Transcriptional regulation of effector CD8+ T cell differentiation and molecular dysfunction during HIV-1 infection

Les cellules T CD8+ jouent un rôle primordial dans le contrôle des infections virales en limitant la dissémination des cellules infectées. Lors de l’infection chronique par le virus HIV, les cellules T CD8+ HIV-spécifiques ne se différencient pas en cellules effectrices fonctionnelles capables de tuer les cellules infectées par le virus ; ces cellules ne sont plus capables de proliférer ou de produire l’ IL-2. Ces cellules expriment PD-1 et l’engagement de PD-1, par son ligand, aboutit a plusieurs de ces déficits fonctionnels des cellules T . Le rôle de PD-1 dans la régulation d'évènements transcriptionnels contrôlant la différentiation et l'obtention des fonction effectrices des cellules T CD8+ reste à démontrer. Id2 joue un rôle central dans la différenciation des cellules T CD8+ effectrices. Nous avons émis l’hypothèse que le défaut de maturation observé chez les cellules T CD8+ PD-1 high HIV-spécifiques (CD8+PD-1hi) au cours de l’infection chronique par le virus HIV pouvait être lié à la diminution d’expression du régulateur Id2. Nous avons ainsi démontré que l'engagement de PD-1 contribuait à une diminution d'expression de Id2 et de ses cibles transcriptionnelles. La surexpression de Id2 de ces cellules a permis de restaurer l'expression de marqueurs tels que Granzyme B et Bcl-2 et diminuir l’expression du marqueur de maturation de CD27. La famille des cytokines à chaine gamma joue un rôle clef dans la survie et l’homéostasie des cellules T. Dans ce travail, nous avons démontré que l’IL-15 était unique grâce à ses capacités de stimulation de l’expression d’Id2 et ses propriétés favorisant la survie ainsi que la différenciation des cellules T CD8+ effectrices. l’IL-15 induit la prolifération de toutes les populations de cellules T mémoires provenant de donneurs sains. L’addition de cette cytokine aux sous-populations cellulaires Ttm et Tem a permis leur différenciation en cellules effectrices capables de produire Granzyme B alors que la stimulation par l’IL-15 des cellules Tcm ne favorise pas leur différenciation. Un test de cytotoxicitié par cytométrie en flux nous a permis de confirmer que la stimulation de cellules T CD8+ HIV spécifiques par l’IL-15 favorisait l’expression de Id2 et restaurait les fonctions cytotoxiques des cellules T CD8+ HIV spécifiques. En conclusion, nous avons pour la première fois dans cette thèse défini les mécanismes moléculaires impliqués dans la modulation de l’expression du régulateur transcriptionnel Id2 par l’IL-15. Nous avons également révélé comment l’engagement de PD-1 conduisait a une altération de l’expression et de la fonction d’Id2 et favorisait la diminution des fonctions effectrices des cellules T CD8-HIV spécifiques. Une perspective de traitement avec des agents tels que l’IL-15 ou le bloquage de PD-1, en combinaison avec les traitements conventionnels, pourrait contribuer à une meilleure stimulation des réponses immunes favorisant ainsi la réactivation des cellules T CD8+ et permettant la destruction de cellules T CD4+ infectées de manière latente.

Self-reported Body Changes and Associated Factors in Persons Living with HIV

The study aimed at verifying the associated factors of self-perceived body changes in adults living with HIV in highly-active antiretroviral therapy (HAART) in the city of Sao Paulo, Brazil. This cross-sectional study was conducted among people living with HIV on HAART for at least three months. A standardized questionnaire was used for assessing self-perceived body changes. Associated factors relating to self-reported body changes in people living with HIV (PLHIV) were assessed with Student`s t-test and chi-square test. In total, 507 patients were evaluated. The mean time since diagnosis was 6.6 years [standard deviation (SD)+/-4.1], and the mean duration of HAART was 5.1 years (SD+/-3.3). Self-perceived body changes were reported by 79.5% of the participants and were associated with viral load and duration of HAART. Fibre intake was lower among males who gained in abdominal fat (p=0.035). HAART-related body changes were reported by the large majority of the population and were associated with demographic and clinical variables.

