Importance of vaccination against human papillomavirus in a rural settlement in Terenos, Mato Grosso do Sul.

OBJECTIVE: To understand health professionals' perceptions about vaccination against human papillomavirus (HPV) in the Santa Mônica rural settlement in Terenos, Mato Grosso do Sul. METHODS: Quantitative and qualitative methodologies, consultations on vaccination cards, records of community health agents and the focus group technique were used. The main factors of hesitation and vaccine refusal were analyzed, as well as the health team's strategies for the process of immunization against HPV, from June to August 2018. RESULTS: Of 121 children and adolescents, 81 (66.94%) received the complete vaccination schedule. Complete vaccination coverage for women was 73.17% (60/82) and for men, 53.8% (21/39). It was observed that, although strategies are adopted for vaccine promotion, such as mobile actions, the public is resistant due to superficial knowledge about the vaccine and its use in an early age group, showing itself to be susceptible to the negative influence of the media and to society's taboos. In addition, difficulties regarding the use of the Unified Health System card and the shortage of professionals were also observed. CONCLUSION: The results explain the immunization coverage below the target and reinforce the need to strengthen the family health strategy, as well as the permanent and continuing education of professionals, in order to increase parental confidence and adherence to vaccination.

Cytokine profiling of samples positive for Chlamydia trachomatis and Human papillomavirus.

Persistent human papillomavirus (HPV) infection is closely associated with cervical carcinoma. Co-infection in the endocervical environment with other microorganisms, such as Chlamydia trachomatis, may increase the risk of HPV infection and neoplastic progression. While in some individuals, Chlamydia trachomatis infection is resolved with the activation of Th1/IFN-γ-mediated immune response, others develop a chronic infection marked by Th2-mediated immune response, resulting in intracellular persistence of the bacterium and increasing the risk of HPV infection. This work aimed to quantify cytokines of the Th1/Th2/Th17 profile in exfoliated cervix cells (ECC) and peripheral blood (PB) of patients positive for Chlamydia trachomatis DNA, patients positive for Papillomavirus DNA, and healthy patients. Cytokine levels were quantified by flow cytometry in ECC and PB samples from patients positive for C. trachomatis DNA (n = 18), patients positive for HPV DNA (n = 30), and healthy patients (n = 17) treated at the Hospital de Amor, Campo Grande-MS. After analysis, a higher concentration of IL-17, IL-6, and IL-4 (p <0.05) in ECC; INF-γ and IL-10 (p <0.05) in PB was found in samples from patients positive for C. trachomatis DNA compared to samples from healthy patients. When comparing samples from patients positive for HPV DNA, there was a higher concentration of cytokines IL-17, IL-10, IL-6, and IL-4 (p <0.05) in ECC and IL-4 and IL-2 (p <0.05) in PB of patients positive for C. trachomatis DNA. These results suggest that induction of Th2- and Th17 mediated immune response occurs in patients positive for C. trachomatis DNA, indicating chronic infection. Our results also demonstrate a high concentration of pro-inflammatory cytokines in ECC of patients positive for C. trachomatis DNA.

Importance of vaccination against human papillomavirus in a rural settlement in Terenos, Mato Grosso do Sul / Importância da vacinação contra o papilomavírus humano em um assentamento rural em Terenos, Mato Grosso do Sul

ABSTRACT OBJECTIVE To understand health professionals' perceptions about vaccination against human papillomavirus (HPV) in the Santa Mônica rural settlement in Terenos, Mato Grosso do Sul. METHODS Quantitative and qualitative methodologies, consultations on vaccination cards, records of community health agents and the focus group technique were used. The main factors of hesitation and vaccine refusal were analyzed, as well as the health team's strategies for the process of immunization against HPV, from June to August 2018. RESULTS Of 121 children and adolescents, 81 (66.94%) received the complete vaccination schedule. Complete vaccination coverage for women was 73.17% (60/82) and for men, 53.8% (21/39). It was observed that, although strategies are adopted for vaccine promotion, such as mobile actions, the public is resistant due to superficial knowledge about the vaccine and its use in an early age group, showing itself to be susceptible to the negative influence of the media and to society's taboos. In addition, difficulties regarding the use of the Unified Health System card and the shortage of professionals were also observed. CONCLUSION The results explain the immunization coverage below the target and reinforce the need to strengthen the family health strategy, as well as the permanent and continuing education of professionals, in order to increase parental confidence and adherence to vaccination.

