INFLUENCE OF BCG (BACILLUS CALMETTE-GUÉRIN) ON THE MATURATION OF HUMAN DENDRITIC CELL (DCS) DIFFERENTIATED FROM MONOCYTES
Background and aims: DCs are obtained in vitro through monocyte differentiation, previous studies show the maturation of DCs by live BCG, but for models targeting immunosuppressed patients, the maturation of DCs by dead BCG needs to be investigated. In this work we evaluated the phenotypic and funct...
| Main Authors: | , , , , , , |
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| Format: | Article |
| Language: | English |
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Elsevier
2023-10-01
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| Series: | Hematology, Transfusion and Cell Therapy |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S2531137923019417 |
| _version_ | 1797654685879894016 |
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| author | TS Lima BF Gonçalves ASC Cypriano C Nogueira RT Pinho HDS Dutra A Maiolino |
| author_facet | TS Lima BF Gonçalves ASC Cypriano C Nogueira RT Pinho HDS Dutra A Maiolino |
| author_sort | TS Lima |
| collection | DOAJ |
| description | Background and aims: DCs are obtained in vitro through monocyte differentiation, previous studies show the maturation of DCs by live BCG, but for models targeting immunosuppressed patients, the maturation of DCs by dead BCG needs to be investigated. In this work we evaluated the phenotypic and functional characteristics of human DCs in vitro stimulated by dead BCG. Methods: DCs were obtained from the isolation of monocytes from the blood of healthy donors. After the isolation step, monocytes were cultured with IL-4 and GM-CSF for differentiation into immature DCs. For terminal maturation, TNF-α and IFN-α (DC-TNF) or killed BCG (DC-BCG) were added. After maturation, DC-TNF and DC-BCG were co-cultured for 5 days with autologous and allogeneic lymphocytes previously stained with CFSE fluorochrome. To phenotypically characterize DCs, after their terminal maturation, some markers were investigated. The strain used was the lyophilized Moreau-RJ. Results: DC-TNF and DC-BCG cultures showed rates of cells expressing HLA-DR: 98% and 98%; CD1A: 69% and 72.37%, CD86: 97% and 98%,CD80: 39% and 36%, CD40: 21% and 15%, PD-1: 18% and 14% and CD83: 55% and 47%, respectively. There was a significant difference in the expression of percentage values only for CD80. MFI (mean fluorescence intensity) levels in DC-TNF and DC-BCG were high in HLA-DR: 9251 and 6867, in CD86: 12975 and 10772, and in CD1A: 6242 and 5694, respectively. DC-TNF and DC-BCG had MFI for PD-1: 755 and 1156; CD40: 877 and 745, in CD80: 753 and 598, and in CD83: 624 and 1529, respectively. There was a significant difference in HLA-DR MFI levels. In lymphocyte proliferation DC-BCG showed greater proliferation of autologous lymphocytes: 11% and 21% allogeneic, while DC-TNF cells: 5% autologous and 16% allogeneic. Discussion and conclusion: The use of dead BCG promotes the maturation of DCs at levels equivalent to TNF. But BCG potentiates lymphocyte proliferation at a higher rate. To interpret the results obtained, a study of cytokine production will be performed. |
| first_indexed | 2024-03-11T17:03:06Z |
| format | Article |
| id | doaj.art-35dc576f52304b46ad682e11be4cb694 |
| institution | Directory Open Access Journal |
| issn | 2531-1379 |
| language | English |
| last_indexed | 2024-03-11T17:03:06Z |
| publishDate | 2023-10-01 |
| publisher | Elsevier |
| record_format | Article |
| series | Hematology, Transfusion and Cell Therapy |
