RESUMEN

Cervical cancer (CC) is a significant health problem, especially in low-income countries. Functional studies on the human papillomavirus have generated essential advances in the knowledge of CC. However, many unanswered questions remain. This mini-review discusses the latest results on CC pathogenesis, HPV oncogenesis, and molecular changes identified through next-generation technologies. Interestingly, the percentage of samples with HPV genome integrations correlates with the degree of the cervical lesions, suggesting a role in the development of CC. Also, new functions have been described for the viral oncoproteins E5, E6, and E7, resulting in the acquisition and maintenance of cancer hallmarks, including proliferation, immune response evasion, apoptosis, and genomic instability. Remarkably, E5 oncoprotein affects signaling pathways involved in the expression of interferon-induced genes and EGFR-induced proliferation, while E6 and E7 oncoproteins regulate the DNA damage repair and cell cycle continuity pathways. Furthermore, next-generation technologies provide vast amounts of information, increasing our knowledge of changes in the genome, transcriptome, proteome, metabolome, and epigenome in CC. These studies have identified novel molecular traits associated with disease susceptibility, degree of progression, treatment response, and survival as potential biomarkers and therapeutic targets.

RESUMEN

BACKGROUND: Men who have sex with men (MSM) are at high risk for oral human papillomavirus (HPV infection). There are no specific screening guidelines to facilitate the identification of people at risk for oral HPV infection. We aimed to estimate the prevalence of oral high-risk HPV and create a risk score to identify MSM at higher risk for prevalent oral HPV. METHODS: We collected baseline data from a clinical trial from a subsample of 500 MSM attending sexually transmitted disease treatment clinics; they provided an oral gargle sample for high-risk HPV detection. We calculated oral high-risk HPV prevalence and 95% confidence intervals (CIs), used a logistic regression model to identify factors associated with high-risk HPV infection, and created a risk score. RESULTS: The prevalence of any oral high-risk HPV among MSM was 11.1% (95% CI: 8.6-14.2), with a higher prevalence observed among men living with HIV (14.8%). Factors independently associated with oral high-risk HPV were age ≥40 years (OR = 2.71, 95% CI: 1.28-5.73 compared to <40 years), being HIV-positive with CD4 count 200-499 (OR = 2.76, 95% CI: 1.34-5.65 compared to HIV-negative), and recent recreational use of vasodilators (poppers/sildenafil) (OR = 2.02, 95% CI: 1.02-2.97). The risk score had good discriminatory power (AUC = 0.70, 95% CI: 0.63-0.77). CONCLUSIONS: MSM have specific predictors for prevalent oral high-risk HPV, and a risk score could be used by clinicians to target men with vaccine recommendations and counseling, and identify those who could benefit from primary interventions given the available resources, or for referral to dental services for follow-up when available.

Asunto(s)

Infecciones por VIH , Enfermedades de la Boca , Infecciones por Papillomavirus , Minorías Sexuales y de Género , Masculino , Humanos , Adulto , Homosexualidad Masculina , Infecciones por VIH/complicaciones , Infecciones por Papillomavirus/complicaciones , Virus del Papiloma Humano , Prevalencia , México/epidemiología , Papillomaviridae , Factores de Riesgo , Enfermedades de la Boca/epidemiología

RESUMEN

Human papillomavirus (HPV) infection is associated with precancerous lesions and cancer of the genital tract both in women and men. The high incidence of cervical cancer worldwide focused the research on this infection mainly in women and to a lesser extent in men. In this review, we summarized epidemiological, immunological, and diagnostic data associated with HPV and cancer in men. We presented an overview of the main characteristics of HPV and infection in men that are associated with different types of cancer but also associated with male infertility. Men are considered important vectors of HPV transmission to women; therefore, identifying the sexual and social behavioral risk factors associated with HPV infection in men is critical to understand the etiology of the disease. It is also essential to describe how the immune response develops in men during HPV infection or when vaccinated, since this knowledge could help to control the viral transmission to women, decreasing the incidence of cervical cancer, but also could reduce other HPV-associated cancers among men who have sex with men (MSM). Finally, we summarized the methods used over time to detect and genotype HPV genomes, as well as some diagnostic tests that use cellular and viral biomarkers that were identified in HPV-related cancers.

