CTLA4.FasL promotes the engraftment of xenogeneic hepatic oval cells in rat spleen

doi:10.12307/2022.674

Abstract

Abstract: BACKGROUND: Cytotoxicn lymphocyte antigen 4.fasrymndrit ligand (CTLA4.FasL) can effectively inhibit alloreactive and autoreactive T lymphocytes. However, the inhibitory effect of CTLA4.FasL on xenogeneic T lymphocytes in hepatic oval cell transplantation via the spleen remains unclear.
OBJECTIVE: To investigate the effect of CTLA4.FasL on the proliferation of xenogeneic lymphocytes, and to monitor the survival and engraftment of CTLA4.FasL-hepatic oval cells in rat spleen.
METHODS: The recombinant lentiviral vectors carrying CTLA4.FasL fusion gene (Lv/CTLA4.FasL) and polybrene were added to the rat hepatic oval cell suspension. We used hepatic oval cells that were not infected with lentivirus and infected blank lentivirus-hepatic oval cells as controls, and then observed the viability of hepatic oval cells and the expression of red fluorescent protein lentiviral vector under a fluorescence microscope. Sprague-Dawley rats undergoing 2/3 hepatectomy were transplanted with CTLA4.FasL-hepatic oval cells, blank lentivirus-hepatic oval cells, hepatic oval cells, or phosphate buffered saline (without hepatic oval cells) through the spleen. At 1, 5, 14, and 21 days after hepatectomy, the concentration of CTLA4.FasL in rat serum was determined by enzyme linked immunosorbent assay, and the spleen of the recipient rats was taken for CK-19 immunohistochemical staining. C57BL/6 mouse lymphocytes were used as stimulator cells, and Sprague-Dawley rat lymphocytes as response cells. In the mixed lymphocyte reaction, CTLA4.FasL-hepatic oval cells, blank lentivirus-hepatic oval cells, and hepatic oval cells were inoculated, or CTLA4.FasL-hepatic oval cells (a mass concentration of 10, 50, and 100 μg/L), blank lentivirus-hepatic oval cells, and hepatic oval cell supernatant. After 96 hours of culture, 5-bromodeoxyuridine method was used to detect the proliferation of lymphocytes in each group.
RESULTS AND CONCLUSION: Both CTLA4.FasL-hepatic oval cells and the culture supernatant could effectively inhibit the proliferation of rat lymphocytes in the xenogeneic mixed lymphocyte culture system. Spleen lymphocytes from the recipient rats transplanted with CTLA4.FasL-hepatic oval cells had a low immune response to mouse lymphocytes, while the spleen cells derived from the recipient rats transplanted with blank lentivirus-hepatic oval cells and hepatic oval cells showed strong responses. CK-19 positive cells were detected in rat spleen parenchyma of the CTLA4.FasL-hepatic oval cell group but not in the phosphate buffered saline group, hepatic oval cell group, and blank lentivirus-hepatic oval cell group at 21 days after transplantation. The results show that CTLA4.FasL can promote the survival and engraftment of hepatic oval cells in the rat spleen parenchyma through inhibiting the proliferation of xenogeneic lymphocytes.

Key words: cytotoxicn lymphocyte antigen 4.fasrymndrit ligand, xenogeneic, hepatic oval cell, spleen, engraftment

CLC Number:

Wan Zhen, Wang Xuzhen, Zhang Xiaogang. CTLA4.FasL promotes the engraftment of xenogeneic hepatic oval cells in rat spleen[J]. Chinese Journal of Tissue Engineering Research, 2022, 26(23): 3728-3732.

