Research ArticleHepatologyImmunology Open Access | 10.1172/JCI163291

Xinjun Lu,1 Shanshan Deng,2 Jiejie Xu,3 Benjamin L. Green,4 Honghua Zhang,1 Guofei Cui,2 Yi Zhou,2,5 Yi Zhang,2,6 Hongwei Xu,2,7 Fapeng Zhang,1 Rui Mao,8 Sheng Zhong,2 Thorsten Cramer,9,10 Matthias Evert,11 Diego F. Calvisi,11 Yukai He,8 Chao Liu,1 and Xin Chen2,4

1Department of Biliary-Pancreatic Surgery, Sun Yat-sen Memorial Hospital, Sun Yat-sen University, Guangzhou, China.

2Department of Bioengineering and Therapeutic Sciences, University of California, San Francisco, California, USA.

3Department of Hearing and Speech Science, Guangzhou Xinhua University, Guangzhou, China.

4University of Hawaii Cancer Center, Honolulu, Hawaii, USA.

5Department of Infectious Diseases, The First Affiliated Hospital of Xi’an Jiaotong University, Xi’an, China.

6Key Laboratory of Biorheological Science and Technology, Ministry of Education, College of Bioengineering, Chongqing University, Chongqing, China.

7Department of Liver Surgery, Center of Liver Transplantation, West China Hospital of Sichuan University, Chengdu, China.

8Department of Medicine, Medical College of Georgia, Augusta University, Augusta, Georgia, USA.

9Department of General, Visceral and Transplantation Surgery, RWTH University Hospital, Aachen, Germany.

10Department of Surgery, Maastricht University Medical Center, Maastricht, The Netherlands.

11Institute of Pathology, University of Regensburg, Regensburg, Germany.

Address correspondence to: Xin Chen, University of Hawaii Cancer Center, Honolulu, Hawaii 96813, USA. Phone: 808.564.3804; Email: xinchen3@hawaii.edu or xin.chen@ucsf.edu. Or to: Chao Liu, Department of Biliary-Pancreatic Surgery, Sun Yat-sen Memorial Hospital, Sun Yat-sen University, 510120 Guangzhou, China. Phone: 86.020.8133.2602; Email: Liuchao3@mail.sysu.edu.cn.

Authorship note: XL, SD, JX, and BLG contributed equally to this work.

Find articles by Lu, X. in: JCI | PubMed | Google Scholar

1Department of Biliary-Pancreatic Surgery, Sun Yat-sen Memorial Hospital, Sun Yat-sen University, Guangzhou, China.

2Department of Bioengineering and Therapeutic Sciences, University of California, San Francisco, California, USA.

3Department of Hearing and Speech Science, Guangzhou Xinhua University, Guangzhou, China.

4University of Hawaii Cancer Center, Honolulu, Hawaii, USA.

5Department of Infectious Diseases, The First Affiliated Hospital of Xi’an Jiaotong University, Xi’an, China.

6Key Laboratory of Biorheological Science and Technology, Ministry of Education, College of Bioengineering, Chongqing University, Chongqing, China.

7Department of Liver Surgery, Center of Liver Transplantation, West China Hospital of Sichuan University, Chengdu, China.

8Department of Medicine, Medical College of Georgia, Augusta University, Augusta, Georgia, USA.

9Department of General, Visceral and Transplantation Surgery, RWTH University Hospital, Aachen, Germany.

10Department of Surgery, Maastricht University Medical Center, Maastricht, The Netherlands.

11Institute of Pathology, University of Regensburg, Regensburg, Germany.

Address correspondence to: Xin Chen, University of Hawaii Cancer Center, Honolulu, Hawaii 96813, USA. Phone: 808.564.3804; Email: xinchen3@hawaii.edu or xin.chen@ucsf.edu. Or to: Chao Liu, Department of Biliary-Pancreatic Surgery, Sun Yat-sen Memorial Hospital, Sun Yat-sen University, 510120 Guangzhou, China. Phone: 86.020.8133.2602; Email: Liuchao3@mail.sysu.edu.cn.

Authorship note: XL, SD, JX, and BLG contributed equally to this work.

Find articles by Deng, S. in: JCI | PubMed | Google Scholar

1Department of Biliary-Pancreatic Surgery, Sun Yat-sen Memorial Hospital, Sun Yat-sen University, Guangzhou, China.

2Department of Bioengineering and Therapeutic Sciences, University of California, San Francisco, California, USA.

3Department of Hearing and Speech Science, Guangzhou Xinhua University, Guangzhou, China.

4University of Hawaii Cancer Center, Honolulu, Hawaii, USA.

5Department of Infectious Diseases, The First Affiliated Hospital of Xi’an Jiaotong University, Xi’an, China.

6Key Laboratory of Biorheological Science and Technology, Ministry of Education, College of Bioengineering, Chongqing University, Chongqing, China.

7Department of Liver Surgery, Center of Liver Transplantation, West China Hospital of Sichuan University, Chengdu, China.

