Heat, irradiation-stressed, non-adherent cells induce changes in phenotype and function of human dendritic cells during maturation process in vitro (2006)
- Authors:
- Autor USP: BARBUTO, JOSE ALEXANDRE MARZAGAO - ICB
- Unidade: ICB
- Assunto: IMUNOLOGIA
- Language: Inglês
- Abstract: Introduction & Objectives: Dendritic cells (DCs) are the most potent APCs and have the unique ability of presenting Ags to naïve lymphocytes and to induce primary immune responses. Data from murine cells show that peripheral immune tolerance is also induced by processing and presentation of self-tissues that die during physiologic tissue turnover (Annu Rev Immunol, 21;685-711, 2003). Therefore, we aimed to study the tolerogenic potential of human DCs and the apoptotic cells' effect on DCs maturation and functional activity. Methods & Results: PBMC were obtained by apheresis from healthy donors and plated to adhere in 12-well plates for 2h (1x107/well). Non-adherent cells (N-Adh) were transferred to culture flask until day 3. Plateadherent PBMC (Adh) were cultured with GM-CSF and IL-4 for 7 days (iDCs) and TNF-á in the last 2 days (mDCs). In day 3 N-Adh were: 1. kept at 37oC (37ni); 2. heated for 30 min at 43oC (H); 3. 137Cs-irradiated ( APO ); 4. H and 137Cs-irradiated (HAPO).Apoptosis kynetics was followed incubating cells of each group with propidium iodide (PI) 0, 24 and 48h after irradiating N-Adh to assess DNA fragmentation by FACS. APO had a mean apoptosis rate of 80.6% ± 11.4 (p<0.01) 48h after irradiation, while HAPO rate (23.9% ± 8) was similar to control cells rate (37ni=21.5% ± 10.8, n=6). These results suggest that heating N-Adh at 43oC for 30 min before irradiation may protect them from irradiation-inducedapoptosis. Then, we added 10% (3-6x105) of APO or HAPO to DCs (mean yield = 4.5x106 ± 1.4 cells). In day 7, DCs were phenotyped by FACS and assayed in 6-day MLR at a 1:30 rate (DC:lymphocytes). Allostimulation was measured by CFSE-dilution of labeled lymphocytes, by FACS. There was an association between the percentage of mDC+HAPO expressing CD86/CD80 and alloestimulation induced, with the highest stimulation indexes (>5) in 3 of the 6 experiments done. Conclusions: The presence of HAPO cells during human DC differentiation seems to induce their activation, while APO cells did not affect DC phenotype. However, these are preliminary data and more experiments need to be done before any conclusions are reached.
- Imprenta:
- Source:
- Título do periódico: Abstracts
- Conference titles: Meeting of the Brazilian Society for Immunology
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ABNT
CARMO, J P M et al. Heat, irradiation-stressed, non-adherent cells induce changes in phenotype and function of human dendritic cells during maturation process in vitro. 2006, Anais.. São Paulo: Instituto de Ciências Biomédicas, Universidade de São Paulo, 2006. . Acesso em: 25 abr. 2024. -
APA
Carmo, J. P. M., Neves, A. R., Tomiyoshi, M. Y., González, R. P., Bergami-Santos, P. C., & Barbuto, J. A. M. (2006). Heat, irradiation-stressed, non-adherent cells induce changes in phenotype and function of human dendritic cells during maturation process in vitro. In Abstracts. São Paulo: Instituto de Ciências Biomédicas, Universidade de São Paulo. -
NLM
Carmo JPM, Neves AR, Tomiyoshi MY, González RP, Bergami-Santos PC, Barbuto JAM. Heat, irradiation-stressed, non-adherent cells induce changes in phenotype and function of human dendritic cells during maturation process in vitro. Abstracts. 2006 ;[citado 2024 abr. 25 ] -
Vancouver
Carmo JPM, Neves AR, Tomiyoshi MY, González RP, Bergami-Santos PC, Barbuto JAM. Heat, irradiation-stressed, non-adherent cells induce changes in phenotype and function of human dendritic cells during maturation process in vitro. Abstracts. 2006 ;[citado 2024 abr. 25 ] - Imunidade celular
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