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Although antiretroviral therapy (ART) is effective in suppressing viral replication, HIV-1 persists in reservoirs and rebounds after ART has been stopped. However, a very few people (eg, elite and post-treatment controllers) are able to maintain viral loads below detection limits without ART, constituting a realistic model for long-term HIV remission. Here, we describe the HIV control mechanisms of an individual who showed exceptional post-treatment control for longer than 15 years.We report the case of a Hispanic woman aged 59 years with sexually acquired acute HIV infection, who was included in an immune-mediated primary HIV infection trial involving a short course of ciclosporine A, interleukin-2, granulocyte macrophage colony-stimulating factor, and pegylated interferon alfa, followed by analytical treatment interruption. We did the following viral assays: total and integrated HIV-1 DNA in CD4 T cells and rectal tissue, quantitative viral outgrowth assay, HIV-1 infectivity in peripheral blood mononuclear cells and CD4 T-cell cultures and viral inhibitory activity by natural killer (NK) and CD8 T cells. NK and T-cell phenotypes were determined by flow cytometry. HLA, killer cell immunoglobulin-like receptors, Δ32CCR5, and NKG2C alleles were genotyped.After ART and immunomodulatory treatment, the person maintained undetectable plasma viral load for 15 years. HIV-1 subtype was CFR_02AG, CCR5-tropic. We found progressive reductions in viral reservoir during the 15-year treatment interruption: total HIV DNA (from 4573·50 copies per 10We described long-term remission in a woman aged 59 years who was treated during primary HIV infection and has maintained undetectable viral load for 15 years without ART. Replication-competent HIV-1 was isolated. NKG2C-memory-like NK cells and γδ T cells were associated with the control viral replication. Strategies promoting these cells could bring about long-term HIV remission.Fondo Europeo para el Desarrollo Regional (FEDER), SPANISH AIDS Research Network (RIS), Fondo de Investigación Sanitaria (FIS), HIVACAT, Institut d'Investigacions Biomèdiques August Pi i Sunyer (IDIBAPS), CERCA Programme/Generalitat de Catalunya, la Caixa Foundation, and Centro de Investigación Biomédica en Red de Enfermedades Infecciosas (CIBERINFEC).For the Spanish translation of the abstract see Supplementary Materials section.

The anionic carbosilane (CBS) dendrimer with sulfonate groups G2-S16 is a promising compound for the preparation of a microbicide gel to prevent HIV infection. However, until now its synthesis required aggressive conditions. Hence, a reliable synthetic procedure is very important to face GMP conditions and clinical trials. In this study, G2-S16 has been prepared by a new approach that involves the addition of an amine-terminated dendrimer to ethenesulfonyl fluoride (C2H3SO3F, ESF) and then transformation to the sulfonate dendrimer by treatment with a base. This strategy also makes feasible the synthesis of a labelled sulfonate dendrimer (G2-S16-FITC) to be used as a molecular probe for in vivo experiments. Interestingly, G2-S16-FITC enters into human peripheral blood mononuclear cells (PBMCs).

Nanotechnology is a science that covers a great number of fields. One of the most prominent is the field of human health. Among the great variety of nanoparticles used in therapeutics, the role of dendrimers should be highlighted. Dendrimers have stimulated wide interest in the fields of chemistry and biology, especially in applications like drug delivery, gene therapy, and chemotherapy, but also many other applications such as image diagnosis, acting as mimics of proteins, use as excipients of drugs, or in the cosmetics industry. Advances in their use as antimicrobials are giving encouraging results. In this sense, dendrimers seem to stand out in treatment against HIV-1, not only for their targeting capacity but also for their own activity in the synthesis of a therapeutic vaccine or to serve as antilatents. This review gives information about dendrimers’ properties, synthesis, and application in drug delivery, diagnosis, and therapy.

Amides from indole-3-glyoxylic acid and 4 - benzoyl-2-methyl-piperazine, which are related to entry inhibitors developed by Bristol-Myers Squibb (BMS), have been synthesized with aliphatic chains located at the C7 position of the indole ring. These spacers contain an azido group suitable for the well-known Cu(I)-catalyzed (3+2)-cycloaddition or an activated triple bond for the less known nucleophilic addition of thiols under physiological conditions. Reaction with polyols (β-cyclodextrin and hyperbranched polyglycerol) decorated with complementary click partners has afforded polyol-BMS-like conjugates that are not cytotoxic (TZM.bl cells) and retain the activity against R5-HIV-1 NLAD8 isolates. Thus, potential vaginal microbicides based on entry inhibitors, which can be called of 4th generation, are reported here for the first time.

Since the success of combined antiretroviral therapy, HIV-1-infected individuals are now living much longer. This increased life expectancy is accompanied by a higher prevalence of HIV-1 associated neurocognitive disorders. Rising too is the incidence in these patients of pathological hallmarks of Alzheimer's disease such as increased deposition of amyloid beta protein (Aβ). Although neurons are major sources of Aβ in the brain, astrocytes are the most numerous glial cells, therefore, even a small level of astrocytic Aβ metabolism could make a significant contribution to brain pathology. Neprilysin (NEP) is a decisive/crucial regulator of Aβ levels. We evaluated the effects of HIV-1 on Aβ deposition and the expression and activity of NEP in primary human astrocytes. Specifically, no differences in intracellular amyloid deposits were found between infected and control cells. However, primary cultures of infected astrocytes showed more extracellular Aβ levels compared to controls. This was accompanied by reduced expression of NEP and to a significant decrease in its activity. These results indicate that the presence of HIV-1 in the brain could contribute to the increase in the total burden of cerebral Aβ.

Cell-to-cell transmission is the most effective pathway for the spread of human immunodeficiency virus (HIV-1). Infected cells expose virus-encoded fusion proteins on their surface as a consequence of HIV-1 replicative cycle that interacts with noninfected cells through CD4 receptor and CXCR4 coreceptor leading to the formation of giant multinucleated cells known as syncytia. Our group previously described the potent activity of dendrimers against CCR5-tropic viruses. Nevertheless, the study of G1-S4, G2-S16, and G3-S16 dendrimers in the context of X4-HIV-1 tropic cell-cell fusion referred to syncytium formation remains still unknown. These dendrimers showed a suitable biocompatibility in all cell lines studied and our results demonstrated that anionic carbosilane dendrimers G1-S4, G2-S16, and G3-S16 significantly inhibit the X4-HIV-1 infection, as well as syncytia formation, in a dose dependent manner. We also demonstrated that G2-S16 and G1-S4 significantly reduced syncytia formation in HIV-1 Env-mediated cell-to-cell fusion model. Molecular modeling and in silico models showed that G2-S16 dendrimer interfered with gp120-CD4 complex and demonstrated its potential use for a treatment.

