Immunology of cervical cancer

ABSTRACT Optimal function of the immune system allows the recognition and elimination of infected and tumor cells. However, these cells can develop mechanisms to evade the cellular immune response. In human papillomavirus (HPV) infection, dysregulation of major histocompatibility complex Class I molecules and other components of the innate immune system promote the survival of infected cells by allowing the infection to persist which, in turn, favors the development of cancer. Further, tumor cells possess inherent mechanisms designed to block the recognition and activation of cytotoxic lymphocytes: particularly, HPV proteins such as E1 and E2 and oncoproteins E5, E6, and E7 that inhibit immune mechanisms and/or stimulate the expression of immunosuppressive cytokines. These mechanisms include a decrease in receptor activation and costimulating molecules on the surface of immune cells, as well as the constitutive expression of molecules that inhibit their function, which allow HPV persistence and tumor progression. Immunotherapy-based therapeutic options are positioned as excellent candidates for the treatment of cervical cancer.

Immunonutrition in cervical cancer: immune response modulation by diet

ABSTRACT In the development of cervical cancer (CC), the immune response plays an essential role, from the elimination of human papillomavirus (HPV) infection to the response against the tumor. For optimal function of the immune response, various factors are required, one of the most important being an adequate nutrition. The complex interaction between nutrients and microbiota maintains the immune system in homeostasis and in case of infection, it provides the ability to fight against pathogen invasion, as occurs in HPV infection. The purpose of this article is to describe the role of diet, food, and specific nutrients in the immune response from the onset of infection to progression to precancerous lesions and CC, as well as the role of diet and nutrition during oncological treatment. The immunomodulatory role of microbiota is also discussed. A detailed analysis of the evidence leads us to recommend a nutritional pattern very similar to the Mediterranean diet or the prudent diet for an optimal immune response. Moreover, pre- and probiotics favorably modulate the microbiota and induce preventive and therapeutic effects against cancer.

Relationships of interleukin-10 with the regulatory T cell ratio and prognosis of cervical cancer patients

OBJECTIVE: This study investigated serum interleukin-10 (IL-10) levels, changes in peripheral blood CD4+CD25+ regulatory T cell (PBCDT) ratios, and the prognosis of cervical cancer (CC) patients. METHODS: Seventy patients with CC composed the observation group, and 70 healthy subjects composed the control group. The PBCDT ratios in the CC patients and healthy subjects were calculated. Serum IL-10 levels were detected with a double antibody sandwich enzyme-linked immunosorbent assay (ELISA). RESULTS: The PBCDT ratio was higher in the patients with active CC [12.16±2.41%] than in the control subjects [6.34±1.05%]. Serum IL-10 levels were higher in the patients with CC [384±106 pg/ml] than in the control subjects [104±50 pg/ml]; the differences in both PBCDT ratio and IL-10 level were statistically significant (p<0.01). Serum IL-10 levels were positively correlated with PBCDT ratios (r=0.375, p<0.05). The 5-year patient survival rate was significantly higher in the low serum IL-10 group (64.2%) than in the high serum IL-10 group (42.8%, p=0.012). CONCLUSIONS: PBCDT ratios and serum IL-10 levels are related to CC activity. These factors are reciprocally related and influence one another, and both are involved in the development and progression of CC. Low IL-10 expression is beneficial regarding the survival of patients with CC.

Glucocorticoid-induced tumor necrosis factor receptor expression in patients with cervical human papillomavirus infection

Introduction The progression of human papillomavirus (HPV) infection in the anogenital tract has been associated with the involvement of cells with regulatory properties. Evidence has shown that glucocorticoid-induced tumor necrosis factor receptor (GITR) is an important surface molecule for the characterization of these cells and proposes that GITR ligand may constitute a rational treatment for many cancer types. We aimed to detect the presence of GITR and CD25 in cervical stroma cells with and without pathological changes or HPV infection to better understand the immune response in the infected tissue microenvironment. Methods We subjected 49 paraffin-embedded cervical tissue samples to HPV DNA detection and histopathological analysis, and subsequently immunohistochemistry to detect GITR and CD25 in lymphocytes. Results We observed that 76.9% of all samples with high GITR expression were HPV-positive regardless of histopathological findings. High GITR expression (77.8%) was predominant in samples with ≥1,000 RLU/PCB. Of the HPV-positive samples negative for intraepithelial lesion and malignancy, 62.5% had high GITR expression. High GITR expression was observed in both carcinoma and high-grade squamous intraepithelial lesion (HSIL) samples (p = 0.16). CD25 was present in great quantities in all samples. Conclusions The predominance of high GITR expression in samples with high viral load that were classified as HSIL and carcinoma suggests that GITR+ cells can exhibit regulatory properties and may contribute to the progression of HPV-induced cervical neoplasia, emphasizing the importance of GITR as a potential target for immune therapy of cervical cancer and as a disease evolution biomarker. .

