Immune Cell Restoration Monitoring After Hematopoietic Stem Cell Transplantation And Its Clinical Significance

Objective: To evaluate ites clinical significance by analyse the variations of cell subset numbers after allogeneic hematopoietic stem cell transplantation(allo-HSCT).Methods:1 A total number of 21 patients underwent allo-HSCT from 2013-7 to 2014-12, in the Blood and Marrow Transplanation Center, Department of Hematology, The Second Hospital of Hebei Medical University.2 The clinical condition of patients. Age:the median age of patients was 35 years old(13-57 years old).Gender: 9 cases were male, 12 cases were femal. Diagnosis and staging: 8 cases were AML( 6 of them were CR1, 2 of them were CR2), 6 cases were ALL( 5 of them were CR1, 1 of them was CR2), 6 cases were MDS(all of them were MDS-RAEB, International Prognostic Scoring Syetem(IPSS): 1 of them was Int-1, 5 of them were Int-2), 1 case was Lymphoma cell leu-kemia(LCL),1 case was SAA.Observation time:1 to 12 months after transplantation.Transplantation mode: 2 cases underwent MUD,10 cases underwent MSD,9 cases underwent Haplo-HSCT.Unrelated donors use peripheral stem cells,haplo-HSCT/MSD use bone marrow and peripheral stem cells.Healthy controls were 7 healthy donors.Sample collection:at the time of 1,2,3,4.5,6,9,12 months after transplantation,collected EDTA anticoagulated peripheral blood 4ml.The control group collected peripheral blood before using G-CSF.3 FACS Cantoll flow cytometry was used to analyse immune cell Ⅱsubsets’ changes. Antibody combination: Tlymphocytes(CD3+, CD3+CD4+, CD3+CD8+,CD4+CD25+Fox P3+), B cells(CD19+),NK cells(CD16+CD56+), m DC(CD123-CD11c+), p DC(CD123+CD11c-). Measurement data apply two independent sample t-test.Results: 1 20 of the patients had rapid hematopoietic resconstitution. The median time when absolute neutrophil counts(ANC) were more than 0.5×109 /L were 12 days(11-19days),the median time of platelet count more than 20×109/L were 12days(11-16days). 2 Recovery of immune cells 2.1 CD3+ cells count at 1 month post transplant were significantly lower than control group(896.69±124.08/ul vs 1426.14±226.85/ul,P<0.001). At 4.5 month after transplantation, the number of CD3+ cells count in transplantation group vs that in control group(1202.50±235.35/ul vs 1426.14±226.85/ul,P=0.159). CD3+CD4+ cells count recover slowly in transplantation group, at 1 month after transplantation it was lower than that in control group(157.85±47.50/ul vs 889.43±148.18/ul, P < 0.001),at 12 months after transplantation it was still below than that in control group(691.50±97.55/ul vs 889.43±148.18/ul, P=0.018).At 1 month after transplantation the CD3+CD8+ cells count in transplantation group vs that in control group(429.27±171.29/ul vs 553.29±111.59/ul,P=0.110). Treg cells count in transplantation group at 12 months after transplantation was still significantly lower than that in control group(10.13±3.02/ul vs 17.86±7.91/ul,P=0.046).CD3+CD8+ cells count recovery quickly,CD3+ cells count in transplantation group vs that in normal at 4.5 month post transplant.While,CD3+CD4+ and Treg cells counts recovery slowly, until 12 month after transplantation it were still lower than that in control group. 2.2 CD19+ cells count in transplantation at 1 month after transplantation was significantly lower than that in control group( 23.68±6.84/ul vs 238.14±62.51/ul, P < 0.001), to 12 months returned to normal(214.17±60.18/ul vs 238.14±62.51/ul,P=0.381). 2.3 NK cells count recover rapidly after transplantation,at 1 month after transplantation recover to the same level as that in control group(249.77±64.70/ul vs 293.64±64.59/ul,P=0.604). 2.4 The m DC cells count recover slowly, at 6 month after transplantation it was still below than that in control group(7.23±3.22/ul vs 15.07±5.10/ul,P=0.006),in 12 month approaching the same level as that of normal group(12.29±3.40/ul vs 15.07±5.10/ul,P=0.412).The p DC cells recover slowly, in 12 months after transplantation near to normal. 3 Recovery of immune cells associated with acute GVHD. 5 of the 21 patients(23.80%) developed severe a GVHD.In the absolute number of CD3+ and CD3+CD8+ cells in patients with a GVHD were significantly higher than that in those without severe a GVHD(P<0.05),and CD3+CD4+ and Treg cells in patients with a GVHD were significantly lower than that in whose without severe a GVGD(P<0.05). 4 Recovery of immune cells and the possibility of relapse.In 21 patients,3 cases of them recurrence(14.29%), at the time of 30,124,135 days after transplantation. 2 cases of them monitored T lymphocyte to 3 months. CD3+,CD3+CD8+ cells in relapse group were lower than the average levels in remission group at three monitored time.CD3+CD4+、Treg cells were in relapse group were higer than the average levels of patients without ralapse at 1 and 2 month. 5 Recovery of immune cells and HLA-indentical degree. At 1 to 3 months after transplantation CD3+CD4+, CD3+CD8+, Treg, NK cells have significantly difference between MUD/MSD and haplo-HSCT(P <0.05).Compared MUD/MSD with haplo-HSCT, CD3+CD4+, Treg cells in MUD/MSD were significantly high than that in haplo-HSCT, CD8+ cells in MUD/MSD were significantly lower than that in haplo-HSCT. B and NK cells in MUD/MSD were significantly higher than that in haplo-HSCT. At the pointed of CD3+CD4+, Treg, CD19+, NK cells in MUD/MSD recover more quickly than that in haplo-HSCT.Conclusion:1 CD3 + cells recovered to normal at 4.5 months after allo-HSCT. CD3 + CD8 + cell recovery faster than CD3 + CD4 + and Treg cells. CD19 + B cells and DC cells returned to normal at 12 months after transplantation. CD16 + CD56 + NK cells were quickly restored, at 1 month after transplantation, it become to normal level.2 Patients who have severe acute GVHD,CD3+CD8+ cells were higer than those who did not.CD3+CD4+ and Treg cells were on the contrary.3 Compared MUD/MSD with haplo-HSCT,1 to 3 months after transplantation, CD3+CD4+, CD3+CD8+, Treg, NK cells have significantly difference between MUD/MSD and haplo-HSCT(P<0.05).Among them CD3+CD8+ cells in MUD/MSD were decreased when comparing with that in haplo-HSCT group(P<0.05).