CAJ | 학술논문

目的探讨角质细胞生长因子(KGF)在小鼠异基因脐血移植(UCBT)后免疫重建中的作用及其机制.方法取孕19.5 d胎鼠外周血作为脐血移植物.第1批UCBT实验中32只BALB/c小鼠随机分为4组,每组8只,分别输注PBS、1×10~6、2×10~6、3×10~6个脐血单个核细胞(MNC).第2批UCBT实验中24只BALB/c小鼠随机分为3组,每组8只,分别输注1×10~6、2×10~6、3×10~6个脐血MNC,所有小鼠+8 d输注1×10~9个血小板支持.第3批UCBT实验中16只BALB/c小鼠随机分为2组,每组8只,KGF组小鼠TBI前注射KGF,对照组注射PBS,两组都输注2×10~6个脐血MNC,+8 d输注血小板支持.以生存期、脾脏淋巴细胞亚群、胸腺输出功能为观察指标.结果第1批UCBT实验中PBS组小鼠+13.5 d内全部死亡,输注1×10~6、2×10~6、3×10~6个MNC三组小鼠+100 d分别生存2只、3只、3只.第2批UCBT中输注1×10~6、2×10~6、3×10~6个MNC三组小鼠+100 d分别生存7只、8只、8只.第3批UCBT实验中,移植后对照组小鼠脾T细胞、NKT细胞、NK细胞和B细胞数分别为(9.57±0.74)×10~6、(0.64±0.06)×10~6、(1.43±0.10)×10~6、(19.13±1.50)×10~6个.KGF输注组分别为(13.47±0.74)×10~6、(0.89±0.03)×10~6、(1.79±0.04)×10~6、(20.50±0.91)×10~6个.KGF输注组T细胞、NKT细胞、NK细胞较对照组高(P＜0.05);对照组小鼠sjTREC水平为(182.2±10.7)拷贝/105个细胞;KGF输注组为(224.2±9.6)拷贝/105个细胞,KGF输注组明显高于PBS对照组(P=0.019).结论孕19.5 d胎鼠外周血富含造血干细胞,+8 d输注血小板浓缩物可建立异基因UCBT小鼠模型.KGF具有增强小鼠异基因UCBT后胸腺输出功能及促进T细胞免疫重建作用.
목적 탐토각질세포생장인자(KGF)재소서이기인제혈이식(UCBT)후면역중건중적작용급기궤제.방법 취잉19.5 d태서외주혈작위제혈이식물.제1비UCBT실험중32지BALB/c소서수궤분위4조,매조8지,분별수주PBS、1×10~6、2×10~6、3×10~6개제혈단개핵세포(MNC).제2비UCBT실험중24지BALB/c소서수궤분위3조,매조8지,분별수주1×10~6、2×10~6、3×10~6개제혈MNC,소유소서+8 d수주1×10~9개혈소판지지.제3비UCBT실험중16지BALB/c소서수궤분위2조,매조8지,KGF조소서TBI전주사KGF,대조조주사PBS,량조도수주2×10~6개제혈MNC,+8 d수주혈소판지지.이생존기、비장림파세포아군、흉선수출공능위관찰지표.결과제1비UCBT실험중PBS조소서+13.5 d내전부사망,수주1×10~6、2×10~6、3×10~6개MNC삼조소서+100 d분별생존2지、3지、3지.제2비UCBT중수주1×10~6、2×10~6、3×10~6개MNC삼조소서+100 d분별생존7지、8지、8지.제3비UCBT실험중,이식후대조조소서비T세포、NKT세포、NK세포화B세포수분별위(9.57±0.74)×10~6、(0.64±0.06)×10~6、(1.43±0.10)×10~6、(19.13±1.50)×10~6개.KGF수주조분별위(13.47±0.74)×10~6、(0.89±0.03)×10~6、(1.79±0.04)×10~6、(20.50±0.91)×10~6개.KGF수주조T세포、NKT세포、NK세포교대조조고(P＜0.05);대조조소서sjTREC수평위(182.2±10.7)고패/105개세포;KGF수주조위(224.2±9.6)고패/105개세포,KGF수주조명현고우PBS대조조(P=0.019).결론 잉19.5 d태서외주혈부함조혈간세포,+8 d수주혈소판농축물가건립이기인UCBT소서모형.KGF구유증강소서이기인UCBT후흉선수출공능급촉진T세포면역중건작용.
Objective To explore the impact and mechanism of keratinocyte growth factor (KGF) on immune reconstitution post murine allogeneic umbilical cord blood transplantation (UCBT).Methods Perpheral blood (PB) from 19.5-day embryos post-conception(E 19.5 d) mice was used as umbilical cord blood (UCB) graft.Thirty-two BALB/c mice were randomly assigned to 4 groups with 8 mice each in the first cohort UCBT.Mice were infused with PBS(control group) or 1×10~6 (group 1A),2×10~6 (group 1B),3×10~6 UCB mononuclear cells (MNCs)(group 1C),respectively.Twenty-four BALB/c mice were randomly assigned to 3 groups with 8 mice each in the second cohort UCBT.Mice were injected with 1×10~6 (group 2A),2×10~6 (group 1 B) or 3×10~6 (UCB) M NCs (group 2C).All mice received platelet transfusion on + 8d.Sixteen BALB/c mice were randomly assigned to 2 groups with 8 mice each in the third cohort UCBT.Mice were injected s.c.with KGF(group 3) or PBS (control group) before TBI.All mice were injected with 2×10~6 UCB MNCs and were supported with platelet transfusion on + 8 d.The survival time,splenic lymphoid cell subsets,sjTREC assay were observed after UCBT.Results The 100-day survival of mice were 2,3 and 3 in group 1A,1B,1C and 7,8,8 in group 2A,2B,2C,respectively.The splenic T,NKT,NK and B cell counts on +35dwere (9.57±0.74)×10~6,(0.64±0.06)×10~6,(1.43±0.10)×10~6 and (19.13±1.50)×106 in control group,respectively;while were (13.47±0.74)×10~6,(0.89±0.03)×10~6,(1.79±0.04)×10~6 and (20.50±0.91)×10~6 in group 3,respectively,being significantly higher than in control group.The sjTREC level was 182.2±10.7 copies per 105 cells in control group;while was 224.2±9.6 copied per 105 cells in group 3,being significantly higher than in control group (P = 0.019).Conclusions Peripheral blood from E19.5d is rich in hematopoietic stem cells.A murine allogeneic UCBT model with platelet support on + 8 d is established.KGF treatment can enhance thymic output and improve T cell immune reconstitution after UCBT.