Lympho-myeloid mikrokörnyezet = Lympho-myeloid microenvironment

A madarak B lymphocyta éréséért felelős szervének a bursa Fabriciinek epithelialis telepe ectodermalis eredetű. Heparin-beadeket testosteronba vagy növekedési faktorokba mártva lokálisan implantálva az embryoba, befolyásolni tudjuk az egyes szervek növekedését és differenciálódásáért. CD45 positive hemopoietikus sejtek nemcsak a lymphomyeloid szerveket, hanem a teljes embryo medenchymáját kolonizálják és lokálisan MHC class II antigént expresszálnak. A bélidegrendszer ganglionsejtjeinek vándorlása a kapilláris endothel sejtek VEGR receptorainak gátlásával bénítható. Csirke embryok somatopleurája a végtagbimbók között önálló vér-és érképzést mutat. A thymus 74.3 positive dentritikus sejtjei hemopoietikus eredetűek, de mikroszkópos phenotypusuk meghatározása eddig sikertelen volt. A 74.3 mAb alakalmas eszköz a II. tipusú pneumocyták monitorizálására. BoA1 mAb antitestünk faji restrikció nélkül felismeri a csirke, pulyka, gyöngytyúk, fürj és az obese csirkék chB6 alloantigénjét. A korábbi és a jelen OTKA támogatásával a GALT két új tagját fedeztük fel, melyeket, mint oesophagealis és pylorikus tonsillákat írtuk le. A madárlép residens reticulum sejtjei két forrásból; hemopoietikus és mesenchymalis származnak. A hemopoietikus eredetű reticulum sejtek által termelt extracelluláris mátrix súlyosan károsódik IBDV infekciót követően. Két új monoclonális antitestünket is karakterizáltuk, melyek kizárólag gyöngytyúk thrombocytákat ismernek fel. | The epithelial anlage of the bursa of Fabricius develops from the ectoderm. Heparin-beads, soaked in testosteron or growth-factors, and locally implanted into embryo can influence the organ differentiation without interfering the proliferation and differentiation the of the whole embryo. CD45 positive hemopoietic cells colonize the entire mesenchyme of the embryo and after colonization they start to express MHC class II antigen. Formation of intestinal nervous system can be inhibited by preventing the binding of the ligand to the VEGF receptor of capillary endothel. The somatopleura between the anterior and posterior limb buds is capable of own hematopoiesis and vasculogenesis. 74.3 positive thymic dendritic cells are of hemopoietic origin. The 74.3 mAb is a convenient tool for monitoring the condition of the type II. pneumocytes. Our novel mAb, designated BoA1, recognizes chB6 alloantigen in different domestic birds obese chicken. By support of the former and recent OTKA grants we discovered two new members of the gut-associated lymphoid tissue (GALT), which were named as oesophageal and pyloric tonsils. The splenic reticular cells develop from two anlages: hemopoietic and mesenchymal. The hemopoietic reticular cells occur in the antigen trapping zone of the spleen. The extracellular matrix of this region is highly sensitive for infectious bursa disease virus infection. The panel of our mAb(s) revealed two mAb(s), which recognize exclusivelly guinea fowl’s thrombocytes

Circadian ritmusos biológiai folyamatok mechanizmusának vizsgálata madár tobozmirigy modellen. = Mechanisms of circadian rhythmic biological processes - studies on avian pineal gland.

