Impact de l'hypotension chez le rat avec encéphalopathie hépatique due à la maladie de foie chronique : implication pour les complications neurologiques suivant la transplantation hépatique

L’encéphalopathie hépatique (EH) est une complication neuropsychiatrique de la maladie de foie telle que la cirrhose, caractérisée par des dysfonctions cognitives et motrices. Le seul traitement curatif est la transplantation hépatique (TH). Historiquement, l’EH est considérée comme un désordre métabolique réversible et il est attendu qu’il soit résolu suivant la TH. Cependant, il a été démontré que des complications neurologiques persistent chez 47% des patients transplantés. La TH est une opération chirurgicale complexe accompagnée de stress péri-opératoire telle que la perte sanguine et l’hypotension. L’hypothèse de ce projet d’étude est que l’EH minimale (EHm) rend le cerveau plus susceptible à une perte neuronale suite à une insulte hypotensive. Nous avons donc caractérisé un modèle d’hypotension chez des rats cirrhotiques avec ligation de la voie biliaire (BDL) dans lequel une hypovolémie de l’artère fémorale a été faite. Avec ce modèle, nous avons étudié l’impact de différentes pressions sanguines de 120 minutes sur le compte neuronal. Nos résultats démontrent que les BDL hypotendus à une pression artérielle moyenne de 60 mmHg et 30 mmHg ont une diminution du compte neuronal et que les neurones mourraient par apoptose (observée par la présence de caspase-3 clivée). Nous avons également déterminé que le flot sanguin cérébral était altéré chez les rats cirrhotiques BDL. Le second objectif était d’évaluer si le traitement de l’EHm par l’ornithine phénylacétate (OP) permettait d’éviter la perte neuronale chez les BDL hypotendus. Nos résultats ont démontrés que l’OP permettait de partiellement rétablir les fonctions cognitives chez les rats BDL. De plus, les rats BDL traités avec l’OP peuvent éviter la mort neuronale. Cependant, le processus apoptotique est toujours enclenché. Ce résultat suggère la possibilité de mort cellulaire retardée par l’OP. Ces résultats suggèrent que les patients cirrhotiques avec EHm sont plus susceptibles à une mort neuronale induite par hypotension. La combinaison de l’EHm et l’hypotension permet d’expliquer les complications neurologiques rencontrées chez certains patients. Le diagnostic et le traitement de ce syndrome doit donc être fait chez les patients cirrhotiques pour éviter ces complications post-TH.

Étude du rôle de l'interleukine-32 dans l'infection à VIH-1

Les progresseurs lents du VIH-1 sont de rares sujets asymptomatiques pendant plusieurs années sans thérapie antirétrovirale. Parmi ces sujets à progression lente vers le SIDA, il est possible qu’un sous-groupe perde le contrôle de leur infection après plusieurs années de contrôle. Notre laboratoire a analysé l’expression différentielle de différentes protéines et voies moléculaires associées à la perte de contrôle de l’infection: l’interleukine-32 (IL-32) est une cytokine pro-inflammatoire dont le niveau des isoformes alpha et delta a significativement diminué chez les progresseurs lents lors de la perte de contrôle. Par ailleurs, des études antérieures ont attribué, de façon intrigante, à l’IL-32 aussi bien des propriétés anti-VIH-1 que des propriétés immunosuppressives induisant un environnement propice à la réplication du VIH-1. Ce projet de maitrise s’est penché sur l’implication de l’IL-32 dans la progression de l’infection à VIH-1 avec un accent particulier sur les progresseurs lents. Nous avons principalement mesuré les niveaux d’IL-32 des sujets séropositifs comparativement aux sujets VIH négatif et estimé les fonctions de cette cytokine à travers des études longitudinales et de corrélation. Nous avons observé que l’IL-32 total demeure plus élevé chez les séropositifs comparativement aux sujets VIH négatif. Également, l’infection par le VIH-1 entraine une augmentation du niveau d’IL-32 total. De plus, après une année de thérapie antirétrovirale, les taux plasmatiques d’IL-32 total demeurent significativement plus élevés que ceux des sujets VIH négatif. Comme attendu, le taux d’IL-32 total augmente lors de la perte de contrôle de l’infection chez les progresseurs lents. Une forte concentration plasmatique d’IL-32 total coïncide avec: 1) une augmentation du taux plasmatique de sCD14 et de la cytokine pro-inflammatoire IL-6, 2) une baisse du compte cellulaire CD4 et une augmentation de la charge virale. Un taux plasmatique élevé de CCL5 pourrait prédire une faible concentration d’IL-32 total. L’isoforme alpha de l’IL-32 est plus élevée dans le plasma des sujets VIH négatif tandis que l’IL-32 gamma semble induire un environnement pro-inflammatoire et immunosuppressif. Il ressort à l’issue de ces observations que l’augmentation de l’IL-32 total est associée à la progression de l’infection à VIH-1 et pourrait constituer un biomarqueur permettant d’apprécier le pronostic de cette infection.