Expressão de proteínas RAP1 recombinantes e produção de anticorpos anti-RAP1: potencial uso como biomarcador no diagnóstico de tumores

Alterações imunofenotípicas qualitativas e quantitativas na expressão da proteína RAP1, uma pequena GTPase da superfamília RAS, estão presentes em diversos tipos de cânceres, tais como carcinomas de células escamosas de orofaringe, câncer papilar da tireóide, câncer de mama, carcinoma de células renais, leucemia,melanoma, neoplasias intraepiteliais e câncer cervical. Entretanto, para a utilização de RAP1 como biomarcador para auxiliar no diagnóstico imuno-histoquímico de tumores, especialmente do câncer cervical, são necessários anticorpos anti-RAP1 abaixo custo, uma vez que, atualmente, os anticorpos disponíveis no Brasil são importados e de custo elevado, tornando inviável sua utilização no diagnóstico de rotina. Assim, este trabalho tem como objetivos a expressão de proteínas RAP1 recombinantes (rRAP1) em sistema bacteriano, e a produção de anticorpos monoclonais e policlonais anti-rRAP1, visando a sua aplicação no diagnóstico de diversos tumores por imuno-histoquímica.

Dois genes RAP1 sintéticos codificantes para as proteínas rRAP1A e rRAP1AB foram desenhados, sintetizados e subclonados no plasmídeo de expressão bacteriano pQE9 e sua expressões obtidas na linhagem hospedeira E.coli M15. Após indução com IPTG, as proteínas rRAP1 foram purificadas por cromatografia líquida em coluna de afinidade de quelato de níquel,obtendo-se o rendimento, por litro de cultura bacteriana, de 185,6 mg/L de rRAP1A e103,9 mg/L de rRAP1AB. As proteínas rRAP1 purificadas foram inoculadas em animais para a produção de anticorpos monoclonais e policlonais anti-rRAP1A e antirRAP1AB. Ensaios imuno-histoquímicos foram realizados em tecidos de pacientes com neoplasia cervical para a avaliação da reatividade dos anticorpos anti-rRAP1com a proteína RAP1 humana. Uma intensa imunorreatividade foi verificada com o anticorpo anti-rRAP1A (policlonal produzido em coelhos) considerado, até o momento, o melhor candidato para uso na detecção da expressão de RAP1 em ensaios imuno-histoquímicos, o que pode auxiliar no diagnóstico de várias neoplasias, especialmente, do câncer do colo uterino.

Expressão de proteínas RAP1 recombinantes e produção de anticorpos anti-RAP1: potencial uso como biomarcador no diagnóstico de tumores / RAP1 recombinant protein Expression and production of anti-RAP1 antibodies: potential use as a biomarker in the diagnosis of tumors

Alterações imunofenotípicas qualitativas e quantitativas na expressão da proteína RAP1, uma pequena GTPase da superfamília RAS, estão presentes em diversos tipos de cânceres, tais como carcinomas de células escamosas de orofaringe, câncer papilar da tireóide, câncer de mama, carcinoma de células renais, leucemia,melanoma, neoplasias intraepiteliais e câncer cervical. Entretanto, para a utilização de RAP1 como biomarcador para auxiliar no diagnóstico imuno-histoquímico de tumores, especialmente do câncer cervical, são necessários anticorpos anti-RAP1 abaixo custo, uma vez que, atualmente, os anticorpos disponíveis no Brasil são importados e de custo elevado, tornando inviável sua utilização no diagnóstico de rotina. Assim, este trabalho tem como objetivos a expressão de proteínas RAP1 recombinantes (rRAP1) em sistema bacteriano, e a produção de anticorpos monoclonais e policlonais anti-rRAP1, visando a sua aplicação no diagnóstico de diversos tumores por imuno-histoquímica...