RESUMEN

Antibodies against the Human Papillomavirus (HPV) L1 protein are associated with past infections and related to the evolution of the disease, whereas antibodies against L1 Virus-Like Particles (VLPs) are used to follow the neutralizing antibody response in vaccinated women. In this study, serum antibodies against conformational (VLPs) and linear epitopes of HPV16/18 L1 protein were assessed to distinguish HPV-vaccinated women from those naturally infected or those with uterine cervical lesions. The VLPs-16/18 were generated in baculovirus, and L1 proteins were obtained from denatured VLPs. Serum antibodies against VLPs and L1 proteins were evaluated by ELISA. The ELISA-VLPs and ELISA-L1 16/18 assays were validated with a vaccinated women group by ROC analysis and the regression analysis to distinguish the different populations of female patients. The anti-VLPs-16/18 and anti-L1-16/18 antibodies effectively detect vaccinated women (AUC = 1.0/0.79, and 0.94/0.84, respectively). The regression analysis showed that anti-VLPs-16/18 and anti-L1-16/18 antibodies were associated with the vaccinated group (OR = 2.11 × 108/16.50 and 536.0/49.2, respectively). However, only the anti-L1-16 antibodies were associated with the high-grade lesions and cervical cancer (CIN3/CC) group (OR = 12.18). In conclusion, our results suggest that anti-VLPs-16/18 antibodies are effective and type-specific to detect HPV-vaccinated women, but anti-L1-16 antibodies better differentiate the CIN3/CC group. However, a larger population study is needed to validate these results.

RESUMEN

HPV E5 is an oncoprotein mainly expressed in premalignant lesions, which makes it an important target for a vaccine to prevent or cure cervical cancer (CC). In this study, we evaluated whether E5 targeted to DEC-205, present in dendritic cells (DCs), could induce a therapeutic protection against HPV16-induced tumor cells in a mouse model. The HPV-16 E5 (16E5) protein was cross-linked to a monoclonal antibody (mAb) specific to mouse DEC-205 (anti-DEC-205:16E5) or to an isotype control mAb (isotype:16E5). Rotavirus VP6 was cross-linked to the mouse anti-DEC-205 mAb (anti-DEC-205:VP6) as a non-specific antigen control. BALB/c mice were inoculated subcutaneously (s.c.) with the 16E5-expressing BMK-16/myc tumor cells, and 7 and 14 days later the mice were immunized s.c. with the conjugates, free 16E5 or PBS in the presence of adjuvant. Tumor growth was monitored to evaluate protection. A strong protective immune response against the tumor cells was induced when the mice were inoculated with the anti-DEC-205:16E5 conjugate, since 70% of the mice controlled the tumor growth and survived, whereas the remaining 30% developed tumors and died by day 72. In contrast, 100% of the mice in the control groups died by day 30. The anti-DEC-205:16E5 conjugate was found to induce 16E5-specific memory T cells, with a Th1/Th17 profile. Both CD4+ and CD8+ T cells contributed to the observed protection. Finally, treating mice that had developed tumors with an anti-PD-1 mAb, delayed the tumor growth for more than 20 days. These results show that targeting 16E5 to DEC-205, alone or combined with an immune checkpoint blockade, could be a promising protocol for the treatment of the early stages of HPV-associated cancer.

Asunto(s)

Células Dendríticas/inmunología , Papillomavirus Humano 16/inmunología , Neoplasias/etiología , Neoplasias/terapia , Proteínas Oncogénicas Virales/inmunología , Infecciones por Papillomavirus/complicaciones , Animales , Anticuerpos Monoclonales/inmunología , Anticuerpos Antivirales/inmunología , Biomarcadores de Tumor , Vacunas contra el Cáncer/administración & dosificación , Vacunas contra el Cáncer/inmunología , Línea Celular Tumoral , Células Dendríticas/metabolismo , Modelos Animales de Enfermedad , Femenino , Humanos , Inmunización , Memoria Inmunológica , Linfocitos Infiltrantes de Tumor/inmunología , Linfocitos Infiltrantes de Tumor/metabolismo , Ratones , Neoplasias/diagnóstico , Infecciones por Papillomavirus/virología , Linfocitos T/inmunología , Linfocitos T/metabolismo