“Effects of monospecific and mixed algal diets on survival, development and biochemical composition of Penaeus monodon larvae”

The main objective of the present work is to acquire information regarding the growth responses of P. monodon larvae (from PZ1 upto PL1) to various mono specific and mixed diets. Evaluate the nutritional quality of selected species of micro algae viz. Chaetoceros calcitrans, Dunaliella salina, Isochrysis galbana and Nannochloropsis salina, larvae at three cell concentrations 10x104 cells/ml, 25x104 cells/ml and 50x104 cells/ml. The P. monodon larvae were transported, at the Nauplius stage, to the laboratory. The larvae were stocked at density of 150 larvae per litre in 5 litre FRP tanks with 3 litres of sea water. The algal cell density given to the larvae varied. The larval stages were fed with increasing densities of algae to evaluate the relationship between the food densities, ingestion rates, development and growth of the larvae. The water quality parameters, the percentage of survival rate, the growth estimation and the algal cell count were done. Each experiment was carried out in triplicate with a control group of larvae fed with Chaetoceros calcitrans. For the estimation standard procedures were used.to P. monodon

Einfluss der Mensch-Tier-Beziehung auf die Eutergesundheit von Milchkühen

In der vorliegenden Arbeit wurde der Einfluss der Mensch-Tier-Beziehung sowie verschiedener Managementmaßnahmen auf die Eutergesundheit der Milchviehherden von 46 Schweizer Betrieben, die am Bestandsbetreuungsprogramm mit Schwerpunkt Eutergesundheit „pro-Q“ teilnahmen, untersucht. Für die Charakterisierung der Mensch-Tier-Beziehung wurden Melkereinstellungen mittels Fragebögen, Melker- und Kuhverhalten beim Melken und die Ausweichdistanz der Kühe im Laufstall erfasst. Die Darstellung der Eutergesundheit erfolgte anhand der Milchleistungsprüfungsergebnisse der Betriebe über ein Jahr als durchschnittlicher Somatic Cell Score (SCS) und als Neuinfektionsrate. Zudem wurde über zytobakteriologische Analysen von Viertelgemelksproben aller laktierenden Kühe der Herden zeitnah am Untersuchungstag der Mensch-Tier-Beziehungserfassung der Herdenanteil zellzahlerhöhter Viertel (>100.000 Zellen /ml), der Anteil Mastitisviertel (>100.000 Zellen /ml und bakteriologisch positiv), und der Anteil Viertel mit Staphylococcus aureus-Befund errechnet. Hinsichtlich des Managements wurden 35 Faktoren, generiert aus Fragebögen und Stall- und Melkarbeitsbewertungen, in die Auswertungen einbezogen. Nach einer univariablen Vorauswahl von Faktoren wurden multivariate Regressionsmodelle und ein Multilevel-Regressionsmodell zu Analysen der Einflüsse auf die Ausweichdistanz und die fünf Eutergesundheitsvariablen berechnet. Als signifikant mit einer geringeren Ausweichdistanz der Kühe assoziierte Faktoren zeigten sich (1) die positive Einstellung der Melker zur Bedeutung des Kontaktes zum Tier, (2) die Bedingung, dass alle Melker alle Kühe kannten, (3) die Selektion auf Umgänglichkeit als Zuchtstrategie, (4) eine großzügige Dimensionierung der Liegefläche und (5) eine längere allgemeine Kontaktzeit zum Tier innerhalb der Arbeitsroutine. Einflüsse auf die Eutergesundheitsvariablen zeigten verschiedene Kombinationen von Faktoren: Unter den Mensch-Tier-Beziehungsvariablen stellte sich der Anteil positiver Interaktionen des Melkers mit den Kühen beim Melken als häufigster Prädiktor für eine bessere Eutergesundheit (SCS sowie Anteil zellzahlerhöhter und Mastitisviertel) heraus. Zudem zeigte eine geringere Ausweichdistanz signifikante Zusammenhänge mit einem geringeren Anteil zellzahlerhöhter Viertel, aber auch, entgegen der Arbeitshypothese, mit einem höheren Anteil Viertel mit Staphylococcus aureus-Befund. Ausschlagen der Kühe im Melkstand trat signifikant häufiger in Betrieben mit einer höheren Neuinfektionsrate auf. Eine längere Kontaktzeit pro Tier und Tag in der Arbeitsroutine war mit einem geringeren Anteil von Vierteln mit Staphylococcus aureus-Befund verbunden. Darüber hinaus zeigten sich folgende Tier- und Managementfaktoren als signifikante Risikofaktoren für die Eutergesundheit: (1) Rasse, insbesondere Holstein, für SCS, Neuinfektionsrate und Anteil Mastitisviertel, (2) Alter der Kühe anhand der Laktationsnummern für SCS und Anteil Mastitisviertel, (3) Anzahl Neuinfektionen einer Kuh für den SCS, (4) Luftsaugen beim Melkzeug-Anhängen und (5) eine fehlende Separierung kranker Kühe für die Neuinfektionsrate, (6) eine großzügig dimensionierte Liegefläche für den Anteil Mastitisviertel, (7) das aktive Treiben trächtiger Rinder durch den Melkstand zur Gewöhnung und (8) die Verwendung von demselben Euterreinigungsmaterial für mehrere Kühe für den Anteil Viertel mit Staphylococcus aureus-Befund. Die Mensch-Tier-Beziehung ist somit relevant für die Eutergesundheit, insbesondere für die Höhe der Zellzahlen, während das Risiko für Neuinfektionen, die zu Euterentzündungen führen, vornehmlich durch Managementfaktoren beeinflusst wird. Folglich wäre es sinnvoll, den Themenkomplex der Mensch-Tier-Beziehung zukünftig vermehrt in die Beratung von Bestandesbetreuungsprogrammen einzubeziehen.