Dois genes RAP1 sintéticos codificantes para as proteínas rRAP1A e rRAP1AB foram desenhados, sintetizados e subclonados no plasmídeo de expressão bacteriano pQE9 e sua expressões obtidas na linhagem hospedeira E.coli M15. Após indução com IPTG, as proteínas rRAP1 foram purificadas por cromatografia líquida em coluna de afinidade de quelato de níquel,obtendo-se o rendimento, por litro de cultura bacteriana, de 185,6 mg/L de rRAP1A e103,9 mg/L de rRAP1AB. As proteínas rRAP1 purificadas foram inoculadas em animais para a produção de anticorpos monoclonais e policlonais anti-rRAP1A e antirRAP1AB. Ensaios imuno-histoquímicos foram realizados em tecidos de pacientes com neoplasia cervical para a avaliação da reatividade dos anticorpos anti-rRAP1com a proteína RAP1 humana. Uma intensa imunorreatividade foi verificada com o anticorpo anti-rRAP1A (policlonal produzido em coelhos) considerado, até o momento, o melhor candidato para uso na detecção da expressão de RAP1 em ensaios imuno-histoquímicos, o que pode auxiliar no diagnóstico de várias neoplasias, especialmente, do câncer do colo uterino...

Analysis of NK cells in peripheral blood and tumor infiltrating lymphocytes in cervical cancer patients / Análisis de la población de células asesinas naturales (NK) en sangre periférica y en linfocitos infiltrantes de tumor en pacientes con cáncer de cuello uterino

Objective: To understand the biologic and clinical importance of intratumoral natural killer cells CD16+CD56+CD3 and NKT CD16+CD56+CD3 cells in immune surveillance against cervical cancer. Methods: To understand the significance of NK (CD16+CD56+CD3-) and NKT (CD16+CD56+CD3-) in immune surveillance against cervical cancer, we analysed 39 peripheral blood and 30 biopsy samples from cervical cancer patients, and 40 peripheral blood and 5 biopsy samples from healthy women with normal cytology. The frequencies of NK and NKT and HLA-I expression in keratinocytes were analysed by flow cytometry. Results: In peripheral blood, a higher frequency of NK was observed in the patient group compared with the controls (p=0.002). However, this increase was not reflected in TILs (p=0,095). A significant reduction of HLA-I expression was observed in the patient group compared to the control group (p=0.019). A low number of NK infiltrated was observed in tumors of patients with HLA-I down regulation, but it was not significant (p=0.374). A low number of NK infiltration was associated with shorter survival, but it was not significant (p=0.275). Conclusions: Our results show that although in peripheral blood an increase in NK population was observed in patient group, this increase was not reflected in TILs. It is possible that this inefficient migration of NK´s into the tumor milieu could be related to the expression of immunosuppressive cytokines, in particular IL-10.

Objetivo: Entender la importancia biológica y clínica de las células intratumorales natural killer (NK) CD16+CD56+CD3- y de las células natural killer T (NKT) CD16+CD56+CD3- en la inmunovigilancia del cáncer de cuello uterino (CCU). Métodos: Para comprender el papel de las NK (CD16+CD56+CD3-) y de las células natural killer T (NKT) (CD16+CD56+CD3-) en la inmunovigilancia del CCU, se analizaron 39 muestras de sangre periférica (SP) y 30 biopsias de pacientes con CCU, así como de 40 muestras de SP y 5 biopsias de cuello uterino de mujeres con citología normal. Las frecuencias de NK y NKT y la expresión de HLA-I se analizaron por citometría de flujo. Resultados: Se observó una mayor frecuencia de NK en SP en el grupo de pacientes comparado con el grupo control (p = 0,002). Sin embargo, este aumento no se reflejó en TIL (p = 0,095). Una reducción significativa de HLA-I se observó en el grupo de pacientes (p = 0,019). Esta disminución se asoció una disminución en el número de NK, pero no fue significativa (p = 0,374). Un bajo número de NK se asoció con una menor supervivencia, pero no fue significativo (p = 0,275). Conclusiones: Nuestros resultados muestran que aunque en SP se observa un incremento de NK, este no se refleja en los TIL. Es posible que este tráfico ineficiente de células NK hacia el tumor esté alterado por la expresión de citoquinas inmunosupresoras, en particular IL-10.