RESUMEN

The human papillomavirus (HPV) is recognized as the main etiologic agent associated with cervical cancer. HPVs are epitheliotropic, and the ones that infect the mucous membranes are classified into low-risk (LR) and high-risk (HR) types. LR-HPVs produce benign lesions, whereas HR-HPVs produce lesions that may progress to cancer. HR-HPV types 16 and 18 are the most frequently found in cervical cancer worldwide. E6 and E7 are the major HPV oncogenic proteins, and they have been profusely studied. Moreover, it has been shown that the HPV16 E5 (16E5) oncoprotein generates transformation, although the molecular mechanisms through which it carries out its activity have not been well defined. In contrast to E6 and E7, the E5 open reading frame is lost during the integration of the episomal HPV DNA into the cellular genome. This suggests that E5 acts at the early stages of the transformation process. In this review, we focused on the biochemical characteristics and functions of the HPV E5 oncoprotein, mainly on its association with growth factor receptors and other cellular proteins. Knowledge of the HPV E5 biology is important to understand the role of this oncoprotein in maintaining the viral cycle through the modulation of proliferation, differentiation, and apoptosis, as well as the alteration of other processes, such as survival, adhesion, migration, and invasion during early carcinogenesis. Finally, we summarized recent research that uses the E5 oncoprotein as a therapeutic target, promising a novel approach to the treatment of cervical cancer in its early stages.

Asunto(s)

Transformación Celular Neoplásica/metabolismo , Papillomavirus Humano 16/patogenicidad , Proteínas Oncogénicas Virales/metabolismo , Neoplasias del Cuello Uterino/genética , Secuencia de Aminoácidos , Animales , Femenino , Humanos , Ratones , Ratones Desnudos , Neoplasias del Cuello Uterino/patología

RESUMEN

The influenza A virus infection continues to be a threat to the human population. The seasonal variation of the virus and the likelihood of periodical pandemics caused by completely new virus strains make it difficult to produce vaccines that efficiently protect against this infection. Antibodies (Abs) are very important in preventing the infection and in blocking virus propagation once the infection has taken place. However, the precise protection mechanism provided by these Abs still needs to be established. Furthermore, most research has focused on Abs directed to the globular head domain of hemagglutinin (HA). However, other domains of HA (like the stem) and other proteins are also able to elicit protective Ab responses. In this article, we review the current knowledge about the role of both neutralizing and non-neutralizing anti-influenza proteins Abs that play a protective role during infection or vaccination.

Asunto(s)

Anticuerpos Antivirales/inmunología , Antígenos Virales/inmunología , Virus de la Influenza A/inmunología , Vacunas contra la Influenza/inmunología , Gripe Humana/inmunología , Epítopos de Linfocito B/inmunología , Humanos

RESUMEN

Hypervirulent Klebsiella pneumoniae have been rarely described in Latin America. This work describes the characterization of hypervirulent K. pneumoniae isolates capsular serotype K2 belonging to sequence types 86 and 380. The assays showed the hypervirulent K. pneumoniae highly virulent, which is determined by the plasmid borne virulence genes. At this time, the hypervirulent K. pneumoniae clinical isolates in Mexico are extensively susceptible to antibiotics.

Asunto(s)

Genotipo , Infecciones por Klebsiella/microbiología , Infecciones por Klebsiella/patología , Klebsiella pneumoniae/clasificación , Klebsiella pneumoniae/patogenicidad , Serogrupo , Animales , Adhesión Celular , Modelos Animales de Enfermedad , Femenino , Humanos , Klebsiella pneumoniae/aislamiento & purificación , Dosificación Letal Mediana , Masculino , México , Ratones Endogámicos BALB C , Pruebas de Sensibilidad Microbiana , Modelos Biológicos , Plásmidos/análisis , Factores de Virulencia/genética