8Department of Medicine, Medical College of Georgia, Augusta University, Augusta, Georgia, USA.

9Department of General, Visceral and Transplantation Surgery, RWTH University Hospital, Aachen, Germany.

10Department of Surgery, Maastricht University Medical Center, Maastricht, The Netherlands.

11Institute of Pathology, University of Regensburg, Regensburg, Germany.

Address correspondence to: Xin Chen, University of Hawaii Cancer Center, Honolulu, Hawaii 96813, USA. Phone: 808.564.3804; Email: xinchen3@hawaii.edu or xin.chen@ucsf.edu. Or to: Chao Liu, Department of Biliary-Pancreatic Surgery, Sun Yat-sen Memorial Hospital, Sun Yat-sen University, 510120 Guangzhou, China. Phone: 86.020.8133.2602; Email: Liuchao3@mail.sysu.edu.cn.

Authorship note: XL, SD, JX, and BLG contributed equally to this work.

Find articles by Xu, J. in: JCI | PubMed | Google Scholar

1Department of Biliary-Pancreatic Surgery, Sun Yat-sen Memorial Hospital, Sun Yat-sen University, Guangzhou, China.

2Department of Bioengineering and Therapeutic Sciences, University of California, San Francisco, California, USA.

3Department of Hearing and Speech Science, Guangzhou Xinhua University, Guangzhou, China.

4University of Hawaii Cancer Center, Honolulu, Hawaii, USA.

5Department of Infectious Diseases, The First Affiliated Hospital of Xi’an Jiaotong University, Xi’an, China.

6Key Laboratory of Biorheological Science and Technology, Ministry of Education, College of Bioengineering, Chongqing University, Chongqing, China.

7Department of Liver Surgery, Center of Liver Transplantation, West China Hospital of Sichuan University, Chengdu, China.

8Department of Medicine, Medical College of Georgia, Augusta University, Augusta, Georgia, USA.

9Department of General, Visceral and Transplantation Surgery, RWTH University Hospital, Aachen, Germany.

10Department of Surgery, Maastricht University Medical Center, Maastricht, The Netherlands.

11Institute of Pathology, University of Regensburg, Regensburg, Germany.

Address correspondence to: Xin Chen, University of Hawaii Cancer Center, Honolulu, Hawaii 96813, USA. Phone: 808.564.3804; Email: xinchen3@hawaii.edu or xin.chen@ucsf.edu. Or to: Chao Liu, Department of Biliary-Pancreatic Surgery, Sun Yat-sen Memorial Hospital, Sun Yat-sen University, 510120 Guangzhou, China. Phone: 86.020.8133.2602; Email: Liuchao3@mail.sysu.edu.cn.

Authorship note: XL, SD, JX, and BLG contributed equally to this work.

Find articles by Green, B. in: JCI | PubMed | Google Scholar |

1Department of Biliary-Pancreatic Surgery, Sun Yat-sen Memorial Hospital, Sun Yat-sen University, Guangzhou, China.

2Department of Bioengineering and Therapeutic Sciences, University of California, San Francisco, California, USA.

3Department of Hearing and Speech Science, Guangzhou Xinhua University, Guangzhou, China.

4University of Hawaii Cancer Center, Honolulu, Hawaii, USA.

5Department of Infectious Diseases, The First Affiliated Hospital of Xi’an Jiaotong University, Xi’an, China.

6Key Laboratory of Biorheological Science and Technology, Ministry of Education, College of Bioengineering, Chongqing University, Chongqing, China.

7Department of Liver Surgery, Center of Liver Transplantation, West China Hospital of Sichuan University, Chengdu, China.

8Department of Medicine, Medical College of Georgia, Augusta University, Augusta, Georgia, USA.

9Department of General, Visceral and Transplantation Surgery, RWTH University Hospital, Aachen, Germany.

10Department of Surgery, Maastricht University Medical Center, Maastricht, The Netherlands.

11Institute of Pathology, University of Regensburg, Regensburg, Germany.

Address correspondence to: Xin Chen, University of Hawaii Cancer Center, Honolulu, Hawaii 96813, USA. Phone: 808.564.3804; Email: xinchen3@hawaii.edu or xin.chen@ucsf.edu. Or to: Chao Liu, Department of Biliary-Pancreatic Surgery, Sun Yat-sen Memorial Hospital, Sun Yat-sen University, 510120 Guangzhou, China. Phone: 86.020.8133.2602; Email: Liuchao3@mail.sysu.edu.cn.

Authorship note: XL, SD, JX, and BLG contributed equally to this work.

Find articles by Zhang, H. in: JCI | PubMed | Google Scholar

1Department of Biliary-Pancreatic Surgery, Sun Yat-sen Memorial Hospital, Sun Yat-sen University, Guangzhou, China.

2Department of Bioengineering and Therapeutic Sciences, University of California, San Francisco, California, USA.

3Department of Hearing and Speech Science, Guangzhou Xinhua University, Guangzhou, China.