Local lymphocytes and nitric oxide synthase in the uterine cervical stroma of patients with grade III cervical intraepithelial neoplasia

OBJECTIVES: Precancerous and cancerous cells can trigger an immune response that may limit tumor development and can be used as a prognostic marker. The aims of the present study were to quantify the presence of B and T lymphocytes, macrophages and cells expressing inducible nitric oxide synthase (iNOS) in the cervical stroma of women with grade III cervical intraepithelial neoplasia (CIN III) or in the intratumoral and peritumoral tissue of women with stage I invasive carcinoma. METHODS: Cervical tissue specimens were obtained from 60 women (20 each from control tissues, CIN III and invasive carcinomas). The average ages in the control, CIN III and invasive groups were 43.9 (± 4.3), 35.5 (± 9.5), and 50 (± 11.2) years, respectively. The specimens were immunohistochemically labeled with antibodies to identify T lymphocytes (CD3), cytotoxic lymphocytes (CD8), B lymphocytes (CD20), macrophages (CD68) and iNOS. We evaluated the markers in the stroma above the squamocolumnar junction (control), at the intraepithelial lesion (CIN cases), and in the nfiltrating tumor. Two independent observers performed the immunohistochemical analysis. RESULTS: T lymphocytes, B lymphocytes, macrophages and iNOS were present more frequently (P<0.05) in the stroma of peritumoral invasive tumors compared to the controls and intratumoral invasive cancer samples. CD3+ and CD20+ lymphocytes were present more frequently in CIN III patients compared to samples from patients with intratumoral invasive cancer (P<0.05). CONCLUSION: High numbers of T and B lymphocytes, macrophages and iNOS-expressing cells in the peritumoral stroma of the invasive tumors were observed. Cell migration appeared to be proportional to the progression of the lesion.

Antibody detection against HPV16 E7 & GP96 fragments as biomarkers in cervical cancer patients.

Background & objectives: Cervical cancer is the second most frequent cancer among females worldwide, especially human papilloma viruses (HPV) types 16 and 18. In viral systems the identification of serological markers would facilitate the diagnosis of HPV infections and virus-related disease. The aim of the present investigation was to determine and search for serologic markers in cervical cancer patients associated with HPV. Methods: A total of 58 Iranian women with invasive cervical carcinoma including adenocarcinoma and squamous cell carcinoma (SCC) were included. Serum antibody response to HPV infections in patients was detected by Western blot and ELISA techniques based on recombinant HPV16E7 and the N-terminal and C-terminal fragments of gp96 (NT-gp96 and CT-gp96) proteins. These recombinant proteins were expressed in Escherichia coli as a His-tag protein and purified using affinity chromatography. Results: The ELISA results indicated that patients with high antibody response to HPV16E7 had significant seroreactivity to CT-gp96 fragment. In Western blot analysis, a strong association between anti-E7, anti-NT-gp96 and anti-CT-gp96 reactivity and cervical cancer was obtained using purified recombinant proteins. In adenocarcinoma cases, no significant difference was observed in seroreactivities between normal and patients. Interpretation & conclusions: The evaluation of cervical cancer patients' seroreactivities against three recombinant proteins (rE7, rNT-gp96 and rCT-gp96) showed significantly higher levels of these markers in SCC only, but not in adenocarcinoma and control groups. Also, the usage of both techniques (ELISA and Western blotting) can provide more reliable tools for diagnosis of cervical cancer.

Cell intrinsic & extrinsic factors in cervical carcinogenesis.

Human papillomavirus (HPV) infection is a common sexually transmitted infection which a majority of infected women are able to clear by mounting an effective immune response. Individuals with a suboptimal immune response may be at increased risk of persistent HPV infection leading to sequelae of various grades of dysplasias and / or associated malignancy. Both cell intrinsic and extrinsic phenomena work in concert to bring about oncogenesis. Cell intrinsic factors for cervical carcinogenesis are: integration of the viral genome into the genome of the host’s cell which correlates with the progression of low grade lesions into high grade ones, inactivation of tumor suppressor genes like p53 and pRB by HPV oncoproteins particularly E6 and E7, deregulation of cell cycle regulators, host DNA synthesis and apoptosis. Cell extrinsic elements include factors contributing towards immune tolerance; some incriminated in the multistep carcinogenesis of HPV induced cervical cancer are: immunoregulatory enzyme indoleamine 2,3-dioxygenase expressing antigen presenting cells, low numbers of invariant Natural Killer T cells, anergic cytotoxic T lymphocytes, regulatory T cells (Tregs), an immunoregulatory microenvironment comprising of increased IL10, TGFβ and reduced IL2; reduced intralesional ratios of effectors (CD4 and CD8) vs. Tregs; and different types of Tregs in the lesions of invasive squamous cell carcinoma. Notch signaling plays a crucial role in regulating T cell differentiation and activation including induction of Tregs. Increased expression of Notch receptor-Jagged 1 and number of Tregs were seen in invasive disease when compared to precancer in cervical cancer. Tregs impart their function either through cytokines or by cell to cell contact. Investigation of the consequences of interference of Notch signaling in terms of the dynamics of intratumoral Tregs in cervical cancer would be interesting.