The interaction of a double stranded small interference RNA (siRNA Nef) with cationic carbosilane dendrimers of generations 1-3 with two different ammonium functions at the periphery ([-NMe2R]+, R=Me, (CH2)2OH) has been studied by experimental techniques (zeta potential, electrophoresis, single molecule pulling experiments) and molecular dynamic calculations. These studies state the presence of different forces on dendriplex formation, depending on generation and type of ammonium group. Whilst for higher dendrimers electrostatic forces mainly drive the stability of dendriplexes, first generation compounds can penetrate into siRNA strands due to the establishment of hydrophobic interactions. Finally, in the particular case of first generation dendrimer [G1O3(NMe2(CH2)2OH))6]6+; the presence of hydroxyl groups reinforces dendriplex stability by hydrogen bonds formation. However, since these small dendrimers do not cover the RNA, only higher generation derivatives protect RNA from degradation.

Amyloid fibrils, which are present in semen, were considered to be a cause of topical vaginal gel ineffectiveness in vivo after microbicides failed as HIV-1 prophylaxis. Therefore, it was necessary to determine whether a dendrimer was suitable for further evaluation in an in vitro model of semen-enhanced viral infection (SEVI). We demonstrated that SEVI in TZM.bl cell cultures increased the infectivity of R5-HIV-1NL(AD8), pTHRO.c and pCH058.c isolates, causing higher IC50 values for two polyanionic carbosilane dendrimers, G2-STE16 and G3-S16. However, both dendrimers maintained protection rates of 90% at non-toxic concentrations. When dendrimers were combined with Tenofovir/Maraviroc (TDF/MVC), the anti-HIV-1 effect remained at a minimum IC50 increase between 1- and 7-fold in the presence of amyloid fibrils. In peripheral blood mononuclear cells (PBMC), IC50 values were slightly influenced by the presence of semen. In brief, dendrimers combined with antiretrovirals showed a synergistic effect. This result plays a crucial role in new microbicide formulations, as it overcomes the negative effects of amyloid fibrils.

Gold nanoparticles (AuNPs) and polycationic macromolecules are used as gene carriers. Their behaviour is dependent on several factors, such as the size and type of the framework, charge, etc. We have combined both types of systems and prepared AuNPs covered with cationic carbosilane dendrons with the aim to evaluate their biocompatibility. Water soluble dendronized cationic AuNPs were prepared following a straightforward procedure from dendrons, a gold precursor and a reducing agent in water and were characterized by 1H NMR, transmission electron microscopy (TEM), dynamic light scattering (DLS), thermogravimetric analysis (TGA), ultraviolet spectroscopy (UV), and zeta potential (ZP). The biological properties of dendrons and AuNPs were determined by hemolysis, platelet aggregation and lymphocyte proliferation. These assays reflect modification of dendron properties when covering nanoparticles. For dendrons, hemolysis and platelet aggregation are generation dependent whilst, for AuNPs these properties are related to the bigger size of NPs. On the other hand, none of the systems induced lymphocyte proliferation. Selected cationic dendrons and AuNPs were chosen for gene delivery experiments employing a small interference RNA (siRNA Nef) against human immunodeficiency virus (HIV).

Acquired immune deficiency syndrome (AIDS) due to human immunodeficiency virus type-1 (HIV-1) represents one of the most important sexually transmitted infections (STI) worldwide. Great international efforts have been made to stop new infections but, to date, several compounds failed as microbicides at different stages of clinical trials. The quest to design new molecules that could prevent these infections is essential. In this work, we synthesized the first, second and third generations of anionic dendrimers having carboxylate and sulfonate terminal groups, respectively named G1C, G2C, G3C and G1S, G2S, and G3S, starting from a family of poly(alkylideneamine) dendrimers with nitrile termini. The anionic terminal groups of these dendrimers were expected to prompt them to act against HIV-1 infection. All dendrimers were fully characterized by 1H- and 13C-NMR, FTIR, MS and zeta potential techniques. Importantly, they were able to remain stable in the solid state and aqueous solutions at least for one and a half years. Screening of these six new dendrimers was then performed to shed light on their potential anti-HIV-1 activity and their mechanism of action. Results showed that the dendrimers were cytocompatible and that G1C and G1S dendrimers had important activity against R5-HIV-1NLAD8 and X4-HIV-1NL4.3 isolates by acting directly on viral particles and blocking their entry in host cells. Additionally, G1C and G1S dendrimers maintained their inhibitory effect at different pH values. Through a vaginal irritation assay carried out in BALB/c mice, the safety of these new dendrimers for topical application was also shown. Taken together, our results clearly show that G1C and G1S dendrimers are strong candidates for developing an effective microbicide to prevent HIV-1 new infections.

[Aim] To research the synergistic activity of G2-S16 dendrimer and dapivirine (DPV) antiretroviral as microbicide candidate to prevent HIV-1 infection., [Materials & methods] We assess the toxicity of DPV on cell lines by MTT assay, the anti-HIV-1 activity of G2-S16 and DPV alone or combined at several fixed ratios. Finally, their ability to inhibit the bacterial growth in vitro was assayed. The analysis of combinatorial effects and the effective concentrations were performed with CalcuSyn software., [Conclusion] Our results represent the first proof-of-concept study of G2-S16/DPV combination to develop a safe microbicide.

Background Current national immunisation schedules differ between countries in terms of vaccine formulation, timing of vaccinations and immunisation programme funding and co-ordination. As a result, some HIV infected paediatric population may be left susceptible to vaccine preventable infections. Vaccines used in healthy population should be subjected to high quality ethical research and be explicitly validated for use in children with special vaccination needs such as those infected with HIV. This survey was completed to assess current vaccination practices and attitudes toward vaccination among pediatricians who care for vertically HIV infected children. Methods An online questionnaire was completed by 46 experts in paediatric HIV-infection from the Paediatric European Network for Treatment of AIDS (PENTA). Data were collected between November 2013 and March 2014. Results 46 units looking after 2465 patients completed the questionnaire. The majority of units (67%) reported that common childhood immunisation were administered by the family doctor or local health services rather than in the HIV specialist centre. Vaccination histories were mostly incomplete and difficult to obtain for 40% of the studied population. Concerns were reported regarding the use of live attenuated vaccines, such as varicella and rotavirus, and these were less frequently recommended (61% and 28% of the units respectively). Monitoring of vaccine responses was employed in a minority of centres (41%). A range of different assays were used resulting in diverse units of measurement and proposed correlates of protection. Conclusion Vaccination practices for perinatally HIV-infected children vary a great deal between countries. Efforts should be made to improve communication and documentation of vaccinations in healthcare settings and to harmonise recommendations relating to additional vaccines for HIV infected children and the use of laboratory assays to guide immunisation. This will ultimately improve coverage and vaccine induced immunity in this vulnerable patient group.

