不同时间输注受者动员脾细胞对小鼠单倍体相合造血干细胞移植后移植物抗宿主病的影响
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摘要
目的:研究小鼠单倍体相合造血干细胞移植后不同时间输注G-CSF动员的自体脾细胞对移植物抗宿主病(GVHD)的影响,并初步探讨其可能机制。方法:将40只造血干细胞移植后的小鼠随机分为4组(n=10):GVHD阳性对照组(control group)、移植后1 d受者细胞输注组(+1 d group)、移植后4 d受者细胞输注组(+4 d group)、移植后7 d受者细胞输注组(+7 d group)。输注3×10~7的G-CSF动员后的受者脾细胞,观察GVHD临床体征及病理变化,并检测各组外周血中CD3~+CD4~+、CD3~+CD8~+细胞亚群及其FasL的表达变化。结果:移植后4 d组的GVHD发生率明显降低,中位存活时间>60 d,显著高于对照(24 d)、移植后1 d(21 d)和移植后7 d组(28 d)(P<0.01),而外周血中T淋巴细胞Fasl的表达显著低于其他3组(P<0.05)。结论:单倍体相合的造血干细胞移植后4 d,输注G-CSF动员的受者脾细胞能抑制供者T淋巴细胞的FasL的表达,显著减少GVHD的发生。
Objective: To explore the effect of infusing G-CSF mobilized recipient spleen cells at different time after haploidentical stem cell transplantation( HSCT) on graft-versus-host disease( GVHD) in mice and its possible mechanism.Methods: Forty mice after HSCT were randomly divided into 4 groups( n = 10) : GVHD positive control group( control group),1st d recipient cell infusion group after transplantation(+1 d group),4th d recipient cell infusion group after transplantation(+4 d group),7th d recipient cell infusion group after transplantation(+7 d group). The mice in control group were injected the normal saline of same equivalent with experimental group which were given the same amount of G-CSF-mobilized recipient spleen cells. The general manifestation and pathological change of GVHD were observed. The expression changes of CD3~+CD4~+,CD3~+CD8~+cell subsets and FasL in peripheral blood were detected by flow cytometry. Results: The incidence of GVHD was significantly decreased in+4 d group and the median survival time was longer than 60 days,which was significantly higher than that of control group( 24 d),+1 d group( 21 d),+7 d group( 28 d).( P < 0. 01,P < 0. 01,P < 0. 01). The Fasl expression of peripheral blood T lymphocytes in+4 d group were significantly lower than that in the other 3 groups( P < 0. 05). Conclusion: The+4 d infusion of G-CSF mobilized recipient spleen cells on 4th day after haploidentical HSC transplantation can inhibit the expression of FasL in donor T lymphocytes,and significantly reduce the incidence of GVHD.
Objective: To explore the effect of infusing G-CSF mobilized recipient spleen cells at different time after haploidentical stem cell transplantation( HSCT) on graft-versus-host disease( GVHD) in mice and its possible mechanism.Methods: Forty mice after HSCT were randomly divided into 4 groups( n = 10) : GVHD positive control group( control group),1st d recipient cell infusion group after transplantation(+1 d group),4th d recipient cell infusion group after transplantation(+4 d group),7th d recipient cell infusion group after transplantation(+7 d group). The mice in control group were injected the normal saline of same equivalent with experimental group which were given the same amount of G-CSF-mobilized recipient spleen cells. The general manifestation and pathological change of GVHD were observed. The expression changes of CD3~+CD4~+,CD3~+CD8~+cell subsets and FasL in peripheral blood were detected by flow cytometry. Results: The incidence of GVHD was significantly decreased in+4 d group and the median survival time was longer than 60 days,which was significantly higher than that of control group( 24 d),+1 d group( 21 d),+7 d group( 28 d).( P < 0. 01,P < 0. 01,P < 0. 01). The Fasl expression of peripheral blood T lymphocytes in+4 d group were significantly lower than that in the other 3 groups( P < 0. 05). Conclusion: The+4 d infusion of G-CSF mobilized recipient spleen cells on 4th day after haploidentical HSC transplantation can inhibit the expression of FasL in donor T lymphocytes,and significantly reduce the incidence of GVHD.