Resposta imune nas lesões intra-epiteliais do colo uterino / Immune response to the cervical intraepithelial lesions

A resposta imune às neoplasias intra-epiteliais cervicais é muito peculiar, visto que se trata de um processo iniciado por um vírus e pelo próprio microambiente no qual a lesão se inicia - o aparelho reprodutor feminino. Nesta revisão, serão discutidos os principais aspectos da resposta imune envolvidos na infecção pelo HPV e nas neoplasias intra-epiteliais cervicais, além de tentar-se dar uma visão para melhor entender os futuros tratamentos que estão sendo propostos para as neoplasias causadas por esta infecçao.

CTL responses to a DNA vaccine encoding E7 gene of human papillomavirus type 16 from an Iranian isolate

Cervical cancer is the most prevalent tumor in developing countries and the second most frequent cancer among female population worldwide. Specific human papillomaviruses and, most notably, HPV types 16 and 18 are recognized as being causally associated with cervical carcinomas. The early HPV type 16 genes, E6 and E7, directly participate in the in vitro transformation of primary human keratinocytes and represent an excellent target for immune therapy of HPV related disease. The aim of this study was the evaluation of the efficacy of a DNA vaccine containing human papillomaviruse type 16 E7 gene [Iranian isolate] in induction of CTL responses in an animal model. In this study, the expression vector containing HPV type 16 E7 gene was constructed and chosen as a model antigen in the development of a therapeutic DNA vaccine in an animal model. CTL responses, cytokine assay, lymphocyte stimulation test, CD4 and CD8 staining and flowcytometry were done for evaluating of the immune responses. Our findings indicate that the target DNA vaccine can induce an E7-specific CTL response, which is important in the lysis of infected tumor cells, compared to negative control [p < 0.005] after in vivo immunization in the mouse system. The developed vaccine may be promising as an anti-cancer vaccine

Role of IL-10 in immune suppression in cervical cancer.

Cervical cancer is the second most common cancer in the women worldwide and the most frequent in developing countries, including India. Human papilloma virus (HPV) is the major etiological factor in cervical cancer patients. Host factors are also critical in regulating tumor growth and cytokines that modulate immunologic control may be of particular importance. In the present study, we investigated the correlation between the presence of HPV and type of cytokines expressed in cervical carcinomas and attempted to elucidate the possible reasons for the immune suppression. Cytokines investigated were type-1 cytokine IFN-gamma (shows immunostimulatory function and capable of limiting tumor growth) and type-2 cytokines IL-4, IL-10 and IL-6 (show immunosuppressive function and capable of stimulating tumor growth). Our data demonstrated the presence of HPV sub-types 16 and 18 in 86% and 13.8% of cervical tumor biopsies, respectively. The cervical tumor biopsies showed increased presence for mRNA for IL-10 and IL-1alpha, while none of the biopsies showed expression for IFN-gamma. A correlation was observed between the presence of HPV in cervical tumor biopsies and mRNA for IL-10. Increased percentages of CD4+CD25+ regulatory T cells (Tregs) were observed in circulation in cervical cancer patients, providing evidence for increased immune suppression. IL-10 may play a key role in maintenance of Tregs and explains the immunosuppressive state of cervical cancer patients.

Major histocompatibility complex class II polymorphisms and risk of cervical cancer in Puerto Rican women

Association between HLA antigens and cervical squamous cell carcinoma has been described in several populations. To verify whether HLA-DRB1 and DQB1 diversity is related to cervical cancer in Puerto Rican women, 40 cases and 50 controls were HLA typed. DRB1*16 (POR=2.89) and DRB1*11 (POR=1.74) were positively associated with cervical cancer. A negative association was found with DRB1*01 (POR=0.52), DRB1*04 (POR=0.60), DRB1*14 (POR=0.33), DRB1*15 (POR=0.65), DQB1*04 (POR=0.33), DQB1*05 (POR=0.64) and DQB1*06 (POR=0.65). We suggest that HLA Class H polymorphisms are involved in genetic susceptibility to cervical cancer in Puerto Rican women. These results should be confirmed in studies with larger sample size to preclude the possibility of false positive observations.

