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.

Human papillomavirus infection and cervical cancer precursor lesions in women living by Amazon rivers: investigation of relations with markers of oxidative stress / Infecção por papilomavírus humano e lesões precursoras do câncer cervicouterino em Ribeirinhas da Amazônia: avaliação da relação com marcadores de estresse oxidativo

ABSTRACT Objective To investigate the relation between oxidative stress markers, human papillomavirus infection and cervical cancer precursor lesions. Methods The study comprised women aged 14 to 60 years living in communities located by Amazon rivers in the state of Pará (Itaituba, Limoeiro do Ajuru and Bragança, 126, 68 and 43 women respectively). Papanicolau smears and polymerase chain reaction tests for human papillomavirus DNA detection were performed. Blood samples were collected to test malondialdehyde, total and oxidized glutathione levels. Results Malondialdehyde, total and oxidized glutathione concentrations did not differ significantly (p>0.05) between women with and without low-grade squamous intraepithelial lesions across communities. Malondialdehyde levels (8.02nmols/mL) were almost five times higher in human papillomavirus-positive compared to human papillomavirus-negative women (1.70nmols/mL) living in Itaituba (statistically significant difference; p<0.05). Malondialdehyde levels did not differ significantly (p>0.05) between human papillomavirus-positive and human papillomavirus-negative women living in remaining communities. Significant (p<0.05) differences in total glutathione levels between human papillomavirus-positive and human papillomavirus-negative women (8.20μg/mL and 1.47μg/mL, respectively) were limited to those living in Bragança. Conclusion Malondialdehyde and total glutathione levels were significantly associated with human papillomavirus infection. However, lack of similar associations with squamous lesions suggest oxidative stress alone does not explain correlations with cervical carcinogenesis. Other factors may therefore be involved.

RESUMO Objetivo Avaliar a relação de marcadores de estresse oxidativo com a infecção pelo papilomavírus humano e as lesões precursoras do câncer cervical. Métodos Foram incluídas mulheres ribeirinhas da Amazônia (a saber: 126 do município de Itaituba, 68 de Limoeiro do Ajuru e 43 de Bragança), todas do Estado do Pará, de 14 a 60 anos. Foram realizados testes Papanicolau e de reação em cadeia de polimerase para a detecção de DNA do papilomavírus humano. Coleta de amostras de sangue foi realizada para a avaliação do malondialdeído e a determinação da glutationa total e oxidada. Resultados As concentrações oxidativas do malondialdeído e os teores de glutationa total e oxidada não apresentaram diferença significativa (p>0,05) nas mulheres com e sem lesão intraepitelial escamosa de baixo grau, em todas as comunidades pesquisadas. Mulheres com papilomavírus humano em Itaituba apresentaram níveis de malondialdeído (8,02nmols/mL) quase cinco vezes maior quando relacionadas àquelas sem o HPV (1,70nmols/mL), com diferença estatística significativa (p<0,05). As outras comunidades não mostraram diferença estatística significativa entre as concentrações de malondialdeído nas mulheres com e sem papilomavírus humano (p>0,05). Os teores de glutationa total mostraram diferença significativa (p<0,05) somente em Bragança naquelas com (8,20µg/mL) e sem a infecção pelo papilomavírus humano (1,47µg/mL). Conclusão Houve associação significativa da resposta oxidativa do malondialdeído e da glutationa total com a infecção pelo papilomavírus humano, porém não houve diferença quando associada à lesão escamosa, sugerindo que o estresse oxidativo isoladamente não explica a relação com a carcinogênese do colo uterino, que deve ser influenciada ainda por outros fatores.

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.

