La citometria amb enfocament acústic permet el diagnòstic de patologies relacionades amb els glòbuls vermells

L'estructura habitual dels eritròcits és un disc bicòncau. Moltes patologies són generades per una distorsió en aquesta estructura per diferents causes com un defecte en l'arquitectura de la membrana. La citometria amb enfocament acústic permet l'avaluació de partícules no esfèriques. El grup de Citòmica Funcional a l'Institut de Recerca de Leucèmia Josep Carreras junt amb altres grups ha estudiat aquest avantatge i ha demostrat la seva utilitat per avaluar les diferents causes de les distorsions o la qualitat dels eritròcits després de la donació de sang de manera immediata.La estructura habitual de los eritrocitos es un disco cóncavo. Sin embargo, muchas patologías son generadas por una distorsión en esta estructura por diferentes causas como por ejemplo un defecto en la arquitectura de la membrana. La citometría con enfoque acústico permite la evaluación de partículas no esféricas. Por esta razón, el grupo de Citómica Funcional a l'Institut de Recerca de Leucèmia Josep Carreras en colaboración con otros grupos ha estudiado esta ventaja de la citometría acústica y ha demostrado su utilidad para evaluar las diferentes causas que pueden provocar distorsiones a los glóbulos rojos o la calidad de los eritrocitos después de la donación de sangre de manera inmediata.The normal shape for a red blood cell is a bi-concave disc. However, a lot of pathologies are generated by a distortion in this shape due to different causes like a defects in the cell membrane skeletal architecture. Acoustic cytometry allows the study of the orientation of non-spherical particles. That is why the Citómica Funcional group in l'Institut de Recerca de Leucèmia Josep Carreras in collaboration with other groups has studied this advantage and has proven its utility to evaluate the different causes of the red blood cells distortions or to evaluate immediately the quality of erythrocytes following blood donation

Més a prop de la quantificació de la resposta immunitària específica front al càncer i les malalties infeccioses

En els tractaments contra el càncer o de les malalties infeccioses és molt important poder determinar l'activitat citotòxica cel·lular, és a dir, la capacitat defensiva que té el sistema immunitari enfront d'una cèl·lula alterada. No obstant això, la manipulació de mostres biològiques, a més de tedioses, comprometen el correcte funcionament de les cèl·lules en el moment de ser estudiades. En aquest article Jordi Petriz presenta un nou mètode que permet estudiar mostres recentment extretes minimitzant l'impacte del seu maneig, a més de reduir el temps a només dues hores.En los tratamientos contra el cáncer o de las enfermedades infecciosas es muy importante poder determinar la actividad citotóxica celular, es decir, la capacidad defensiva que tiene el sistema inmunitario frente a una célula alterada. Sin embargo, la manipulación de muestras biológicas, además de tediosas, comprometen el correcto funcionamiento de las células en el momento de ser estudiadas. En este artículo Jordi Petriz presenta un nuevo método que permite estudiar muestras recién extraídas minimizando el impacto de su manejo, además de reducir el tiempo a sólo dos horas

Flow cytometric significance of cellular alkaline phosphatase activity in acute myeloid leukemia

In this prospective hospital-based cohort study that included 43 newly diagnosed patients with acute myeloid leukemia, flow cytometric cellular alkaline phosphatase (ALP) activity within primitive leukemic cells allowed us to identify two groups of patients at diagnosis according to the numbers of leukemic blasts expressing ≥ 12% of ALP+ cells (27 patients, Group A) and less than 12% of ALP+ cells (16 patients, Group B). Differences in outcome for complete response, relapse or treatment resistance, and exitus were statistically analyzed and were significant, when comparing the two groups. The overall survival (OS) and event-free survival (EFS) differences between Group A and B were statistically significant. The survival of Group A patients was significantly shorter than those for Group B. No significant relationship was detected in outcome when comparing ELN prognostic-risk group based on cytogenetic and molecular profile (patients in the favorable, intermediate, and adverse risk groups). Flow cytometric cellular ALP activity at diagnosis may be used to estimate relapses and disease persistence more accurately. The limitations of our study include the small number of patients enrolled and a short follow-up, due to its prospective nature

Flow-cytometry-based protocols for human blood/marrow immunophenotyping with minimal sample perturbation

This protocol provides instructions to improve flow cytometry analysis of marrow/peripheral blood cells by avoiding erythrolytic solutions, density gradients, and washing steps. We describe two basic approaches for identifying cell surface antigens with minimal sample perturbation, which have been successfully used to identify healthy and pathologically rare cells. The greatest advantage of these approaches is that they minimize the unwanted effect caused by sample preparation, allowing for improved study of live cells at the point of analysis. For complete details on the use and execution of this protocol, please refer to Petriz et al. (2018)

Unmasking the expression of PD-L1 in Myeloid Derived Suppressor Cells : a case study in lung cancer to discover new drugs with specific on-target efficacy

Altres ajuts: We thank CERCA Programme/Generalitat de Catalunya and The Josep Carreras Foundation for institutional support. The authors are also very grateful for their advice and technical support to Sergio Ramón, Víctor Querol, Clara Streiff, Paola Paglia, and Lluís Sainz from Thermo Fisher for all his comments and discussions on earlier work in this research field. Jordi Petriz also acknowledges the financial support from The Obra Social La Caixa and Thermo Fisher Scientific

