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THE IMMUNOREGULATORY FUNCTIONS OF EXOSOME DERIVED FROM MESENCHYMAL STEM CELLS (MSCS) THAT GENETICALLY MODIFIED BY ADENO-ASSOCIATED VIRUS (AAV)
Author(s): ,
Li Liu
Affiliations:
Hematopoietic Stem Cell Transplantation Center,National Clinical Research Center for Blood Diseases, State Key Laboratory of Experimental Hematology, Institute of Hematology & Blood Diseases Hospital, Chinese Academy of Medical Sciences & Peking Union Medical College,Tianjin,China
,
Sizhou Feng
Affiliations:
Hematopoietic Stem Cell Transplantation Center,National Clinical Research Center for Blood Diseases, State Key Laboratory of Experimental Hematology, Institute of Hematology & Blood Diseases Hospital, Chinese Academy of Medical Sciences & Peking Union Medical College,Tianjin,China
Xiaolei Pei
Affiliations:
Hematopoietic Stem Cell Transplantation Center,National Clinical Research Center for Blood Diseases, State Key Laboratory of Experimental Hematology, Institute of Hematology & Blood Diseases Hospital, Chinese Academy of Medical Sciences & Peking Union Medical College,Tianjin,China
EHA Library. Liu L. 06/09/21; 325488; EP728
Li Liu
Li Liu
Contributions
Abstract
Presentation during EHA2021: All e-poster presentations will be made available as of Friday, June 11, 2021 (09:00 CEST) and will be accessible for on-demand viewing until August 15, 2021 on the Virtual Congress platform.

Abstract: EP728

Type: E-Poster Presentation

Session title: Gene therapy, cellular immunotherapy and vaccination - Biology & Translational Research

Background
There exist kinds of limitations of MSCs during its application, while exosome derived from MSCs has shown promising future.

Aims

To verify the effects and mechanisms of natural MSCs-exosome in treating acute GVHD in mice, then explore and establish a method for targeted modification of MSCs-exosome and verify the functions of the modified MSCs-exosome.

Methods

In different doses of MSCs-exosome groups and MSCs group, the weight loss of acute GVHD mice was observed, then the proliferation levels of activated T cells were measured through T cell activation experiment in vitro and OVA antigen specific T cell activation experiment in vivo. AAV2YF3 mutants carrying PD-L1 and PD-L1-ITGB1 was obtained after the construction of recombinant expression vectors and then was applied to infect human MSCs to modify their exosome. The immunoregulatory functions of the modified MSCs-exosome were measured with methods as above.

Results

1) Mouse MSCs-exosome (300ug*3 times) and MSCs (1e6*3 times) effectively alleviated the weight loss of acute GVHD mice. 2) Compared with IL2, 10ug, 25ug and 50ug human MSCs-exosome inhibited the proliferation of activated T cells in vitro, respectively 86.01% (IL2), 40%, 39.61% and 42.84%; compared with PBS, 50ug, 100ug and 200ug mouse MSCs-exosome inhibited the proliferation of antigen-specifically activated OT-1 cells in vivo, respectively 42.6%, 33.1%, 14.2% and 10.6%. 3) After the infection of AAV2YF3 mutant carrying PD-L1 or PD-L1-ITGB1, the positive proportion of MSCs-exosome exceeds 40% and 60%, respectively. 4) Compared with the natural state, human MSCs-exosome modified by PD-L1 or PD-L1-ITGB1 showed better proliferation inhibitory effect in vivo, and increased the proportion of Treg cells in vitro.

Conclusion

MSCs-exosome exhibited similar immunomodulatory effects with MSCs. MSCs-exosome after PD-L1 and PD-L1-ITGB1 targeted modification effectively inhibited the proliferation of activated T cells and increased the proportion of Treg cells.

Keyword(s): AAV, Graft-versus-host disease (GVHD), Mesenchymal stem cell

Presentation during EHA2021: All e-poster presentations will be made available as of Friday, June 11, 2021 (09:00 CEST) and will be accessible for on-demand viewing until August 15, 2021 on the Virtual Congress platform.

Abstract: EP728

Type: E-Poster Presentation

Session title: Gene therapy, cellular immunotherapy and vaccination - Biology & Translational Research

Background
There exist kinds of limitations of MSCs during its application, while exosome derived from MSCs has shown promising future.

Aims

To verify the effects and mechanisms of natural MSCs-exosome in treating acute GVHD in mice, then explore and establish a method for targeted modification of MSCs-exosome and verify the functions of the modified MSCs-exosome.

Methods

In different doses of MSCs-exosome groups and MSCs group, the weight loss of acute GVHD mice was observed, then the proliferation levels of activated T cells were measured through T cell activation experiment in vitro and OVA antigen specific T cell activation experiment in vivo. AAV2YF3 mutants carrying PD-L1 and PD-L1-ITGB1 was obtained after the construction of recombinant expression vectors and then was applied to infect human MSCs to modify their exosome. The immunoregulatory functions of the modified MSCs-exosome were measured with methods as above.

Results

1) Mouse MSCs-exosome (300ug*3 times) and MSCs (1e6*3 times) effectively alleviated the weight loss of acute GVHD mice. 2) Compared with IL2, 10ug, 25ug and 50ug human MSCs-exosome inhibited the proliferation of activated T cells in vitro, respectively 86.01% (IL2), 40%, 39.61% and 42.84%; compared with PBS, 50ug, 100ug and 200ug mouse MSCs-exosome inhibited the proliferation of antigen-specifically activated OT-1 cells in vivo, respectively 42.6%, 33.1%, 14.2% and 10.6%. 3) After the infection of AAV2YF3 mutant carrying PD-L1 or PD-L1-ITGB1, the positive proportion of MSCs-exosome exceeds 40% and 60%, respectively. 4) Compared with the natural state, human MSCs-exosome modified by PD-L1 or PD-L1-ITGB1 showed better proliferation inhibitory effect in vivo, and increased the proportion of Treg cells in vitro.

Conclusion

MSCs-exosome exhibited similar immunomodulatory effects with MSCs. MSCs-exosome after PD-L1 and PD-L1-ITGB1 targeted modification effectively inhibited the proliferation of activated T cells and increased the proportion of Treg cells.

Keyword(s): AAV, Graft-versus-host disease (GVHD), Mesenchymal stem cell

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