RESUMEN

CD4+ T cells are major players in the immune response against several diseases; including AIDS, leishmaniasis, tuberculosis, influenza and cancer. Their activation has been successfully achieved by administering antigen coupled with antibodies, against DC-specific receptors in combination with adjuvants. Unfortunately, most of the adjuvants used so far in experimental models are unsuitable for human use. Therefore, human DC-targeted vaccination awaits the description of potent, yet nontoxic adjuvants. The nontoxic cholera B subunit (CTB) can be safely used in humans and it has the potential to activate CD4+ T cell responses. However, it remains unclear whether CTB can promote DC activation and can act as an adjuvant for DC-targeted antigens. Here, we evaluated the CTB's capacity to activate DCs and CD4+ T cell responses, and to generate long-lasting protective immunity. Intradermal (i.d.) administration of CTB promoted late and prolonged activation and accumulation of skin and lymphoid-resident DCs. When CTB was co-administered with anti-DEC205-OVA, it promoted CD4+ T cell expansion, differentiation, and infiltration to peripheral nonlymphoid tissues, i.e., the skin, lungs and intestine. Indeed, CTB promoted a polyfunctional CD4+ T cell response, including the priming of Th1 and Th17 cells, as well as resident memory T (RM) cell differentiation in peripheral nonlymphoid tissues. It is worth noting that CTB together with a DC-targeted antigen promoted local and systemic protection against experimental melanoma and murine rotavirus. We conclude that CTB administered i.d. can be used as an adjuvant to DC-targeted antigens for the induction of broad CD4+ T cell responses as well as for promoting long-lasting protective immunity.

Asunto(s)

Adyuvantes Inmunológicos/administración & dosificación , Toxina del Cólera/administración & dosificación , Células Dendríticas/inmunología , Lectinas Tipo C/antagonistas & inhibidores , Receptores de Superficie Celular/antagonistas & inhibidores , Vacunación/métodos , Animales , Antígenos CD/inmunología , Línea Celular Tumoral/trasplante , Modelos Animales de Enfermedad , Femenino , Humanos , Inyecciones Intradérmicas , Lectinas Tipo C/inmunología , Activación de Linfocitos/inmunología , Masculino , Melanoma/inmunología , Melanoma/prevención & control , Ratones , Ratones Endogámicos BALB C , Ratones Endogámicos C57BL , Ratones Transgénicos , Antígenos de Histocompatibilidad Menor/inmunología , Receptores de Superficie Celular/inmunología , Rotavirus/inmunología , Infecciones por Rotavirus/inmunología , Infecciones por Rotavirus/prevención & control , Infecciones por Rotavirus/virología , Células TH1/inmunología , Células Th17/inmunología , Resultado del Tratamiento

RESUMEN

E5 oncoprotein activity from high risk human papillomaviruses (HPVs) is associated with growth factor receptor signaling, but the function of this protein is not well understood. In this study, we investigated the role of HPV-16 E5 on the cell cycle progression during EGF-stimulation. Wild-type and NIH 3T3 cells over-expressing human EGF-receptor were transfected with HPV-16 E5 gene and the cell cycle progression was characterized. This analysis showed that the E5-expressing cells increased DNA synthesis (S-phase) by around 40%. Cell cycle protein analysis of E5-expressing cells showed a reduction in the half-life of p27(Kip1) protein as compared to control cells (18.4 vs. 12.7 h), an effect that was enhanced in EGF-stimulated cells (12.8 vs. 3.6 h). Blockage of EGF-receptor activity abrogated E5 signals as well as p27(Kip1) down-regulation. These results suggest that E5 and the EGF-receptor cooperate to enhance cell cycle entry and progression through regulating p27(Kip1) expression at protein level.

Asunto(s)

Inhibidor p27 de las Quinasas Dependientes de la Ciclina/genética , Receptores ErbB/fisiología , Papillomavirus Humano 16/fisiología , Papillomavirus Humano 16/patogenicidad , Proteínas Oncogénicas Virales/fisiología , Animales , Secuencia de Bases , Ciclo Celular/fisiología , Inhibidor p27 de las Quinasas Dependientes de la Ciclina/fisiología , Cartilla de ADN/genética , Regulación hacia Abajo , Receptores ErbB/genética , Genes Virales , Interacciones Huésped-Patógeno/genética , Interacciones Huésped-Patógeno/fisiología , Papillomavirus Humano 16/genética , Humanos , Ratones , Células 3T3 NIH , Proteínas Oncogénicas Virales/genética , Proteínas Recombinantes/genética , Proteínas Recombinantes/metabolismo , Transducción de Señal/fisiología

