consequences for endocytosis. [less ▲]Detailed reference viewed: 47 (8 ULg)
Regulation of p63 Isoforms by Snail and Slug Transcription Factors in Human Squamous Cell Carcinoma.
Herfs, Michael ; Hubert, Pascale ; Suarez-Carmona, Meggy et al
in American Journal of Pathology (2010), 176(4), 1941-1949
TP63 is a p53-related gene that contains two alternative promoters, which give rise to transcripts that encode proteins with (TAp63) or without (DeltaNp63) an amino-transactivating domain. Whereas the ... [more ▼]
TP63 is a p53-related gene that contains two alternative promoters, which give rise to transcripts that encode proteins with (TAp63) or without (DeltaNp63) an amino-transactivating domain. Whereas the expression of p63 is required for proper development of epithelial structures, the role of p63 in tumorigenesis remains unclear. Here, we investigated the role of Snail and Slug transcription factors, known to promote epithelial-to-mesenchymal transitions during development and cancer, in the regulation of p63 isoforms in human squamous cell carcinoma (SCC). In the present study, we observed that the expressions of DeltaN and TAp63 isoforms were, respectively, down- and up-regulated by both Snail and Slug. However, the induction of TAp63 was not directly caused by these two transcription factors but resulted from the loss of DeltaNp63, which acts as dominant-negative inhibitor of TAp63. In SCC cell lines and cancer tissues, high expression of Snail and Slug was also significantly associated with altered p63 expression. Finally, we showed that DeltaNp63 silencing reduced cell-cell adhesion and increased the migratory properties of cancer cells. These data suggest that the disruption of p63 expression induced by Snail and Slug plays a crucial role in tumor progression. Therefore, p63 and its regulating factors could constitute novel prognosis markers in patients with SCC and attractive targets for the therapeutic modulation of neoplastic cell invasiveness. [less ▲]Detailed reference viewed: 79 (18 ULg)
Implication of plasmacytoid dendritic cells in the malignant transformation of cervical epithelial metaplasia
Demoulin, Stéphanie ; Somja, Joan ; Herfs, Michael et al
Poster (2010)Detailed reference viewed: 9 (0 ULg)
Fish and chips
DELVENNE, Philippe ; Deprez, Manuel ; BISIG, Bettina et al
in Revue Médicale de Liège (2010), 65 Spec no.
Academic hospital laboratories should offer patients the possibility to have the most accurate diagnosis by the development of new analyses, such as molecular biology tests including FISH (Fluorescent In ... [more ▼]
Academic hospital laboratories should offer patients the possibility to have the most accurate diagnosis by the development of new analyses, such as molecular biology tests including FISH (Fluorescent In Situ Hybridization) and chips (microarrays,...). The purpose of this article is to describe the principles and the potential applications of these techniques. [less ▲]Detailed reference viewed: 52 (9 ULg)
Local applications of GM-CSF induce the recruitment of immune cells in cervical low-grade squamous intraepithelial lesions.