Determinación del tipo de anemia y su relación con la ingesta alimentaria y marcadores bioquímicos en pacientes con cáncer cérvico uterino / Determination of the type of anemia and its relationship to food consumption and biochemical markers in patients with cervical cancer

Introduction: Anemia is a syndrome frequently present in cancer patients and it effects their quality of life. In cervical cancer, anemia develops in most of the patients, without knowledge of its possible cause: transvaginal bleeding presents in advanced stages, or a diet low in iron and other micronutrients can also contribute to this phenomenon. Objective: To determine the type of anemia, serum levels of iron, ferrite, transferring, folic acid and vitamin B12 and relate them with dietary intake. Material and methods: We conducted a transversal study in 55 cervical cancer patients without neither active transvaginal bleeding nor oncologic treatment. A blood sample was taken in order to determine the biochemical markers from which we classified the type of anemia and serum micronutrient levels. Diet was evaluated using a semi quantitative food frequency questionnaire. Results and conclusions: The most frequent type of anemia as normocytic norm chromic, which can be associated to the pathology per se. Patients with microcytic anemia showed allow iron consumption, as well as low iron serum levels. Overall, daily-recommended allowances were not met; this can contribute to nutritional alterations.

Introducción: La anemia es un síndrome que los pacientes con cáncer presentan frecuentemente, repercutiendo este fenómeno en su calidad de vida. En el cáncer cérvicouterino (CaCu), la anemia se desarrolla en la mayoría de las pacientes sin saber la causa que la produce; como el sangrado que se presenta en estadios avanzados o por el tipo de dieta baja en hierro y otros micronutrimentos. Objetivo: Determinar el tipo de anemia, las concentraciones séricas de hierro sérico, ferritina, transferrina, ácido fólico y vitamina B12 y relacionarlo con la ingestión dietética. Sujetos y métodos: Se realizó un estudio transversal, con 55 pacientes con diagnóstico de cáncer cérvico uterino sin sangrado transvaginal presente ni tratamiento oncológico. Se tomó una muestra de sangre a fin de determinar parámetros bioquímicos a partir de los cuales se determinó la presencia de anemia y el tipo de ella, así como concentraciones de micronutrimentos hematopoyéticos. Se aplicó una frecuencia alimentaria semicuantitativa para evaluar la ingestión dietética. Resultados y conclusiones: La anemia de mayor frecuencia fue de tipo normocítica normocrómica, misma que puede asociarse a la patología en curso. Las pacientes con anemia microcítica mostraron concentraciones bajas de hierro sérico, así como un bajo consumo del mismo. En general, no se cubrieron las recomendaciones de ingestión de micronutrimentos, aspecto que puede favorecer las alteraciones nutricionales.

Lipid peroxidation and antioxidants in different stages of cervical cancer: Prognostic significance.

Background: Free radical Injury is associated with cancer, but how the extent of oxidative stress correlates with the FIGO (International Federation of Gynecology and Obstetrics) stage in Carcinoma Cervix (Ca Cx), and its significance as a prognostic marker, is not clear and needs an in-depth study. Aim: To correlate the blood levels of Lipid Peroxidation (LPO), Reduced Glutathione (GSH), Superoxide Dismutase (SOD), and Vitamin A and E levels with the clinical stage in Ca Cx. Settings and Design: This is a Prospective Case Control Study. Materials and Methods: LPO, SOD, reduced GSH were estimated by Bio Chemical Assays and Vitamins by High Performance Liquid Chromatography (HPLC). Statistical Analysis: The cases and controls were compared using One Way ANOVA and different stages over different time periods were individually compared by Repeated Measure Analysis of Variance. Results: The results indicated a statistically significant increase of LPO vis-a-vis the FIGO stage of Ca Cx and control, while the antioxidant status as depicted by GSH and SOD decreased. Vitamin A and E levels were significantly lower in cancer cases as compared to the control. Conclusion: Increased LPO and reduced antioxidant levels may be taken as associated predictive markers, thus suggesting that Ca Cx cases should get nutritive supplements to contain the blood LPO level and maintain a positive balance of antioxidants for a better outcome in terms of delayed recurrence and better Quality of Life (QOL).