Técnica in útero vs ex útero para recolección de sangre de cordón umbilical como fuente de células madres

La técnica de recolección es el primer paso para obtener una muestra de sangre de cordón umbilical de óptima calidad. Dicho proceso es esencial en los eventos que conducen al éxito del trasplante de células madre. Aunque no existe consenso internacional sobre el procedimiento de recolección, dos técnicas son las principales: la técnica in utero y la técnica ex utero. La técnica in utero aporta ventajas en cuanto al volumen sanguíneo y conteo celular. El parto por cesárea, si se realiza por razones obstétricas es benéfico para la recolección con técnica in utero. Es necesaria la realización de estudios que permitan mayor nivel de evidencia.

Avaliação da relação entre contagens de células somáticas e os agentes mais prevalentes de mastite em explorações leiteiras da região de entre Douro e Vouga

Quando um microrganismo patogénico invade a glândula mamária, através do canal do teto, pode ocorrer infecção intramamária, desencadeando uma resposta inflamatória – a mastite. Esta possui diversas etiologias, sendo normalmente de origem bacteriana. Na resposta inflamatória, actuam as células somáticas presentes no leite. O presente estudo teve como principal objectivo avaliar a relação entre as contagens de células somáticas e os agentes causadores de mastite em amostras de leite. Foram utilizados dados referentes aos resultados de análises de leite para diagnóstico de mastite realizadas em amostras de explorações de Entre Douro e Vouga. Os resultados consistem em identificação dos agentes e respectivo antibiograma, CCS, sendo ainda registado o objectivo da análise (Secagem ou Tratamento). Determinaram-se as prevalências dos agentes, avaliando de seguida a relação entre as seguintes variáveis: agentes e CCS; CCS e objectivo da análise; e entre agentes e objectivo da análise. Concluiu-se que os Staphylococcus coagulase-negativa são os mais prevalentes, 14,5%, seguindo, Streptococcus uberis, 7,6%; Escherichia coli, 6,3%; Lactococcus garviae, 5,8% e Staphylococcus aureus, 4,1%. Concluiu-se ainda, a existência de diferenças estatisticamente significativas nas análises realizadas, confirmando a relação entre as demais variáveis.