RESUMO OBJETIVO Compreender as percepções dos profissionais de saúde acerca da vacinação contra o papilomavírus humano (HPV) no Complexo de assentamentos Santa Mônica, em Terenos, Mato Grosso do Sul. MÉTODOS Foram utilizadas metodologias quanti-qualitativas, consultas em cartões vacinais, registros de agente comunitários de saúde e a técnica de grupo focal. Foram analisados os principais fatores de hesitação e recusa vacinal, bem como as estratégias da equipe de saúde para o processo de imunização contra o HPV, de junho a agosto de 2018. RESULTADOS De 121 crianças e adolescentes, 81 (66,94%) receberam o esquema vacinal completo. A cobertura vacinal completa feminina foi de 73,17% (60/82) e a masculina de 53,8% (21/39). Observou-se que, embora sejam adotadas estratégias para a promoção da vacina, como ações volantes, o público encontra-se resistente devido ao conhecimento superficial sobre a vacina e sua utilização em faixa etária precoce, mostrando-se suscetível à influência negativa da mídia e aos tabus da sociedade. Além disso, dificuldades quanto ao uso do cartão do Sistema Único de Saúde e a escassez de profissionais também foram observadas. CONCLUSÃO Os resultados justificam a cobertura vacinal abaixo da meta e reforçam a necessidade de fortalecimento da estratégia de saúde da família, bem como da educação permanente e continuada dos profissionais, a fim de aumentar a confiança dos pais e a adesão à vacinação.

Correlation between melatonin concentration and cytokines in cervical mucus in positive samples for the presence of human papillomavirus.

The human papillomavirus (HPV) is the main causative agent of cervical cancer, characterized by neoplastic lesions in the cervix. Based on the morphology of the cells of the uterine cervix, the findings are classified as negative intraepithelial lesions for malignancies, low-grade squamous intraepithelial lesions, high-grade squamous intraepithelial lesions (HSILs), atypical squamous cells of undetermined significance and atypical squamous cells of undetermined significance without excluding HSILs (ASCs-H). The progression of neoplastic lesions is related to the cervix's microenvironmental inflammatory process and mediated by the expression and stimulation of cytokines. Cervical mucus is a viscous liquid secretion composed of proteins, inorganic components, pro-and anti-inflammatory agents, and an important protective barrier. This study aimed to quantify and correlate cytokines IL-6, IL-8, and IL-10 and Melatonin in cervical mucus. According to the results, a decrease in MLT was observed in LSIL, HSI, and ASC-H groups than in the NILM group. The cytokines IL-6 and IL-8 showed greater expression in the LSIL and HSIL groups than the NILM group. HSIL group showed a negative correlation between the MLT and IL-6 and IL-8 concentrations. In the ASC-US group, IL8 level was positively correlated to MLT levels. We suggest that IL-6, IL-8, and MLT levels in HSIL groups are decisive for the progression of neoplastic lesions in HPV infections. New cervical cancer treatment strategies may include cytokine and melatonin control targets for effective immunotherapy.

IL-6 and IL-10 in the serum and exfoliated cervical cells of patients infected with high-risk human papillomavirus.

Persistent infection by high-risk human papillomavirus (HR-HPV) is the main cause of cervical cancer and its precursor lesions. While some cytokines help immune cells in virus clearance, others contribute to the persistence of infection and neoplastic progression. Here, the levels of interferon (IFN)-γ, tumor necrosis factor (TNF)-α, interleukin (IL)-10, IL-6, IL-4, and IL-2 were quantified in the serum and exfoliated cervical cells (ECCs) of patients with HR-HPV, and the presence of IL-6+ cells was investigated in uterine cervix biopsies. Cytokine levels in the serum and ECCs of 26 HR-HPV DNA-positive patients and 18 HPV DNA-negative patients were measured using flow cytometry. Fifteen uterine cervix biopsy samples embedded in paraffin were subjected to immunohistochemical analysis for the detection of IL-6+ cells. HR-HPV-positive patients showed increased IL-6 and IL-10 in the ECCs and serum, respectively. Compared with HPV DNA-positive patients, HPV DNA-negative patients had higher levels of IL-6 in ECCs. Patients with multiple infections of HPV had higher levels of IL-6 in their ECCs than those with a single infection. Immunostaining of uterine cervix biopsy samples revealed no differences in IL-6 expression between the different classes of histopathological lesions. However, differences were observed in the expression levels of IL-6 and IL-10 at the systemic and local levels in HR-HPV-positive patients without cervical lesions. Considering the functional characteristics of these cytokines, it can be inferred that such patients are prone to persistent HPV infection.

