Sérhæfingarmynstur þekjuvefsfrumna úr berkju í frumurækt

The respiratory system epithelium, including the bronchi and lungs, consists of several different types of epithelial cells. The upper respiratory tract has a pseudostratified epithelium and the epithelium grows thinner in the more distal respiratory tract ending in a simple epithelium in the alveoli where the gas exchange occurs. The epithelium of the upper respiratory tract is composed of various types of epithelial cells such as ciliated, goblet and basal cells. The basal cells are thought to be stem cells in the airways and the more differentiated cells of the epithelium arise from them. The basal cells serve as a reserve cell population that divides asymmetrically in response to injury. Research on the attributes of epithelial cells in vitro is important as the results can give information on how the cells of the bronchi and lungs develop. In these studies it is beneficial to use both primary cells and established cell lines. Primary cells are cultured from donor tissue and are thought to represent the cell of origin better than established cell lines that have been genetically altered. Established cell lines have been grown for a long time in artificial environments, both in cell culture media and in two dimensional culture. Primary cells have been a part of a living being and reflect in part the state of the body they are taken from. However, they have a limited lifespan, as they have not been immortalized like the established cell lines and reproducing results can be difficult because of variations between donors. In this study, primary epithelial cells were cultured from 19 bronchial tissue samples. The marker expression showed considerable donor variation and indicated a change in marker expression by time and passage. There seems to be a tendency for enrichment of p63 positive cells after first passage and mucin expression that identifies goblet cells does not seem to be present in passages after the initial one. The VA10 cell line is a bronchial epithelial cell line that has a marker expression corresponding to basal cells. The VA10 cells are p63 positive, a basal cell marker, generating branching bronchioalveolar-like structures and pseudostratified epithelium in 3D culture and air-liquid culture, respectively. The VA10 cells are a heterogeneous group with a broad surface marker expression and a part of the project was to test if the cell line could be enriched for more basal cell properties by using a cell sorting technique based on antibodies against surface proteins expressed on basal cells. Subgroups of cells that had a high expression of basal cell markers NGFR and β4 integrin respectively were isolated. They showed a reduction in growth and a marker expression profile similar to that of the parental VA10 cells. The VA10 cells were immortalized with the E6 and E7 genes, which degrade and bind p53 and retinoblastoma protein respectively. The original VA10 cell line forms branching structures with small cavities but their lumen formation is flawed. The E6 and E7 genes were knocked down with a shRNA construct in an effort to make VA10 cells that would have an improved lumen formation in branching structures in Matrigel. The knockdown of E6 and E7 was successful but when the shRNA was activated, the branching formation in Matrigel was severely reduced. This study describes the characterization of freshly isolated primary cells from the human bronchus. The results underline the importance of using primary cells from multiple donors because of donor variance. Additionally, the cells are quite dynamic when it comes to surface marker expression, potentially changing as time passes. It is important to try and further our understanding of human primary cells to make them more useful as research tools. At the same time it would be beneficial to understand the strengths and limitations of both primary cells and cells lines since the best experiments would undoubtedly be conducted by combining the strengths of both for the most accurate conclusions