4University of Hawaii Cancer Center, Honolulu, Hawaii, USA.

5Department of Infectious Diseases, The First Affiliated Hospital of Xi’an Jiaotong University, Xi’an, China.

6Key Laboratory of Biorheological Science and Technology, Ministry of Education, College of Bioengineering, Chongqing University, Chongqing, China.

7Department of Liver Surgery, Center of Liver Transplantation, West China Hospital of Sichuan University, Chengdu, China.

8Department of Medicine, Medical College of Georgia, Augusta University, Augusta, Georgia, USA.

9Department of General, Visceral and Transplantation Surgery, RWTH University Hospital, Aachen, Germany.

10Department of Surgery, Maastricht University Medical Center, Maastricht, The Netherlands.

11Institute of Pathology, University of Regensburg, Regensburg, Germany.

Address correspondence to: Xin Chen, University of Hawaii Cancer Center, Honolulu, Hawaii 96813, USA. Phone: 808.564.3804; Email: xinchen3@hawaii.edu or xin.chen@ucsf.edu. Or to: Chao Liu, Department of Biliary-Pancreatic Surgery, Sun Yat-sen Memorial Hospital, Sun Yat-sen University, 510120 Guangzhou, China. Phone: 86.020.8133.2602; Email: Liuchao3@mail.sysu.edu.cn.

Authorship note: XL, SD, JX, and BLG contributed equally to this work.

Find articles by Cui, G. in: JCI | PubMed | Google Scholar

1Department of Biliary-Pancreatic Surgery, Sun Yat-sen Memorial Hospital, Sun Yat-sen University, Guangzhou, China.

2Department of Bioengineering and Therapeutic Sciences, University of California, San Francisco, California, USA.

3Department of Hearing and Speech Science, Guangzhou Xinhua University, Guangzhou, China.

4University of Hawaii Cancer Center, Honolulu, Hawaii, USA.

5Department of Infectious Diseases, The First Affiliated Hospital of Xi’an Jiaotong University, Xi’an, China.

6Key Laboratory of Biorheological Science and Technology, Ministry of Education, College of Bioengineering, Chongqing University, Chongqing, China.

7Department of Liver Surgery, Center of Liver Transplantation, West China Hospital of Sichuan University, Chengdu, China.

8Department of Medicine, Medical College of Georgia, Augusta University, Augusta, Georgia, USA.

9Department of General, Visceral and Transplantation Surgery, RWTH University Hospital, Aachen, Germany.

10Department of Surgery, Maastricht University Medical Center, Maastricht, The Netherlands.

11Institute of Pathology, University of Regensburg, Regensburg, Germany.

Address correspondence to: Xin Chen, University of Hawaii Cancer Center, Honolulu, Hawaii 96813, USA. Phone: 808.564.3804; Email: xinchen3@hawaii.edu or xin.chen@ucsf.edu. Or to: Chao Liu, Department of Biliary-Pancreatic Surgery, Sun Yat-sen Memorial Hospital, Sun Yat-sen University, 510120 Guangzhou, China. Phone: 86.020.8133.2602; Email: Liuchao3@mail.sysu.edu.cn.

Authorship note: XL, SD, JX, and BLG contributed equally to this work.

Find articles by Zhou, Y. in: JCI | PubMed | Google Scholar

1Department of Biliary-Pancreatic Surgery, Sun Yat-sen Memorial Hospital, Sun Yat-sen University, Guangzhou, China.

2Department of Bioengineering and Therapeutic Sciences, University of California, San Francisco, California, USA.

3Department of Hearing and Speech Science, Guangzhou Xinhua University, Guangzhou, China.

4University of Hawaii Cancer Center, Honolulu, Hawaii, USA.

5Department of Infectious Diseases, The First Affiliated Hospital of Xi’an Jiaotong University, Xi’an, China.

6Key Laboratory of Biorheological Science and Technology, Ministry of Education, College of Bioengineering, Chongqing University, Chongqing, China.

7Department of Liver Surgery, Center of Liver Transplantation, West China Hospital of Sichuan University, Chengdu, China.

8Department of Medicine, Medical College of Georgia, Augusta University, Augusta, Georgia, USA.

9Department of General, Visceral and Transplantation Surgery, RWTH University Hospital, Aachen, Germany.

10Department of Surgery, Maastricht University Medical Center, Maastricht, The Netherlands.

11Institute of Pathology, University of Regensburg, Regensburg, Germany.

Address correspondence to: Xin Chen, University of Hawaii Cancer Center, Honolulu, Hawaii 96813, USA. Phone: 808.564.3804; Email: xinchen3@hawaii.edu or xin.chen@ucsf.edu. Or to: Chao Liu, Department of Biliary-Pancreatic Surgery, Sun Yat-sen Memorial Hospital, Sun Yat-sen University, 510120 Guangzhou, China. Phone: 86.020.8133.2602; Email: Liuchao3@mail.sysu.edu.cn.

Authorship note: XL, SD, JX, and BLG contributed equally to this work.