Vitamin B12 status of Thai women with neoplasia of the cervix uteri.

The vitamin B12 statuses of Thai women with high- and low-grade cervical dysplasia were studied and compared with women with normal cytological smears. Serum vitamin B12 and vitamin B12 intakes were assessed, as well as demographic characteristics, sexual behavior, reproductive and menstrual history, exogenous hormone use, personal and familial medical history, smoking habit, and other risk factors. The presence or absence of genital HPV DNA was determined by polymerase chain reaction (PCR). Serum vitamin B12 levels in women with normal cytological smears were significantly higher than those with both high- and low-grade cervical dysplasia (p<0.001). Low vitamin B12 serum levels were significantly statistically associated with increased low-grade (OR = 4.08; 95% CI = 1.41-11.79; p < 0.05) and increased high-grade cervical dysplasia risk (OR = 3.53; 95% CI = 1.24-10.04; p< 0.05) for the highest vs lowest quartiles of serum vitamin B12. This study indicated a relationship between low vitamin B12 status and increased risk of cervical cancer.

Detection of cancer cervix by estimation of sialic acid.

The study was undertaken to determine the sialic acid concentration of cervical tissue and serum of women with healthy and unhealthy cervix. The sialic acid value in local tissue of cervix and in serum showed slight elevation in benign inflammatory lesions, moderate elevation was found in severe dysplasia and preinvasive carcinoma and marked elevation in invasive carcinoma cervix. The values, both in the serum and in the cervical tissue, increased gradually with the advancement of grading of frank malignancy. The elevation of sialic acid in the serum was slow to start with but persisted for a longer period, wereas the concentration in local tissue was found to be increasing with the pathological process.

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

Apoptotic activity in lymphocytes from radiation-treated cervical cancer patients.

PURPOSE: The per cent apoptotic activity in lymphocytes from pre-treated cervical cancer patients was cross-sectionally compared with post-treated patients at 1 month, 3 months, 1 year, and 5 years after completing the standard radiation therapeutic regimen. In addition, the differences in the per cent apoptotic activities among various stages of cervical cancer were simultaneously analyzed. MATERIAL AND METHOD: Blood samples were collected from five patients in each stage of cervical cancer before treatment, and at 1 month, 3 months, 1 year, and 5 years after completing the radiation therapy. The control samples were collected from healthy and aged-match female blood donors. The lymphocytes were separated and exposed to 0.5 Gy Co irradiation to induce apoptosis. The apoptotic cells were detected by in situ terminal deoxynucleotidyl transferase (TdT assay) and counted under a fluorescent microscope. Both the apoptotic index and per cent apoptotic activity were calculated. RESULTS: The per cent apoptotic activities in lymphocytes from all pre-treated patients with stage II and III cervical cancer were significantly lower than the controls (p = 0.001). The apoptotic activity in normal control, however, was not significantly different from the pre-treated stage I cervical cancer group. Following radiation therapy, the apoptotic activities at 1 month, 3 month and 1 year were increased in all stages. The per cent apoptotic activity, in all stages of cervical cancer at 5 years after treatment, was statistically higher than that of the pre-treated groups. CONCLUSION: There was a decrease of per cent apoptotic activity in lymphocytes from all pre-treated cervical cancer patients in the present study, the change of which was reversed to normal after treatment in non-recurrent cases.

Serum copper levels in carcinoma of ovary and cervix.

