Establishment of an In Vitro Assay to Evaluate Indirect T-cell Allo-Response Using Donor PBMC-Pulsed Dendritic Cells

K. Iwasaki¹, T. Tomosugi², T. Sekiya³, H. Hamana⁴, Y. Miwa¹, K. Futamura², M. Okada², T. Hiramitsu², N. Goto², S. Narumi², Y. Watarai², H. Kishi⁴, M. Okumura⁵, S. Ashimine⁵, K. Ishiyama⁵, T. Kobayashi⁵

¹Department of Kidney Diseases and Transplant Immunology, Aichi Medical University School of Medicine, Nagakute, Japan, ²Department of Transplant Surgery, Japanese Red Cross Aichi Medical Center Nagoya Daini Hospital, Nagoya, Japan, ³Department of Immune Regulation, The Research Center for Hepatitis and Immunology, Ichikawa, Japan, ⁴Department of Immunology, University of Toyama, Toyama, Japan, ⁵Department of Renal Transplant Surgery, Aichi Medical University School of Medicine, Nagakute, Japan

Meeting: 2022 American Transplant Congress

Abstract number: 915

Keywords: Allorecognition, Antigen presentation, CD4, HLA antibodies

Topic: Basic Science » Basic Science » 03 - Antigen Presentation / Allorecognition / Dendritic Cells

Session Information

Session Name: Antigen Presentation / Allorecognition / Dendritic Cells

Session Type: Poster Abstract

Date: Sunday, June 5, 2022

Session Time: 7:00pm-8:00pm

Presentation Time: 7:00pm-8:00pm

Location: Hynes Halls C & D

*Purpose: Conventional MLR-based methods which evaluate donor-specific immune response via direct T-cell recognition pathway have been reported to be useful in predicting acute TCMR. However, since DSA is produced through the activation of B-cells via indirect T-cell allo-response, monitoring methods have yet to be established. In this study, dendritic cells (DC) differentiated from recipient PBMC-derived CD14⁺ monocytes were used to set up a method for assessing indirect allo-response and to evaluate the patient’s sensitization status in kidney transplant patients.

*Methods: Fourteen pre-formed DSA, 15 unsensitized, and 11 de novo DSA positive patients were examined before and after kidney transplantation. CD14⁺ cells were isolated, mixed with irradiated donor PBMCs, and cultured with IL-4/GM-CSF for 4 days and IL-1β/TNFα for additional 2 days to be used as pulsed DC. IL-21 was confirmed by ELSIPOT assay after CD4 T-cells were divided into naïve and memory cells.

*Results: In a study among unsensitized patients, pulsed DC preferentially enhanced naïve response than memory CD4 T-cells. Preformed DSA positive patient showed the IL-21 production from memory CD4 T-cells as well as naïve one. Furthermore, even in patients who showed only naive responses before transplantation, memory response was observed when de novo DSA became positive (p<0.05).

*Conclusions: Donor PBMC-pulsed DC-CD4 assay was established to evaluate the indirect response associated with DSA production. Whether CD4 T-cell activation detected prior to kidney transplantation are associated with early de novo DSA production is currently under investigation.

To cite this abstract in AMA style:

Iwasaki K, Tomosugi T, Sekiya T, Hamana H, Miwa Y, Futamura K, Okada M, Hiramitsu T, Goto N, Narumi S, Watarai Y, Kishi H, Okumura M, Ashimine S, Ishiyama K, Kobayashi T. Establishment of an In Vitro Assay to Evaluate Indirect T-cell Allo-Response Using Donor PBMC-Pulsed Dendritic Cells [abstract]. Am J Transplant. 2022; 22 (suppl 3). https://atcmeetingabstracts.com/abstract/establishment-of-an-in-vitro-assay-to-evaluate-indirect-t-cell-allo-response-using-donor-pbmc-pulsed-dendritic-cells/. Accessed May 18, 2025.

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