Find articles by Zhang, Y. in: JCI | PubMed | Google Scholar

1Department of Biliary-Pancreatic Surgery, Sun Yat-sen Memorial Hospital, Sun Yat-sen University, Guangzhou, China.

2Department of Bioengineering and Therapeutic Sciences, University of California, San Francisco, California, USA.

3Department of Hearing and Speech Science, Guangzhou Xinhua University, Guangzhou, China.

4University of Hawaii Cancer Center, Honolulu, Hawaii, USA.

5Department of Infectious Diseases, The First Affiliated Hospital of Xi’an Jiaotong University, Xi’an, China.

6Key Laboratory of Biorheological Science and Technology, Ministry of Education, College of Bioengineering, Chongqing University, Chongqing, China.

7Department of Liver Surgery, Center of Liver Transplantation, West China Hospital of Sichuan University, Chengdu, China.

8Department of Medicine, Medical College of Georgia, Augusta University, Augusta, Georgia, USA.

9Department of General, Visceral and Transplantation Surgery, RWTH University Hospital, Aachen, Germany.

10Department of Surgery, Maastricht University Medical Center, Maastricht, The Netherlands.

11Institute of Pathology, University of Regensburg, Regensburg, Germany.

Address correspondence to: Xin Chen, University of Hawaii Cancer Center, Honolulu, Hawaii 96813, USA. Phone: 808.564.3804; Email: xinchen3@hawaii.edu or xin.chen@ucsf.edu. Or to: Chao Liu, Department of Biliary-Pancreatic Surgery, Sun Yat-sen Memorial Hospital, Sun Yat-sen University, 510120 Guangzhou, China. Phone: 86.020.8133.2602; Email: Liuchao3@mail.sysu.edu.cn.

Authorship note: XL, SD, JX, and BLG contributed equally to this work.

Find articles by Xu, H. in: JCI | PubMed | Google Scholar

1Department of Biliary-Pancreatic Surgery, Sun Yat-sen Memorial Hospital, Sun Yat-sen University, Guangzhou, China.

2Department of Bioengineering and Therapeutic Sciences, University of California, San Francisco, California, USA.

3Department of Hearing and Speech Science, Guangzhou Xinhua University, Guangzhou, China.

4University of Hawaii Cancer Center, Honolulu, Hawaii, USA.

5Department of Infectious Diseases, The First Affiliated Hospital of Xi’an Jiaotong University, Xi’an, China.

6Key Laboratory of Biorheological Science and Technology, Ministry of Education, College of Bioengineering, Chongqing University, Chongqing, China.

7Department of Liver Surgery, Center of Liver Transplantation, West China Hospital of Sichuan University, Chengdu, China.

8Department of Medicine, Medical College of Georgia, Augusta University, Augusta, Georgia, USA.

9Department of General, Visceral and Transplantation Surgery, RWTH University Hospital, Aachen, Germany.

10Department of Surgery, Maastricht University Medical Center, Maastricht, The Netherlands.

11Institute of Pathology, University of Regensburg, Regensburg, Germany.

Address correspondence to: Xin Chen, University of Hawaii Cancer Center, Honolulu, Hawaii 96813, USA. Phone: 808.564.3804; Email: xinchen3@hawaii.edu or xin.chen@ucsf.edu. Or to: Chao Liu, Department of Biliary-Pancreatic Surgery, Sun Yat-sen Memorial Hospital, Sun Yat-sen University, 510120 Guangzhou, China. Phone: 86.020.8133.2602; Email: Liuchao3@mail.sysu.edu.cn.

Authorship note: XL, SD, JX, and BLG contributed equally to this work.

Find articles by Zhang, F. in: JCI | PubMed | Google Scholar

1Department of Biliary-Pancreatic Surgery, Sun Yat-sen Memorial Hospital, Sun Yat-sen University, Guangzhou, China.

2Department of Bioengineering and Therapeutic Sciences, University of California, San Francisco, California, USA.

3Department of Hearing and Speech Science, Guangzhou Xinhua University, Guangzhou, China.

4University of Hawaii Cancer Center, Honolulu, Hawaii, USA.

5Department of Infectious Diseases, The First Affiliated Hospital of Xi’an Jiaotong University, Xi’an, China.

6Key Laboratory of Biorheological Science and Technology, Ministry of Education, College of Bioengineering, Chongqing University, Chongqing, China.

7Department of Liver Surgery, Center of Liver Transplantation, West China Hospital of Sichuan University, Chengdu, China.

8Department of Medicine, Medical College of Georgia, Augusta University, Augusta, Georgia, USA.

9Department of General, Visceral and Transplantation Surgery, RWTH University Hospital, Aachen, Germany.

10Department of Surgery, Maastricht University Medical Center, Maastricht, The Netherlands.

11Institute of Pathology, University of Regensburg, Regensburg, Germany.