Local roles of TGF-beta superfamily members in the control of ovarian follicle development

Members of the transforming growth factor-beta (TGF-beta) superfamily have wide-ranging influences on many tissue and organ systems including the ovary. Two recently discovered TGF-beta superfamily members, growth/differentiation factor-9 (GDF-9) and bone morphogenetic protein-15 (BMP-15; also designated as GDF-9B) are expressed in an oocyte-specific manner from a very early stage and play a key role in promoting follicle growth beyond the primary stage. Follicle growth to the small antral stage does not require gonadotrophins but appears to be driven by local autocrine/paracrine signals from both somatic cell types (granulosa and theca) and from the oocyte. TGF-beta superfamily members expressed by follicular cells and implicated in this phase of follicle development include TGF-beta, activin, GDF-9/9B and several BMPs. Acquisition of follicle-stimulating hormone (FSH) responsiveness is a pre-requisite for growth beyond the small antral stage and evidence indicates an autocrine role for granulosa-derived activin in promoting granulosa cell proliferation, FSH receptor expression and aromatase activity. Indeed, some of the effects of FSH on granulosa cells may be mediated by endogenous activin. At the same time, activin may act on theca cells to attenuate luteinizing hormone (LH)-dependent androgen production in small to medium-size antral follicles. Dominant follicle selection appears to depend on differential FSH sensitivity amongst a growing cohort of small antral follicles. Activin may contribute to this selection process by sensitizing those follicles with the highest "activin tone" to FSH. Production of inhibin, like oestradiol, increases in selected dominant follicles, in an FSH- and insulin-like growth factor-dependent manner and may exert a paracrine action on theca cells to upregulate LH-induced secretion of androgen, an essential requirement for further oestradiol secretion by the pre-ovulatory follicle. Like activin, BMP-4 and -7 (mostly from theca), and BMP-6 (mostly from oocyte), can enhance oestradiol and inhibin secretion by bovine granulosa cells while suppressing progesterone secretion; this suggests a functional role in delaying follicle luteinization and/or atresia. Follistatin, on the other hand, may favor luteinization and/or atresia by bio-neutralizing intrafollicular activin and BMPs. Activin receptors are expressed by the oocyte and activin may have a further intrafollicular role in the terminal stages of follicle differentiation to promote oocyte maturation and developmental competence. In a reciprocal manner, oocyte-derived GDF-9/9B may act on the surrounding cumulus granulosa cells to attenuate oestradiol output and promote progesterone and hyaluronic acid production, mucification and cumulus expansion.(C) 2003 Elsevier Science B.V. All rights reserved.

Profiling of composition and metabolic activities of the colonic microflora of growing pigs fed diets supplemented with prebiotic oligosaccharides

It is evident that quantitative information on different microbial groups and their contribution in terms of activity in the gastrointestinal (GI) tract of humans and animals is required in order to formulate functional diets targeting improved gut function and host health. In this work, quantitative information on levels and spatial distributions of Bacteroides spp, Eubacterium spp, Clostridium spp, Escherichia coli, Bifidobacterium spp and Lactobacillus/Enterococcus spp. along the porcine large intestine was investigated using 16S rRNA targeted probes and fluorescent in situ hybridisation (FISH). Caecum, ascending colon (AC) and rectum luminal digesta from three groups of individually housed growing pigs fed either a corn-soybean basal diet (CON diet) or a prebiotic diet containing 10 g/kg oligofructose (FOS diet) or trans-galactooligosaccharides (TOS diet) at the expense of cornstarch were analysed. DAPI staining was used to enumerate total number of cells in the samples. Populations of total cells, Bacteroides, Eubacterium, Clostridium and Bifidobacterium, declined significantly (P < 0.05) from caecum to rectum, and were not affected by dietary treatments. Populations of Lactobacillus/ Enterococcus and E coli did not differ throughout the large intestine. The relative percent (%) contribution of each bacterial group to the total cell count did not differ between caecum and rectum, with the exception of Eubacterium that was higher in the AC digesta. FISH analysis showed that the sum of all bacterial groups made up a small percentage of the total cells, which was 12.4%, 21.8% and 10.3% in caecum, AC and rectum, respectively. This supports the view that in swine, the diversity of GI microflora might be higher compared to other species. In terms of microflora metabolic activity, the substantially higher numerical trends seen in FOS and TOS treatments regarding total volatile fatty acid, acetate concentrations and glycolytic activities, it could be postulated that FOS and TOS promoted saccharolytic activities in the porcine colon. (c) 2006 Elsevier Ltd. All rights reserved.