Synergistic effect and ultrastructural changes in Trypanosoma cruzi caused by isoobtusilactone A in short exposure of time.

Butanolides have shown a variety of biological effects including anti-inflammatory, antibacterial, and antiprotozoal effects against certain strains of Trypanosoma cruzi. Considering the lack of an effective drug to treat T. cruzi infections and the prominent results obtained in literature with this class of lactones, we investigated the anti-T. cruzi activity of five butanolides isolated from two species of Lauraceae, Aiouea trinervis and Mezilaurus crassiramea. Initially, the activity of these compounds was evaluated on epimastigote forms of the parasite, after a treatment period of 4 h, followed by testing on amastigotes, trypomastigotes, and mammalian cells. Next, the synergistic effect of active butanolides against amastigotes was evaluated. Further, metacyclogenesis inhibition and infectivity assays were performed for the most active compound, followed by ultrastructural analysis of the treated amastigotes and trypomastigotes. Among the five butanolides studied, majoranolide and isoobtusilactone A were active against all forms of the parasite, with good selectivity indexes in Vero cells. Both butanolides were more active than the control drug against trypomastigote and epimastigote forms and also had a synergic effect on amastigotes. The most active compound, isoobtusilactone A, which showed activity against all tested strains inhibited metacyclogenesis and infection of new host cells. In addition, ultrastructural analysis revealed that this butanolide caused extensive damage to the mitochondria of both amastigotes and trypomastigotes, resulting in severe morphological changes in the infective forms of the parasite. Altogether, our results highlight the potential of butanolides against the etiologic agent of Chagas disease and the relevance of isoobtusilactone A as a strong anti-T. cruzi drug, affecting different events of the life cycle and all evolutionary forms of parasite after a short period of exposure.

Cytokine profile in Leishmania-positive blood donors.

Serum levels of interleukin 2 (IL-2), interleukin 4 (IL-4), interleukin 6 (IL-6), interleukin 10 (IL-10), interleukin 17 (IL-17), interferon gamma (IFN-γ), tumor necrosis factor α (TNF-α), and interleukin 1ß (IL-1ß), cytokines involved in the immune response, were investigated in 75 Leishmania-positive blood donors living in endemic areas. Based on their status in 2011 and 2015, the subjects were clustered into three groups: positive for at least one diagnostic method in both years, but lacking clinical progression to disease (G1); positive on at least one method in 2011 but negative in 2015 (G2); negative on all methods in both years (G3). Donors were interviewed for sociodemographic data collection and underwent clinical evaluation and laboratory tests. Serum cytokines were quantified using a CBA Flex set (BD Biosciences). Significant differences were found for all the cytokines evaluated, with lower concentrations in consistently Leishmania-negative individuals. The exception was IFN-γ, with similar levels among all donors. No changes consistent with active disease were observed in the laboratory results for Leishmania-positive donors who underwent clinical evaluation, none of whom progressed to disease. This suggests that infection control is associated with serum IL-17 levels. Resolution of Leishmania infection in positive donors may be related to high levels of IL-17 and low levels of IL-10, highlighting the role played by IL-17 in asymptomatic Leishmania-infected individuals.

Oncogenic high-risk human papillomavirus in patients with full denture.