Address correspondence to: Xin Chen, University of Hawaii Cancer Center, Honolulu, Hawaii 96813, USA. Phone: 808.564.3804; Email: xinchen3@hawaii.edu or xin.chen@ucsf.edu. Or to: Chao Liu, Department of Biliary-Pancreatic Surgery, Sun Yat-sen Memorial Hospital, Sun Yat-sen University, 510120 Guangzhou, China. Phone: 86.020.8133.2602; Email: Liuchao3@mail.sysu.edu.cn.

Authorship note: XL, SD, JX, and BLG contributed equally to this work.

Find articles by Mao, R. in: JCI | PubMed | Google Scholar

1Department of Biliary-Pancreatic Surgery, Sun Yat-sen Memorial Hospital, Sun Yat-sen University, Guangzhou, China.

2Department of Bioengineering and Therapeutic Sciences, University of California, San Francisco, California, USA.

3Department of Hearing and Speech Science, Guangzhou Xinhua University, Guangzhou, China.

4University of Hawaii Cancer Center, Honolulu, Hawaii, USA.

5Department of Infectious Diseases, The First Affiliated Hospital of Xi’an Jiaotong University, Xi’an, China.

6Key Laboratory of Biorheological Science and Technology, Ministry of Education, College of Bioengineering, Chongqing University, Chongqing, China.

7Department of Liver Surgery, Center of Liver Transplantation, West China Hospital of Sichuan University, Chengdu, China.

8Department of Medicine, Medical College of Georgia, Augusta University, Augusta, Georgia, USA.

9Department of General, Visceral and Transplantation Surgery, RWTH University Hospital, Aachen, Germany.

10Department of Surgery, Maastricht University Medical Center, Maastricht, The Netherlands.

11Institute of Pathology, University of Regensburg, Regensburg, Germany.

Address correspondence to: Xin Chen, University of Hawaii Cancer Center, Honolulu, Hawaii 96813, USA. Phone: 808.564.3804; Email: xinchen3@hawaii.edu or xin.chen@ucsf.edu. Or to: Chao Liu, Department of Biliary-Pancreatic Surgery, Sun Yat-sen Memorial Hospital, Sun Yat-sen University, 510120 Guangzhou, China. Phone: 86.020.8133.2602; Email: Liuchao3@mail.sysu.edu.cn.

Authorship note: XL, SD, JX, and BLG contributed equally to this work.

Find articles by Zhong, S. in: JCI | PubMed | Google Scholar

1Department of Biliary-Pancreatic Surgery, Sun Yat-sen Memorial Hospital, Sun Yat-sen University, Guangzhou, China.

2Department of Bioengineering and Therapeutic Sciences, University of California, San Francisco, California, USA.

3Department of Hearing and Speech Science, Guangzhou Xinhua University, Guangzhou, China.

4University of Hawaii Cancer Center, Honolulu, Hawaii, USA.

5Department of Infectious Diseases, The First Affiliated Hospital of Xi’an Jiaotong University, Xi’an, China.

6Key Laboratory of Biorheological Science and Technology, Ministry of Education, College of Bioengineering, Chongqing University, Chongqing, China.

7Department of Liver Surgery, Center of Liver Transplantation, West China Hospital of Sichuan University, Chengdu, China.

8Department of Medicine, Medical College of Georgia, Augusta University, Augusta, Georgia, USA.

9Department of General, Visceral and Transplantation Surgery, RWTH University Hospital, Aachen, Germany.

10Department of Surgery, Maastricht University Medical Center, Maastricht, The Netherlands.

11Institute of Pathology, University of Regensburg, Regensburg, Germany.

Address correspondence to: Xin Chen, University of Hawaii Cancer Center, Honolulu, Hawaii 96813, USA. Phone: 808.564.3804; Email: xinchen3@hawaii.edu or xin.chen@ucsf.edu. Or to: Chao Liu, Department of Biliary-Pancreatic Surgery, Sun Yat-sen Memorial Hospital, Sun Yat-sen University, 510120 Guangzhou, China. Phone: 86.020.8133.2602; Email: Liuchao3@mail.sysu.edu.cn.

Authorship note: XL, SD, JX, and BLG contributed equally to this work.

Find articles by Cramer, T. in: JCI | PubMed | Google Scholar |

1Department of Biliary-Pancreatic Surgery, Sun Yat-sen Memorial Hospital, Sun Yat-sen University, Guangzhou, China.

2Department of Bioengineering and Therapeutic Sciences, University of California, San Francisco, California, USA.

3Department of Hearing and Speech Science, Guangzhou Xinhua University, Guangzhou, China.

4University of Hawaii Cancer Center, Honolulu, Hawaii, USA.

5Department of Infectious Diseases, The First Affiliated Hospital of Xi’an Jiaotong University, Xi’an, China.

6Key Laboratory of Biorheological Science and Technology, Ministry of Education, College of Bioengineering, Chongqing University, Chongqing, China.

7Department of Liver Surgery, Center of Liver Transplantation, West China Hospital of Sichuan University, Chengdu, China.

8Department of Medicine, Medical College of Georgia, Augusta University, Augusta, Georgia, USA.