The role of serum copper level (SCL) as a diagnostic and prognostic tool in genital tract malignancies was evaluated. SCL was employed as a marker for response to treatment (surgical/radiotherapy). 129 women attending gynaecology outpatient department or admitted in the gynecology ward were studied. Of these 77 patients in the disease (study) group were proven cases of genital tract malignancies and 52 served as controls. Outcome measures studied were: SCL levels estimated before initiation of any treatment i.e. surgery/radiotherapy; thereafter, at two weeks after completion of treatment. Follow up of the study subjects was done between 4-8 weeks and 8-10 weeks, when the patients were evaluated for any recurrence of disease and SCLs were also estimated. Kruskal-Wallis one-way analysis of variance determined whether values varied significantly among the different groups studied. Mean SCLs were found to be significantly elevated in cases of Ca ovary (n = 15), early CaCx (n = 14) and late CaCx (n = 48), as compared to the control group, comprising of women with no signs and symptoms of malignancy (n = 52). SCLs decreases significantly (P < 0.001) after treatment of Ca ovary and CaCx. These results indicate a possible clinical usefulness of estimating serum copper levels in women with genital tract cancer and suggest a role for SCL in the evaluation of the disease activity and as a prognostic tool in the management of genital malignancies.

Serum tissue polypeptide specific antigen (TPS) in locoregional failure and distant metastasis of cervical carcinoma.

Tissue polypeptide specific antigen (TPS) was measured by the ELISA Technique in the sera of 51 patients with locoregional failure and metastasis of squamous cell carcinoma of the cervix in order to evaluate the serum level of TPS in known cases of metastasis and recurrence. There were 32 cases of local residual or recurrent disease and 19 cases of distant metastasis, including lymph nodes (paraaortic and supraclavicular lymph node) and visceral metastasis. The range of TPS levels in the locoregional failure group were 38.2 355.2 micro/l with a mean of 312.5 and 35.7 - 4822 micro/l with a mean of 833.36 micro/l in the metastatic group. With the cut-off value of 90 micro/l, the rates of TPS elevation were 27 in 32 cases (84.37%) of the loco-regional failure group and 16 in 19 cases (84.21%) of the metastatic group. Among the metastatic group, the mean of TPS level in visceral metastasis was much higher than the group of lymph node metastasis (1518.4 micro/l vs 215.1 micro/l). TPS level might be used as the follow-up guide for prediction of locoregional failure and metastasis in squamous cell carcinoma of the cervix after the completion of the treatment. In patients with a significantly high level of serum TPS, the distant metastases or local recurrence should be searched for. However, a prospective study of TPS levels in cervical cancer patients after completion of treatment should be done in order to evaluate the sensitivity and specificity of this tumor marker.

ADN del virus del papiloma humano en plasma de pacientes con carcinoma cervicouterino: un potencial marcador de enfermedad residual mínima / DNA-human paillomavirus in serum of patients with cervical carcinoma: a potential marker in minimum residual disease

Antecedentes. El carcinoma cervicouterino es la primera causa de muerte por cáncer en mujeres mexicanas. Estudios moleculares han demostrado que el virus del papiloma humano (VHP) es el principal agente etiológico de esta enfermedad. En este tumor no existe un marcador sensible y específico que pudiera utilizarse como marcador de enfermedad residual mínima y para prevenir recaídas tempranamente. Dado que la amplificación en plasma de secuencias de ADN específicas a tumores se ha utilizado con este fin, en este trabajo se determinó la factibilidad de amplificar, mediante reacción en cadena de la polimerasa en el ADN del plasma, secuencias de VPH en pacientes con cáncer cervicouterino. Material y métodos. Se analizaron 34 pacientes con carcinoma cervicouterino invasor con diferentes estadios clínicos sin tratamiento previo y 24 mujeres con infección cervical por VPH pero sin lesiones invasoras. La extracción del ADN se realizó mediante técnicas estándar: digestión con proteinasa K, extracción con fenol-cloroformo y precipitación con etanol. Para verificar la calidad del ADN, se amplificó un fragmento del gen de la ß-globina. Las secuencias del VPH se amplificaron con los ligonucleótidos generales LICI-LIC2. Las muestras positivas también se amplificaron con oligonucleótidos específicos para VPH tipos 16 y 18. Resultados. ADN del VPH se encontró en el 70 por ciento de los casos de pacientes con cáncer y no se amplificó en los 24 controles. No se encontró correlación entre el resultado de la reacción en caderna de la polimerasa con el estadio clínico ni con el tipo viral, peso se observó menor frecuencia de positividad en los tumores adenoescamosos. Conclusión. Los resultados sugieren que la detección del ADN del VPH en el plasma de las pacientes es específica de aquéllas con cáncer invasor y que esto podría constituir un marcador de enfermedad residual mínima y predictor temprano de recaídas

Plasma level of antioxidant nutrients (retinol and alphatocopherol) in cases with different grades of cervical carcinoma.