Human Papillomavirus (HPV) has considerable tropism for epithelial and mucosal tissues and can therefore be found in several anatomical sites, including the oral cavity. This study aimed to investigate the presence of HPV-DNA and the most frequent viral types in patients using full dentures, compare to patients not using full dentures and to associate its presence with socio-epidemiological and behavioral factors. The study consisted of 90 patients with or without full dentures at the time of collection, treated at a public dental clinic. The samples were obtained by exfoliating the oral cavity, and analyzed for HPV-DNA using the nested PCR with PGMY09/11 (450-bp), and general primers GP5+/GP6+ (150-bp). Genotyping was performed by specific-type PCR to HPV 6, 11, 16, 18, 31, 33, and 45; and Restriction Fragment Length Polymorphism (RFLP). Pearson's Chi-square test (x 2 ) or Fisher's exact test were applied and significant variables in these tests were analyzed by multinomial logistic regression analysis to estimate odds ratio (OR). HPV-DNA was detected in 27.7% of samples and, among those obtained from patients using full dentures, positivity for HPV-DNA was 41.9% (p = 0.025). The most frequent viral types were low-risk HPV 6 and 11, and high-risk HPV 31 and 45. Patients who used full dentures had an odds ratio of 2.1 to be positive for HPV DNA. Our results indicate the need for periodic dental follow-up of patients with full dentures in order to preserve the basic conditions of oral health, and also to monitor the appearance of lesions with malignant potential.

Interleukin-17 expression in the serum and exfoliated cervical cells of patients infected with high-risk oncogenic human papillomavirus.

Persistent infection by high-risk oncogenic human papillomavirus (HR-HPV) is the main cause of cervical cancer and its precursor lesions, and both the systemic and local immunological responses play an important role in eliminating or maintenance this infection. Th17 cells, as well as interleukin (IL)-17, are related to tumor growth and persistence of viral infection. Thus, this study aimed to quantify IL-17 in the serum and exfoliated cervical cells of HR-HPV-infected patients and healthy patients as well as identify CD4+IL17+ cells and IL-17 production in uterine cervix biopsies to better understand the behavior of this cytokine in HPV infections. IL-17 was quantified (pg/mL) in the serum and exfoliated cervical cells of 26 HR-HPV-infected patients, and in 18 healthy patients, using flow cytometry. Fifteen paraffin-embedded biopsy samples from the uterine cervix were subjected to immunohistochemistry to detect CD4+IL-17+ and IL-17+ cells. There was a significant increase in the concentration of IL-17 in HR-HPV-positive patients' serum when compared to that in samples of exfoliated cervical cells (p < 0.05). Likewise, when compared with that in healthy patients, the IL-17 concentration was still higher in HR-HPV-positive patients sera (p < 0.05). We did not find differences in the amount of CD4+IL-17+ cells and other IL-17-secreting cells between different histopathological lesions. Our results suggest that HR-HPV infection predominantly stimulates systemic IL-17 production along with less localized expression.

Self-Collection and Molecular Diagnosis for Detection of Human Papillomavirus: Why Incorporate It?

PURPOSE OF REVIEW: Cervical cancer, the third cause of death by cancer among Brazil's women, is associated with human papillomavirus (HPV) infection. In some countries of South America, North America, Europe, and Oceania, initial screening for HPV DNA and subsequent follow-up with HPV-positive patients using colposcopy and cytological testing are used as preventative measures. RECENT FINDINGS: For HPV DNA detection, it is necessary to obtain cervical cells by conventional clinical collection method or self-collection of the cells that flake off from the uterine cervix and vaginal canal. Self-collection has been shown to be a viable option for obtaining samples and is a less invasive method that is more accepted by women. Thus, it can potentially decrease the limitations of the conventional clinical collection methods. The efficiency of the self-collection method aligned with the implementation of HPV molecular testing, if adopted by public and private health care systems, may extend the reach of current cervical cancer prevention efforts. In addition, considering all phases from triage to treatment, this method may reduce health care costs and the time spent by patients and health care teams to conduct examinations and collect samples.

Detection of regulatory T cell phenotypic markers and cytokines in patients with human papillomavirus infection.

