이종 신장이식에서 인터페론-감마를 이용한 자연살해세포의 면역학적 평가 = Immunological Evaluation of NK Cells Using IFN-γ in Xenogeneic Kidney Transplantation

다국어 초록 (Multilingual Abstract)

Xenogeneic kidney transplantation from genetically modified pigs to Non-human primates represents a promising strategy to mitigate the global organ shortage crisis[Montgomery et al., 2022]. Nevertheless, hyperacute and acute rejection mechanisms, mediated by both innate and adaptive immune responses, continue to impede successful long-term graft survival[O’Connell, 2005]. The relative contribution of innate immunity, particularly NK cell-mediated responses, to xenogeneic rejection remains incompletely characterized. [Chen et al., 2006].
In this study, I performed immunophenotypic analysis on PBMCs collected over time from xenotransplant recipients throughout the xenotransplantation period. Flow cytometry analysis revealed a correlation between T cell (CD3⁺), NK cell (NKG2D⁺), and IFN-γ-secreting cell populations and CRP concentration. Additionally, to confirm in vitro, human NK92mi cells and porcine kidney cells, PK15, were co-cultured to confirm the xenogeneic cell stimulation ability.
The analysis results showed that T cells and NK cells were suppressed in the early post-transplantation period (POD7) and recovered in the late transplantation period (Necropsy). In particular, IFN-γ⁺ cells were confirmed to increase as the late period (Necropsy) approached, and showed a high correlation with CRP concentration. In vitro experiments also confirmed that IFN-γ secretion by human NK cells in response to porcine kidney cell stimulation increased significantly over time.
This study confirmed that the immune response observed in xenotransplantation, despite the use of immunosuppressive agents to reduce immune rejection, was NK cell-mediated innate immune activation and IFN-γ-mediated inflammatory responses. These results highlight the importance of NK cells, like T cells, in modulating immune rejection in future xenotransplantation, providing a mechanistic basis for strategies targeting both innate and adaptive immunity.

목차 (Table of Contents)

• I. 서론 1
• II. 재료 및 방법 7
• 2.1. 연구 윤리 승인 7
• 2.2. 실험동물 7
• 2.3. 신장 이식 수술 9
• 2.4. 면역억제, 항염증 치료 및 지지요법 9
• 2.5. 채혈 및 PBMC 분리 9
• 2.6. 혈액학적, 혈액생화학적 분석 10
• 2.7. Flow cytometry를 이용한 T/NK 세포 분석 10
• 2.8. NK92-mi, PK15 공동배양 실험 14
• 2.9. 통계분석 14
• III. 결과 16
• 3.1. 이식 수혜 게잡이 원숭이 생존율 16
• 3.2. 혈액학적 변화 18
• 3.3. 혈액 생화학적 변화 18
• 3.4. C-반응성 단백질(CRP)의 시간 변화 21
• 3.5. 말초혈액 단핵세포(PBMC) 분석: T 세포와 NK 세포의 동역학 24
• 3.6. 시간에 따른 IFN-γ⁺ 세포 변화 분석 35
• 3.7. CRP와 NK세포 활성화 간의 관계 39
• 3.8. 체내 : 이종항원 자극에 따른 NK세포 매개 IFN-γ 분비 검증 41
• IV. 고찰 43
• V. 결론 46
• VI. 참고 문헌 47