Resumen Introduccion Los avances en el tratamiento antirretroviral han mejorado la esperanza de vida de ninos con infeccion por VIH por transmision vertical. Sin embargo, han aparecido nuevos retos. Planteamos este estudio con el objetivo de determinar los aspectos psicosociales y el conocimiento sobre su enfermedad en una cohorte de adolescentes con infeccion por VIH por transmision vertical. Metodos Se incluyeron pacientes con infeccion por VIH por transmision vertical con edades comprendidas entre 12-19 anos. Los datos se obtuvieron mediante entrevista semiestructurada y el Strengths and Difficulties Questionnaire para cribado de trastornos emocionales y de conducta. Resultados Se evaluaron 96 pacientes (58% mujeres) con mediana de edad de 15 anos (11-19,1) y mediana de edad del diagnostico de 1,70 anos (0-12,2). La mediana de CD4 en el momento del corte fue 626 cels/mm3 (132-998); el 72% de los pacientes presentaban una carga viral Conclusion Se objetivan dificultades psicosociales en un elevado porcentaje de pacientes (conocimiento de la enfermedad, relacion con pares, fracaso escolar...) que podrian tener impacto en su incorporacion a la vida adulta. Son necesarios mas estudios para profundizar en el origen y evolucion de las dificultades observadas, asi como intervenir para prevenir y modificar esta situacion.

Enrique Vacas-Córdoba,1–3 Marta Galán,3,4 Francisco J de la Mata,3,4 Rafael Gómez,3,4 Marjorie Pion,1–3 M Ángeles Muñoz-Fernández1–3 1Laboratorio InmunoBiología Molecular, Hospital General Universitario Gregorio Marañón, Madrid, Spain; 2Instituto de Investigación Sanitaria del Gregorio Marañón, Madrid, Spain; 3Networking Research Center on Bioengineering, Biomaterials and Nanomedicine, (CIBER-BBN), Madrid, Spain; 4Dendrimers for Biomedical Applications Group (BioInDen), University of Alcalá, Madrid, Spain Abstract: Self-administered topical microbicides or oral preexposure prophylaxis could be very helpful tools for all risk groups to decrease the human immunodeficiency virus (HIV)-1 infection rates. Up until now, antiretrovirals (ARVs) have been the most advanced microbicide candidates. Nevertheless, the majority of clinical trials has failed in HIV-1 patients. Nanotechnology offers suitable approaches to develop novel antiviral agents. Thereby, new nanosystems, such as carbosilane dendrimers, have been shown to be safe and effective compounds against HIV with great potential as topical microbicides. In addition, because most of the attempts to develop effective topical microbicides were unsuccessful, combinatorial strategies could be a valid approach when designing new microbicides. We evaluated various combinations of anionic carbosilane dendrimers with sulfated (G3-S16) and naphthyl sulfonated (G2-NF16) ended groups with different ARVs against HIV-1 infection. The G3-S16 and G2-NF16 dendrimers showed a synergistic or additive activity profile with zidovudine, efavirenz, and tenofovir in the majority of the combinations tested against the X4 and R5 tropic HIV-1 in cell lines, as well as in human primary cells. Therefore, the combination of ARVs and polyanionic carbosilane dendrimers enhances the antiviral potency of the individual compounds, and our findings support further clinical research on combinational approaches as potential microbicides to block the sexual transmission of HIV-1. Keywords: microbicides, HIV, carbosilane dendrimer, antiretroviral, synergy

Here we synthesized carbosilane, generation 1 to 3, and PEG-based dendrons functionalized at the periphery with NHBoc groups and at the focal point with azide and alkyne moieties, respectively. The coupling of these two types of dendrons via click chemistry led to the formation of new hybrid dendrimers with two distinct moieties, the hydrophobic carbosilane and the hydrophilic PEG-based dendron. The protected dendrimers were transformed into cationic ammonium dendrimers. These unique amphiphilic dendrimers were studied as vectors for gene therapy against HIV in peripheral blood mononuclear cells (PBMC) and their performance was compared with that of a PEG-free carbosilane dendrimer. The presence of the PEG moiety afforded lower toxicities and evidenced a weaker interaction between dendrimers and siRNA when compared to the homodendrimer analogous. Both features, lower toxicity and lower dendriplex strength, are key properties for use of these vectors as carriers of nucleic material., Comunidad de Madrid, Ministerio de Sanidad y Consumo, Ministerio de Economía y Empresa

The synthesis and characterization of four families of anionic carbosilane dendrimers bearing carboxylate, phospho- nate, naphthylsulfonate, and sulfate terminal groups prepared by cycloaddition of azide-alkyne catalyzed by copper (CuAAC) are presented here. For the preparation of these anionic carbosi- lane dendrimers, two strategies starting from azide-terminated carbosilane dendrimers were followed: (i) click coupling of neu- tral alkynes followed by derivatization into anionic moieties or (ii) click coupling of anionic alkynes. Both strategies require dif- ferent reaction conditions in order to accommodate the different substrate polarities. These anionic dendrimers, in general, do not present cell toxicity in vitro until concentration up to 20 mM. Therefore, they can be used in inhibition experiments in concentrations below this limit. We have observed that den- drimers bearing phosphonate groups possess poor anti-HIV capabilities in vitro in PBMCs, while carboxylate dendrimers can reduce HIV infection levels moderately. On the other hand, sul- fate and naphthylsulfonate dendrimers are powerful anti-HIV agents and their antiviral activity is generation and concentra- tion dependent. V C 2014 Wiley Periodicals, Inc. J. Polym. Sci., Part A: Polym. Chem. 2014, 00, 000-000

Age-related biological deterioration also includes immune system deterioration and, in consequence, a rise in the incidence and prevalence of infections and cancers, as well as low responses to vaccination strategies. Out of all immune cell subsets, T-lymphocytes seem to be involved in most of the age-related defects. Since T-lymphocytes mature during their passage through the thymus, and the thymus shows an age-related process of atrophy, thymic regression has been proposed as the triggering event of this immune deterioration in elderly people. Historically, it has been accepted that the young thymus sets the T-lymphocyte repertoire during the childhood, whereupon atrophy begins until the elderly thymus is a non-functional evolutionary trace. However, a rising body of knowledge points toward the thymus functioning during adulthood. In the elderly, higher thymic function is associated with a younger immune system, while thymic function failure is associated with all-cause mortality. Therefore, any new strategy focused on the improvement of the elderly quality of life, especially those trying to influence the immune system, should take into account, together with peripheral homeostasis, thymus function as a key element in slowing down age-related decline.

