Myo1f has an essential role in γδT intraepithelial lymphocyte adhesion and migration

γδT intraepithelial lymphocyte represents up to 60% of the small intestine intraepithelial compartment. They are highly migrating cells and constantly interact with the epithelial cell layer and lamina propria cells. This migratory phenotype is related to the homeostasis of the small intestine, the control of bacterial and parasitic infections, and the epithelial shedding induced by LPS. Here, we demonstrate that Myo1f participates in the adhesion and migration of intraepithelial lymphocytes. Using long-tailed class I myosins KO mice, we identified the requirement of Myo1f for their migration to the small intestine intraepithelial compartment. The absence of Myo1f affects intraepithelial lymphocytes’ homing due to reduced CCR9 and α4β7 surface expression. In vitro, we confirm that adhesion to integrin ligands and CCL25-dependent and independent migration of intraepithelial lymphocytes are Myo1f-dependent. Mechanistically, Myo1f deficiency prevents correct chemokine receptor and integrin polarization, leading to reduced tyrosine phosphorylation which could impact in signal transduction. Overall, we demonstrate that Myo1f has an essential role in the adhesion and migration in γδT intraepithelial lymphocytes

Lrba participates in the differentiation of IgA+ B lymphocytes through TGFβR signaling

Introduction Lrba is a cytoplasmic protein involved in vesicular trafficking. Lrba -deficient ( Lrba-/- ) mice exhibit substantially higher levels of IgA in both serum and feces than wild-type (WT) mice. Transforming growth factor β1 (TGFβ1) and its receptors (TGFβR I and II) is essential for differentiating IgA+ B cells. Furthermore, increased IgA production suggests a potential connection between Lrba and the TGFβR signaling pathway in IgA production. However, the specific function of Lrba in B cell biology remains unknown. Aim Given the increased IgA levels in Lrba -/- mice, the goal in this work was to explore the lymph organs where the switch to IgA occurs, and if TGFβR function is affected. Methods Non-immunized Lrba-/- mice were compared with Lrba+/+ mice. IgA levels in the serum and feces, as well as during peripheral B cell development, were determined. IgA+ B cells and plasma cells were assessed in the small intestine and secondary lymphoid organs, such as the spleen, mesenteric lymph nodes, and Peyer’s patches. The TGFβR signaling pathway was evaluated by determining the expression of TGFβR on B cells. Additionally, SMAD2 phosphorylation was measured under basal conditions and in response to recombinant TGFβ. Finally, confocal microscopy was performed to investigate a possible interaction between Lrba and TGFβR in B cells. Results Lrba-/- mice exhibited significantly higher levels of circulating IgA, IgA+ B, and plasma cells than in peripheral lymphoid organs those in WT mice. TGFβR expression on the membrane of B cells was similar in both Lrba-/- and Lrba+/+ mice. However, intracellular TGFβR expression was reduced in Lrba-/- mice. SMAD2 phosphorylation showed increased levels under basal conditions; stimulation with recombinant TGFβ elicited a poorer response than in that in Lrba+/+ B cells. Finally, we found that Lrba colocalizes with TGFβR in B cells. Conclusion Lrba is essential in controlling TGFβR signaling, subsequently regulating SMAD2 phosphorylation on B cells. This mechanism may explain the increased differentiation of IgA+ B cells and production of IgA-producing plasma cells

Selective Determination of Lysine in Dry-Cured Meats Using a Sensor Based on Lysine-α-Oxidase Immobilised on a Nylon Membrane

An enzymatic sensor employing lysine oxidase (LOx) with the immobilised enzyme system by crosslinking with glutaraldehyde using an immunodyne ABC nylon membrane, in combination with an oxygen electrode, has been optimised to determine the lysine content in dry-cured ham and dry-fermented sausage at different cured times. The amperometric signal obtained due to the oxygen depletion (consumed oxygen) during the lysine oxidation was recorded at 5 s in the immobilised enzyme sensor, and the reaction rates (slope) were related to the lysine content. A linear relationship between the consumed oxygen as a function of time (mg O2/l/s) and the lysine concentration in the range 10–250 μM (R2 = 0.9946) for the immobilised enzyme system was found. The immobilised enzyme sensor showed a high specificity and sensibility. Nevertheless, the stability of the immobilised enzyme at the assay temperature was very poor, and thus, a new membrane was required for each analysis. The analysis of lysine with the immobilised enzyme system in cured meat samples revealed very good agreement with the determination performed through standard HPLC methodology, which validated the use of this sensor as an alternative technique to evaluate cured meat quality.This study was funded by Grant Prometeo 2012/001 from Conselleria d’Educació of Generalitat Valenciana (Valencia, Spain) and the scholarship to F. A. Jadán Piedra from Secretaria de Educación Superior, Ciencia, Tecnología e Innovación (SENESCYT) of Ecuador.Peer reviewe