Engraftment Potential of Adipose Tissue-Derived Human Mesenchymal Stem Cells After Transplantation in the Fetal Rabbit

Due to their favorable intrinsic features, including engraftment, differentiation, and immunomodulatory potential, adult mesenchymal stem cells (MSCs) have been proposed for therapeutic in utero intervention. Further improvement of such attributes for particular diseases might merely be achieved by ex vivo MSC genetic engineering previous to transplantation. Here, we evaluated for the first time the feasibility, biodistribution, long-term engraftment, and transgenic enhanced green fluorescent protein (EGFP) expression of genetically engineered human adipose tissue-derived MSCs (EGFP+-ASCs) after intra-amniotic xenotransplantation at E17 of gestation into our validated pregnant rabbit model. Overall, the procedure was safe (86.4% survival rate; absence of anatomical defects). Stable, low-level engraftment of EGFP+-ASCs was confirmed by assessing the presence of the pWT-EGFP lentiviral provirus in the young transplanted rabbit tissues. Accordingly, similar frequencies of provirus-positive animals were found at both 8 weeks (60%) and 16 weeks (66.7%) after in utero intervention. The presence of EGFP+-ASCs was more frequent in respiratory epithelia (lung and trachea), according to the route of administration. However, we were unable to detect EGFP expression, neither by real-time polymerase chain reaction nor by immunohistochemistry, in the provirus-positive tissues, suggesting EGFP transgene silencing mediated by epigenetic events. Moreover, we noticed lack of both host cellular immune responses against xenogeneic ASCs and humoral immune responses against transgenic EGFP. Therefore, the fetal microchimerism achieved by the EGFP+-ASCs in the young rabbit hosts indicates induction of donor-specific tolerance after fetal rabbit xenotransplantation, which should boost postnatal transplantation for the early treatment/prevention of many devastating congenital disorders

Phagocytic Activity Is Impaired in Type 2 Diabetes Mellitus and Increases after Metabolic Improvement

OBJECTIVE: 1) To evaluate whether peripheral blood mononuclear cells (PBMCs) from type 2 diabetic patients present an impairment of phagocytic activity; 2) To determine whether the eventual impairment in phagocytic activity is related to glycemic control and can be reversed by improving blood glucose levels. METHODS: 21 type 2 diabetic patients and 21 healthy volunteers were prospectively recruited for a case-control study. In addition, those patients in whom HbA1c was higher than 8% (n = 12) were hospitalized in order to complete a 5-day intensification treatment of blood glucose. Phagocytic activity was assessed by using a modified flow cytometry procedure developed in our laboratory based on DNA/RNA viable staining to discriminate erythrocytes and debris. This method is simple, highly sensitive and reproducible and it takes advantage of classic methods that are widely used in flow cytometry. RESULTS: Type 2 diabetic patients showed a lower percentage of activated macrophages in comparison with non-diabetic subjects (54.00±18.93 vs 68.53±12.77%; p = 0.006) Significant negative correlations between phagocytic activity and fasting glucose (r = -0.619, p = 0.004) and HbA1c (r = -0.506, p = 0.019) were detected. In addition, multiple linear regression analyses showed that either fasting plasma glucose or HbA1c were independently associated with phagocytic activity. Furthermore, in the subset of patients who underwent metabolic optimization a significant increase in phagocytic activity was observed (p = 0.029). CONCLUSIONS: Glycemic control is related to phagocytic activity in type 2 diabetes. Our results suggest that improvement in phagocytic activity can be added to the beneficial effects of metabolic optimization

The acute gravitational stress of parabolic flight affects red blood cell aggregation and functionality of circulating immune cells

Postprint (published version

Effects of rapid gravity load changes on immunophenotyping and leukocyte function of human peripheral blood after parabolic flight

One of the biological systems that suffers a physiological de-conditioning in space is the immune system. It is in charge of defending the body against pathogens and other aggressions. The aim of this work is to assess if there are any relevant changes in the aggregation of erythrocytes, cell count, immunophenotyping and functionality after parabolic flight. This effect has been assessed ex vivo using human peripheral blood, which was drawn from the radial vein (n=6 healthy volunteers) and anticoagulated with heparin and EDTA. Blood samples were split into two aliquots and maintained in two identical thermally isolated boxes; one stayed on the ground whereas the other one was subjected to parabolic flight. The parabolic flight consisted of 15 parabolas performed with a Mudry CAP-10B acrobatic aircraft. Each parabola consists of 8 seconds of hypogravity preceded and followed by 2 seconds of hypergravity. Any of the biological parameters measured showed no statistically significant differences. Altered gravity could increase aggregation of red blood cells, as demonstrated by a decrease in the number of single cells after parabolic flight exposure. No counting changes in haemoglobin concentration were observed when comparing the two different groups. Furthermore, potential functional alterations of monocytes and neutrophils cannot be rejected. Although these possible changes could be associated with hypogravity, other factors such as hypergravity and acceleration or deceleration cannot be ruled out. Our findings indicate that, under this specific experimental setup, there was no significant alteration in leukocyte immunophenotyping and functional capacity when using ex vivo blood samples and short exposure to altered gravity.Peer ReviewedObjectius de Desenvolupament Sostenible::3 - Salut i BenestarPostprint (published version