An overview of human papillomaviruses and current vaccine strategies.

Cervical cancer is one of the most common cancers in women worldwide, particularly in developing countries. The viral origin of cervical cancer has been proven beyond any reasonable doubt. Persistent infection with certain subsets of human papillomaviruses is recognized as a necessary cause for the development of cervical cancer. Persistence of oncogenic HPVs, immunodeficiency, high HPV viral load and cofactors like smoking, multiple sex partners and poor nutrition predispose to cervical cancer. Prophylactic vaccines using HPV virus-like particles containing capsid protein L1 have shown protection against disease in animals and are currently undergoing clinical trials. Therapeutic vaccines using HPV E6 and E7 proteins are also being investigated for their ability to remove residual infection.

Terapia génica con citocinas contra cáncer cervicouterino / Gene therapy with cytokines against cervical cancer

La terapia génica es una excelente alternativa para el tratamiento de muchas enfermedades. La capacidad para manipular el DNA ha permitido dirigir la terapia génica para corregir la función de un gen alterado, aumentar la expresión de un gen o activar la respuesta inmune. Así, se puede proponer el uso del DNA como un medicamento capaz de controlar, corregir o curar una enfermedad. La terapia génica contra cáncer tiene un potencial enorme, y en la última década se han obtenido resultados muy alentadores del uso del DNA para controlar diversas neoplasias en modelos animales, lo cual ha permitido su aplicación en protocolos experimentales en humanos. Esta revisión concentra una reseña de los fundamentos de la terapia génica y su aplicación en cáncer cervical, desde el punto de vista de las alteraciones de la respuesta inmune enfocadas al microambiente tumoral y el uso de las citocinas como moduladores de la respuesta inmune.

Immune response in cervical dysplasia induced by human papillomavirus: the influence of human immunodeficiency virus-1 co-infection - review

Human immunodeficiency virus (HIV-1) has become an important risk factor for human papillomavirus (HPV) infection and the development of HPV associated lesions in the female genital tract. HIV-1 may also increase the oncogenicity of high risk HPV types and the activation of low risk types. The Center for Disease Control and Prevention declared invasive cervical cancer an acquired immunodeficience virus (AIDS) defining illness in HIV positive women. Furthermore, cervical cancer happens to be the second most common female cancer worldwide. The host's local immune response plays a critical factor in controlling these conditions, as well as in changes in the number of professional antigen-presenting cells, cytokine, and MHC molecules expression. Also, the production of cytokines may determine which arm of the immune response will be stimulated and may influence the magnitude of immune protection. Although there are many studies describing the inflammatory response in HPV infection, few data are available to demonstrate the influence of the HIV infection and several questions regarding the cervical immune response are still unknown. In this review we present a brief account of the current understanding of HIV/HPV co-infection, emphasizing cervical immune response.

Asociación entre la presencia de anticuerpos anti-Ras y anti-VPH16 E4/E7 y lesiones intraepiteliales del cérvix / Association between anti-Ras and anti-HPV16 E4/E7 antibodies with cervical intraepithelial lesions

OBJETIVO: Determinar si anticuerpos séricos contra E4, E7 y Ras pueden ser utilizados como marcadores de lesiones tempranas del cérvix uterino asociadas al virus del papiloma humano. MATERIAL Y MÉTODOS: Entre marzo de 1999 y abril de 2000 se realizó un estudio sero-epidemiológico de casos y controles en la clínica de displasias del Hospital General Doctor Gea González, en la Ciudad de México, en 116 muestras de suero para evaluar la presencia de anticuerpos anti-E4, E7 y Ras utilizando un ELISA de captura. Se estimaron razones de momios e intervalos de confianza de 95 por ciento RESULTADOS: Anticuerpos anti-E7 se asociaron a mujeres con lesiones NIC III, mientras que anticuerpos anti-E4 y anti-Ras fueron más frecuentes en lesiones NIC I-II. Al evaluar el perfil de anticuerpos que presentaron las mujeres, encontramos que a) anticuerpos contra dos proteínas predicen la existencia de una lesión NIC I-II, y b) la presencia de tres anticuerpos predicen una lesión NIC III. CONCLUSIONES: La detección de anticuerpos séricos contra E4, E7 y Ras en combinación con otras técnicas de diagnóstico, podrían ser de utilidad para detectar oportunamente a mujeres con lesiones tempranas asociadas al Virus del Papiloma Humano y en riesgo de desarrollar cáncer