Estudo da resposta imune em lesões intraepiteliais anais em pacientes infectados e não infectados pelo HIV e HPV / Study of immune response in intraepithelial lesions in infected and non-infected patients by HIV and HPV

RESUMO

O Papiloma Vírus Humano (HPV) é o principal agente etiológico do câncer do trato anogenital. Uma maior prevalência e incidência de desenvolvimento do carcinoma e doenças associadas ao HPV têm sido observadas em indivíduos infectados pelo HIV. A história natural da infecção pelo HPV ainda não está totalmente elucidada, assim como aresposta imune que ocorre na co-infeção pelo HIV/HPV,principalmente na mucosa anal.Objetivo do presente estudo foi avaliar a resposta imune “in situ” em amostras de biópsias de indivíduos infectados pelo HIV em acompanhamento no Instituto Nacional de Infectologia/FIOCRUZ, RJ. Um total de 114 biópsias foi analisado através de TissueMicro-Array, sendo 15 de indivíduos Não infectados pelo HIV, todos sem lesão, e 99 deindivíduos Infectados pelo HIV 21 sem lesão, 39 com NIA1, e 39 com NIA2/3. Foirealizado PCR e sequenciamento para identificação do DNA de HPV e imunohistoquímicapara análise dos marcadores imunes CD4, CD8, Foxp3, T-bet e IL-10 e SLPI. A análiseestatística foi feita no software SPSS 15.0 aplicando os testes Kruskall-Wallis, Teste Quiquadradoe Teste Exato de Fisher. Pacientes Infectados pelo HIV com NIA 2/3apresentaram nadir CD4+ <50 cél/mm³ comparados aos pacientes normais (p = 0,01).Quanto aos marcadores imunes, indivíduos Infectados pelo HIV apresentaram uma maiorexpressão de Foxp3 e IL-10 de acordo com a gravidade da lesão (p= 0,002)...

ABSTRACT

Human Papillomavirus (HPV) is the main etiologic agent for lower genital tract cancers. Ahigher incidence and prevalence of these cancers has been reported in HIV infectedindividuals. The natural history of HPV infection is still unclear and the immune responsewith HIV/HPV co-infection, particularly in the anal mucosa, is poorly understood. The aimof this study was to evaluate the immune response in situ in ano-rectal biopsies fromHIV-infected patients followed up at the National Institute of Infectious Diseases /FIOCRUZ, RJ. A total of 114 biopsies were analyzed by Tissue Micro-Array 15 were fromHIV negative individuals without any lesions and 99 from HIV-positive individuals 21without lesions, 39 with AIN 1 and 39 with AIN 2/3. PCR and sequencing was run toidentify HPV DNA. CD4, CD8, Foxp3, T-bet, IL-10 and SLPI were analyzed byimmunohistochemistry. Statistical analysis was carried out using SPSS 15.0. TheKruskal-Wallis, chi-square and Fisher's exact tests were applied. HIV-positive patients withAIN 2/3 showed a nadir count of CD4 + <50 cells/mm³ compared to normal subjects (p =0.01). HIV positive individuals showed a higher expression of FoxP3 and IL-10 accordingto the severity of the lesion (p = 0.002). A positive coefficient correlation was foundbetween FoxP3 and IL-10 (0.34; p = 0.27). The analysis showed that 93.4 percent (101/107) ofthe samples had HPV DNA, and the most common types were HPV 16 (26.9 percent), followedby HPV 6 (15.7 percent), HPV 59 (13 percent) and HPV 18 (10.2 percent). Interestingly, samples fromindividuals with high-risk oncogenic HPV DNA were negative for SLPI and there was lessexpression in AIN 2/3 compared to the no-lesion group of HIV + individuals, showing aninverse association with HPV type and degree of AIN...