HIV/AIDS e estilo de vida: parâmetros imunológicos, virológicos, síndrome lipodistrófica e sua relação com a prática de exercícios

This descriptive study aimed to investigate the relationship between expression of immunological (TCD4 +) and virological (viral load) parameters, lipodystrophy syndrome and lifestyle variables of people living with HIV who underwent a program of physicalexercise. Initially, the sample was composed by 17 persons, recorded at the Department of Giselda Trigueiro Hospital Care (GTH), Natal-RN . With the passing of the stages of intervention (physical training program- PTP) the number of participants has changed (17, 9, 8 and 7) as the training phases (I, II, III and IV). Data collected were on total cholesterol, High Density Lipoprotein (HDL) and triglycerides. The % fat (% F), body mass index (BMI), waist-hip ratio (WHR), lean mass (LM), central fat (BF), total (TF) and peripheral (PF), weight and height were used to measure the morphological parameters. For control of variables (TCD4 +, viral load, Triglycerides, Cholesterol, HDL), the information contained in the blood tests every four months were investigated. After Phase I and II, it was used a structured interview. Then sampling was carried out considering the pre-and post-tests 1, 2, 3 and 4 (after 16, 32, 48 and 64 weeks of training, respectively). Daily, the intensity of the work was checked by the scale of perceived exertion for exercises adapted to resistance34. Procedures used were descriptive statistics (dispersion, absolute and relative frequencies, means, standard deviations and minimum and maximum values) as well as Spearman linear correlation adopting a significance level of p ≤ 0.05. Positive changes were observed for TCD4 + and viral load in all phases of the PTP. For the morphological components, the loss of central subcutaneous fat (CSF) and total subcutaneous fat (TSF) for both sexes and the decrease in % BF among women were the most dramatic results after the training phase I. For men, results were more significant to % F and LM in Phases I and II and peripheral subcutaneous fat (PSF) declined in all phases of the exercise. As for women, results were more expressive for % F and TM in the phases I and II and the PSF decreased in all phases of the exercise. Whereas for women, the waist/hip ratio (WCQ) and 0% F decreased and showed a positive association with triglycerides (WHR r * 0.82, p 0.042, r 0.88 TSF *, p 0.019 and r 1.00 ** CSF, p <0.001) and among men with limb subcutaneous fat (LSF)* r 0.65, p 0.029). The PTP provided improvement in the health, self-esteem and quality of life, proving to be a possible strategy to positively influence the expression of immunological parameters (TCD4 +) and virological (viral load) and morphological components of people living with HIV causing no deleterious effects in these parameters

Paracoccidioidomycosis: an unusual presentation in a young girl disclosing an unnoted HIV-infection

The association of paracoccidioidomycosis with AIDS is apparently less frequent than expected. The authors present an unusual case of paracoccidioidomycosis in a 13-year-old female student which was later found to be the first opportunistic infection in the course of the patient's HIV-infection. The clinical presentation followed an accidental incised wound on the palmar region initially described as a 'sporotrichotic-chancre'. After good response under sulfamethoxazole-trimethoprin, the patient relapsed and presented an associated oral candidiasis. HIV-infection was documented and additional investigation showed CD4(+) T-cells=22/mm(3), CD8(+)=280 cell/mm(3) and viral load=4,043 log. This case report presents an uncommon dermatological-clinical picture in the youngest patient in which such association has been reported to date.

Intestinal parasitic infections in HIV/AIDS patients: epidemiological, nutritional and immunological aspects