RESUMEN

This study used an in vivo mouse model to analyze the response of dendritic cells (DCs) in Peyer's patches (PPs) within the first 48 h of infection with the wild-type murine rotavirus EDIM (EDIM(wt)). After the infection, the absolute number of DCs was increased by 2-fold in the PPs without a modification of their relative percentage of the total cell number. Also, the DCs from PPs of infected mice showed a time-dependent migration to the subepithelial dome (SED) and an increase of the surface activation markers CD40, CD80, and CD86. This response was more evident at 48 h postinfection (p.i.) and depended on viral replication, since DCs from PPs of mice inoculated with UV-treated virus did not show this phenotype. As a result of the activation, the DCs showed an increase in the expression of mRNA for the proinflammatory cytokines interleukin-12/23p40 (IL-12/23p40), tumor necrosis factor alpha (TNF-alpha), and beta interferon (IFN-beta), as well as for the regulatory cytokine IL-10. These results suggest that, a short time after rotavirus infection, the DCs from PPs play a critical role in controlling the infection and, at the same time, avoiding an excessive inflammatory immune response.

Asunto(s)

Células Dendríticas/inmunología , Ganglios Linfáticos Agregados/inmunología , Infecciones por Rotavirus/inmunología , Animales , Antígeno B7-1/metabolismo , Antígeno B7-2/metabolismo , Secuencia de Bases , Antígenos CD40/metabolismo , Movimiento Celular/inmunología , Citocinas/genética , Cartilla de ADN/genética , Células Dendríticas/patología , Modelos Animales de Enfermedad , Femenino , Mediadores de Inflamación/metabolismo , Ratones , Ratones Endogámicos BALB C , Ganglios Linfáticos Agregados/patología , ARN Mensajero/genética , ARN Mensajero/metabolismo , Infecciones por Rotavirus/patología , Infecciones por Rotavirus/virología

RESUMEN

Cervical cancer (CC) is a major public health problem in developing countries and its most significant etiological risk factor is infection by the human papillomavirus (HPV). The main approach to date for the prevention of CC has been through screening programs, using the cervical smear (PAP test) to detect precursory lesions. The sensitivity and specificity of the PAP smear depend on the skills of the observer to recognize and classify a variety of cellular abnormalities. The development of early diagnoses to detect HPV infection has been a problem as cytology and colposcopy identify the lesion at an advanced stage. Therefore, molecular approaches have become more successful for early CC diagnosis. These molecular techniques recognize HPV DNA sequences by DNA hybridization, PCR-RFLP, hybrid capture and reverse line blot systems. Unfortunately, these systems cannot determine whether the HPV infection is active, latent or persistent. Thus, immunological techniques such as Western blot and ELISA have been designed to follow the immune response against the virus, and they can also be used to identify the stage of the infection. Several companies have developed, manufactured and merchandised gene-based testing systems for the screening, monitoring and diagnosis of HPV. Our review and comments focus on the critical analysis of existing products and their use in clinical practice as well as on immunological systems used mainly in research, but that may be applied in large population screening programs.

Asunto(s)

Infecciones por Papillomavirus/complicaciones , Infecciones por Papillomavirus/diagnóstico , Neoplasias del Cuello Uterino/microbiología , Femenino , Humanos , Pruebas Inmunológicas , Técnicas de Diagnóstico Molecular

RESUMEN

Cervical cancer (CC) is a major public health problem in developing countries and its most significant etiological risk factor is infection by the human papillomavirus (HPV). The main approach to date for the prevention of CC has been through screening programs, using the cervical smear (PAP test) to detect precursory lesions. The sensitivity and specificity of the PAP smear depend on the skills of the observer to recognize and classify a variety of cellular abnormalities. The development of early diagnoses to detect HPV infection has been a problem as cytology and colposcopy identify the lesion at an advanced stage. Therefore, molecular approaches have become more successful for early CC diagnosis. These molecular techniques recognize HPV DNA sequences by DNA hybridization, PCR-RFLP, hybrid capture and reverse line blot systems. Unfortunately, these systems cannot determine whether the HPV infection is active, latent or persistent. Thus, immunological techniques such as Western blot and ELISA have been designed to follow the immune response against the virus, and they can also be used to identify the stage of the infection. Several companies have developed, manufactured and merchandised gene-based testing systems for the screening, monitoring and diagnosis of HPV. Our review and comments focus on the critical analysis of existing products and their use in clinical practice as well as on immunological systems used mainly in research, but that may be applied in large population screening programs.

