HoloMonitor i Nature / Scientific Reports

Bloggen nås av information att ungerska forskare,mestadels från PHI´s partner Semmelweis University,har fått en forskningsrapport publicerad i Natures Scientific Reports.

Rapporten offentligjordes igår och handlar om Trophoblastceller, hur och varför tumörer uppkommer i samband med graviditet.Forskarna har kommit ett steg närmare problematiken sen den förra rapporten som kom för 1 år sen.

Published online:03 April 2018

The impact of circulating preeclampsia-associated extracellular vesicles on the migratory activity and phenotype of THP-1 monocytic cells

 Abstract
Intercellular communication via extracellular vesicles (EVs) and their target cells, especially immune cells, results in functional and phenotype changes that consequently may play a significant role in various physiological states and the pathogenesis of immune-mediated disorders. 
Monocytes are the most prominent environment-sensing immune cells in circulation, skilled to shape their microenvironments via cytokine secretion and further differentiation. Both the circulating monocyte subset distribution and the blood plasma EV pattern are characteristic for preeclampsia, a pregnancy induced immune-mediated hypertensive disorder. We hypothesized that preeclampsia-associated EVs (PE-EVs) induced functional and phenotypic alterations of monocytes. 
First, we proved EV binding and uptake by THP-1 cells. Cellular origin and protein cargo of circulating PE-EVs were characterized by flow cytometry and mass spectrometry. 
An altered phagocytosis-associated molecular pattern was found on 12.5 K fraction of PE-EVs: an elevated CD47 “don’t eat me” signal (p < 0.01) and decreased exofacial phosphatidylserine “eat-me” signal (p < 0.001) were found along with decreased uptake of these PE-EVs (p < 0.05). The 12.5 K fraction of PE-EVs induced significantly lower chemotaxis (p < 0.01) and cell motility but accelerated cell adhesion of THP-1 cells (p < 0.05). The 12.5 K fraction of PE-EVs induced altered monocyte functions suggest that circulating EVs may have a role in the pathogenesis of preeclampsia.

Introduction
Extracellular vesicles (EVs) are nanosized particles enclosed by a phospholipid bilayer membrane. EVs been shown to mediate intercellular communication. EVs are significant players of autocrine, paracrine as well as endocrine signalling. EVs are produced by living cells and can be detected in all biological fluids tested so far.
Trophoblast-derived EVs originate from the feto-maternal interface and are present in the circulating EV pool. They can be detected by several methods, including flow cytometry (FC), confocal microscopy, mass spectrometry and transmission electron microscopy.

https://en.wikipedia.org/wiki/Trophoblast

 Methods

The HoloMonitor M4 (Phase Holographic Imaging, Lund, Sweden) was used as an incubator-proof time-lapse and cell tracking system for adherent cells. In the present experiments the chemokinetic effects were evaluated by holographic tracking, measurement and calculation of the following parameters: (1) migration: the shortest direct distance from the starting point to the end point (μm); (2) motility: the actual way travelled from the starting point to the end point (μm). THP-1 cells (4 × 10⁴) were seeded on a Petri dish (35 mm diameter) and cultured for 24 h. After the incubation resting THP-1 cells were monitored for cell migration and motility for 2 hours (control THP-1 cells). Next, 12.5 K fraction of HP-EVs or PE-EVs were added to the resting cells and migration and motility activities of the cells were documented (HP-EVs or PE-EVs treated cells). Cell migration was normalized to the untreated resting THP-1 cells (Suppl. Fig. 11). In time-lapse tracking the settings were: total time = 2 h; interval: 30 sec. For evaluation of images of time-lapse automatic background thresholding method was used with “Minimum error sets” algorithm (adjustment = 128) using the minimum error histogram-based threshold method; in object definition, the minimum error object size was 36. The number of evaluated cells/image: 50; the total number of evaluated images: 240. For manual cell identification, objects of the marginal zone were abandoned. For holographic microscopy HoloStudio M4 2.5 program was used to analyse data.

Min kommentar
En oerhört viktig forskning som är betydelsefull i flera aspekter.
Vid en graviditet med ovanstående förlopp riskerar både modern som fostret att avlida.
Kan forskarna nå insikter om hur sjukdomsförloppet uppstår är man närmare att kunna bota.
HoloMonitor visar än en gång hur förtjänstfullt den kan användas i forskarnas händer.