9Department of General, Visceral and Transplantation Surgery, RWTH University Hospital, Aachen, Germany.

10Department of Surgery, Maastricht University Medical Center, Maastricht, The Netherlands.

11Institute of Pathology, University of Regensburg, Regensburg, Germany.

Address correspondence to: Xin Chen, University of Hawaii Cancer Center, Honolulu, Hawaii 96813, USA. Phone: 808.564.3804; Email: xinchen3@hawaii.edu or xin.chen@ucsf.edu. Or to: Chao Liu, Department of Biliary-Pancreatic Surgery, Sun Yat-sen Memorial Hospital, Sun Yat-sen University, 510120 Guangzhou, China. Phone: 86.020.8133.2602; Email: Liuchao3@mail.sysu.edu.cn.

Authorship note: XL, SD, JX, and BLG contributed equally to this work.

Find articles by Evert, M. in: JCI | PubMed | Google Scholar

1Department of Biliary-Pancreatic Surgery, Sun Yat-sen Memorial Hospital, Sun Yat-sen University, Guangzhou, China.

2Department of Bioengineering and Therapeutic Sciences, University of California, San Francisco, California, USA.

3Department of Hearing and Speech Science, Guangzhou Xinhua University, Guangzhou, China.

4University of Hawaii Cancer Center, Honolulu, Hawaii, USA.

5Department of Infectious Diseases, The First Affiliated Hospital of Xi’an Jiaotong University, Xi’an, China.

6Key Laboratory of Biorheological Science and Technology, Ministry of Education, College of Bioengineering, Chongqing University, Chongqing, China.

7Department of Liver Surgery, Center of Liver Transplantation, West China Hospital of Sichuan University, Chengdu, China.

8Department of Medicine, Medical College of Georgia, Augusta University, Augusta, Georgia, USA.

9Department of General, Visceral and Transplantation Surgery, RWTH University Hospital, Aachen, Germany.

10Department of Surgery, Maastricht University Medical Center, Maastricht, The Netherlands.

11Institute of Pathology, University of Regensburg, Regensburg, Germany.

Address correspondence to: Xin Chen, University of Hawaii Cancer Center, Honolulu, Hawaii 96813, USA. Phone: 808.564.3804; Email: xinchen3@hawaii.edu or xin.chen@ucsf.edu. Or to: Chao Liu, Department of Biliary-Pancreatic Surgery, Sun Yat-sen Memorial Hospital, Sun Yat-sen University, 510120 Guangzhou, China. Phone: 86.020.8133.2602; Email: Liuchao3@mail.sysu.edu.cn.

Authorship note: XL, SD, JX, and BLG contributed equally to this work.

Find articles by Calvisi, D. in: JCI | PubMed | Google Scholar

1Department of Biliary-Pancreatic Surgery, Sun Yat-sen Memorial Hospital, Sun Yat-sen University, Guangzhou, China.

2Department of Bioengineering and Therapeutic Sciences, University of California, San Francisco, California, USA.

3Department of Hearing and Speech Science, Guangzhou Xinhua University, Guangzhou, China.

4University of Hawaii Cancer Center, Honolulu, Hawaii, USA.

5Department of Infectious Diseases, The First Affiliated Hospital of Xi’an Jiaotong University, Xi’an, China.

6Key Laboratory of Biorheological Science and Technology, Ministry of Education, College of Bioengineering, Chongqing University, Chongqing, China.

7Department of Liver Surgery, Center of Liver Transplantation, West China Hospital of Sichuan University, Chengdu, China.

8Department of Medicine, Medical College of Georgia, Augusta University, Augusta, Georgia, USA.

9Department of General, Visceral and Transplantation Surgery, RWTH University Hospital, Aachen, Germany.

10Department of Surgery, Maastricht University Medical Center, Maastricht, The Netherlands.

11Institute of Pathology, University of Regensburg, Regensburg, Germany.

Address correspondence to: Xin Chen, University of Hawaii Cancer Center, Honolulu, Hawaii 96813, USA. Phone: 808.564.3804; Email: xinchen3@hawaii.edu or xin.chen@ucsf.edu. Or to: Chao Liu, Department of Biliary-Pancreatic Surgery, Sun Yat-sen Memorial Hospital, Sun Yat-sen University, 510120 Guangzhou, China. Phone: 86.020.8133.2602; Email: Liuchao3@mail.sysu.edu.cn.

Authorship note: XL, SD, JX, and BLG contributed equally to this work.

Find articles by He, Y. in: JCI | PubMed | Google Scholar |

1Department of Biliary-Pancreatic Surgery, Sun Yat-sen Memorial Hospital, Sun Yat-sen University, Guangzhou, China.

2Department of Bioengineering and Therapeutic Sciences, University of California, San Francisco, California, USA.

3Department of Hearing and Speech Science, Guangzhou Xinhua University, Guangzhou, China.

4University of Hawaii Cancer Center, Honolulu, Hawaii, USA.