The neuraminidase (NA) epitope from the Mexican AH1N1 influenza virus was identified by using sequences registered at the GenBank during the peak of a pandemic (from April 2009 to October 2010). First, NA protein sequences were submitted for multiple alignment analysis, and their three-dimensional models (3-D) were then built by using homology modeling. The most common sequence (denominated wild-type) and its mutants were submitted to linear and nonlinear epitope predictors, which included the major histocompatibility complex type II (MHC II) and B-cell peptides. The epitope prediction was in accordance with evolutionary behavior and some protein structural properties. The latter included a low NA mutation rate, NA 3-D surface exposure, and the presence of high hindrance side chain residues. After selecting the epitope, docking studies and molecular dynamics (MD) simulations were used to explore interactions between the epitope and MHC II. Afterward, several experimental assays were performed to validate the theoretical study by using antibodies from humans (infected by pandemic H1N1) and rabbits (epitope vaccination). The results show 119 complete sequences that were grouped into 28 protein sequences according to their identity (one wild-type and 27 representative mutants (1–5 mutations)). The predictors yielded several epitopes, with the best fit being the one located in the C-terminal region. Theoretical Methods: demonstrated that the selected epitope reached the P4, P6, P7, and P9 pockets of MHC II, whereas the experimental evidence indicates that the epitope is recognized by human antibodies and also by rabbit antibodies immunized with the peptide.

In order to improve the efficiency of the anti-inflammatory drug ibuprofen, cationic carbosilanedendrimers and dendrons with ibuprofen at their periphery or at their focal point, respectively, havebeen synthesized and the release of the drug was studied using HPLC. Macrophages were used toevaluate the anti-inflammatory effect of the ibuprofen-conjugated dendritic systems and comparedwith mixtures of non-ibuprofen dendritic systems in the presence of the drug. The cationic ibuprofenconjugateddendron was the compound that showed higher anti-inflammatory properties. It reduces the LPS-induced COX-2 expression, decreases the release of several inflammatory cytokines such as TNFalfa, IL-1beta, IL-6 and CCL3. These results open new perspectives in the use of these compounds asdrug carriers., Ministerio de Economía y Empresa

Background Initiating ART during acute/recent HIV-1 infection reduces viral reservoir formation. It has been proposed that, during this phase, the size of the viral reservoir could be further reduced by the association of immunomodulatory therapy with ART. Contradictory results have emerged, however, from two trials evaluating the impact on immune recovery and the viral reservoir of adding cyclosporine A to ART during primary HIV-1 infection. Patients and methods Twenty patients with acute/recent HIV-1 infection were randomized to receive ART alone (tenofovir, emtricitabine and lopinavir/ritonavir) or associated with 8 weeks of cyclosporine A (0.3-0.6 mg/kg twice daily). The impact on viral load, immune response and integrated and non-integrated DNA viral reservoir at 0, 8 and 36 weeks of treatment was evaluated. Results The estimated median time from HIV-1 infection to ART onset was 63 days (IQR 53; 79.5) with 90% of patients at Fiebig V stage. No significant differences were observed in viral load decay, CD4 T cell recovery, immune response markers or the evolution of integrated DNA at week 8 (end of cyclosporine A) and week 36 between groups. However, non-integrated DNA significantly increased in the cyclosporine A arm between weeks 0 and 36. Cyclosporine A was well tolerated. Conclusions Adding cyclosporine A to ART during acute/recent infection did not improve immune recovery. However, unintegrated DNA increased in the cyclosporine A group, suggesting an anti-integration effect, a point warranting further research (ClinicalTrials.gov Identifier: NCT00979706).

Background: CD8 T cells are crucial in the immune responses against HIV infection, but HIV-infected adults suffer a naive CD8 T-cell depletion and accelerated senescence caused by chronic antigen stimulation. Although HIV-infected children preserve a better immune reconstitution capacity their CD8 responses are defective. We wanted to know, whether HIV vertical transmission produces a premature aging of the CD8 population due to antigen exposition to HIV from birth and persistent chronic activation. Methods: We conducted a multicentre cross-sectional study that compared vertically HIV-infected children with detectable (viremic) or undetectable (aviremic) viral load and age-matched healthy children. Using multiparameter flow cytometry, we studied within the CD8 population the frequencies of naive, memory, effector memory (effector memory), and TemRA subsets and measured markers of senescence, activation, and proliferation in these cells. Results: We found that naive subset in viremic children was markedly decreased and had a replicative senescence phenotype. Furthermore, viremic children showed increased frequencies of memory, TEM and TemRA CD8 T cells, with a more activated and replicative senescence phenotype. We found that HIV-infected children with undetectable viral load have an increased senescence in memory and effector CD8 T cells, but the frequencies and phenotype of the CD8 subsets analyzed are comparable to healthy children. Conclussions: Our study shows that CD8 T cells of HIV-infected children have a more senescent phenotype when compared with age-matched healthy children. Interestingly enough, our results support the importance of maintaining undetectable viral load in HIV-infected children to avoid the premature ageing and dysfunction of CD8 T cells. © 2012 by Lippincott Williams & Wilkins.

We evaluated the 2G-NN16-carbosilane dendrimer activities in Th17 response as a potential therapy for Th17 deregulated pathologies. IL17A, IL17F, IL22, IL23 and other interleukins secreted by Th17 cells CD4+ cells were down regulated when cells were cultured in the presence of this dendrimer. Furthermore, IL17F and IL17A protein levels in splenocytes from mice pretreated with 2G-NN16 dendrimer in a Th17 induction mouse model were lower than those corresponding to PBS treated mice. Treatment of mice with 2G-NN16 inhibited the Th17 response causing much more pathogenicity as indicated by the increase in the number of Candida albicans colonies in the kidneys as compared to PBS-treated mice. All these results suggest a potential pharmacological application for this dendrimer in the therapy of Th17-mediated diseases.