A study was conducted on 23 patients on various grades of cervical cancer, selected purposely from Dhaka Medical College Hospital. Vitamin A and E level of this histologically proved cases showed mean serum level of vitamin A was 0.41 +/- 0.19 and that vitamin E was 4.21 +/- 2.15, and these low level was constant in all Histopathological grading of cervical cancer. Contraceptive user and smoker had very low level of vitamin A and E. Majority (51%) of the cases, were from lower middle class and 49% were from lower class, seventy seven percent were illiterate, 97% were married before 18 years, 57% had more than 6 children, 71% did not use contraceptive. The availability of vitamin A rich food were within reach of lower income group, than the food source of vitamin E which are usually of animal origin. It is concluded that in cervical cancer had low level of antioxidant vitamins A and E. Further study is indicated for understanding detail antioxidant nutrients role in aetiopathology of cervical cancer in Bangladesh.

Proliferación de leucocitos de sangre periférica en presencia de sueros de pacientes con cáncer cervicouterino e interleucina-2 recombinante / Proliferation of peripheral blood lymphocytes in presence of sera from patients with cervical cancer and interleukin-2

Diversos estudios han centrado su atención en el establecimiento de una técnica que permita activar y hacer responder de forma específica a linfocitos del paciente contra su propio tumor. Esto se ha conseguido en ocasiones activando linfocitos con mitógenos endógenos, como la interleucina-2 (IL-2). Sin embargo, se ha encontrado que algunos tumores secretan al torrente sanguíneo factores inhibidores de este tipo de respuesta inmune. Con la finalidad de determinar si las células de cáncer de cérvix (CaCu) secretan al torrente sanguíneo factores inhibidores de la respuesta proliferadora de linfocitos, en este trabajo se utilizaron sueros de 12 pacientes con CaCu en cultivos de leucocitos de sangre periférica (LSP). Para evaluar la posible inhibición de estos sueros en la activación mediada por la IL-2, se utilizaron tanto LSP de pacientes con CaCu como de donadores normales. Los resultados obtenidos muestran que in vitro no existe inhibición de la activación a la proliferación de LSP provenientes de pacientes con CaCu por la rIL-2, tanto en presencia de sueros normales como de pacientes con CaCu. En consecuencia, estos resultados indican que las células malignas que constituyen al CaCu probablemente no secretan factores inhibidores de la activación linfocitaria y que los LSP de estos pacientes no han perdido la capacidad de ser activados por la IL-2.

Serum fucose levels in gynaecological disorders including carcinoma cervix.

Serum protein bound fucose levels and the ratio between serum protein bound fucose and total serum protein were determined in 40 healthy females, 60 patients with benign gynaecological disorders and 50 patients with previously untreated cases of carcinoma cervix. The mean serum fucose level in healthy subjects was 8.96 mg% and the ratio between serum protein bound fucose and total serum protein was 1.29 x 10(-3). Mean serum fucose level in patients with benign gynaecological disorders was 9.42 mg% and the ratio between serum protein bound fucose and total serum protein was 1.42 x 10(-3). In stage I and II carcinoma cervix patients mean serum fucose was 11.92 mg% and the ratio between serum protein bound fucose and total serum protein was 1.68 x 10(-3). In stage III and IV carcinoma cervix patients the mean serum fucose was 17.76 mg% and the ratio between serum protein bound fucose and total serum protein was 2.64 x 10(-3).