Figures/Tables 4

References

[1] KO S, RUSSELL JO, MOLINA LM, et al. Liver progenitors and adult cell plasticity in hepatic injury and repair: knowns and unknowns. Annu Rev Pathol. 2020;15:23-50.
[2] LEE CA, SINHA S, FITZPATRICK E, et al. Hepatocyte transplantation and advancements in alternative cell sources for liver-based regenerative medicine. J Mol Med (Berl). 2018;96(6):469-481.
[3] MAKDASI E, AMSILI S, ARONIN A, et al. Toxicology and pharmacokinetic studies in mice and nonhuman primates of the nontoxic, efficient, targeted hexameric FasL: CTLA4-FasL. Mol Cancer Ther. 2020;19(2):513-524.
[4] PODOJIL JR, MILLER SD. Targeting the B7 family of co-stimulatory molecules: successes and challenges. Biodrugs. 2013;27(1):1-13.
[5] WANG F, YU JY, WANG Y, et al. Combination therapy with TNFR-Fc and CTLA4-FasL using the recombinant adeno-associated virus potently suppresses adjuvant-induced arthritis in rats.Appl Microbiol Biotechnol. 2015;99(15):6327-6337.
[6] LI WL, SU J, YAO YC, et al. Isolation and characterization of bipotent liver progenitor cells from adult mouse. Stem Cells. 2006;24(2):322-332.
[7] ADDANTE A, RONCERO C, LAZCANOITURBURU N, et al. A Signaling crosstalk between BMP9 and HGF/c-Met regulates mouse adult liver progenitor cell survival. Cells. 2020;9(3):752.
[8] GE JY, ZHENG YW, TSUCHIDA T, et al. Hepatic stellate cells contribute to liver regeneration through galectins in hepatic stem cell niche. Stem Cell Res Ther. 2020;11(1):425.
[9] BELASCHK E, ROHN S, MUKIIBI R, et al. Isolation, characterization and cold storage of cells isolated from diseased explanted livers. Int J Artif Organs. 2017;40(6):294-306.
[10] JULICH-HAERTEL H, TIWARI M, MEHRFELD C, et al. Isolation and enrichment of liver progenitor subsets identified by a novel surface marker combination. J Vis Exp. 2017;120:55284.
[11] DAI H, ZHENG Y, THOMSON AW, et al. Transplant tolerance induction: insights from the Liver. Front Immuno. 2020;11:1044.
[12] LI P, ZHANG YY, DENGJ. PDL1Ig gene-loaded BMSCs Induce liver transplantation immune tolerance. Eur Rev Med Pharmacol Sci. 2018;22(10):3214-3223.
[13] CHEN QY, YOU Y, ZHANG YJ, et al. Hepatocyte growth factor mediates a novel form of hepatic stem/progenitor cell-induced tolerance in a rat xenogeneic liver rejection model. Int Immunopharmacol. 2021;90:107180.
[14] ZHANG W, WANG F, WANG B, et al. Intraarticular gene delivery of CTLA4-FasL suppresses experimental arthritis. Int Immunol. 2012;24(6):379-388.
[15] FAGG WS, LIU NY, YANG MJ, et al. Magnetic targeting of stem cell derivatives enhances hepatic engraftment into structurally normal liver. Cell Transplant. 2017; 26(12):1868-1877.
[16] VISHWAKARMA SK, BARDIA A, LAKKIREDDY C, et al. Intraperitoneal transplantation of bioengineered humanized liver grafts supports failing liver in acute condition. Mater Sci Eng C Mater Biol Appl. 2019;98:861-873.
[17] MAS VR, MALUF DG. In Utero cell transplantation as a tool for fixing liver-based inherited metabolic disorders: early technical evidence supporting potential feasibility. Transplantation.2019;103(7):1300-1301.
[18] WANG LT, WANG CM, WANG ZZ, et al. Transforming the spleen into a liver-like organ in vivo. Sci Adv.2020;6(24):eaaz9974.
[19] MUTHIAH MD, HUANG DQ, ZHOU L, et al. A murine model demonstrating reversal of structural and functional correlates of cirrhosis with progenitor cell transplantation. Sci Rep. 2019;9(1):15446.
[20] IRUDAYASWAMY A, MUTHIAH M, ZHOU L, et al. Long-term fate of human fetal liver progenitor cells transplanted in injured mouse livers. Stem Cells. 2018;36(1): 103-113.
[21] HERRERO A, PRIGENT J, LOMBARD C, et al. Adult-derived human liver stem/progenitor cells infused 3 days postsurgery improve liver regeneration in a mouse model of extended hepatectomy. Cell Transplant. 2017;26(2):351-364.