Kimutattuk, hogy a csirke tobozmirigyben a filogenetikailag ismertebb óragének többsége, a Clock, Bmal1, Bmal2, Per1, Per2, Per3, Cry1 és Cry2 expresszálódik. A Cry1, Cry2, Bmal2 és a Clock óragének mRNS-eit sikerült kimutatni 15 napos csirke embriók tobozmirigyeiből is, valamint a kikelt állatokban mért mRNS ritmushoz fázisban hasonló mintázatot mértünk 18 napos csirke embriókban is. Az óragének expressziós mintázatát mind az éjszaka alkalmazott megvilágítás, mind az in vitro alkalmazott PACAP kezelés megváltoztatta. Kimutattuk, hogy a MT és cAMP koncentrációjának napszakos ritmusa a 16-18. embrionális napokon jelenik meg in vitro körülmények között is. A ritmikus szekréció kialakulása azonban periodikus ingerek nélkül késik. Igazoltuk, hogy bár a tobozmirigy fény-receptorai már a 14. embrionális napon is működőképesek, a fényingernek még nincs direkt, in vitro a corpus pinealéra gyakorolt szinkronizáló hatása. Ugyanakkor a 18. embrionális naptól kezdve a csirke tobozmirigy már a rendkívül alacsony intenzitású (10 lux) megvilágítással is vezérelhető. Eredményeink szerint mind a PACAP, mind a VIP már a 13. embrionális napon fokozza a cAMP és a melatonin termelést, de a ritmus fejlődését, illetve annak fázisát nem változtatja meg. Kimutattuk, hogy a tobozmirigy in vitro MT szekrécióját a környezeti hőmérséklet periodikusan alkalmazott, mindössze három órás megváltoztatása is módosítja, a hőmérséklet változására adott válasz azonban az állat életkorának függvénye. | We have demonstrated that most of the known, philogenetically conserved clock genes (Clock, Bmal1, Bmal2, Per1, Per2, Per3, Cry1 and Cry2) are expressed in the chicken pineal gland. Expression of Cry1, Cry2, Bmal2 and Clock mRNA-s were shown also in the pineal glands of chicken embryos. Similar 24-h pattern of clock gene expression were detected in 18 day old chicken embryos and in post hatched chickens (6 weeks old). The expression patterns of clock genes were altered by light exposure at subjective night and also by in vitro PACAP administration. It was shown, that the circadian rhythm of both MT secretion and cAMP efflux appears on the 16-18th embryonic days, even in vitro. Without periodic environmental stimuli, the development of the MT rhythm was delayed. Although the light receptors of the pineal gland were found to be responsive to light from the 14th embryonic day, the illumination did not have a synchronizing effect on the in vitro MT secretion at this age. However, from the 18th day, the pineal MT secretion could be controlled by illumination with even the very low intensity (10 lux). Our results showed that both PACAP and VIP induced an increase in both cAMP and MT secretion already from the 13th day of embryonic life, but none of these drugs influenced the development or the phase of the MT rhythm. It was presented that the in vitro MT secretion could be modified age-dependently by only 3 hour-long daily alterations of environmental temperature

In and Out of the Bursa-The Role of CXCR4 in Chicken B Cell Development

In contrast to mammals, early B cell differentiation and diversification of the antibody repertoire in chickens do not take place in the bone marrow but in a specialized gut associated lymphoid tissue (GALT), the bursa of Fabricius. During embryonic development, B cell precursors migrate to the bursa anlage, where they proliferate and diversify their B cell receptor repertoire. Around hatch these diversified B cells start to emigrate from the bursa of Fabricius and populate peripheral lymphoid organs, but very little is known how the migratory processes are regulated. As CXCL12 (syn. SDF-1) and CXCR4 were shown to be essential for the control of B cell migration during the development of lymphoid tissues in mammals, we analyzed expression and function of this chemokine/chemokine-receptor pair in the chicken bursa. We found a strong variation of mRNA abundance of CXCL12 and CXCR4 in different stages of bursa development, with high abundance of CXCL12 mRNA in the bursa anlage at embryonic day 10 (ED10).In situhybridization demonstrated disseminated CXCL12 expression in the early bursa anlage, which condensed in the developing follicles and was mainly restricted to the follicle cortex post-hatch. Flow cytometric analysis detected CXCR4 protein already on early B cell stages, increasing during bursal development. Post-hatch, a subpopulation with the hallmarks of emigrating B cells became detectable, which had lower CXCR4 expression, suggesting that downregulation of CXCR4 is necessary to leave the CXCL12-high bursal environment.In vivoblockade of CXCR4 using AMD3100 at the time of B cell precursor immigration strongly inhibited follicle development, demonstrating that CXCL12 attracts pre-bursal B cells into the bursal anlage. Altogether, we show that CXCL12 and its receptor CXCR4 are important for both populating the bursa with B cells and emigration of mature B cells into the periphery post hatch, and that CXCR4 function in primary B cell organs is conserved between mammals and birds