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El cáncer cervical (CC) es el mayor problema de salud pública en países en vías de desarrollo, al ser la infección por el virus del papiloma humano (HPV) el factor etiológico más importante de esta enfermedad. Actualmente, el principal acercamiento para la prevención del CC ha sido a través de programas de detección oportuna del cáncer, lo cual se ha realizado a través del estudio citológico del Papanicolaou (Pap) para la detección de lesiones precursoras. Sin embargo, la sensibilidad y especificidad de la prueba de Pap depende de la destreza del observador en el reconocimiento y clasificación de diferentes anormalidades en las células. El desarrollo de sistemas de diagnóstico temprano para la detección de la infección por HPV ha sido problemático debido a que tanto la citología como la colposcopía identifican la lesión cervical en estadios muy avanzados. De esta forma, la aplicación de las pruebas moleculares ha sido exitosa en el diagnóstico temprano del CC. Estas técnicas moleculares se basan en el reconocimiento de secuencias de ADN de HPV por medio de hibridación del ADN, PCR-RFLP, captura de híbridos (hybrid capture) y el sistema de línea reversa (reverse line blot). Desafortunadamente, estos sistemas no pueden identificar si se trata de una infección activa, latente o persistente. Por esta razón, las técnicas inmunológicas como el Western blot y el ELISA han sido diseñadas para realizar el seguimiento de la respuesta inmune contra el virus. Por ello, aquí se hace una revisión de las técnicas moleculares utilizadas para detectar el HPV, como factor de riesgo del CC, así como las técnicas inmunológicas desarrolladas para el seguimiento de la respuesta inmune contra este virus. Existen diferentes compañías que han desarrollado, manufacturado y comercializado pruebas diagnósticas basadas en identificación de genes para el tamizaje, monitoreo y diagnóstico de HPV. Nuestra revisión se enfoca en el análisis de productos que existen en el mercado, así como en los sistemas inmunológicos usados en investigación y en programas de tamizaje de extensas poblaciones.

Asunto(s)

Femenino , Humanos , Infecciones por Papillomavirus/complicaciones , Infecciones por Papillomavirus/diagnóstico , Neoplasias del Cuello Uterino/microbiología , Pruebas Inmunológicas , Técnicas de Diagnóstico Molecular

RESUMEN

Human Papillomavirus (HPV) infection is the main etiologic agent of cervical cancer and HPV E6 and E7 oncogenes trans-regulate many cellular genes. An association between TGF-beta1 gene expression and cervical cancer development has been suggested; however, the mechanisms by which HPV influences TGF-beta1 expression remain unclear. In the present study we analyzed the mechanism through which HPV-16 E6 and E7 oncoproteins regulate the TGF-beta1 promoter in cervical tumor cells. Our results showed that E6 and E7 increased TGF-beta1 promoter activity. Furthermore, we identified a specific DNA sequence motif in the TGF-beta1 core promoter that is responsible for trans-activation and that corresponds to the Sp1e-binding site associated with HPV-16 E6 and E7 oncoproteins. Mutational analysis showed that the Sp1e recognition site abolished the trans-activation caused by E6 and E7. These results suggest a physical interaction and functional cooperation between viral oncoproteins and cellular regulatory elements of the TGF-beta1 promoter, and may explain the contribution of HPV-16 to TGF-beta1 gene expression in cervical cancer.

Asunto(s)

Proteínas de la Matriz Extracelular/química , Proteínas de la Matriz Extracelular/metabolismo , Papillomavirus Humano 16/patogenicidad , Proteínas Oncogénicas Virales/metabolismo , Proteínas Represoras/metabolismo , Factor de Transcripción Sp1/metabolismo , Activación Transcripcional , Factor de Crecimiento Transformador beta/química , Factor de Crecimiento Transformador beta/metabolismo , Secuencia de Bases , Línea Celular Tumoral , Proteínas de la Matriz Extracelular/genética , Papillomavirus Humano 16/metabolismo , Humanos , Datos de Secuencia Molecular , Proteínas E7 de Papillomavirus , Regiones Promotoras Genéticas , Factor de Crecimiento Transformador beta/genética