Infection with human papillomavirus (HPV) is the main cause of cervical cancer. Viral persistence is considered the main risk factor for neoplastic progression and evidence suggests that regulatory T cells (Treg) play an important role in the failure of viral elimination. The aim of this study was to detect phenotypic markers of Treg and cytokines interleukin (IL)-10 and transforming growth factor (TGF)-ß, in the cervical microenvironment of HPV-infected patients. One hundred and one samples of uterine cervix embedded in paraffin were analyzed. We used immunohistochemistry to examine the coexpression of the CD25/FOXP3 and CD4/TGF-ß markers, and the expression of GITR and IL-10 in cells present in the cervical stroma. We detected a microenvironment composed of high proportions of CD25+ FOXP3+ , CD4+ TGFß+ , IL-10+ , and GITR+ cells in samples with high viral loads and severe lesions of HPV-infected patients. The abundance of these markers, indicative of the presence of Treg cells and immunosuppressive cytokines, was significantly associated with severe lesions and elevated viral loads in the examined samples. These results suggest that Treg cells may be involved in maintaining a microenvironment favorable for viral persistence and neoplastic progression. Our findings support those of previous studies that suggested that these markers could be used to predict HPV persistence and neoplastic progression, and as potential targets for immune response modulation.

Non-polarized cytokine profile of a long-term non-progressor HIV infected patient

ABSTRACT The HIV-1 initial viral infection may present diverse clinical and laboratory course and lead to rapid, intermediate, or long-term progression. Among the group of non-progressors, the elite controllers are those who control the infection most effectively, in the absence of antiretroviral therapy (ART). In this paper, the TH1, TH2 and TH17 cytokines profiles are described, as well as clinical and laboratory aspects of an HIV-infected patient with undetectable viral load without antiretroviral therapy. Production of IL-6, IL-10, TNF-α, IFN-γ, and IL-17 was detected; in contrast IL-4 was identified. Host-related factors could help explain such a level of infection control, namely the differentiated modulation of the cellular immune response and a non-polarized cytokine response of the TH1 and TH2 profiles.

Non-polarized cytokine profile of a long-term non-progressor HIV infected patient.

The HIV-1 initial viral infection may present diverse clinical and laboratory course and lead to rapid, intermediate, or long-term progression. Among the group of non-progressors, the elite controllers are those who control the infection most effectively, in the absence of antiretroviral therapy (ART). In this paper, the TH1, TH2 and TH17 cytokines profiles are described, as well as clinical and laboratory aspects of an HIV-infected patient with undetectable viral load without antiretroviral therapy. Production of IL-6, IL-10, TNF-α, IFN-γ, and IL-17 was detected; in contrast IL-4 was identified. Host-related factors could help explain such a level of infection control, namely the differentiated modulation of the cellular immune response and a non-polarized cytokine response of the TH1 and TH2 profiles.

Human papillomavirus and coinfections with Chlamydia trachomatis, Gardnerella vaginalis, and Trichomonas vaginalis in self-collected samples from female sex workers in the Central-Western region of Brazil / Papilomavírus humano e coinfecção por Chlamydia trachomatis, Gardnerella vaginalis e Trichomonas vaginalis em amostras autocoletadas de mulheres profissionais do sexo da região Centro-Oeste do Brasil

ABSTRACT Introduction: Human papillomavirus (HPV) is intimately associated with cervical cancer, and the presence of coinfections, such as with Chlamydia trachomatis, Gardnerella vaginalis and Trichomonas vaginalis, may potentiate or facilitate HPV infection. Female sex workers are considered vulnerable to the acquisition of these infections due to exposure to risk factors. Objective: To determine HPV infection, viral types and coinfections in self-collected samples from female sex workers. Methods: Self-collected samples from female sex workers, of vaginal canal and uterine cervix, were subjected to HPV-deoxyribonucleic acid (DNA) detection, viral genotyping by type-specific polymerase chain reaction (PCR), restriction fragment length polymorphism (RFLP), and the detection of coinfection. Results: HPV-DNA was detected in 19.4% of the samples, and HPV 31, 6, and 53 were the most frequently detected types. There was a predominance of high-risk oncogenic HPV (HR-HPV) and a strong presence of simultaneous infections with multiple HPV types (84.6%). Coinfections with both HPV and C. trachomatis, and HPV and G. vaginalis were detected. The variables that were statistically associated with HPV infection and the presence of multiple infections were non-use of condoms and non-compliance with regular cervical cytology screening. Conclusion: The results highlight the importance of more comprehensive studies among vulnerable populations, aiming to establish measures to raise awareness about the risks of contracting sexually transmitted infections, as well as to support future studies for introducing HPV vaccines with wider coverage of viral types.