Yfirtjáning Oct4 og POU5F1P1 í frumum af blöðruhálskirtilsuppruna

Krabbamein í blöðruhálskirtli er sjúkdómur sem hrjáir marga karlmenn en það er algengasta krabbameinið sem greint var á Íslandi á árunum 2005-2009. Blöðruhálskirtilskrabbamein greinist seint en meðalaldur við greiningu á Íslandi er 71 ár. Þessi hái aldur við greiningu veldur því að sumir einstaklingar kjósa þiggja ekki meðferð. Sterkustu áhættuþættirnir fyrir blöðruhálskirtilskrabbameini eru aldur, kynþáttur, fjölskyldusaga og erfðir en margir einkirnisbreytileikar hafa fundist sem tengjast auknum líkum á að fá meinið. Markmið þessa verkefnis var að yfirtjá tvö gen með veiruferju í frumulínunni PZHPV7 sem er af blöðruhálskirtilsuppruna til að sjá hvort yfirtjáningin myndi hafa áhrif á svipgerð frumnanna eða lífeðlisfræðilega eiginleika. Genin sem voru valin eru Oct4 og POU5F1P1, það fyrrnefnda er umritunarþáttur sem stjórnar tjáningu gena í frumum á stofnfrumustigi og hefur verið tjáður í kjarna krabbameinsfrumna. Hið síðarnefnda er gervigen af Oct4 sem er að jafnaði ekki tjáð en rannsóknir hafa sýnt fram á að það er einnig tjáð í kjarna krabbameinsfrumna úr ýmsum líffærum. Ástæðan fyrir vali þessara próteina eru tengsl þeirra við krabbamein. Ástæðan fyrir vali blöðruhálskirtilskrabbameins er sú að POU5F1P1 er staðsett á svæði í erfðamenginu þar sem margir erfðabreytileikar tengdir krabbameini, bæði í blöðruhálskirtli og annars staðar, liggja. Margar rannsóknir tengdar Oct4 hafa verið gerðar. Samspil meginumritunarþáttanna sem viðhalda fjölhæfu ástandi í fósturfrumum, Oct4, Sox2 og Nanog, hefur verið skoðað í grunninn, Oct4 er tengt krabbameinsmyndandi frumum sem viðhalda krabbameinsæxlum og það er einn af Yamanaka þáttunum sem voru notaðir til að búa til fyrstu framkölluðu stofnfrumurnar (e. induced pluripotent stem cells, iPSC). POU5F1P1 hefur minna verið rannsakað en tengsl þess við Oct4 sem gervigen þess, tjáning þess í krabbameinum og staðsetning gervigensins í nálægð við erfðafjölbreytileikana sem tengdir hafa verið við krabbamein gera það að mjög áhugaverðu rannsóknarefni. Í þessari ritgerð verða kynntar fyrstu niðurstöður úr þessu verkefni en til stendur að halda vinnunni við það áfram. Frumur voru ræktaðar upp eftir smitun og litaðar með mótefnum. GFP ljómun var skoðuð en grænljómandi frumur eru til marks um að smitið hafi tekist. Mótefnalitunin bendir til þess að yfirtjáningin hafi tekist þó enn eigi eftir að staðfesta það. GFP ljómandi þyrpingar má finna í öllum frumulínum, þó almenn ljómun sé minnst í POU5F1P1 línunni. Frumur voru ræktaðar upp í matrigeli og fyrstu niðurstöður um vaxtarhæfni þeirra og svipgerð benda til þess genin hafi einhver áhrif. Það virðist ljóst að veirusmitið sjálft hefur áhrif á frumurnar en vera má að þau áhrif minnki með tímanum. Ræktunartíma frumnanna í matrigeli er þó ekki lokið og þessar niðurstöður eru eingöngu vísbendingar um hegðun frumnanna

Effective treatment with balneophototherapy and narrowband UVB monotherapy reduces skin homing Th17/Tc17 and Th22/Tc22 effector cells in peripheral blood in patients with psoriasis.

To access publisher's full text version of this article click on the hyperlink belowLandspitali University Hospital Research Fund Icelandic Technology Development Fund Blue Lagoon Clini

Detailed Multiplex Analysis of SARS-CoV-2 Specific Antibodies in COVID-19 Disease

This work was supported in part by The Student Innovation Fund. The funding body had no role in the design of the study, collection, analysis, interpretation of data or writing of the manuscript. Funding Information: We would like to thank Runólfur Pálsson and Elías Eyþórsson for their assistance with the patient demographics. The Department of Clinical Microbiology for their assistance and last but not least the patients. Publisher Copyright: © Copyright © 2021 Brynjolfsson, Sigurgrimsdottir, Einarsdottir, Bjornsdottir, Armannsdottir, Baldvinsdottir, Bjarnason, Gudlaugsson, Gudmundsson, Sigurdardottir, Love, Kristinsson and Ludviksson.A detailed understanding of the antibody response against SARS-CoV-2 is of high importance, especially with the emergence of novel vaccines. A multiplex-based assay, analyzing IgG, IgM, and IgA antibodies against the receptor binding domain (RBD), spike 1 (S1), and nucleocapsid proteins of the SARS-CoV-2 virus was set up. The multiplex-based analysis was calibrated against the Elecsys® Anti-SARS-CoV-2 assay on a Roche Cobas® instrument, using positive and negative samples. The calibration of the multiplex based assay yielded a sensitivity of 100% and a specificity of 97.7%. SARS-CoV-2 specific antibody levels were analyzed by multiplex in 251 samples from 221 patients. A significant increase in all antibody types (IgM, IgG, and IgA) against RBD was observed between the first and the third weeks of disease. Additionally, the S1 IgG antibody response increased significantly between weeks 1, 2, and 3 of disease. Class switching appeared to occur earlier for IgA than for IgG. Patients requiring hospital admission and intensive care had higher levels of SARS-CoV-2 specific IgA levels than outpatients. These findings describe the initial antibody response during the first weeks of disease and demonstrate the importance of analyzing different antibody isotypes against multiple antigens and include IgA when examining the immunological response to COVID-19.Peer reviewe