| spelling | doaj.art-35dc576f52304b46ad682e11be4cb6942023-10-20T06:48:04ZengElsevierHematology, Transfusion and Cell Therapy2531-13792023-10-0145S983S984INFLUENCE OF BCG (BACILLUS CALMETTE-GUÉRIN) ON THE MATURATION OF HUMAN DENDRITIC CELL (DCS) DIFFERENTIATED FROM MONOCYTESTS Lima0BF Gonçalves1ASC Cypriano2C Nogueira3RT Pinho4HDS Dutra5A Maiolino6Universidade Federal do Rio de Janeiro (UFRJ), Rio de Janeiro, RJ, BrazilUniversidade Federal do Rio de Janeiro (UFRJ), Rio de Janeiro, RJ, BrazilUniversidade Federal do Rio de Janeiro (UFRJ), Rio de Janeiro, RJ, BrazilUniversidade Federal do Rio de Janeiro (UFRJ), Rio de Janeiro, RJ, BrazilFundação Oswaldo Cruz (Fiocruz), Rio de Janeiro, RJ, BrazilUniversidade Federal do Rio de Janeiro (UFRJ), Rio de Janeiro, RJ, BrazilUniversidade Federal do Rio de Janeiro (UFRJ), Rio de Janeiro, RJ, BrazilBackground and aims: DCs are obtained in vitro through monocyte differentiation, previous studies show the maturation of DCs by live BCG, but for models targeting immunosuppressed patients, the maturation of DCs by dead BCG needs to be investigated. In this work we evaluated the phenotypic and functional characteristics of human DCs in vitro stimulated by dead BCG. Methods: DCs were obtained from the isolation of monocytes from the blood of healthy donors. After the isolation step, monocytes were cultured with IL-4 and GM-CSF for differentiation into immature DCs. For terminal maturation, TNF-α and IFN-α (DC-TNF) or killed BCG (DC-BCG) were added. After maturation, DC-TNF and DC-BCG were co-cultured for 5 days with autologous and allogeneic lymphocytes previously stained with CFSE fluorochrome. To phenotypically characterize DCs, after their terminal maturation, some markers were investigated. The strain used was the lyophilized Moreau-RJ. Results: DC-TNF and DC-BCG cultures showed rates of cells expressing HLA-DR: 98% and 98%; CD1A: 69% and 72.37%, CD86: 97% and 98%,CD80: 39% and 36%, CD40: 21% and 15%, PD-1: 18% and 14% and CD83: 55% and 47%, respectively. There was a significant difference in the expression of percentage values only for CD80. MFI (mean fluorescence intensity) levels in DC-TNF and DC-BCG were high in HLA-DR: 9251 and 6867, in CD86: 12975 and 10772, and in CD1A: 6242 and 5694, respectively. DC-TNF and DC-BCG had MFI for PD-1: 755 and 1156; CD40: 877 and 745, in CD80: 753 and 598, and in CD83: 624 and 1529, respectively. There was a significant difference in HLA-DR MFI levels. In lymphocyte proliferation DC-BCG showed greater proliferation of autologous lymphocytes: 11% and 21% allogeneic, while DC-TNF cells: 5% autologous and 16% allogeneic. Discussion and conclusion: The use of dead BCG promotes the maturation of DCs at levels equivalent to TNF. But BCG potentiates lymphocyte proliferation at a higher rate. To interpret the results obtained, a study of cytokine production will be performed.http://www.sciencedirect.com/science/article/pii/S2531137923019417 |
| spellingShingle | TS Lima BF Gonçalves ASC Cypriano C Nogueira RT Pinho HDS Dutra A Maiolino INFLUENCE OF BCG (BACILLUS CALMETTE-GUÉRIN) ON THE MATURATION OF HUMAN DENDRITIC CELL (DCS) DIFFERENTIATED FROM MONOCYTES Hematology, Transfusion and Cell Therapy |
| title | INFLUENCE OF BCG (BACILLUS CALMETTE-GUÉRIN) ON THE MATURATION OF HUMAN DENDRITIC CELL (DCS) DIFFERENTIATED FROM MONOCYTES |
| title_full | INFLUENCE OF BCG (BACILLUS CALMETTE-GUÉRIN) ON THE MATURATION OF HUMAN DENDRITIC CELL (DCS) DIFFERENTIATED FROM MONOCYTES |
| title_fullStr | INFLUENCE OF BCG (BACILLUS CALMETTE-GUÉRIN) ON THE MATURATION OF HUMAN DENDRITIC CELL (DCS) DIFFERENTIATED FROM MONOCYTES |
| title_full_unstemmed | INFLUENCE OF BCG (BACILLUS CALMETTE-GUÉRIN) ON THE MATURATION OF HUMAN DENDRITIC CELL (DCS) DIFFERENTIATED FROM MONOCYTES |
| title_short | INFLUENCE OF BCG (BACILLUS CALMETTE-GUÉRIN) ON THE MATURATION OF HUMAN DENDRITIC CELL (DCS) DIFFERENTIATED FROM MONOCYTES |
| title_sort | influence of bcg bacillus calmette guerin on the maturation of human dendritic cell dcs differentiated from monocytes |
| url | http://www.sciencedirect.com/science/article/pii/S2531137923019417 |
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