5Department of Infectious Diseases, The First Affiliated Hospital of Xi’an Jiaotong University, Xi’an, China.

6Key Laboratory of Biorheological Science and Technology, Ministry of Education, College of Bioengineering, Chongqing University, Chongqing, China.

7Department of Liver Surgery, Center of Liver Transplantation, West China Hospital of Sichuan University, Chengdu, China.

8Department of Medicine, Medical College of Georgia, Augusta University, Augusta, Georgia, USA.

9Department of General, Visceral and Transplantation Surgery, RWTH University Hospital, Aachen, Germany.

10Department of Surgery, Maastricht University Medical Center, Maastricht, The Netherlands.

11Institute of Pathology, University of Regensburg, Regensburg, Germany.

Address correspondence to: Xin Chen, University of Hawaii Cancer Center, Honolulu, Hawaii 96813, USA. Phone: 808.564.3804; Email: xinchen3@hawaii.edu or xin.chen@ucsf.edu. Or to: Chao Liu, Department of Biliary-Pancreatic Surgery, Sun Yat-sen Memorial Hospital, Sun Yat-sen University, 510120 Guangzhou, China. Phone: 86.020.8133.2602; Email: Liuchao3@mail.sysu.edu.cn.

Authorship note: XL, SD, JX, and BLG contributed equally to this work.

Find articles by Liu, C. in: JCI | PubMed | Google Scholar

1Department of Biliary-Pancreatic Surgery, Sun Yat-sen Memorial Hospital, Sun Yat-sen University, Guangzhou, China.

2Department of Bioengineering and Therapeutic Sciences, University of California, San Francisco, California, USA.

3Department of Hearing and Speech Science, Guangzhou Xinhua University, Guangzhou, China.

4University of Hawaii Cancer Center, Honolulu, Hawaii, USA.

5Department of Infectious Diseases, The First Affiliated Hospital of Xi’an Jiaotong University, Xi’an, China.

6Key Laboratory of Biorheological Science and Technology, Ministry of Education, College of Bioengineering, Chongqing University, Chongqing, China.

7Department of Liver Surgery, Center of Liver Transplantation, West China Hospital of Sichuan University, Chengdu, China.

8Department of Medicine, Medical College of Georgia, Augusta University, Augusta, Georgia, USA.

9Department of General, Visceral and Transplantation Surgery, RWTH University Hospital, Aachen, Germany.

10Department of Surgery, Maastricht University Medical Center, Maastricht, The Netherlands.

11Institute of Pathology, University of Regensburg, Regensburg, Germany.

Address correspondence to: Xin Chen, University of Hawaii Cancer Center, Honolulu, Hawaii 96813, USA. Phone: 808.564.3804; Email: xinchen3@hawaii.edu or xin.chen@ucsf.edu. Or to: Chao Liu, Department of Biliary-Pancreatic Surgery, Sun Yat-sen Memorial Hospital, Sun Yat-sen University, 510120 Guangzhou, China. Phone: 86.020.8133.2602; Email: Liuchao3@mail.sysu.edu.cn.

Authorship note: XL, SD, JX, and BLG contributed equally to this work.

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Authorship note: XL, SD, JX, and BLG contributed equally to this work.

Published April 11, 2023 - More info

Published in Volume 133, Issue 11 on June 1, 2023
J Clin Invest. 2023;133(11):e163291. https://doi.org/10.1172/JCI163291.
© 2023 Lu et al. This work is licensed under the Creative Commons Attribution 4.0 International License. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/. Published April 11, 2023 - Version history
Received: July 6, 2022; Accepted: April 4, 2023 View PDF Abstract

Many patients with hepatocellular carcinoma (HCC) do not respond to the first-line immune checkpoint inhibitor treatment. Immunization with effective cancer vaccines is an attractive alternative approach to immunotherapy. However, its efficacy remains insufficiently evaluated in preclinical studies. Here, we investigated HCC-associated self/tumor antigen, α-fetoprotein–based (AFP-based) vaccine immunization for treating AFP (+) HCC mouse models. We found that AFP immunization effectively induced AFP-specific CD8+ T cells in vivo. However, these CD8+ T cells expressed exhaustion markers, including PD1, LAG3, and Tim3. Furthermore, the AFP vaccine effectively prevented c-MYC/Mcl1 HCC initiation when administered before tumor formation, while it was ineffective against full-blown c-MYC/Mcl1 tumors. Similarly, anti-PD1 and anti–PD-L1 monotherapy showed no efficacy in this murine HCC model. In striking contrast, AFP immunization combined with anti–PD-L1 treatment triggered significant inhibition of HCC progression in most liver tumor nodules, while in combination with anti-PD1, it induced slower tumor progression. Mechanistically, we demonstrated that HCC-intrinsic PD-L1 expression was the primary target of anti–PD-L1 in this combination therapy. Notably, the combination therapy had a similar therapeutic effect in the cMet/β-catenin mouse HCC model. These findings suggest that combining the AFP vaccine and immune checkpoint inhibitors may be effective for AFP (+) HCC treatment.

