TY  - JOUR
AU  - Nikolovski, Neda
AU  - Paunović, Verica
AU  - Stojanović, Ivana D.
PY  - 2019
UR  - https://www.sciencedirect.com/science/article/pii/S002217591930047X?via%3Dihub
UR  - https://radar.ibiss.bg.ac.rs/handle/123456789/3347
AB  - Polyclonal T regulatory cells (Treg - CD4+CD25+CD127lowFoxp3+) are used in several protocols for the treatment of type 1 diabetes (T1D), multiple sclerosis and graft-versus host disease in clinical trials. However, general opinion is that autoantigen-specific Treg could be more efficient in autoimmunity suppression due to their direct effect on pathogenic autoantigen-specific effector T cells. This study describes isolation and expansion of insulin-specific Treg in vitro. Insulin-specific Treg are uniformly distributed in lymphoid tissues however their number is extremely low. To enrich the proportion of insulin-specific Treg, pure CD4+ cells were co-cultured with insulin B chain peptide-loaded dendritic cells, isolated from mice that develop T1D spontaneously - NOD mice. Insulin-specific CD4+ cell expansion peaked after 48 h of incubation and was in favour of Treg. These cells were then sorted using insulin peptide-loaded MHC class II tetramers and cultured in vitro for 48 h in the presence of TCR stimulators, TGF-β and IL-2. The proportion of gained insulin-specific cells with T regulatory phenotype (CD4+CD25highCD127lowGITR+FoxP3+) was in average between 93% and 97%. These cells have shown potent in vitro suppressive effect on T effector cells, produced IL-10 and TGF-β and expressed PD-1 and CD39. Further proliferation of these insulin-specific Treg required the presence of dendritic cells, anti-CD3 antibody and IL-2. This study provides new, reproducible experimental design for the enrichment and expansion of insulin-specific Treg that can be used for the cell-based therapy of autoimmunity.
T2  - Journal of Immunological Methods
T1  - Isolation and enrichment of mouse insulin-specific CD4+ T regulatory cells.
VL  - 470
DO  - 10.1016/j.jim.2019.04.011
SP  - 46
EP  - 54
ER  - 

@article{
author = "Nikolovski, Neda and Paunović, Verica and Stojanović, Ivana D.",
year = "2019",
abstract = "Polyclonal T regulatory cells (Treg - CD4+CD25+CD127lowFoxp3+) are used in several protocols for the treatment of type 1 diabetes (T1D), multiple sclerosis and graft-versus host disease in clinical trials. However, general opinion is that autoantigen-specific Treg could be more efficient in autoimmunity suppression due to their direct effect on pathogenic autoantigen-specific effector T cells. This study describes isolation and expansion of insulin-specific Treg in vitro. Insulin-specific Treg are uniformly distributed in lymphoid tissues however their number is extremely low. To enrich the proportion of insulin-specific Treg, pure CD4+ cells were co-cultured with insulin B chain peptide-loaded dendritic cells, isolated from mice that develop T1D spontaneously - NOD mice. Insulin-specific CD4+ cell expansion peaked after 48 h of incubation and was in favour of Treg. These cells were then sorted using insulin peptide-loaded MHC class II tetramers and cultured in vitro for 48 h in the presence of TCR stimulators, TGF-β and IL-2. The proportion of gained insulin-specific cells with T regulatory phenotype (CD4+CD25highCD127lowGITR+FoxP3+) was in average between 93% and 97%. These cells have shown potent in vitro suppressive effect on T effector cells, produced IL-10 and TGF-β and expressed PD-1 and CD39. Further proliferation of these insulin-specific Treg required the presence of dendritic cells, anti-CD3 antibody and IL-2. This study provides new, reproducible experimental design for the enrichment and expansion of insulin-specific Treg that can be used for the cell-based therapy of autoimmunity.",
journal = "Journal of Immunological Methods",
title = "Isolation and enrichment of mouse insulin-specific CD4+ T regulatory cells.",
volume = "470",
doi = "10.1016/j.jim.2019.04.011",
pages = "46-54"
}

Nikolovski, N., Paunović, V.,& Stojanović, I. D.. (2019). Isolation and enrichment of mouse insulin-specific CD4+ T regulatory cells.. in Journal of Immunological Methods, 470, 46-54.
https://doi.org/10.1016/j.jim.2019.04.011

Nikolovski N, Paunović V, Stojanović ID. Isolation and enrichment of mouse insulin-specific CD4+ T regulatory cells.. in Journal of Immunological Methods. 2019;470:46-54.
doi:10.1016/j.jim.2019.04.011 .

Nikolovski, Neda, Paunović, Verica, Stojanović, Ivana D., "Isolation and enrichment of mouse insulin-specific CD4+ T regulatory cells." in Journal of Immunological Methods, 470 (2019):46-54,
https://doi.org/10.1016/j.jim.2019.04.011 . .