This study applied a socioeconomic questionnaire designed to evaluate the frequency of intestinal parasites and characterize epidemiological, nutritional, and immunological variables in 105 HIV/AIDS patients - with and without parasitic infections, attending the Day Hospital in Botucatu, UNESP, from 2007 to 2008. Body mass index was calculated and the following tests performed: parasitological stool examinations; eosinophil, IgE, CD4(+) T and CD8(+) T lymphocyte cell counts; albumin test; viral load measure; and TNF-alpha, IFN-gamma, IL-2, IL-5 and IL-10 cytokine levels. Results were positive for parasitic intestinal infections in 12.4% of individuals. Most patients had good socioeconomic conditions with basic sanitation, urban dwellings, treated water supply and sewage, good nutritional and immunological status and were undergoing HAART. Parasites were found at the following frequencies: Entamoeba - five patients (38.5%), Giardia lamblia-four (30.7%), Blastocystis hominis-three (23.0%), Endolimax nana-two (15.4%), and Ascaris lumbricoides - one (7.7%). There were no significant differences between the two groups for eosinophils, albumin, IgE, CD4(+) T and CD8(+) T lymphocytes, INF-gamma, IL-2, or IL-10. Most patients also showed undetectable viral load levels. Significant differences were found for TNF-alpha and IL-5. These results show the importance of new studies on immunodeficient individuals to increase understanding of such variables.

Fatores protetores e de risco envolvidos na transmissão vertical do HIV-1

Este estudo avalia os fatores maternos e fetais envolvidos na transmissão vertical do HIV-1 em 47 pares de mãe e filho. As variáveis comportamentais, demográficas e obstétricas foram obtidas mediante entrevista; os dados referentes ao parto e ao recém-nascido, dos prontuários das maternidades. Durante o terceiro trimestre de gestação foi realizada a contagem da carga viral materna e dos linfócitos T CD4+. A média de idade foi de 25 anos e 23,4% das gestantes eram primigestas, e o fator comportamental mais prevalente foi não usar preservativos. Dentre as gestantes, 48,9% tinham células CD4+ superior a 500 células/mm³ e 93,6% se enquadravam na categoria clínica A; 95,7% submeteram-se à profilaxia com zidovudina durante a gestação ou no parto, a qual foi ministrada a todos os recém-nascidos; 50,0% delas foram submetidas à cesárea eletiva. Apesar de expostas a vários fatores de risco e protetores, nenhuma criança tornou-se infectada. A transmissão vertical resulta de um desequilíbrio entre os fatores, com predomínio dos de risco sobre os protetores.

Avaliação da estrutura organizacional da assistência ambulatorial em HIV/Aids no Brasil

OBJETIVO: No contexto de acesso universal à terapia antiretroviral, os resultados do Programa de Aids dependem da qualidade do cuidado prestado. O objetivo do estudo foi avaliar a qualidade do cuidado dos serviços ambulatoriais que assistem pacientes de Aids. MÉTODOS: Estudo realizado em sete Estados brasileiros, em 2001 e 2002. Foi avaliada a qualidade do atendimento a pacientes com Aids quanto à disponibilidade de recursos e a organização do trabalho de assistência. Um questionário com 112 questões estruturadas abordando esses aspectos, foi enviado a 336 serviços. RESULTADOS: A taxa de resposta foi de 95,8% (322). Os indicadores de disponibilidade de recursos mostram uma adequação maior do que os indicadores de organização do trabalho. Não faltam antiretrovirais em 95,5% dos serviços, os exames de CD4 e Carga Viral estão disponíveis em quantidade adequada em 59 e 41% dos serviços, respectivamente. em 90,4% dos serviços há pelo menos um profissional não médico (psicólogo, enfermeiro ou assistente social). Quanto à organização, 80% não agendavam consulta médica com hora marcada; 40,4% agendavam mais que 10 consultas médicas por período; 17% não possuíam gerentes exclusivos na assistência e 68,6% não realizavam reuniões sistemáticas de trabalho com a equipe. CONCLUSÕES: Os resultados apontam que além de garantir a distribuição mais homogênea de recursos, o programa precisa investir no treinamento e disseminação do manejo do cuidado, conforme evidenciado nos resultados da organização de trabalho.