Introduction

Hepatocellular carcinoma (HCC) accounts for most primary liver tumors and is one of the deadliest malignancies worldwide (1). Most HCCs are diagnosed in the advanced stage when limited therapeutic options are available. For years, the multikinase inhibitors sorafenib and lenvatinib represented the first-line drug regimens, although they only provided limited survival benefits to patients with progressed HCC (2). The success of immune checkpoint inhibitors (ICI), targeting programmed cell death protein 1 (PD1) and its ligand PD-L1 (also called CD274), and cytotoxic T-lymphocyte-associated protein 4 (CTLA4) in the therapy of some solid tumors has sparked significant interest in applying immunotherapy to HCC. Recently, the breakthrough of the Atezolizumab (anti–PD-L1) plus Bevacizumab (anti-VEGF) and tremelimumab (anti-CTLA4) plus durvalumab (anti-PD1) combination treatments in HCC demonstrated the promise of immunotherapy in the treatment of HCC (3, 4).

α-fetoprotein (AFP) is a major plasma protein produced by the yolk sac and the fetal liver during embryonic development. AFP level decreases rapidly after birth and is reexpressed in most HCC tumors, serving as a diagnostic biomarker in the clinic (5). The function of AFP in adults is unclear; nonetheless, it is a promising potential target for HCC immunotherapy (6–9). Butterfield et al. first identified 4 HLA-A*0201–restricted AFP epitopes and developed AFP peptide–based vaccinations for HCC (10, 11). This pioneering work suggested that AFP-recognizing T cells had not been entirely deleted from the T cell repertoire during the negative selection in the thymus. Since modest AFP-specific CD8+ T cell response was detected, these findings established a potential target for AFP-based HCC immunotherapy. A phase I/II clinical trial was conducted to evaluate the immunizing efficiency of these 4 AFP peptides pulsed onto dendritic cells (DC) in patients with HCC. Moderate AFP-specific T cell responses were detected to at least one of the peptides after vaccination (6). However, no significant clinical benefits were observed in these patients, suggesting that additional approaches may be needed to enhance this antitumor response.

Previously, a study reported multiple AFP epitopes that could bind to the mouse major histocompatibility complex–class I (MHC I) molecules based on an Epitoptimizer algorithm (12). Further amino acid modifications were employed to increase the affinity of the peptides to MHC I. The optimized AFP499 and AFP212 peptides induced high AFP antigen-specific T cells. Not surprisingly, followed by an AFP prime and boost immunization strategy, more infiltrating CD8+ T cells appeared in the tumors, and AFP499 and AFP212 peptides significantly delayed the subcutaneous and DEN-induced mouse HCC development (13). These studies suggested that the AFP vaccine could help prevent or delay HCC formation. However, whether this approach has therapeutic efficacy against existing HCC lesions remains unknown.

In the current study, we investigated the therapeutic potency of the AFP vaccination strategy (AFP lentivirus prime plus AFP499 peptide boost) in the c-MYC/Mcl1 and cMet/β-catenin murine HCC models. Our studies suggest that AFP immunization has limited antitumor activities against HCC. However, combined AFP vaccination and anti–PD-L1 antibody strongly inhibited HCC progression, supporting the usefulness of this combination immunotherapy for treating HCC.

Results

AFP immunization elicits functional AFP499+ cytotoxic T lymphocytes. A previous study identified an H2-Kb restricted and optimized AFP peptide (AFP499) that could induce AFP499 peptide-specific CD8+ cytotoxic T lymphocytes (CTLs) (AFP499+ CTLs) by AFP lentivirus prime and AFP499 peptide boost strategy (Figure 1, A and B) (13). As a positive control, we used a similar approach to express a foreign antigen, namely the SV40 Large T antigen, which induced high levels of Large T peptide-specific CD8+ T cells (Figure 1B). Importantly, no cross-reaction of AFP499 and Large T tetramer staining was detected, confirming their specificity. Notably, most peptide-specific CD8+ T cells, induced by the AFP499 or Large T, expressed abundant IFN-γ (Figure 1C), a marker for immune cell activation, indicating these CD8+ T cells were potentially functional. Other effects of the AFP vaccination strategy included increased CD8+ T cells and decreased CD4+ T cells and B cells in the liver (Supplemental Figure 1; supplemental material available online with this article; https://doi.org/10.1172/JCI163291DS1). In summary, the AFP lentivirus prime and AFP499 peptide boost strategy effectively induced AFP499+ CTL in vivo.

Figure 1

AFP or Large T antigen immunization elicits functional peptide-specific CD8+ T cells. (A) Overview of AFP and Large T peptide immunization procedure. (B) Representative results of AFP and Large T peptide-specific CD8+ T cells induced by the corresponding peptides. (C) Both AFP and Large T peptide-specific CD8+ T cells exhibit robust IFN-γ expression. Immu, immunization.

AFP immunization significantly delay