Hubert, Pascale ; Doyen, Jean ; Capelle, Xavier et al
in American Journal of Reproductive Immunology (2010), 64(2), 126-136
Abstract Problem Quantitative alterations of antigen-presenting cells (APC) in (pre)neoplastic lesions of the uterine cervix associated with human papillomavirus (HPV) infection suggest a diminished ... [more ▼]
Abstract Problem Quantitative alterations of antigen-presenting cells (APC) in (pre)neoplastic lesions of the uterine cervix associated with human papillomavirus (HPV) infection suggest a diminished capacity to capture viral antigens and to induce a protective immune response. Method of study To test if a cervical application of GM-CSF could restore an immune response against HPV in women with cervical low-grade squamous intraepithelial lesions (LSIL). We performed two clinical trials with11 healthy women and 15 patients with LSIL. Results GM-CSF applications were well tolerated in all enrolled women and no difference in toxicity between the treated and placebo groups was observed during the follow up (until 30 months). Interestingly, in the GM-CSF treated group, a significant increased APC and cytotoxic T lymphocyte infiltration was observed in the cervical biopsies with no change in regulatory T cell numbers. All the HPV16+ patients exhibited an immune response against HPV16 after GM-CSF applications, as shown by NK and/or T cells producing IFN-γ whereas no cellular immune response was observed before the treatment. Moreover, the anti-VLP antibody titers also increased after the treatment. Conclusion These encouraging results obtained from a limited number of subjects justify further study on the therapeutic effect of APC in cervical (pre)neoplastic lesions. [less ▲]Detailed reference viewed: 137 (55 ULg)
Effects of HPV-16 E5, E6 and E7 Proteins on Survival, Adhesion, Migration and Invasion of Trophoblastic Cells
; ; et al
in Carcinogenesis (2010), 31(3), 473-80
Amongst high-risk human papillomaviruses (HPV), HPV-16 infection is the most prevalent causative factor for cervical cancer. Beside other mucosal targets, HPV-16 was reported to infect the placenta and to ... [more ▼]
Amongst high-risk human papillomaviruses (HPV), HPV-16 infection is the most prevalent causative factor for cervical cancer. Beside other mucosal targets, HPV-16 was reported to infect the placenta and to replicate in trophoblastic cells. Since these cells share invasive properties of tumoral cells, they represent an ideal model to investigate several oncogenic processes. In the present work, we analyzed the impacts of HPV-16 E5, E6 and E7 oncoproteins on the trophoblastic model. Our results showed that E5 impaired the viability of trophoblastic and cervical cell lines but E6 and E7, favouring cell growth, neutralised the E5 cytotoxic effect. In addition, E5 decreased the adhesiveness of trophoblastic cells to the tissue culture plastic and to endometrial cells similarly as previously described for E6 and E7. E5 and E6 plus E7 increased also their migration and their invasive properties. Cells expressing HPV-16 early proteins under the control of the LCR endogenous promoter displayed growth advantage and were also more motile and invasive compared to control cells. Interestingly, the E-cadherin was down regulated in trophoblastic cells expressing E5, E6 and E7. NF-kB and AP-1 activities were also enhanced. In conclusion, HPV-16 early proteins enhanced trophoblastic growth and intensify the malignant phenotype by impairing cell adhesion leading to increased cellular motile and invasive properties. HPV-16 E5 participated, with E6 and E7, in these changes by impairing Ecadherin expression, a hallmark of malignant progression. [less ▲]Detailed reference viewed: 70 (6 ULg)
Progression model tissue microarray (TMA) for the study of uterine carcinomas.
; Boniver, Jacques ; Delvenne, Philippe
in Disease Markers (2010), 28(5), 267-72
Cervical and endometrial uterine carcinomas are heterogeneous groups of cancers, which are preceded by preneoplastic lesions. More accurate tools are needed to improve the diagnosis and to define markers ... [more ▼]
Cervical and endometrial uterine carcinomas are heterogeneous groups of cancers, which are preceded by preneoplastic lesions. More accurate tools are needed to improve the diagnosis and to define markers which may be relevant for the diagnosis, prediction of disease progression and therapeutic response.High throughput technologies for testing and validating molecular targets in cancer lesions and in their precursors are presently available. Among them, the tissue microarray (TMA) presents the advantage of a morphological control of the analyzed tissue fragment. In this article, we review the different aspects of the TMA technology with a special consideration to a uterine carcinogenesis model. [less ▲]Detailed reference viewed: 11 (3 ULg)
Current concepts in the pathology and epigenetics of endometrial carcinoma.
; Somja, Joan ; Dehan, Pierre et al
in Pathology (2010), 42(7), 613-7
In the Western world, endometrial carcinoma is the most common malignant tumour of the female genital tract and is the fourth most common cancer in women. Two different clinicopathological subtypes are ... [more ▼]
In the Western world, endometrial carcinoma is the most common malignant tumour of the female genital tract and is the fourth most common cancer in women. Two different clinicopathological subtypes are recognised: the oestrogen-related (type I, endometrioid) and the non-oestrogen related (type II, non-endometrioid). This article reviews the epidemiology, risk factors, genetic alterations during endometrial carcinogenesis, features of tumours and precursors and early detection of the disease. Insights into the epigenetic alterations, with emphasis on DNA methylation during endometrial carcinogenesis, and their diagnostic value are also provided. [less ▲]Detailed reference viewed: 64 (13 ULg)
Increase in viral load, viral integration, and gain of telomerase genes during uterine cervical carcinogenesis can be simultaneously assessed by the HPV 16/18 MLPA-assay.