Effects of drying methods and conditions on antimicrobial activity of edible chitosan films enriched with galangal extract

The aim of this work was to study the effects of drying methods and conditions (i.e., ambient drying, hot air drying at 40 degrees C, vacuum drying and low-pressure superheated steam drying within the temperature range of 70-90 degrees C at an absolute pressure of 10 kPa) as well as the concentration of galangal extract on the antimicrobial activity of edible chitosan films against Staphylococcus aureus. Galangal extract was added to the film forming solution as a natural antimicrobial agent in the concentration range of 0.3-0.9 g/100 g. Fourier transform infrared (FTIR) spectra and swelling of the films were also evaluated to investigate interaction between chitosan and the galangal extract. The antimicrobial activity of the films was evaluated by the disc diffusion and viable cell count method, while the morphology of bacteria treated with the antimicrobial films was observed via transmission electron microscopy (TEM). The antimicrobial activity, swelling and functional group interaction of the antimicrobial films were found to be affected by the drying methods and conditions as well as the concentration of the galangal extract. The electron microscopic observations revealed that cell wall and cell membrane of S. aureus treated by the antimicrobial films were significantly damaged. (C) 2009 Elsevier Ltd. All rights reserved.

Emergence of variants with altered survival properties in stationary phase cultures of Campylobacter jejuni

During the stationary phase of Campylobacter jejuni NCTC 11351 viable numbers fluctuate in a characteristic fashion. After reaching the maximum cell count (ca. 2 X 10(9) CFU/ml) in early stationary phase (denoted phase 1), viable numbers subsequently decrease to about 10(6) CFU/ml after 48 h and then increase again to about 10(8) CFU/ml (denoted phase 2) before decreasing once more to a value intermediate between the previous maximum and minimum values. To investigate whether the increase in viable numbers following the initial decline was due to the emergence of a new strain with a growth advantage in stationary phase analogous to the 'GASP' phenotype described in Escherichia coli [Science 259 (1993) 1757], we conducted mixed culture experiments with cells from the original culture and antibiotic-resistant marked organisms isolated from the re-growth phase. In many experiments of this type, strains isolated from phase 2 failed to out-compete the original strain and we have thus been unable to demonstrate a convincing GASP phenotype. However, strains isolated from phase 2 showed a much lower rate of viability loss in early stationary phase and a small increase in resistance to aeration, peroxide challenge and heat, indicating that the emergent strain was different from the parent. These results support the view that dynamic population changes occur during the stationary phase of C jejuni that may play a role in the survival of this organism. (C) 2003 Published by Elsevier B.V.

The use of gaseous ozone and gas packaging to control populations of Salmonella infantis and Pseudomonas aeruginosa on the skin of chicken portions

Chilled breasts of chicken were inoculated with Salmonella infantis or Pseudomonas aeruginosa and then given one of the following treatments: (i) exposure to gaseous ozone (>2000 ppm for up to 30 min); (ii) storage under 70% CO2:30% N-2; and (iii) exposure to gaseous ozone (>2000 ppm for 15 min) followed by storage under 70% CO2:30% N-2; all storage at 7degreesC. Gaseous ozone reduced the counts of samnonellae by 97(Y,, and pseudomonads by 95%, but indigenous coliforms were unaffected. Under the modified atmosphere, the cell count of S. infantis was reduced by 72% following initial exposure and then stabilised, coliforms grew, but Ps. aeruginosa behaved like S. infantis-initial reduction (58%) followed by stability. Exposure to gaseous ozone followed by gas packaging allowed survival of S. infantis, Ps. aeruginosa and coliforms over 9 days at 7degreesC, but there was no evidence of any sensory deterioration. It is proposed that the latter treatment could, in a modified form perhaps, be used to reduce the contamination of chicken carcasses with salmonellae and improve their shelf-life. (C) 2004 Elsevier Ltd. All rights reserved.

Production of Bovine Hand-Made Cloned Embryos by Zygote-Oocyte Cytoplasmic Hemi-complementation