引文
1 Magenau J,Runaas L,Reddy P.Advances in understanding the pathogenesis of graft-versus-host disease.Br J Haematol,2016;173(2):190-205.
2 Stikvoort A,Gertow J,Sundin M,et al.Chimerism patterns of long-term stable mixed chimeras posthematopoietic stem cell transplantation in patients with nonmalignant diseases:follow-up of longterm stable mixed chimerism patients.Biol Blood Marrow Transplant,2013;19(5):838-844.
3 Ringden O,Erkers T,Aschan J,et al.A prospective randomized toxicity study to compare reduced-intensity and myeloablative conditioning in patients with myeloid leukaemia undergoing allogeneic haematopoietic stem cell transplantation.J Intern Med,2013;274(2):153-162.
4 Frassoni F,Strada P,Sessarego M,et al.Mixed chimerism after allogeneic marrow transplantation for leukaemia:correlation with dose of total body irradiation and graft-versus-host disease.Bone Marrow Transplant,1990;5(4):235-240.
5 Cooke KR,Kobzik L,Martin TR,et al.An experimental model of idiopathic pneumonia syndrome after bone marrow transplantation:I.The roles of minor H antigens and endotoxin.Blood,1996;88(8):3230-3239.
6 Billingham RE.The biology of graft-versus-host reactions.Harvey Lect,1966-1967;62:21-78.
7 Choi SW,Reddy P.Current and emerging strategies for the prevention of graft-versus-host disease.Nat Rev Clin Oncol,2014;11(9):536-547.
8 Briones J,Novelli S,Sierra J.T-cell costimulatory molecules in acute-graft-versus host disease:therapeutic implications.Bone Marrow Res,2011;2011:976793.
9 Burroughs L,Storb R.Low-intensity allogeneic hematopoietic stem cell transplantation for myeloid malignancies:separating graft-versusleukemia effects from graft-versus-host disease.Curr Opin Hematol,2005;12(1):45-54.
10 Balon J,Halaburda K,Bieniaszewska M,et al.Early complete donor hematopoietic chimerism in peripheral blood indicates the risk of extensive graft-versus-host disease.Bone Marrow Transplant,2005;35(11):1083-1088.
11 Ildstad ST,Sachs DH.Reconstitution with syngeneic plus allogeneic or xenogeneic bone marrow leads to specific acceptance of allografts or xenografts.Cah Rev The,1984;307(5947):168-170.
12 邓晓辉,梁辉,严云,等.移植物与受体T细胞的平衡对移植物抗宿主病影响的实验研究.上海医学,2005;28(4):312-314.
13 魏家臣.Fas/Fas L介导凋亡细胞机制及其病理生理意义(文献综述).国外医学.外科学分册,2003;30(1):21-23.
14 Via CS,Nguyen P,Shustov A,et al.A major role for the Fas pathway in acute graft-versus-host disease.J Immunol,1996;157(12):5387-5393.
15 Shustov A,Nguyen P,Finkelman F,et al.Differential expression of Fas and Fas ligand in acute and chronic graft-versus-host disease:up-regulation of Fas and Fas ligand requires CD8+T cell activation and IFN-gamma production.J Immunol,1998;161(6):2848-2855.
16 Braun MY,Lowin B,French L,et al.Cytotoxic T cells deficient in both functional fas ligand and perforin show residual cytolytic activity yet lose their capacity to induce lethal acute graft-versus-host disease.J Exp Med,1996;183(2):657-661.
17 冯晓勤,周淑芸,刘启发,等.T细胞表达Fas L与急性移植物抗宿主病相关性研究.中华血液学杂志,2002;23(10):542-543.
18 Leskovar P,Pretnar G.Prevention of the graft-versus-host disease(Gv HD)by a controlled host-versus-graft reaction(Hv GR)or by specially premanipulated third-party effector cells.Some advantages of the combined allogeneic and autologous bone marrow transplantation,as deduced from animal experiments.Hampshire:Stock Ton Press,1998:35-36.
19 Hardy NM,Hakim F,Steinberg SM,et al.Host T cells affect donor T cell engraftment and graft-versus-host disease after reduced-intensity hematopoietic stem cell transplantation.Biol Blood Marrow Transplant,2007;13(9):1022-1030.
