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Evaluation of THP-1 cells as a source of in vitro differentiated Langerhans cells
Malmö University, Faculty of Health and Society (HS), Department of Biomedical Science (BMV).
2022 (English)Independent thesis Basic level (degree of Bachelor), 10 credits / 15 HE creditsStudent thesis
Abstract [en]

Langerhans cells are antigen-presenting immune cells in the epidermis, of myeloid origin. They control inflammatory responses and maintain homeostasis in the skin through a constant exchange of signals with keratinocytes, melanocytes, and local regulatory T cells. Like dendritic cells they can migrate to lymph nodes and prime immature T-cells but are in fact tissue-resident macrophages. To study skin immunology and the function of Langerhans cells there is a need for readily available cells expressing key proteins of Langerhans cells, such as the endosomal recycling complex known as Birbeck granules, and the antigen-presenting molecule CD1a. The immortalized monocytic leukemia cell line THP-1 was subjected to three experiments of differentiation using Phorbol 12-myristate-13-acetate and the cytokines transforming growth factor β, granulocyte/macrophage colony-stimulating factor, interleukin 4 and tumor necrosis factor alpha. The resulting protein expression were measured using seven CD markers expressed by macrophages, Langerhans cells and dendritic cells. The migratory abilities of the differentiated cells were evaluated using Boyden chamber assay and the chemo-attractants CCL19 and CCL21 that serves as ligands for CCR7, expressed on cells migrating to lymph nodes for antigen presentation. The THP-1 cells were partly able to differentiate into Langerhans cells and dendritic cells, and showed an ability to migrate toward the chemokine gradient. The differentiated cells did not express Langerhans unique proteins to the desired extent.

Abstract [sv]

Langerhanska celler är antigenpresenterande immunceller i epidermis, med myeloid ursprung. De styr inflammatoriska svar och upprätthåller homeostas i huden genom ett konstant utbyte av signaler med keratinocyter, melanocyter och lokala regulatoriska T-celler. Liksom dendritiska celler kan de migrera till lymfkörtlar och aktivera omogna T-celler, men är i själva verket vävnadsmakrofager. För att studera immunologi och Langerhanska cellers funktion i hud behövs lättillgängliga celler som uttrycker proteiner centrala för Langerhanska celler, såsom det endosomala återvinningskomplexet Birbecks granula och den antigenpresenterande molekylen CD1a. Den imortaliserade monocytiska leukemicellinjen THP-1 genomgick tre differentieringsexperiment där Phorbol 12-myristat-13-acetat och cytokinerna transformerande tillväxtfaktor β, granulocyt/makrofagkolonistimulerande faktor, interleukin 4 och tumörnekrosfaktor alfa användes. Det resulterande proteinuttrycket mättes med hjälp av av sju CD-markörer som uttrycks av makrofager, Langerhanska celler och dendritiska celler. Migrationsförmågan hos de differentierade cellerna utvärderades med ’Boyden chamber’-analys och två kemokiner; CCL19 och CCL21 som är ligander för CCR7, vanligen uttryckt av celler som migrerar till lymfkörtlar för antigenpresentation. THP-1-cellerna kunde delvis differentiera till Langerhanska celler och dendritiska celler, och uppvisade förmåga att kunna migrera mot kemokingradienten. De differentierade cellerna uttryckte däremot inte Langerhanska cellers unika proteiner i önskad utsträckning.

Place, publisher, year, edition, pages
2022. , p. 25
Keywords [en]
CCR7, CD markers, dendritic cells, differentiation, Langerhans cells, migration assay, THP-
Keywords [sv]
CCR7, CD-markörer, dendritiska celler, differentiering, Langerhanska celler, migrationsanalys, THP-1
National Category
Cell Biology
Identifiers
URN: urn:nbn:se:mau:diva-52724OAI: oai:DiVA.org:mau-52724DiVA, id: diva2:1668816
Educational program
HS Biomedicinsk laboratorievetenskap
Presentation
2022-05-30, 15:15 (Swedish)
Supervisors
Examiners
Available from: 2022-06-22 Created: 2022-06-13 Last updated: 2022-06-22Bibliographically approved

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