PMID- 34508853 OWN - NLM STAT- MEDLINE DCOM- 20220217 LR - 20220217 IS - 1878-5492 (Electronic) IS - 0966-3274 (Linking) VI - 69 DP - 2021 Dec TI - Effects of serum from mismatched patients with solid organ transplantation on the activation of microvascular cultures isolated from adipose tissues. PG - 101462 LID - S0966-3274(21)00102-7 [pii] LID - 10.1016/j.trim.2021.101462 [doi] AB - BACKGROUND: Aggregating the human leukocyte antigen (HLA) Class I antigens on the endothelial membrane has been known to elicit an activation, an underlying mechanism of chronic rejection in organ transplant recipients. The current study aims at examining the endothelial responses using HLA typed microvascular cultures from human adipose tissues upon exposure to the serum that contain corresponding antibodies collected from mismatched transplant recipients. METHODS: We have successfully cultured 30 microvascular cultures and typed their HLAs. They are functionally competent to respond to inflammatory TNF-alpha stimulation and the aggregating monoclonal antibody against HLA Class I. The post-transplantation serum was collected either from the recipients with pathologically diagnosed chronic rejection or from the recipients without rejection. We determined their activation either by double-staining the endothelial cells in crude cultures with flow cytometry or by quantifying cytokine releases in purified endothelial cells using ELISA. RESULTS: Under our current protocol, adipose tissue cultures are functionally intact in regard to its responses to TNF-alpha and anti-HLA Class I antibody. We observed that the post-transplantation serum with rejection contained the pathogenic antibodies and led to proinflammatory activation, as demonstrated by not only increased CD54+/CD31+ and CD106+/CD31+ cell counts but also inflammatory cytokine releases including MCP-1, IL-8 and RANTES. CONCLUSION: This methodological study provides the feasibility of examining the pathogenicity of the alloantibodies in mis-transplant serum. Potentially, the endothelial activation elicited as a result of exposure can be used as an alternative readout for chronic rejection. SIGNIFICANCE: We prototype an ex vivo model that enables us to examine whether allogenic antibodies from the recipient can functionally activate microvascular endothelial cells from the donor adipose tissues. This system can be further developed as crossmatch using cellular responses as readouts for chronic rejection for post-transplant surveillance. CI - Copyright (c) 2021 Elsevier B.V. All rights reserved. FAU - Shi, Qiang Sebastian AU - Shi QS AD - Minnie & Max T. Voelcker Laboratory, Tianjin International Joint Academy of Biomedicine, S1515 Room, 220 Dongting Road, TEDA, Tianjin, China; Minnie & Max T. Voelcker Laboratory LLC, 1120 Piedmont Lane, Richardson, TX 75080, USA. Electronic address: shi_qiang@voelckerlab.com. FAU - Li, Dai-Hong AU - Li DH AD - Transplant Unit, Department of Blood Bank, Tianjin First Central Hospital, 24 Fukang Road, Nankai District, Tianjin, China. FAU - Wu, Cheng-Yu AU - Wu CY AD - Transplant Immunology Laboratory, Central Texas Baylor Scott & White Health, 2401 South 31st Street, Temple, TX 76508, United States of America. Electronic address: Chengyu.Wu@BSWHealth.org. FAU - Liu, Da-Zhen AU - Liu DZ AD - Department of Urology, General Hospital, Tianjin Medical University, 154 Anshan Street, Heping District, Tianjin, China. FAU - Hu, Jun AU - Hu J AD - Department of Colorectal Cancer Surgery, Tianjin Medical University Cancer Institute and Hospital, West Huanhu Road, Hexi District, Tianjin 300060, China. Electronic address: junhu@tmu.edu.cn. FAU - Cui, Yun-Long AU - Cui YL AD - Department of Colorectal Cancer Surgery, Tianjin Medical University Cancer Institute and Hospital, West Huanhu Road, Hexi District, Tianjin 300060, China. FAU - Zhao, Na AU - Zhao N AD - Minnie & Max T. Voelcker Laboratory, Tianjin International Joint Academy of Biomedicine, S1515 Room, 220 Dongting Road, TEDA, Tianjin, China; Minnie & Max T. Voelcker Laboratory LLC, 1120 Piedmont Lane, Richardson, TX 75080, USA. FAU - Chen, Li AU - Chen L AD - Transplant Unit, Department of Blood Bank, Tianjin First Central Hospital, 24 Fukang Road, Nankai District, Tianjin, China; Minnie & Max T. Voelcker Laboratory LLC, 1120 Piedmont Lane, Richardson, TX 75080, USA. FAU - Askar, Medhat AU - Askar M AD - Transplant Immunology, Baylor University Medical Center, 3500 Gaston Ave, 4th Floor of the Y Wing, RM# L-0470, Dallas, TX 75246, United States of America. Electronic address: Medhat.Askar@BSWHealth.org. LA - eng PT - Journal Article PT - Research Support, Non-U.S. Gov't DEP - 20210908 PL - Netherlands TA - Transpl Immunol JT - Transplant immunology JID - 9309923 RN - 0 (HLA Antigens) RN - 0 (Isoantibodies) SB - IM MH - Adipose Tissue MH - *Endothelial Cells MH - Graft Rejection MH - HLA Antigens MH - Humans MH - Isoantibodies MH - *Organ Transplantation OTO - NOTNLM OT - Antibody analysis OT - Cell-based assay OT - Endothelial activation OT - HLA antigens/immunology OT - Organ transplantation EDAT- 2021/09/12 06:00 MHDA- 2022/02/19 06:00 CRDT- 2021/09/11 20:10 PHST- 2021/07/23 00:00 [received] PHST- 2021/08/26 00:00 [revised] PHST- 2021/09/01 00:00 [accepted] PHST- 2021/09/12 06:00 [pubmed] PHST- 2022/02/19 06:00 [medline] PHST- 2021/09/11 20:10 [entrez] AID - S0966-3274(21)00102-7 [pii] AID - 10.1016/j.trim.2021.101462 [doi] PST - ppublish SO - Transpl Immunol. 2021 Dec;69:101462. doi: 10.1016/j.trim.2021.101462. Epub 2021 Sep 8.