Current advances in neonatology have improved survival among preterm and low-birth-weight infants. However, the risk of neonatal death in preterm infants is much greater than in full-term neonates and is frequently associated with infections.Little is known about the immune status of preterm neonates; therefore, we analyzed the frequency and absolute counts of different immune populations in 211 cord blood samples taken from very-preterm to full-term neonates.We found that absolute counts of all the immune subsets analyzed (i.e., monocytes, granulocytes, B cells, natural killer (NK) cells, CD4(+), and CD8(+) T cells) were markedly lower in preterm infants than in full-term infants. Surprisingly, we observed that regulatory T cells (Tregs) were the only cell subset that did not decrease in preterm infants, and their frequency was even higher than in full-term infants.Tregs are crucial to maternal-fetal tolerance, but their suppressive role could be also implicated in the leukopenia observed in preterm infants. We did not observe differences in thymic function, but we found that plasma levels of interleukin (IL)-7 and the frequency of its receptor were significantly decreased in preterm infants. Our results could help to identify leukopenia and to implement immune therapies that significantly diminish mortality in preterm neonates.

The virological response after an 8-day maraviroc monotherapy has been proposed to be an alternative method to determine whether an CCR5 antagonist should be prescribed to HIV-infected patients. The frequency of patients eligible for a combined antiretroviral therapy which includes maraviroc on the basis of the result of this clinical test is not well-known at the moment. In the same way, clinical and immunovirological factors associated with the virological response after antagonist exposure need to be determined. Ninety consecutive HIV-infected patients were exposed to an 8-day maraviroc monotherapy. The virological response was considered positive if either a reduction of ≥1-log 10 HIV RNA copies/ml or an undetectable viral load (+ T-cell counts. The CXCR4-tropic virus level was the only variable independently associated with the virological response after short-term maraviroc exposure. Lower CD4 + T-cell strata were associated with higher CXCR4-tropic virus levels. These results support the suggestion that CCR5 antagonists should be an early treatment option before the expansion of CXCR4-tropic strains.

Herein, we describe the synthesis and characterization of a sulfonate-containing N-donor ligand in its sodium salt and acid forms, Na2[(DES)MeN(CH2)2NMe(DES)]·2H2O (1) and [(DES)MeN+H(CH2)2N+HMe(DES)]·H2O (2), and its corresponding metal complexes, [{(DES)MeN(CH2)2NMe(DES)}M(H2O)2]·nH2O [M = Ni (3), Co (4), Cu (5) and Zn (6)]. Treatment of HIV-infected MT-2 cells with Ni, Co and Cu complexes inhibit virus replication up to 50–70 % both in pre- and post-infected cells as a result of dual preventive and therapeutic behaviour.

OBJECTIVES Hepatitis C virus (HCV) antiviral therapy might lead to decreased chronic immune activation and endothelial dysfunction associated with cardiovascular risk. The aim was to evaluate the effect of HCV eradication on serum markers of inflammation and endothelial dysfunction markers in HIV/HCV co-infected patients. METHODS We carried out a retrospective study of 69 HIV/HCV co-infected patients on interferon (IFN)-α plus ribavirin. In addition, 47 HIV-infected subjects were selected as a control group. A sustained virological response (SVR) was defined as an undetectable HCV viral load up to 24 weeks after the end of treatment. Tumour necrosis factor (TNF) receptor-1 (TNF-R1), soluble E-selectin (sE-selectin), soluble P-selectin (sP-selectin), soluble intercellular adhesion molecule 1 (sICAM-1) and soluble vascular cell adhesion molecule 1 (sVCAM-1) were measured using a multiplex immunoassay kit. RESULTS HIV/HCV co-infected patients had higher values of soluble TNF-R1 (sTNF-R1), sE-selectin and sICAM-1 than HIV mono-infected patients (P

Several carbosilane wedges of generations 1–3 have been synthesized, following the divergent method, containing at the focal point a C–Br bond and as peripheral functional groups SiMeCl2, SiMe(C3H5)2, SiMe2Cl, SiMe2H, and ester units SiMe2{(C3H6)N(C2H4CO2Me)2}. The dendrons functionalized with SiMe(C3H5)2 and SiMe2{(C3H6)N(C2H4CO2Me)2} groups were used to synthesize spherical dendrimers derived from 1,3,5-(HO)3C6H3, leaving the outer groups unchanged. The allyl dendrimers thus obtained were used as precursors to prepare new dendrimers functionalized with SiMeCl2, SiMe2Cl, SiMe2H, amine units SiMe2{(C3H6)NH2} and also ester units SiMe2{(C3H6)N(C2H4CO2Me)2}.

One of the primary limitations of RNA interference as a technique for gene regulation is effective delivery of siRNA into the target cells. Dendrimers are nanoparticles that are increasingly being used as oligonucleotide and drug delivery vehicles. We have developed amino-terminated carbosilane dendrimers (CBS) as a means to protect and transport siRNA. Initially, stability studies showed that CBS bind siRNA via electrostatic interactions. Dendrimer-bound siRNA was found to be resistant to degradation by RNase. Cytotoxicity assays of CBS/siRNA dendriplexes with peripheral blood mononuclear cells (PBMC) and the lymphocytic cell line SupT1 revealed a maximum safe dendrimer concentration of 25 microg/ml. Next, utilizing flow cytometry and confocal microscopy, lymphocytes were seen to be successfully transfected by fluorochrome-labeled siRNA either naked or complexed with CBS. Dendriplexes with +/- charge ratio of 2 were determined to have the highest transfection efficiency while maintaining a low level of toxicity in these systems including hard-to-transfect HIV-infected PBMC. Finally, CBS/siRNA dendriplexes were shown to silence GAPDH expression and reduce HIV replication in SupT1 and PBMC. These results point to the possibility of utilizing dendrimers such as CBS to deliver and transfect siRNA into lymphocytes thus allowing the use of RNA interference as a potential alternative therapy for HIV infection.

Introduccion Existen pocos estudios sobre como la coinfeccion por el virus de la inmunodeficiencia humana (VIH) y el virus de la hepatitis C (VHC) alteran su evolucion al coexistir en ninos. Metodos Se estudiaron cinco ninos coinfectados por el VIH y el VHC con tratamiento antirretroviral gran actividad (TARGA) con lopinavir/ritonavir (LPV/r). Su evolucion fue comparada con un grupo control de 72 pacientes con infeccion por el VIH. Resultados Todos los pacientes mantuvieron una recuperacion inmunologica sostenida durante el seguimiento y tambien alcanzaron un mejor control de la carga viral que el grupo control. Ninguno de los cinco ninos coinfectados experimento ningun efecto adverso a LPV/r durante el estudio. Conclusion La coinfeccion por el VIH/VHC no disminuye los beneficios obtenidos con el TARGA.

