Transplantation tolerance induced by regulatory T cells: in vivo mechanisms and sites of action.

The mechanisms by which CD4(+)CD25(+)Foxp3(+) T (Treg) cells regulate effector T cells in a transplantation setting and their in vivo homeostasis still remain to be clarified. Using a mouse adoptive transfer model, we analyzed the in vivo expansion, trafficking, and effector function of alloreactive T cells and donor-specific Treg cells, in response to a full-thickness skin allograft. Fluorescent-labeled CD4(+)CD25(-) and antigen-specific Treg cells were transferred alone or co-injected into syngeneic BALB/c-Nude recipients transplanted with skins from (C57BL/6 x BALB/c) F1 donors. Treg cells divided in vivo, migrated and accumulated in the allograft draining lymph nodes as well as within the graft. The co-transfer of Treg cells did not modify the early activation and homing of CD4(+)CD25(-) T cells in secondary lymphoid organs. However, in the presence of Treg cells, alloreactive CD4(+)CD25(-) T cells produced significantly less IFN-gamma and were present in reduced numbers in the secondary lymphoid organs. Furthermore, time-course studies showed that Treg cells were recruited into the allograft at a very early stage after transplantation and effectively prevented the infiltration of effector T cells. In conclusion, suppression of rejection requires the early recruitment to the site of antigenic challenge of donor-specific Treg cells, which then mainly regulate the effector arm of T cell alloresponses

In vivo mechanisms leading to transplantation tolerance induced by regulatory T cells

Purpose: The mechanisms by which CD4+CD25+Foxp3+ T cells (Tregs) regulate effector T cells in a transplantation setting and their in vivo homeostasis still remain to be clarified. Using a mouse adoptive transfer and skin transplantation model, we analyzed the in vivo expansion, effector function and trafficking of effector T cells and donor-specific Tregs, in response to an allograft. Methods and materials: Antigen-specific Tregs were generated and expanded in vitro by culturing freshly isolated Tregs from BALB/c mice (H2d) with syngeneic dendritic cells pulsed with an allopeptide (here the Kb peptide derived from the MHC class I molecule of allogeneic H2b mice). Fluorescent-labelled CD4+CD25- naive T cells and donor-antigen-specific Tregs were transferred alone or coinjected into syngeneic BALB/c-Nude recipients transplanted with allogeneic C57BL/6xBALB/c donor skin. Results: As opposed to their in vitro hyporesponsiveness, Tregs divided in vivo, migrated and accumulated in the allograft draining lymph nodes (drLN) and within the graft. The co-transfer of Tregs did not modify the early proliferation and homing of CD4+CD25- T cells to secondary lymphoid organs. But, in the presence of Tregs, effector T cells produced significantly less IFN- and IL-2 effector cytokines, while higher amounts of IL-10 were detected in the spleen and drLN of these mice. Furthermore, time-course studies showed that Tregs were recruited into the allograft at a very early stage posttransplantation and prevented infiltration by effector T cells. Conclusion: Overall, our results suggest that suppression of graft rejection involves the early recruitment of donor-specific Tregs at the sites of antigenic challenge and that Tregs mainly regulate the effector arm of T cell alloresponses

Experimental and theoretical modelling of blind-ended vessels within a developing angiogenic plexus

Angiogenic sprouts at the leading edge of an expanding vascular plexus are recognised as major regulators of the structure of the developing network. Early in sprout development, a vascular lumen is often evident which communicates with the parent vessel while the distal tip is blind-ended. Here we describe the temporal evolution of blind-ended vessels (BEVs) in a small wound made in the panniculus carnosus muscle of a mouse viewed in a dorsal skin-fold window-chamber model with intra-vital microscopy during the most active period of angiogenesis (days 5–8 after injury). Although these structures have been mentioned anecdotally in previous studies, we observed BEVs to be frequent, albeit transient, features of plexus formation. Plasma leakage into the surrounding extracellular matrix occurring from these immature conduits could play an important role in preparing hypoxic tissue for vascular invasion. Although sprout growth is likely to be regulated by its flow environment, the parameters regulating flow into and through BEVs have not been characterised in situ. Longitudinal data from individual animals show that the number of BEVs filled with plasma alone peaks at day 7, when they can exceed 150 ?m in length. Additionally, BEVs greater than 40 ?m in length are more likely to be filled with stationary erythrocytes than with plasma alone. Using a mathematical model, we show how the flux of 150kD fluorinated (FITC-) dextran through an individual plasma-filled BEV is related to its geometry being determined primarily by its surface area; by fitting theoretical intensity values to experimental data we assess the permeability of the vessel to FITC-dextran. Plasma skimming provides a mechanistic explanation for the observation that BEVs with larger surface area are more likely to recruit erythrocytes

Phosphorus availability in a low pH highly weathered soil as affected by added salts Disponibilidade de fósforo num solo ácido afetada pela aplicação de sais

Concentration and identity of cations and anions in the soil solution may affect soil P reactions and thus P availability. The magnitude of these reactions was evaluated in this research after application of various salts to a highly weathered low pH soil. Chloride, nitrate, and sulfate salts of Na, NH4, K, Ca, Mg, Sr, or Cu were added to the soil after addition of 360mg P/kg trying to simulate ion concentrations around granules of fertilizers in the soil. Thirty days later, P was determined in the soil solution (Pli) and on the solid phase (Psi). The soil samples of some treatments were leached with water and three days later a new soil solution was displaced. Separate addition of all salts increased Pli, except NaCl at the lowest rate. The increase of Pli was highly associatcd with amount of native cations displaced to the soil solution by the applied salts. Salt solubility, concentration, and sometimes identity of cation and anion also influenced Pli. Some salts decreased Psi, but this was not correlated with any soil property measured. The effects caused by salts on Pli and Psi disappeared after leaching the soil samples.<br>A concentração eletrolítica e o tipo de cations e anions da solução do solo podem afetar as reações do fósforo com possíveis reflexos na disponibilidade de P aos vegetais. Nessa pesquisa quantificou-se o efeito de vários sais nos valores das determinações analíticas que afetam a disponibilidade de fósforo. Sais de nitrato, cloreto e sulfato foram aplicados a amostras de um alfisol ácido após a aplicação de 360mg P/kg, simulando concentrações que ocorrem no solo ao redor de grânulos de fertilizantes. Fósforo lábil (Psi) e P na solução do solo (Pli) foram determinados após 30 dias de incubação, antes e depois de percolar água pelo solo. Todos os sais aumentaram a concentração de P na solução do solo, exceto a menor dose de NaCl. O aumento do Pli foi correlacionado com a quantidade de cations originalmente no solo deslocados para a solução do solo. Solubilidade, concentração, e a espécie dos cations e anions aplicados também exerceram efeito no Pli. Alguns sais diminuíram o Psi porém esse decréscimo não se correlacionou com nenhuma determinação efetuada. A percolação de água eliminou os efeitos ocasionados pelos sais nos valores de fósforo