Utility of CD127 combined with FOXP3 for identification of operational tolerance after liver transplantation.
Loss of cell surface expression of CD127 on CD4(+)CD25(++) regulatory T-cells (Tregs) may be a useful marker to efficiently isolate Tregs. As FOXP3 was specifically used to identify Tregs, combining these two markers could give better identification for patient with operational tolerance (OT) after liver transplantation. To testify this mixed lymphocyte reaction (MLR), the function of circulating CD4(+)CD25(++)CD127(dim) cells (CD127(dim) cells) was examined in immunosuppression (IS)-free pediatric recipients after liver transplantation (LTx) (group operational tolerance: OT) (Gr-tol n=25) compared to recipients who could not stop IS due to clinically overt rejection (group intolerance) (Gr-intol n=18), recipients who were weaning IS (Gr-weaning n=11) and age-matched healthy volunteers (Gr-vol n=11). In addition, the frequencies of CD127(dim) cells vs CD4(+)CD25(++)CD127(dim)FOXP3(+) (CD127(dim)FOXP3(+)) cells were compared in these four groups by FACS analyses. Our results showed that The proliferation of CD4 cells to donor antigens was reduced compared to third-party antigens only in Gr-tol (P=0.022) but not in other groups (P=NS). Depletion of CD127(dim) cells resulted in a donor antigen-specific abrogation of this MLR hyporesponsiveness in Gr-tol (P<0.001) but not other groups (P=NS). This implied that CD127 efficiently isolated donor antigen-specific Tregs. The frequencies of CD127(dim) cells were significantly lower in Gr-intol (5.2%±1.9%) compared to those in Gr-tol (7.8%±1.8%) (P<0.001) as were the frequencies of CD127(dim) FOXP3(+) cells (Gr-tol: 5.4%±1.7% vs Gr-intol: 2.9%±1.0%, P<0.001). Of interest, there were fewer CD127(dim)FOXP3(+) cells in Gr-intol (2.9%±1%) than in Gr-weaning (5.1%±1.8%) (P=0.002), but no difference in CD127(dim) cells (Gr-intol: 5.2%±1.9% vs Gr-weaning: 6.7%±2.0%) (NS). Thus, combining FOXP3 with CD127 for phenotype analysis demonstrated an unequivocal difference between Gr-intol and Gr-weaning that was not detected by CD127 alone. In conclusion CD127 was a useful surface marker to isolate donor-antigen-specific-Tregs in OT after LTx. The additive effect of its combination with FOXP3 is important in phenotypical Treg analyses of OT patients.
Microbiology and Immunology Department, Faculty of Medicine, Assiut University, Assiut 71515, Egypt; Department of Hematology and Immunology, College of Medicine, Umm Al-Qura University, Mecca 50431, Saudi Arabia.
Hanaa
H
Nafady-Hego
Department of Surgery, Graduate School of Medicine, Kyoto University, Kyoto 606-8507, Japan.
Ying
Y
Li
Department of Surgery, Graduate School of Medicine, Kyoto University, Kyoto 606-8507, Japan.
Hidenori
H
Ohe
Department of Anesthesia, Faculty of Medicine, Assiut University, Assiut 71515, Egypt; Department of Anesthesiology, King Abdullah Medical City, Mecca 21955, Saudi Arabia.
Hamed
H
Elgendy
Department of Surgery, Graduate School of Medicine, Kyoto University, Kyoto 606-8507, Japan.
Xiangdong
X
Zhao
World Premier International Research Center, Immunology Frontier Research Center, Osaka University, Osaka 565-0871, Japan.
Shimon
S
Sakaguchi
Transplantation Laboratory, The University of Sydney, Sydney, NSW 2006, Australia.
G Alex
GA
Bishop
Department of Disaster and Comprehensive Medicine, Fukushima Medical University, Fukushima 960-1295, Japan. Electronic address: tkoshiba@fmu.ac.jp.
Takaaki
T
Koshiba
D000293:Adolescent; D000328:Adult; D015415:Biomarkers / Q000378:metabolism*; D002478:Cells, Cultured; D002648:Child; D005260:Female; D051858:Forkhead Transcription Factors / Q000378:metabolism*; D006084:Graft Rejection / Q000175:diagnosis*; D006801:Humans; D007108:Immune Tolerance; D053699:Interleukin-7 Receptor alpha Subunit / Q000378:metabolism*; D007519:Isoantigens / Q000276:immunology; D016031:Liver Transplantation; D007959:Lymphocyte Culture Test, Mixed; D008212:Lymphocyte Depletion; D008297:Male; D050378:T-Lymphocytes, Regulatory / Q000276:immunology*; D055815:Young Adult
D015415:Biomarkers; C418974:FOXP3 protein, human; D051858:Forkhead Transcription Factors; D053699:Interleukin-7 Receptor alpha Subunit; D007519:Isoantigens
Biomarker; Functional analysis; IL-7 receptor; Pediatric liver transplantation; Phenotypical analysis