Immune recovery after prolonged highly active antiretroviral therapy (HAART) with lopinavir/ritonavir has been reported in adults but not in children. Our study aimed at evaluating the long-term use of lopinavir/ritonavir among children in a clinical setting.We carried out a retrospective study on 69 protease inhibitor (PI)-experienced vertically HIV-infected children on HAART containing lopinavir/ritonavir. We analysed the changes in percentage CD4+ cell count (%CD4+) and viral load (VL) and identified prognostic factors to achieve CD4+25% and undetectable VL (uVL) (or=400 copies/mL) by logistic regression.During the first 2 years, we found an increase in the %CD4+ in children with baseline CD4+ between 0% and 15% and those with baseline VL30,000 copies/mL. We found a decrease in VL in all groups of children. From second to fourth year, we found an increase in %CD4+ in all the children who had CD4+25% and in those with baseline VL100,000 copies/mL. We found that %CD4+ at baseline had a strong positive association with achieving CD4+25% at 6, 12, 18, 24, 36 and 48 months of follow-up. We also found that length of PI use had a negative association with reaching CD4+25% at 24 and 48 months and achieving uVL at 12 and 24 months. VL at baseline had a negative association with achieving uVL at 18 and 24 months.Our study demonstrates ongoing immune recovery among children on HAART with lopinavir/ritonavir after 4 years of follow-up. Lopinavir/ritonavir, when given as part of a salvage regimen, is safe and well tolerated in HIV-infected children.

We carried out a retrospective study to analyze the long-term response to highly active antiretroviral therapy of 19 vertically human immunodeficiency virus type 1/hepatitis C virus (HCV-1/HIV) coinfected children. The clinical, immunologic, viral, and biochemical variables were assayed at 0, 1, 2, 3, 4, 5, and 6 years of follow-up. Our data suggest that CD4 T-cell recovery and viral load control during long-term highly active antiretroviral therapy among HIV-1/HCV children were similar to those described in HIV-1 monoinfected children, but hepatic function was significantly altered in HIV-1/HCV children.

We carried out a longitudinal study to analyze the immune recovery of four patients with aggressive HIV-associated lymphoma (HIV+ Ly+) treated with highly active antiretroviral therapy (HAART) and high-dose chemotherapy followed by autologous peripheral blood stem cell transplantation (ASCT). We also studied three control non-HIV-infected patients with lymphoma (HIV-Ly+) and six HIV patients on HAART without lymphoma (HIV+ Ly-). After 12 months of follow-up, the HIV HIV+Ly+ patients reached the pre-ASCT CD4+ levels, despite a transient decrease after the ASCT. All ASCT patients (HIV+Ly+ and HIV-Ly+) showed an increase in CD4+, CD4+ CD45RO+, and CD4+CD28+ T cells/microl. Although HIV+Ly+ patients had values of CD4+, CD4+CD45RO+, and CD4+CD28+ T cells/microl lower than the HIV-Ly+ patients, their recovery rate over the 12 months after ASCT appeared to be better. HIV+Ly+ patients had higher pre-ASCT plasma IL-7 levels than HIV-Ly+, however, these values decreased after ASCT. All ASCT patients showed a slight increase of TCR rearrangement excision circles (TRECs) and they did not have a different pattern of TREC evolution. We could not find differences between HIV+Ly+ patients 12 months after ASCT and HIV+Ly- in DNA-HIV (copies/10(6) cell). Overall, HIV+Ly+ patients showed an appropriate immune reconstitution 12 months after ASCT, and, interestingly, they had an amount of DNA-HIV copies similar to HIV+Ly- control patients in their CD4+ cells.

Background: Virus-specific CD4 T cells play a critical role in antiviral immunity. HIV1-specific CD4 T cells in chronically infected adults are mostly composed of IFN-gsecreting cells, whereas a selective defect in IL-2-secreting CD4 T cells has been demonstrated. HIV-1-specific IL-2-secreting CD4 T cells are key components of effective immunity. Objective: To determine the function of HIV-1-specific CD4 T cells in HIV-1 vertically infected children after antiretroviral treatment (ART). Design: Twenty-three vertically HIV-infected children treated with ART for an extended period (mean 7 years) were retrospectively studied. Methods: The function of HIV-1-specific CD4 T cells was determined by their ability to secrete IL-2 and IFN-g after stimulation with HIV-1 p55 gag protein using polychromatic flow cytometry. Results: Substantial differences in the patterns of CD4 T-cell responses were associated with different conditions of response to ART. Interestingly, children with suppression of viraemia below 50 HIV-1-RNA copies/ml of plasma for at least 2 years showed dominant IL-2 CD4 T-cell responses; children with viraemia below 50 copies but experiencing transient blips of viraemia showed polyfunctional (IL-2 plus IFN-g) CD4 Tcell responses; children with uncontrolled high viraemia levels had dominant IFN-g CD4 T-cell responses. Furthermore, the total frequency of HIV-1-specific CD4 T cells including IL-2 and IFN-g-secreting cells was significantly higher compared with HIVinfected adults with chronic infection. Conclusion: The higher frequency of HIV-1-specific CD4 T cells in children compared with adults and the recovery of IL-2-secreting CD4 T cells after successful ARTmediated suppression of virus replication indicate a greater capacity of immune restoration in children than adults.

Astrocytes are the major glial cell within the central nervous system and have a number of important physiological properties related to brain homeostasis. They provide trophic support to neurons and are immune cells with key roles during states-of-inflammation. The potential for production of proinflammatory cytokines and its consequences has been studied in the context of HIV-1 infection of normal human astrocytes (NHA). NHA express TLR3, TLR4 and TLR5. TLR3 ligation induced the strongest proinflammatory polarizing response, characterized by generation of high levels of TNF-α, IL-6 and IL-8. HIV-1 increased the transient production of key inflammatory mediators and exposure to LPS of HIV-1-infected cells increased significantly the cytokine secretion. We confirmed that it is necessary viral gene expression from the moment of pretreatment with antiretrovirals inhibited totally HIV-1-induced TLR response. The higher response to LPS from HIV-1-infected cells did not correlate with TLR4 or MyD88 increased expression. LPS responsiveness of infected cells parallels MHC class II expression, but not CD14. HIV-1-infected NHA present increased sensitivity to the proinflammatory effects of LPS. If this phenomenon occurs in vivo, it will contribute to the immunopathogenesis of this disease and may ultimately offer novel targets for immunomodulatory therapy.