2 Stikvoort A,Gertow J,Sundin M,et al.Chimerism patterns of long-term stable mixed chimeras posthematopoietic stem cell transplantation in patients with nonmalignant diseases:follow-up of longterm stable mixed chimerism patients.Biol Blood Marrow Transplant,2013;19(5):838-844.
3 Ringden O,Erkers T,Aschan J,et al.A prospective randomized toxicity study to compare reduced-intensity and myeloablative conditioning in patients with myeloid leukaemia undergoing allogeneic haematopoietic stem cell transplantation.J Intern Med,2013;274(2):153-162.
4 Frassoni F,Strada P,Sessarego M,et al.Mixed chimerism after allogeneic marrow transplantation for leukaemia:correlation with dose of total body irradiation and graft-versus-host disease.Bone Marrow Transplant,1990;5(4):235-240.
5 Cooke KR,Kobzik L,Martin TR,et al.An experimental model of idiopathic pneumonia syndrome after bone marrow transplantation:I.The roles of minor H antigens and endotoxin.Blood,1996;88(8):3230-3239.
6 Billingham RE.The biology of graft-versus-host reactions.Harvey Lect,1966-1967;62:21-78.
7 Choi SW,Reddy P.Current and emerging strategies for the prevention of graft-versus-host disease.Nat Rev Clin Oncol,2014;11(9):536-547.
8 Briones J,Novelli S,Sierra J.T-cell costimulatory molecules in acute-graft-versus host disease:therapeutic implications.Bone Marrow Res,2011;2011:976793.
9 Burroughs L,Storb R.Low-intensity allogeneic hematopoietic stem cell transplantation for myeloid malignancies:separating graft-versusleukemia effects from graft-versus-host disease.Curr Opin Hematol,2005;12(1):45-54.
10 Balon J,Halaburda K,Bieniaszewska M,et al.Early complete donor hematopoietic chimerism in peripheral blood indicates the risk of extensive graft-versus-host disease.Bone Marrow Transplant,2005;35(11):1083-1088.
11 Ildstad ST,Sachs DH.Reconstitution with syngeneic plus allogeneic or xenogeneic bone marrow leads to specific acceptance of allografts or xenografts.Cah Rev The,1984;307(5947):168-170.
12 邓晓辉,梁辉,严云,等.移植物与受体T细胞的平衡对移植物抗宿主病影响的实验研究.上海医学,2005;28(4):312-314.
13 魏家臣.Fas/Fas L介导凋亡细胞机制及其病理生理意义(文献综述).国外医学.外科学分册,2003;30(1):21-23.
14 Via CS,Nguyen P,Shustov A,et al.A major role for the Fas pathway in acute graft-versus-host disease.J Immunol,1996;157(12):5387-5393.
15 Shustov A,Nguyen P,Finkelman F,et al.Differential expression of Fas and Fas ligand in acute and chronic graft-versus-host disease:up-regulation of Fas and Fas ligand requires CD8+T cell activation and IFN-gamma production.J Immunol,1998;161(6):2848-2855.
16 Braun MY,Lowin B,French L,et al.Cytotoxic T cells deficient in both functional fas ligand and perforin show residual cytolytic activity yet lose their capacity to induce lethal acute graft-versus-host disease.J Exp Med,1996;183(2):657-661.
17 冯晓勤,周淑芸,刘启发,等.T细胞表达Fas L与急性移植物抗宿主病相关性研究.中华血液学杂志,2002;23(10):542-543.
18 Leskovar P,Pretnar G.Prevention of the graft-versus-host disease(Gv HD)by a controlled host-versus-graft reaction(Hv GR)or by specially premanipulated third-party effector cells.Some advantages of the combined allogeneic and autologous bone marrow transplantation,as deduced from animal experiments.Hampshire:Stock Ton Press,1998:35-36.
19 Hardy NM,Hakim F,Steinberg SM,et al.Host T cells affect donor T cell engraftment and graft-versus-host disease after reduced-intensity hematopoietic stem cell transplantation.Biol Blood Marrow Transplant,2007;13(9):1022-1030.