; ; Herfs, Michael et al
in American Journal of Pathology (2010), 177(4), 2022-33
Oncogenic human papillomavirus (HPV) infection is the most important risk factor in cervical carcinogenesis cases; high viral loads, viral integration into the host genome, and gain of the telomerase ... [more ▼]
Oncogenic human papillomavirus (HPV) infection is the most important risk factor in cervical carcinogenesis cases; high viral loads, viral integration into the host genome, and gain of the telomerase-related genes, TERT and TERC, are all factors associated with progression to cancer. A recently developed multiparameter HPV 16/18 multiplex ligation-dependent probe amplification (MLPA) assay, which allows the simultaneous assessment of these factors, was applied to a series of 67 normal and (pre)malignant frozen uterine cervical samples, as well as to 91 cytological preparations, to test the ability of the MLPA assay to identify high-risk lesions on the basis of these factors. Validation was performed using quantitative PCR, the PapilloCheck and fluorescence in situ hybridization. Only 5 out of 37 normal tissue samples or low-grade cervical lesions (ie, CIN1 and condyloma) showed either an HPV16 viral load higher than 25 copies per cell, viral integration, and/or gain of one of the telomerase-related genes, whereas for the high-grade cervical lesions, one or more of these risk factors was found in 25 of 30 cases. The HPV MLPA assay showed a sensitivity of 83% and a specificity of 86% in frozen cervical specimens. Furthermore, the feasibility of the MLPA assay was shown for cytological samples, where in 57% of high-grade squamous intraepithelial lesion cases, the high-risk factors were detected using this assay. [less ▲]Detailed reference viewed: 20 (3 ULg)
The L1 major capsid protein of HPV16 differentially modulates APC trafficking according to the vaccination or natural infection context.
Herman, Ludivine ; Hubert, Pascale ; Herfs, Michael et al
in European Journal of Immunology (2010), 40(11), 3075-84
Human papillomavirus (HPV) infection, particularly type 16, is causally associated with cancer of the uterine cervix. The progression of cervical lesions suggests that viral antigens are not adequately ... [more ▼]
Human papillomavirus (HPV) infection, particularly type 16, is causally associated with cancer of the uterine cervix. The progression of cervical lesions suggests that viral antigens are not adequately presented to the immune system. The aim of this study was to determine whether HPV16 viral particles can influence the trafficking of human DC/Langerhans cells (LC), either by direct interactions with DC or following incubation with human normal keratinocytes that are in close contact with LC in the squamous epithelium. We first demonstrated that HPV16 L1 major capsid protein, when self-assembled into virus-like particles (VLP), is able to induce in DC an over-expression of CXC receptor 4 (CXCR4) via the activation of the NF-kappaB signaling pathway and to enhance DC motility in the presence of CXCL12, suggesting an ability to migrate towards lymph nodes. We also showed that conditioned media of HPV16 VLP-treated keratinocytes induce a lower LC migration than those from untreated keratinocytes and that prostaglandin E2 (PGE(2)), detected in HPV16 VLP-treated keratinocyte supernatants, may reduce LC recruitment into the squamous epithelium. Taken together, our data demonstrate that HPV16 VLP may differentially regulate the immune protective response according to their target cells. [less ▲]Detailed reference viewed: 34 (10 ULg)
Characterization of hepatitis C virus-induced nasal mucosa remodelling.
El Shazly, Amr ; ; Roncarati, Patrick et al
in Histopathology (2010), 57(3), 488-92Detailed reference viewed: 27 (4 ULg)