The enteric neural crest progressively loses capacity to form enteric nervous system

Cells of the vagal neural crest (NC) form most of the enteric nervous system (ENS) by a colonising wave in the embryonic gut, with high cell proliferation and differentiation. Enteric neuropathies have an ENS deficit and cell replacement has been suggested as therapy. This would be performed post-natally, which raises the question of whether the ENS cell population retains its initial ENS-forming potential with age. We tested this on the avian model in organ culture in vitro (3 days) using recipient aneural chick midgut/hindgut combined with ENS- donor quail midgut or hindgut of ages QE5 to QE10. ENS cells from young donor tissues (<= QE6) avidly colonised the aneural recipient, but this capacity dropped rapidly 2-3 days after the transit of the ENS cell wavefront. This loss in capability was autonomous to the ENS population since a similar decline was observed in ENS cells isolated by HNK1 FACS. Using QE5, 6, 8 and 10 midgut donors and extending the time of assay to 8 days in chorio-allantoic membrane grafts did not produce 'catch up' colonisation. NC-derived cells were counted in dissociated quail embryo gut and in transverse sections of chick embryo gut using NC, neuron and glial marker antibodies. This showed that the decline in ENS-forming ability correlated with a decrease in proportion of ENS cells lacking both neuronal and glial differentiation markers, but there were still large numbers of such cells even at stages with low colonisation ability. Moreover, ENS cells in small numbers from young donors were far superior in colonisation ability to larger numbers of apparently undifferentiated cells from older donors. This suggests that the decline of ENS-forming ability has both quantitative and qualitative aspects. In this case, ENS cells for cell therapies should aim to replicate the embryonic ENS stage rather than using post-natal ENS stem/progenitor cells

Lympho-myeloid és hemopoietikus szervek fejlődésbiológiája: embryomanipuláció = Development of the lympho-myeloid and hemopoietic organs: embryo manipulation

A pályázat fő célja volt a bursai szekréciós dendritikus (BSDC), a follikuláris dendritikus (FDC) és a Langerhans sejtek (LC) eredetének és differenciálódásának vizsgálata. BSDC véreredetű sejtek terminális differenciálódásuk függ az anyai eredetű antigénektől. A A fertőző bursa betegséget okozó vírus először a BSDC-ben mutatható ki, bizonyítva, hogy a BSDC az elsőrendű célsejtje a vírusoknak. 5-fluorouracil okozta heterophil depléció eliminálja a vírusfertőzés okozta mortalitást és a súlyos klinikai tüneteket és emeli a BSDC-k számát. A heterophil granulocyták a BSDC-vel együtt jelentős szerepet játszanak a fertőző bursa betegség (Gumboro) klinikai tüneteinek kialakulásában. Az LC haptén kezelés hatására kivándorolnak az epidermisből és subcután előforduló lymphoid nodulusokban jelennek meg. A lép ellipszoidhoz-asszociált sejtei (EAC) hemopoietikus eredetűek. Az EAC az egyetlen olyan phagocyta tulajdonságú sejt, amely internalizálja a β-galactozidázt ez pedig elindítja az EAC migrációját a T dependens területek (PALS) és a csíracentrumok felé. Monoklonális antitest (E5G12) felismeri az EAC-t és az FDC-t is, mely lehetőséget adott kettős festésre (β-gal hisztokémia és E5G12 immuncytokémia). A kettős festés egyértelmű bizonyítékot szolgáltatott arra, hogy a lép csíracentrumok FDC-jei EAC eredetűek. A csirke embryok vérében perifériás vér fibrocyták jelen vannak, melyek hozzájárulnak a lép organogenesiséhez. Ez a munka vezetett az a lép ellipszoidok körüli Schweigger-Seidel hüvely tokjának felfedezéséhez. A csirkék gastrointestinalis traktusában oesophagealis és pylorus tonsillak kerültek leírásra. | The origin and differentiation of bursal secretory dendritic (BSDC), follicular dendritic (FDC), and Langerhans cells (LC) have been studied. BSDC are blood-borne hemopoietic cells, and their differentiation depends on maternal antigen. The antibody against infectious bursal disease viruses indicates that the BSDC are primary targets of the IBDV. Depletion of heterophil granulocytes by 5-fluorouracil results in elimination of major clinical symptoms and mortality, increase the number of BSDC. After hapten treatment the Langerhans cells migrate out of the epidermis and appear in the subcutaneous lymph nodules. The splenic ellipsoid-associated cells (EAC) are blood-borne cells. They exclusively internalize the β-galactosidase, which initiate their migration to the T dependent region (PALS) and germinal centers. Monoclonal antibody E5G12, recognizes EAC and splenic FDC. Double-staining (β-galactosidase histochemistry) and E5G12 immunocytochemistry provided clearcut evidence that the EAC were precursors of FDC. No double labeled cells were found in the GC of the GALT, which might indicate that the origin of splenic and GALT FDC have had different origin. The presence of the avian peripheral blood fibrocytes in the circulation contribute to the splenic extracellular matrix, which forms arround the ellipsoid a discontinuous sheath, what we called "capsule of Schweigger-Seidel sheath" (CSS). We have identified oesophageal and duodenal tonsils, which are specific for the birds