LIFE QUALITY of PEOPLE INFECTED BY HIV WITH OR WITHOUT ANTIRETROVIRAL TREATMENT

The "HIV/Aids-Quality of Life" (HAT-Qol) is a specific multifunctional instrument used to measure the life quality of HIV infected persons. It is divided into nine domains: general activity, sexual activity, secrecy about HIV seropositivity, concern about health, financial concern, awareness about HIV, satisfaction with life, issues about medications and belief in the doctor. The current study analyzed the life quality of HIV infected individuals-who attended the DST/Aids Program in Maringa city, Parana state-regarding the use or not of antiretroviral therapy (TARV) and their demographic, epidemiological and clinical characteristics. Data were collected by retrospective analysis from 1,200 medical charts of patients registered in the program. The HAT-Qol instrument was applied before routine medical consultation. One hundred and sixty-nine patients, who had HIV infection confirmed, were divided into two groups, G1 with 118 individuals receiving antiretroviral therapy and G2 with 51 individuals who were not under this therapy.Result analysis, regarding social and demographic characteristics, revealed no difference among responses related to gender, educational degree and sexual option. Age influenced satisfaction with sexual activity and marital status. Regarding HIV awareness, the lowest response index or worst quality of life came from, respectively, men between 50 and 69 years old and patients who did not have regular partners compared with the ones who did. Additionally, it was observed that the time of diagnosis influenced general activities, HIV awareness, concern about health and financial issues, satisfaction with life and topics about medications. The variables were compared in both groups. There was no influence on the use or not of antiretroviral therapy regarding age, sexual activity, HIV diagnosis time and the domains that evaluated general activities, financial concern, awareness of HIV and satisfaction with life. In relation to time of diagnosis, there was an influence only in persons who had been diagnosed two or three years before, in which a lower quality of life was observed among individuals who were not under antiretroviral therapy. It was not possible to compare variables about medication use, HIV plasmatic viral rate ant time of diagnosis, because G2 individuals were not receiving the antiretroviral therapy. Furthermore, no comparison was made regarding marital status and HIV awareness, because there were no married individuals in G2. Thus, the analysis of the results showed that the use of antiretroviral treatment did not influence the life quality of HIV patients studied by the HAT-Qol scale.

A comparative evaluation of open loop and closed loop drug administration strategies in the treatment of AIDS.

In recent years, many researchers in the field of biomedical sciences have made successful use of mathematical models to study, in a quantitative way, a multitude of phenomena such as those found in disease dynamics, control of physiological systems, optimization of drug therapy, economics of the preventive medicine and many other applications. The availability of good dynamic models have been providing means for simulation and design of novel control strategies in the context of biological events. This work concerns a particular model related to HIV infection dynamics which is used to allow a comparative evaluation of schemes for treatment of AIDS patients. The mathematical model adopted in this work was proposed by Nowak & Bangham, 1996 and describes the dynamics of viral concentration in terms of interaction with CD4 cells and the cytotoxic T lymphocytes, which are responsible for the defense of the organism. Two conceptually distinct techniques for drug therapy are analyzed: Open Loop Treatment, where a priori fixed dosage is prescribed and Closed Loop Treatment, where the doses are adjusted according to results obtained by laboratory analysis. Simulation results show that the Closed Loop Scheme can achieve improved quality of the treatment in terms of reduction in the viral load and quantity of administered drugs, but with the inconvenience related to the necessity of frequent and periodic laboratory analysis.

Analysis of a sub-optimal scheme of drug dosage in the AIDS treatment

Here the results for CD4+T cells count and the viral load obtained from HIV sero-positive patients are compared with results from numerical simulations by computer. Also, the standard scheme of administration of drugs anti HIV (HAART schemes) which uses constant doses is compared with an alternative sub-optimal teatment scheme which uses variable drug dosage according to the evolution of a quantitative measure of the side effects. The quantitative analysis done here shows that it is possible to obtain, using the alternative scheme, the same performance of actual data but using variable dosage and having fewer side effects. Optimal control theory is used to solve and also to provide a prognosis related to the strategies for control of viraemia.