The aim of this study was to evaluate the effect of the cytoplast type and activation process on development of cloned embryos. Bovine oocytes (MII) or zygotes at the one-cell stage (IVF) were manually bisected and segregated in MII or IVF hemi-cytoplasts or hemi-karyoplasts. Adult skin cells from a bovine female were used as nucleus donors (SC). Experimental groups were composed of IVF embryos; parthenogenetic embryos; handmade cloned (HMC) embryos; and reconstructed HMC embryos using IVF hemi-cytoplast + MII hemi-cytoplast + SC (G-I); IVF hemi-cytoplast + IVF hemi-cytoplast + SC (G-II); MII hemi-cytoplast + IVF hemi-karyoplast (G-III); and IVF hemi-cytoplast + IVF hemi-karyoplast (G-IV). Embryos from G-I to G-IV were allocated to subgroups as sperm-activated (SA) or were further chemically activated (SA + CA). Embryos from all groups and subgroups were in vitro cultured in the WOW system. Blastocyst development in subgroup G-I SA (28.2%) was similar to IVF (27.0%) and HMC (31.4%) controls, perhaps due to a to a more suitable activation process and/or better complementation of cytoplasmic reprogramming factors, with the other groups and subgroups having lower levels of development. No blastocyst development was observed when using IVF hemi-karyoplasts (G-III and G-IV), possibly due to the manipulation process during a sensitive biological period. In summary, the presence of cytoplasmic factors from MII hemi-oocytes and the sperm activation process from hemi-zygotes appear to be necessary for adequate in vitro development, as only the zygote-oocyte hemi-complementation was as efficient as controls for the generation of bovine cloned blastocysts.

Delivery of cloned offspring: experience in Zebu cattle (Bos indicus)

The production of a healthy cloned calf is dependent on a multitude of successful steps, including reprogramming mediated by the oocyte, the development of a functional placenta, adequate maternal-fetal interaction, the establishment of a physiological metabolic setting and the formation of a complete set of well-differentiated cells that will eventually result in well-characterised and fully competent tissues and organs. Although the efficiency of nuclear transfer has improved significantly since the first report of a somatic cell nuclear transfer-derived animal, there are many descriptions of anomalies concerning cloned calves leading to high perinatal morbidity and mortality. The present article discusses some our experience regarding perinatal and neonatal procedures for cloned Zebu cattle (B. indicus) that has led to improved survival rates in Nellore cloned calves following the application of such `labour-intensive technology`.

Influence of age on the development of immunological lung response in intrauterine undernourishment

Objective: We investigated the effect of intrauterine undernourishment on some features of asthma using a model of allergic lung inflammation in rats. The effects of age at which the rats were challenged (5 and 9 wk) were also evaluated. Methods: Intrauterine undernourished offspring were obtained from dams that were fed 50% of the nourished diet of counterparts and were immunized at 5 and 9 wk of age. They were tested for immunoglobulin E anti-ova titers (by passive cutaneous anaphylaxis), cell count in the bronchoal-veolar fluid, leukotriene concentration, airway reactivity, mucus production, and blood corticosterone and leptin concentrations 21 d, after immunologic challenge. Results: Intrauterine undernourishment significantly reduced the antigen-specific immunoglobulin E production, inflammatory cell infiltration into airways, mucus secretion, and production of leukotrienes B-4/C-4 in the lungs in both age groups compared with respective nourished rats. The increased reactivity to methacholine that follows antigen challenge was not affected by intrauterine undernourishment. Corticosterone levels increased with age in the undernourished rats` offspring, but not in the nourished rats` offspring. Undernourished offspring already presented high levels of corticosterone before inflammatory stimulus and were not modified by antigen challenge. Leptin levels increased with challenge in the nourished rats but not in the undernourished rats and could not be related to corticosterone levels in the. undernourished rats. Conclusion: Intrauterine undernourishment has a striking and age-dependent effect on the off spring, reducing lung allergic inflammation. (C) 2008 Elsevier Inc. All rights reserved.

Allogeneic apoptotic thymocyte-stimulated dendritic cells expand functional regulatory T cells

P>Dendritic cells (DCs) play an important role in the clearance of apoptotic cells. The removal of apoptotic cells leads to peripheral tolerance, although their role is still not clear. We show that the uptake of apoptotic thymocytes by DCs converts these cells into tolerogenic DCs resistant to maturation by lipopolysaccharide, modulating the production of interleukin-12 and up-regulating the expression of transforming growth factor-beta(1) latency associated peptide. We also observed that DCs pulsed with apoptotic cells in the allogeneic context were more efficient in the expansion of regulatory T cells (Tregs), and that this expansion requires contact between DCs and the T cell. The Tregs sorted from in vitro culture suppressed the proliferation of splenocytes in vitro in a specific and non-specific manner. In the in vivo model, the transfer of CD4+ CD25- cells to Nude mice induced autoimmunity, with cell infiltrate found in the stomach, colon, liver and kidneys. The co-transfer of CD4+ CD25- and CD4+ CD25+ prevented the presence of cell infiltrates in several organs and increased the total cell count in lymph nodes. Our data indicate that apoptotic cells have an important role in peripheral tolerance via induction of tolerogenic DCs and CD4+ CD25+ Foxp3+ cells that present regulatory functions.