Exosomal small RNA profiling in first-trimester maternal blood explores early molecular pathways of preterm preeclampsia

IntroductionPreeclampsia (PE) is a severe obstetrical syndrome characterized by new-onset hypertension and proteinuria and it is often associated with fetal intrauterine growth restriction (IUGR). PE leads to long-term health complications, so early diagnosis would be crucial for timely prevention. There are multiple etiologies and subtypes of PE, and this heterogeneity has hindered accurate identification in the presymptomatic phase. Recent investigations have pointed to the potential role of small regulatory RNAs in PE, and these species, which travel in extracellular vesicles (EVs) in the circulation, have raised the possibility of non-invasive diagnostics. The aim of this study was to investigate the behavior of exosomal regulatory small RNAs in the most severe subtype of PE with IUGR.MethodsWe isolated exosomal EVs from first-trimester peripheral blood plasma samples of women who later developed preterm PE with IUGR (n=6) and gestational age-matched healthy controls (n=14). The small RNA content of EVs and their differential expression were determined by next-generation sequencing and further validated by quantitative real-time PCR. We also applied the rigorous exceRpt bioinformatics pipeline for small RNA identification, followed by target verification and Gene Ontology analysis.ResultsOverall, &gt;2700 small RNAs were identified in all samples and, of interest, the majority belonged to the RNA interference (RNAi) pathways. Among the RNAi species, 16 differentially expressed microRNAs were up-regulated in PE, whereas up-regulated and down-regulated members were equally found among the six identified Piwi-associated RNAs. Gene ontology analysis of the predicted small RNA targets showed enrichment of genes in pathways related to immune processes involved in decidualization, placentation and embryonic development, indicating that dysregulation of the induced small RNAs is connected to the impairment of immune pathways in preeclampsia development. Finally, the subsequent validation experiments revealed that the hsa_piR_016658 piRNA is a promising biomarker candidate for preterm PE associated with IUGR.DiscussionOur rigorously designed study in a homogeneous group of patients unraveled small RNAs in circulating maternal exosomes that act on physiological pathways dysregulated in preterm PE with IUGR. Therefore, our small RNA hits are not only suitable biomarker candidates, but the revealed biological pathways may further inform us about the complex pathology of this severe PE subtype

Műszaki műanyag forgácsolhatóságának vizsgálata

A műszaki műanyagok felhasználása az utóbbi években, évtizedekben fokozottan növekszik. Ezeknek az anyagoknak a befejező megmunkálása történhet forgácsolással. A dolgozat fő témája egy adott műszaki műanyag, a polioximetilén forgácsolhatóságának vizsgálata esztergálás körülményei között. Az anyag forgácsolhatóságának vizsgálata kísérletterv segítségével történt. A vizsgálatok szárazon és hűtő-kenő közeggel is el lettek végezve. Kimenő paraméterként az iparban is gyakran használt érdességi paraméterek voltak mérve. A dolgozat összehasonlítja, majd minősíti a száraz és hűtőkenő folyadékkal történő forgácsolást ennél az anyagnál