The β-chemokines MIP-1α and RANTES and lipoprotein metabolism in HIV-infected Brazilian patients

HIV patients are predisposed to the development of hypertriglyceridemia and hypercholesterolemia as a result of both viral infection and HIV infection therapy, especially the protease inhibitors. Chemokines and cytokines are present at sites of inflammation and can influence the nature of the inflammatory response in atherosclerosis. We investigated the correlation between biochemical variables and β-chemokines (MIP-1α and RANTES) and the apolipoprotein E genotype in HIV-infected individuals. The apolipoproteins were measured by nephelometry. Triglycerides and total cholesterol were determined by standard enzymatic procedures. The β-chemokines were detected by ELISA. The genetic category of CCR5 and apolipoprotein E were determined by PCR amplification and restriction enzymes. Immunological and virological profiles were assessed by TCD4 + and TCD8 + lymphocyte counts and viral load quantification. Positive correlations were found between apo E and CD8 + (p = 0.035), apo E and viral load (p = 0.018), MIP-1α and triglycerides (p = 0.039) and MIP-1α and VLDL (p = 0.040). Negative correlations were found between viral load and CD4 + (p = 0.05) and RANTES and CD4 + (p = 0.029). The β-chemokine levels may influence lipid metabolism in HIV-infected individuals. © 2005 by The Brazilian Journal of Infectious Diseases and Contexto Publishing. All rights reserved.

Immune reconstitution in HIV-1 infected patients treated for two years with highly active antiretroviral therapy

The aim of this paper was to evaluate the immune reconstitution of HIV-1 patients subjected to highly active antiretroviral therapy (HAART) for two years or more according to CD 45RA and CD 45RO cell count; determination of IL-2, IFN-γ, IL-4, IL-10 and TNF-α serum levels; CD 4 + T and CD 8 + T lymphocyte count; and plasma viral load (VL) determination. For this purpose, a cross sectional study was carried out in the Tropical Diseases Area, Botucatu School of Medicine, São Paulo State University, UNESP, Botucatu, São Paulo, Brazil. Between June 2001 and April 2002, 37 HIV-1 infected patients were evaluated, 13 with treatment indication but untreated (G1), 9 subjected to HAART for 5-7 months (G2), and 15 treated for two years or more (G3); both treated groups used medication regularly and without failure. Forty-nine normal individuals were studied as controls (GC-1 and GC-2). There was a tendency (p<0.10) for the predominance of two nucleoside reverse transcriptase inhibitors (NRTI) associated with one non-nucleoside reverse transcriptase inhibitor (NNRTI) regimen in G2; and two NRTI associated with a protease inhibitor (PI) in G3. Statistical differences between groups were seen for CD 45RA (G1<[G3=GC-2]; p<0.05) and CD 45RO (G1[G2=G3]; p<0.001), TNF-α serum determination ([G1>G3; G2=intermediate]>GC-1; p<0.001), IL-2 (G1<[G2=G3=GC-1]; p<0.01), IFN-γ ([G1=GC-1]>[GC-2=G3]; p<0.001), IL-4 and IL-10 ([G1=G2=G3]>GC-1; p<0.001), serum cytokine profiles, with a higher proportion of subtype 2 in G1 and mature subtype 0 in G2 and G3 (p<0.005). There was no statistical difference for CD 8 + T lymphocyte counts (G1=G2=G3; p<0.50). Consistency was seen between positive correlations of profile 1 definer cytokines (IL-2 and IFN-γ), CD 45RA and CD 45RO cells, and CD 4 + T lymphocyte counts and between positive correlations of profile 2 definer cytokines (IL-4 and IL-10) with TNF-α, and VL. The negative correlations were also consistent as they expressed the inverse of the positives. The variables with the highest number of correlations were IL-2, IFN-γ, and VL, followed by CD 45RA and CD 45RO cells, and IL-10. The variables with the lowest number of correlations were CD 4 + T and CD 8 + T lymphocytes. The results express the partial but important immune reconstitution in HIV-1 infected individuals with the interference of HAART and the importance of cytokines especially IL-2 and IFN-γ, and CD 45RA and CD 45RO cells as surrogate markers of this reconstitution.