RESUMO Introdução: O papilomavírus humano (HPV) está intimamente associado ao câncer cervical, e a presença de coinfecções, como por Chlamydia trachomatis, Gardnerella vaginalis e Trichomonas vaginalis, pode potencializar ou facilitar a infecção por HPV. As mulheres profissionais do sexo são consideradas vulneráveis à aquisição dessas infecções devido à exposição aos fatores de risco. Objetivo: Determinar a infecção por HPV, os tipos virais e as coinfecções em amostras autocoletadas de mulheres profissionais do sexo. Métodos: Amostras autocoletadas de mulheres profissionais do sexo, do canal vaginal e da cérvice uterina, foram submetidas a detecção do HPV-ácido desoxirribonucleico (DNA), genotipagem viral por reação em cadeia da polimerase (PCR) tipo específica e restriction fragment length polymorphism (RFLP) e detecção de coinfecção. Resultados: O HPV-DNA foi detectado em 19,4% das amostras, sendo os tipos HPV 31, 6 e 53 os mais frequentes. Houve predominância de HPV de alto risco (HR-HPV) e elevada presença de infecções múltiplas (84,6%). A presença de coinfecções foi observada tanto para HPV e C. trachomatis quanto para HPV e G. vaginalis. Observou-se também que mulheres profissionais do sexo que não fazem uso de preservativos e aquelas que não realizam o exame citológico rotineiramente estão predispostas à aquisição da infecção causada pelo HPV. Conclusão: Os resultados obtidos ressaltam a importância de estudos mais abrangentes entre as populações vulneráveis, objetivando estabelecer medidas para a conscientização sobre os riscos de aquisição das infecções sexualmente transmitidas, bem como auxiliar estudos futuros para introdução de vacinas contra o HPV com maior cobertura de tipos virais.

Cytokine profile and proviral load among Japanese immigrants and non-Japanese infected with HTLV-1 in a non-endemic area of Brazil.

The lifetime risk of HTLV-1-associated myelopathy/tropical spastic paraparesis (HAM/TSP) development differs among ethnic groups. To better understand these differences, this prospective cohort study was conducted to investigate the cytokine profile and the HTLV-1 proviral load (PVL) in Japanese and non-Japanese populations with HAM/TSP and asymptomatic carriers (ACs). The serum IL-2, IL-4, IL-6, IL-10, IL-17, TNF-α, and IFN-γ levels were quantified using the Cytometric Bead Array in 40 HTLV-1-infected patients (11 HAM/TSP and 29 ACs) and 18 healthy controls (HCs) in Brazil. Among ACs, 15 were Japanese descendants and 14 were non-Japanese. Of 11 patients with HAM/TSP, only one was a Japanese descendant. The HTLV-1 PVL was quantified by real-time PCR. The HTLV-1 PVL was 2.7-fold higher in HAM/TSP patients than ACs. Regardless of the clinical outcome, the PVL was significantly higher in patients younger than 60 years than older patients. The HAM/TSP and ACs had higher IL-10 serum concentrations than that of HCs. The ACs also showed higher IL-6 serum levels than those of HCs. According to age, the IL-10 and IL-6 levels were higher in ACs non-Japanese patients older than 60 years. HAM/TSP patients showed a positive correlation between IL-6 and IL-17 and a negative correlation between the PVL and IL-17 and IFN-γ. In the all ACs, a significant positive correlation was observed between IL-2 and IL-17 and a negative correlation was detected between IL-10 and TNF-α. Only 6.25% of the Japanese patients were symptomatic carriers, compared with 41.67% of the non-Japanese patients. In conclusion, this study showed that high levels of HTLV-1 PVL was intrinsicaly associated with the development of HAM/TSP. A higher HTLV-1 PVL and IL10 levels found in non-Japanese ACs over 60 years old, which compared with the Japanese group depicts that the ethnic background may interfere in the host immune status. More researches also need to be undertaken regarding the host genetic background to better understand the low frequency of HAM/TSP in Japanese HTLV-1-infected individuals.