Anionic carbosilane dendrons decorated with sulfonate functions and one thiol moiety at the focal point have been used to synthesize water-soluble gold nanoparticles (AuNPs) through the direct reaction of dendrons, gold precursor, and reducing agent in water, and also through a place-exchange reaction. These nanoparticles have been characterized by NMR spectroscopy, TEM, thermogravimetric analysis, X-ray photoelectron spectroscopy (XPS), UV/Vis spectroscopy, elemental analysis, and zeta-potential measurements. The interacting ability of the anionic sulfonate functions was investigated by EPR spectroscopy with copper(II) as a probe. Different structures and conformations of the AuNPs modulate the availability of sulfonate and thiol groups for complexation by copper(II). Toxicity assays of AuNPs showed that those produced through direct reaction were less toxic than those obtained by ligand exchange. Inhibition of HIV-1 infection was higher in the case of dendronized AuNPs than in dendrons.

A synthetic strategy has been developed for the preparation of new peripheral amine- or ammonium-terminated carbosilane dendrimers of type nG-[Si(OCH2CH2NMe2)y]x or nG-[Si(OCH2CH2NMe3+I–)y]x, respectively. It consists of the alcoholysis of well-known chlorosilane-terminated dendrimers with N,N-dimethylethanolamine and the subsequent quaternization with MeI. All these systems are susceptible to hydrolysis, although the decomposition depends on concentration and dendrimer generation. Evaluation of dendrimer toxicities by phase-contrast light microscopy and MTT assay were carried out, and evidence of dendrimer/oligonucleotide complex formation was carried out by gel electrophoresis. (© Wiley-VCH Verlag GmbH & Co. KGaA, 69451 Weinheim, Germany, 2006)

Transcriptional induction of cyclooxygenase-2 (COX-2) occurs early after T cell receptor triggering and has functional implications in inflammation. Here, we show that phosphodiesterase (PDE)-4 inhibitors block COX-2 induction and prostaglandin synthesis in activated T cells. COX-2 inhibition by PDE4 inhibitors occurs mainly at the transcriptional level. Two response elements for the nuclear factor of activated T cells (NFAT) in the COX-2 promoter were required for inhibition by these drugs. PDE4 inhibitors did not affect NFAT nuclear translocation upon T cell activation; rather they prevented NFAT binding to DNA and induction of the transactivation function of GAL4-NFAT. These effects seem to be cAMP/PKA independent as they were not mimicked by the permeable analog dBcAMP or by forskolin, neither can be reverted by the PKA inhibitors H89 or KT-5720. These results may explain some of the anti-inflammatory properties of PDE4 inhibitors through the blockade of NFAT-mediated transactivation of pro-inflammatory genes such as COX-2.

Fundamento y objetivo Estudiar la relacion existente entre subpoblaciones de linfocitos T y la carga viral plasmatica (CV) y los circulos de escision del receptor del linfocito T (TREC) en un grupo de ninos infectados por el virus de la inmunodeficiencia humana (VIH). Pacientes y metodo Se realizo un estudio longitudinal retrospectivo durante un ano en 20 ninos infectados por el VIH que recibian tratamiento antirretroviral de gran actividad. Se analizo la relacion existente entre los cambios en las subpoblaciones de linfocitos T con los cambios en la CV y los TREC en sangre periferica mediante un modelo de regresion lineal. Resultados Los incrementos de linfocitos T CD4+ memoria y los linfocitos T CD4+ activados tuvieron una asociacion negativa con el incremento del log10 TREC. Sin embargo, los incrementos de linfocitos T CD4+ virgen tuvieron una relacion positiva con los incrementos del log10 TREC. Por otro lado, los incrementos de linfocitos T CD8+ memoria, linfocitos T CD8+ activados y linfocitos T CD8+ efectores se correlacionaron de forma positiva con el log10 CV. Sin embargo, la subpoblacion de linfocitos T CD8+ virgen tuvo una asociacion negativa con el log10 CV. Conclusiones Las subpoblaciones de linfocitos T CD4+ y T CD8+ varian de distinta forma en respuesta a los cambios producidos por el tratamiento antirretroviral de gran actividad en ninos infectados verticalmente por el VIH. Las subpoblaciones de linfocitos T CD4+ dependen mas de la funcion timica, y las subpoblaciones de linfocitos T CD8+, de la replicacion viral. Asi, la caida de la activacion celular permitiria que el timo produjera mayor numero de linfocitos virgen y que estos no se diferencien a linfocitos de memoria/efectores y mueran por apoptosis.

Fundamento y objetivo: La infeccion por el virus de la inmunodeficiencia humana de tipo 1 (VIH-1) tiene distintos patrones de progresion de la enfermedad con manifestaciones muy heterogeneas. El objetivo de este estudio ha sido analizar las subpoblaciones de celulas T CD8+ en un grupo de ninos infectados por el VIH-1 asintomaticos a largo plazo (ALD) y que tenian recuento de celulas T CD4+ estables (superior 600/µl). Pacientes y metodos: Estudio transversal en ninos infectados verticalmente por el VIH-1, mayores de 7 anos, divididos de acuerdo con su clasificacion clinica e inmunologica en a) ALD: 7 ninos con infeccion por el VIH-1 asintomaticos en A1 y b) PR (progresion rapida): 14 ninos con infeccion por el VIH-1 en C3. El grupo control lo constituyeron 17 ninos sin infeccion. La caracterizacion de subpoblaciones de celulas T CD8+ se hizo por citometria de flujo de 3 colores. Resultados: El grupo ALD mostro recuentos de celulas T CD8+ memoria (CD8+CD45RO+, CD8+CD45RA-CD28+ y CD8+CD45RA-CD62L+) similares a los del control, e inferiores a los del grupo PR. El grupo ALD tambien tuvo valores de celulas T CD8+ virgenes (CD8+CD45RA+CD62L+, CD8+CD45RA+CD28+, CD8+CD45RA+, CD8+CD28+ y CD8+CD62L+) similares a los del control, pero superiores a los del grupo PR. Asimismo presento valores similares de celulas T CD8+CD38+ al control, siendo mas bajos en el grupo PR. Sin embargo, el grupo ALD tuvo valores mas altos de CD8+HLA-DR+CD38+ y CD8+HLA-DR+ que el control. Ademas, presento valores mas elevados de celulas T CD8+ preefectoras que el grupo control, pero valores similares de T CD8+ efectoras. Tambien, el grupo ALD tuvo valores mas bajos de celulas T CD8+CD11b+ que el control. Conclusiones: Los ninos infectados por el VIH-1 asintomaticos tuvieron una distribucion de subpoblaciones CD8+ casi normal, con excepcion de las celulas T CD8+ activadas y preefectoras