Cytokines profile and clinical-metabolic alterations in HIV-1-infected individuals with and without lypodistrophy

Highly active antiretroviral therapy (HAART) has lead to a dramatic decrease in the morbidity of patients infected with HIV. However, metabolic side effects, including lipodystrophy and dyslipidemia, have been reported in patients treated with antiretroviral therapy (HAART). The aim of this study was to analyze the clinical and metabolic alterations and the cytokines TNF-α, IFN-γ, IL-2, IL-10 and TNF-II receptors profile in the serum of treated HIV-1-infected individuals with or without lipodystrophy. Eighty-four adult patients were analyzed, 42 females and 42 males, their mean age was 37 years old, and they received HAART for at least 15 months. These patients were ambulatory outpatients from the Infectious and Parasitary Disease Area of Botucatu School of Medicine, UNESP. Subsequently the individuals were distributed into 2 groups, G1: 42 HIV-infected individuals with lipodystrophy, and G2: 42 HIV-infected individuals without lipodystrophy. Among the antiretrovirals used, stavudine was more associated to the lipodystrophy group and zidovudine to the group without lipodystrophy. CD4, CD8, viral load, glucose, albumin, and the circulating lipid did not present any difference in the group comparison, except for triglyceride that was elevated in the lipodystrophy group and HDL which was present in low concentration in more patients of G1. The cytokines TNF-α, TNF-RII, and IL-10 profile presented high levels in the lipodystrophy group; also it was positively correlated with this group. On the other hand, IL-2 and IFN-γ presented low levels in this group. High levels of TNF-α and its receptor seem to be associated to the development of lipodystrophy in patients receiving HAART.

Bezafibrate for the treatment of hypertriglyceridemia in HIV1-infected patients on highly active antiretroviral therapy

The use of highly active antiretroviral therapy (HAART) in HIV-infected patients has been associated with the development of risk factors for cardiovascular diseases (CD) including dyslipidemia and insulin resistance, hypertriglyceridemia being the most frequent metabolic disturbance in these patients. Fibrates are indicated when hypertriglyceridemia is accentuated and persists for over six months. We evaluated the efficacy and safety of bezafibrate for the treatment of hypertriglyceridemia in HIV-infected individuals on HAART. All patients received 400mg/day of bezafibrate and were evaluated three times: Mo (pre-treatment), M1 (one month after treatment), and M2 (six months after treatment). Fifteen adult individuals, eight males and seven females with mean age = 41.2 ± 7.97 years and triglyceride serum levels ≥400mg/dL were included in the study. Smoking, alcohol ingestion and sedentarism rates were 50%, 6.66% and 60%, respectively. Family history of CD, hypertension and diabetes mellitus was reported in 33.3%, 40% and 46.7% of the cases, respectively, while dyslipidemia was reported by only 13.3%. More than half of the patients were using a protease inhibitor plus a nucleotide analog transcriptase inhibitor. Eutrophy and tendency toward overweight were observed at all three study time points. There were significant reductions in triglyceride serum levels from Mo to M1 and from Mo to M2. No significant changes were observed in the serum levels of creatine phosphokinase, hepatic enzymes, CD4 +, CD8 + and viral load. Therefore, bezafibrate seems to be safe and effective for the reduction of hypertriglyceridemia in HIV-infected patients on HAART. © 2006 by The Brazilian Journal of Infectious Diseases and Contexto Publishing. All rights reserved.

Cytokine profile and clinical metabolic alterations in HIV-1 infected individuals with and without lipodistrophy

The extensive use of Highly Active Antiretroviral Therapy (HAART) has transformed HIV infection into a chronic condition. Thus, metabolic alterations including lipodystrophy and dyslipidemia have been associated with the use of such medications. The objective of the present study was to analyze clinical metabolic alterations and the profile of TNF-α, IFN-γ, IL-2, IL-10, and TNF-α type II soluble receptor in serum of HIV-1 individuals with and without lipodystrophy. Eighty-four adults were evaluated, 42 males and 42 females, mean age 37 years, and HAART time of at least 15 months. Two groups were formed, G1: 42 individuals with lipodystrophy, and G2: 42 without lipodistropy. From the HAART used, stavudine was more associated with the lipodystrophy group and zidovudine with the non-lipodystrophy group. CD4 and CD8 values, viral load, glucose, albumin, and lipids were not different between groups, except for triglycerides, which were high in the lipodystrophy group, and HDL, whose concentration was reduced in G1. TNF-α, TNF-RII, and IL-10 profiles were high and had positive correlation; IL-2 and IFN-γ had reduced levels in the lipodystrophy group. High TNF-α and its receptor levels seem to be associated with lipodystrophy development in individuals under HAART therapy.