Predominant overexpression of CD25/FOXP3, IFN-γ, and suppressive cytokines in high-grade lesion samples infected with human papillomavirus / PPredominância de superexpressão de CD25/FOXP3, IFN-γ e citocinas supressoras em amostras de lesão de alto grau infectadas pelo papilomavírus humano

ABSTRACT Introduction: Human papillomavirus (HPV) persistent infection is the leading cause of cervical cancer and its precursor lesions, and the inappropriate immune response is among the factors that contribute to viral persistence. This may be influenced by regulatory T (Treg) cells and the production of immunosuppressive cytokines, such as transforming growth factor beta (TGF-β) and interleukin-10 (IL-10). Objective: We established the profile of the predominant response, Th1 or immunosuppressive response, in the tissue microenvironment, by detecting interferon-gamma (IFN-γ), TGF-β, and IL-10, as well as the co-expression of IL-2 receptor alpha (CD25) and forkhead box P3 (FOXP3). Methods: Seventy-four samples from uterine cervix biopsies that underwent HPV deoxyribonucleic acid (DNA) detection and histopathology analysis were immunostained to detect CD25/FOXP3, IFN-γ and suppressive cytokines in lymphocytes. Results: The microenvironment of high-grade squamous intraepithelial lesion (HSIL) samples with high numbers of viral particles (≥ 10,000 copies/ml) contained high numbers of CD25/FOXP3+, TGF-β+, IL-10+, and IFN-γ+ cells. Conclusion: The co-expression of CD25/FOXP3 and the expression of TGF-β, and IL-10 in HSIL samples suggest the existence of Treg cells in these locations, although IFN-γ expression was observed in several cells in these samples. Our data suggest that this cytokine could be related to immunosuppressed microenvironment maintenance, favoring the persistent HPV infection and the progression to carcinoma.

RESUMO Introdução: A infecção persistente por papilomavírus humano (HPV) é a principal causa do câncer cervical e suas lesões precursoras, e a resposta imune inadequada está entre os fatores que contribuem para a persistência viral. Isso pode ser influenciado por células T regulatórias (Treg) e pela produção de citocinas imunossupressoras, como o fator de transformação de crescimento beta (TGF-β) e a interleucina 10 (IL-10). Objetivo: Estabelecemos o perfil de resposta predominante, resposta Th1 ou imunossupressora, no microambiente tecidual, pela detecção de interferon gama (IFN-γ), TGF-β, e IL-10, bem como a coexpressão do receptor da cadeia alfa da IL-2 (CD25) e do forkhead box P3 (FOXP3). Método: Setenta e quatro amostras de biópsias de cérvice uterina, submetidas à detecção do ácido desoxirribonucleico (DNA) de HPV e à análise histopatológica, foram utilizadas nas reações de imuno-histoquímica para detectar IFN-γ, TGF-β, IL-10 e CD25/FOXP3 em linfócitos. Resultados: O microambiente das amostras de lesões intraepiteliais escamosas de alto grau (HSIL) com elevado números de partículas virais (≥ 10.000 cópias/ml) continha elevado número de células CD25/FOXP3+, TGF-β+, IL-10+ e IFN-γ+. Conclusão: A coexpressão de CD25/FOXP3 e a expressão de TGF-β nas amostras HSIL sugerem a existência de células Treg nesses locais, embora a expressão de IFN-γ tenha sido observada em várias células. Nossos dados sugerem que essa citocina pode estar relacionada com a manutenção do microambiente imunossuprimido, favorecendo a infecção persistente por HPV e a progressão para carcinoma.

Local immunosuppression induced by high viral load of human papillomavirus: characterization of cellular phenotypes producing interleukin-10 in cervical neoplastic lesions.

A specific immune response to human papillomavirus (HPV) in the cervical microenvironment plays a key role in eradicating infection and eliminating mutated cells. However, high-risk HPVs modulate immune cells to create an immunosuppressive microenvironment, and induce these immune cells to produce interleukin 10 (IL-10). This production of IL-10, in conjunction with HPV infection, contributes to the appearance of cervical neoplastic lesions. We sought to characterize the IL-10-producing cellular phenotype, and investigate the influence of host and HPV factors upon the induction of an immunosuppressive microenvironment. Immunohistochemical analysis demonstrated an increase in IL-10 production by keratinocytes, macrophages and Langerhans cells in high-grade cervical lesions and cervical cancer. This increase was more pronounced in patients older than 30 years, and was also correlated with high viral load, and infection with a single HPV type, particularly high-risk HPVs. Our results indicate the existence of a highly immunosuppressive microenvironment composed of different IL-10-producing cellular phenotypes in cervical cancer samples, and samples classified as high-grade cervical lesions (cervical intraepithelial neoplasia stages II and III). The immunosuppressive microenvironment that developed for these different cellular phenotypes favours viral persistence and neoplastic progression.

