Alterations Of Cytokines And Platelet Function During Hematopoietic Stem Cell Transplantation

Objective:(1)To study alterations of cytokines TNF-α, IL1-βand IFN-γin early phase of allogeneic hematopoietic stem cell transplantation and in the course of conditioning treatment, then to study relationship between cytokines and transplant-related complications acute graft-versus-host disease (aGVHD), thrombosis and infection.(2)To study the changes of platelet aggregation function and platelet membrance glycoprotein after conditioning treatment in patients undergoing hematopoietic stem cell transplantation, then to analyze the significance of these alterations. Methods:1 Detection of cytokines1.1 Patient characteristicsFrom June 2006 to July 2008, 95 patients undergoing Allo-HSCT in our hospital were enrolled in this study. There were 65 males and 30 females, aged from 12 to 60 years old (median age was 32 years), AML in 28 cases, ALL in 24 cases, CML in 29 cases, other diseases in 14 cases (lymphoma, MDS, AA et al). 54 patients received transplants from match sibling donors, 29 from unrelated donors, 12 from partially matched family donors. The condition regimen mainly included modified BU/CY and TBI plus CY according to the type of the donor and disease. All HSCT patients were divided into aGVHD group ( 43 cases), thrombus group (TMA, HVOD, arterial and phlebothrombosis, 5cases) and infection group (31cases).1.2 Specimen collection and cytokines detectionThe serum specimen of all patients undergoing allogeneic hematopoietic stem cell transplant were collected once a week in the morning from preconditioning, 4 days after preconditioning until the time when they were discharged from hospital, 20 healthy adults were detected as normal controls. Alterations of serum concentration of TNF-α, IL1-β, and IFN-γwere evaluated by ELISA during conditioning treatment and each week after transplantation, changes of TNF-α, IL1-β, and IFN-γwere detected during the process of conditioning treatment.2 Detection of platelet aggregation and platelet glycoprotein2.1 Patient characteristicsFrom August 2007 to April 2008, 29 patients undergoing HSCT were enrolled in this study. There were 17 males and 12 females, aged from 14 to 55 years old (median age was 31 years). AML in 10 cases, ALL in 11 cases, CML in 3 cases, other diseases in 5 cases. 13 patients received transplants from match sibling donors, 8 from unrelated donors, 4 from partially matched family donors, 4 received autogenous transplantation. The preconditioning regimen included modified BU/CY, TBI plus CY and BEAM(for autogenous transplantation). Regimen such as CSA+MTX (or plus ATG/MMF) were used to prevent aGVHD.2.2 Method for Detection of platelet aggregation and platelet membrane glycoprotein Platelet glycoprotein GPⅠb, GPⅡb, GPⅢa and CD62P were analyzed by flow cytometry in 29 patients undergoing hematopoietic stem cell transplantation pre and post conditioning treatment, 15 healthy people were detected as as normal controls. Platelet aggregation was detected at the same time, Anticoagulant Adenosine diphosphate (ADP) were utilized to promote platelet aggregation.Results: (1) TNF-αlevels in aGVHD patients undergoing Allo-HSCT were already higher than normal controls before conditioning treatment (P﹤0.01), other patients did not significantly changed during this course, levels of TNF-αduring the conditioning treatment and 1 or 4 weeks after transplantation also increased.TNF-αlevels in all patients were even higher 4 days after conditioning treatment began followed by a tendency of a reduced level during conditioning treatment (P﹤0.05). levels of TNF-αincreased in aGVHD thrombotic and infected patients once complication happened, TNF-αlevels in aGVHD patients increased most significantly, and TNF-αin thrombtic patients increased more significantly than infected patients (P﹤0.05). aGVHD and thrombotic patients began to increase two weeks before complication developed, levels of TNF-αin infected patients did not change at the same time. IL1-βlevels did not change during conditioning treatment. levels of IL1-βin aGVHD and thrombotic patients had already increased 1 and 2 weeks after transplantation. IL1-βin 2-4°aGVHD were most prominent before transplantation and 4 weeks after transplantation.IL1-βincreased in aGVHD, thrombotic and infected patients once complication happened, levels of IL1-βin thrombtic patients increased most significantly, and IL1-βincreased more significantly in aGVHD patients than infected patients (P﹤0.01). aGVHD patients began to increase one week before complication developed and thrombotic patients began to increase two weeks before complication developed. IFN-γlevels did not change in all patients during the process of transplantion.(2)Platelet membrane glycoprotein GPⅡb, GPⅢa and CD62P increase obviously than normal controls and preconditioning levels after conditioning(P﹤0.05), GPⅠb did not changed at the same time. Platelet aggregation to ADP was 56.73%±20.38 % before conditioning, almost the same as normal people. the maximal platelet aggregation decline significantly to 31.38%±25.81% after conditioning (P﹤0.01).Number of platelets decreased significantly during conditioning treatment.Conclusion:(1)Changes of Cytokines happen during the course of allo-HSCT, TNF-αlevels are affected by preconditioning treatment, reflecting the vascular damage and onset of some transplant associated complications. Levels of TNF-αand IL1-βare closely related to aGVHD or thrombotic complications. Monitoring changes of TNF-αand IL1-βlevels contributes to early discovery of aGVHD and thrombotic complications. Increase of TNF-αand IL1-βin the early phase of transplantion plays an important role in aGVHD and thrombosis.(2)Platelet aggregation dysfunction and activation of platelet happen in patients undergoing hematopoietic stem cell transplantation after preconditioning treatment, rearch on platelet function has broad prospects in the future.