To asses the role of interleukin 7 (IL-7) in the thymic reconstitution of CD4 T cells observed in children after successful antiretroviral therapy, a longitudinal study in five vertically HIV-1-infected children was carried out. Thymic function, IL-7 plasma levels, viral load, and T-lymphocytes subsets were determined every 2 or 3 months for about 90 months. In all the children, the drop in CD4+ T cells below 5–10% was associated with a marked increase in IL-7 plasma levels. The drastic decrease in viral load after treatment, led in all the cases to a recover of CD4 to levels higher than 30%, which was associated to an increase in thymic production of T cells and followed by a decrease in IL-7 to the normal levels. We conclude that the drop in CD4 in HIV children would induce an increase of IL-7 as part of a homeostatic mechanism. IL-7 would induce the thymus to produce new T cells to recover the normal levels of CD4 when the viral load was low and so the thymic function was not inhibited. The increase in the thymic production of new T cells recovers the CD4 population, and leads to a normalization of IL-7 levels.

Fundamento y objetivo: Evaluar la capacidad del tratamiento antirretroviral de gran actividad (TARGA) para reconstituir totalmente el sistema inmunologico en un nino infectado por el virus de la inmunodefiencia humana (VIH) con sida grave y profunda inmunodepresion. Paciente y metodo: La respuesta proliferativa y la produccion de citocinas de las celulas mononucleares de sangre periferica (CMSP) frente a fitohemoglotinina (PHA) fueron evaluadas por incorporacion de [3H]-timidina y enzimoinmunoanalisis, respectivamente. Las subpoblaciones linfocitarias T se cuantificaron por citometria de flujo y la produccion timica de celulas T, mediante la cuantificacion de TCR rearrangement excision circles (TREC). Resultados: En sus primeros anos de vida, el nino presento otitis recurrente por Pneumococcus. A los 2-3 anos de edad, las celulas T CD4+ descendieron de 822 a 154 celulas x 106/l, iniciandose tratamiento con zidovudina (AZT), aunque nunca supero las 350 celulas T CD4+ x 106/l. A los 6 anos de edad, la cifra de celulas T CD4+ era inferior a 15 x 106/l, y se administro estavudina (d4T) junto a lamivudina (3TC) sin recuperar las celulas T CD4+. A los 7 anos de edad, la cifra de celulas T CD4+ era del 0,8% y la carga viral (CV) de 32.000 copias/ml. Este estado coincidio con un leiomioma plantar que fue extirpado y una neumonia por Pneumocystis carinii. Tras 6 meses de TARGA (con d4T, 3TC, viracept y efavirenz) alcanzo valores de celulas T CD4+ del 25%, celulas T CD4 de 597 x 106/l, y CV indetectable. Ademas se produjo un incremento del numero absoluto de celulas virgenes y valores de TREC. Tras el TARGA aumentaron la respuesta linfoproliferativa a PHA y la produccion de interferon gamma (IFN-?). Actualmente el paciente esta asintomatico. Conclusion: La recuperacion inmunologica y clinica del nino infectado por el VIH en fases muy avanzadas de la enfermedad fue posible gracias al TARGA, lo que demuestra la plasticidad de su sistema inmunitario.

Fundamento Realizar un analisis de las subpoblaciones T implicadas en la recuperacion delsistema inmunitario en ninos infectados por el VIH-1 con terapia antirretroviral de gran actividad(TARGA) durante mas de 24 meses. Pacientes y metodo Se estudio a 17 ninos infectados por el VIH-1: a) grupo Res (ninos infectadospor el VIH-1 respondedores a TARGA): 10 ninos en categoria C3 al entrar en el estudio y trasmas de 24 meses con TARGA recuperan celulas T CD4+ (> 25% y 500 celulas T CD4+/ml) ycontrolan la replicacion viral, y b) grupo no-Res (ninos infectados por el VIH-1 no respondedoresa TARGA): 7 ninos en categoria C3 al entrar en el estudio y que tras mas de 24 meses con TARGAno recuperan celulas T CD4+ ( Resultados El grupo Res recupero los valores de celulas T CD4+ y CD8+ virgen (CD45RA+CD62L+)y memoria (CD45RO+) hasta alcanzar los valores del grupo control, siendo las diferencias significativascon respecto al grupo no-Res, excepto para las celulas CD8+CD45RO+ de los gruposRes y no-Res, que fueron mas elevadas que el grupo control. Ademas, el grupo Res tuvo valoresmas bajos de celulas CD8+HLA-DR+CD38+ que el grupo no-Res, aunque ambos grupos VIH-1(Res y no-Res) tuvieron valores significativamente mas altos de celulas T CD4+ y CD8+ activadas(HLA-DR+CD38+) que el grupo control. Conclusiones la recuperacion del sistema inmunitario inducida por la TARGA en los ninos infectadospor el VIH-1 parece ser la consecuencia de la disminucion de la activacion cronica delsistema inmunitario y de la recuperacion de las celulas T virgen.

Fundamento Evaluar los cambios producidos en el sistema inmunitario de ninos infectados porel virus de la inmunodeficiencia humana (VIH) tratados con terapia antirretroviral. Pacientes y metodo Se estudio a 23 ninos infectados verticalmente por el VIH: a) N-A1: 10 ninosen categoria A1; b) N-B2: 6 ninos en categoria B2, y c) N-C3: 7 ninos en categoria C3, y13 ninos sanos sin VIH como grupo control. La respuesta proliferativa y la produccion de citocinasde las CMSP frente a mitogenos fue evaluada por incorporacion de [3H]-timidina y enzimoinmunoanalisis,respectivamente. Las subpoblaciones linfocitarias T CD4+ y T CD8+ se cuantificaronpor citometria de flujo. Resultados Los ninos N-A1 tuvieron indices de estimulacion frente a PHA, PWM y anti-CD3+anti-CD28 similares al grupo control y mayores que los ninos N-C3. La produccion de TNF-α porlas CMSP sin estimular de los ninos VIH fue mas elevada que en el grupo control (p Conclusion Nuestros datos demuestran que, a pesar del tratamiento antirretroviral, siguen existiendoimportantes diferencias en el perfil inmunologico de los ninos con infeccion por el VIHen los distintos estadios clinicos de la enfermedad.