Presence of highly oncogenic human papillomavirus in the oral mucosa of asymptomatic men

OBJECTIVES: The aim of this study was to identify highly oncogenic forms of human papillomavirus in the oral mucosa of asymptomatic men. METHODS: In this study, we analyzed samples of exfoliated cells from the oral cavity of 559 asymptomatic men. DNA-human papillomavirus was detected using the consensus primers PGMY09/11; viral genotyping was performed using type-specific PCR and restriction fragment length polymorphism. RESULTS: DNA-human papillomavirus was detected in 1.3% of the study participants and of those 42.8% were infected by more than one type of virus. Viral types included HPV6, 11, 89 (low oncogenic risk), and HPV52, 53 (high oncogenic risk). Increased vulnerability to human papillomavirus infection was observed in individuals aged over 26 years, among those who reported oral sex practices, and in those who have had more than 16 sexual partners since first engaging in sexual intercourse. CONCLUSIONS: There was a low prevalence of human papillomavirus detection in the oral mucosa of asymptomatic men. Highly oncogenic human papillomavirus types and infection by more than one viral type was observed. Oral sex practices and a large number of sexual partners may increase the risk of acquiring human papillomavirus infection. .

Good agreements between self and clinician-collected specimens for the detection of human papillomavirus in Brazilian patients

Women infected with human papillomavirus (HPV) are at a higher risk of developing cervical lesions. In the current study, self and clinician-collected vaginal and cervical samples from women were processed to detect HPV DNA using polymerase chain reaction (PCR) with PGMY09/11 primers. HPV genotypes were determined using type-specific PCR. HPV DNA detection showed good concordance between self and clinician-collected samples (84.6%; kappa = 0.72). HPV infection was found in 30% women and genotyping was more concordant among high-risk HPV (HR-HPV) than low-risk HPV (HR-HPV). HPV16 was the most frequently detected among the HR-HPV types. LR-HPV was detected at a higher frequency in self-collected; however, HR-HPV types were more frequently identified in clinician-collected samples than in self-collected samples. HPV infections of multiple types were detected in 20.5% of clinician-collected samples and 15.5% of self-collected samples. In this study, we demonstrated that the HPV DNA detection rate in self-collected samples has good agreement with that of clinician-collected samples. Self-collected sampling, as a primary prevention strategy in countries with few resources, could be effective for identifying cases of HR-HPV, being more acceptable. The use of this method would enhance the coverage of screening programs for cervical cancer.

Good agreements between self and clinician-collected specimens for the detection of human papillomavirus in Brazilian patients.

Women infected with human papillomavirus (HPV) are at a higher risk of developing cervical lesions. In the current study, self and clinician-collected vaginal and cervical samples from women were processed to detect HPV DNA using polymerase chain reaction (PCR) with PGMY09/11 primers. HPV genotypes were determined using type-specific PCR. HPV DNA detection showed good concordance between self and clinician-collected samples (84.6%; kappa = 0.72). HPV infection was found in 30% women and genotyping was more concordant among high-risk HPV (HR-HPV) than low-risk HPV (HR-HPV). HPV16 was the most frequently detected among the HR-HPV types. LR-HPV was detected at a higher frequency in self-collected; however, HR-HPV types were more frequently identified in clinician-collected samples than in self-collected samples. HPV infections of multiple types were detected in 20.5% of clinician-collected samples and 15.5% of self-collected samples. In this study, we demonstrated that the HPV DNA detection rate in self-collected samples has good agreement with that of clinician-collected samples. Self-collected sampling, as a primary prevention strategy in countries with few resources, could be effective for identifying cases of HR-HPV, being more acceptable. The use of this method would enhance the coverage of screening programs for cervical cancer.