Clinical Significance Of Protein Chip Technology In Detection Of 22 Serum Biomarkers In Patients With AGVHD

Objective:Allogeneic hematopoietic stem cell transplantation(allo-HSCT)is an effective method to treat a variety of hematologic malignancies.Acute graft versus host disease(aGVHD)is a common complication after allogeneic hematopoietic stem cell transplantation,which seriously affects the quality of life and survival rate of patients,which is the main cause of non recurrent death in allo-HSCT patients.At present,the diagnosis of aGVHD mainly depends on the clinical manifestations and pathological examination of the damaged target organs.But its clinical symptoms are various and lack of specificity,and pathological examination often because of various reasons can not be carried out in clinical diagnosis and treatment,so that the timeliness of its diagnosis was affected.Therefore,to explore some of the clinical practical,noninvasive,fast monitoring methods of aGVHD,so that patients in the early of the immune activation state can be diagnosed and treated as early as possible,so that more effective control of aGVHD,which is an urgent need to solve clinical problem.Protein chip technology is a new technology which combines microelectronics,micro mechanics,chemistry and life science.And it has the characteristics of high throughput,high specificity,high sensitivity,miniaturization,automation and so on.In this study,22 serum biomarkers of aGVHD and non aGVHD patients after allogeneic hematopoietic stem cell transplantation were detected by sandwich membrane protein chip technology.And to preliminary screening of molecular markers that related to the development and progression of aGVHD,and to lay the foundation for clinical early intervention and prognosis evaluation of aGVHD.In the initial screening and the occurrence and development of aGVHD and prognosis related molecular markers,and lay the foundation for the clinical early intervention and prognosis of post aGVHD.Method:In 7days before transplantion and 7days,14 days,28 days,56 days,100 days after transplantation,continuous point of collection the morning fasting venous blood of patients with allo-HSCT in November 2014 to November 2015 in the Department of Hematology,Xinqiao Hospital,Third Military Medical University,were observed for a period of 100 days after transplantation.10 patients with aGVHD were selected as the experience group,and 10 patients without aGVHD were randomly selected as the control group.Protein chip technology was used to detect the content of 22 kinds of serum biomarkers(IL-1β,IL-2,IL-4,IL-6,IL-7,IL-8,IL-10,IL-12p40,IL-17 A,IL-21,IL-22,IL-23,MIF,TNF-α,TGF-β,IFN-γ,Elafin,ST2,TLR4,TNFR1,vWF,TF)in two groups of patients at different time points before and after transplantation.Using AAH-CUST data analysis software to detect the results of data pre analysis,and T test,the data expressed as mean ± standard(?)deviation.Results:1.According to protein chip detection,Comparison of aGVHD group and non aGVHD group,there was no significant difference between the two groups in the seventh days before transplantation(P>0.05);Seventh days after transplantation,the levels of IL-2,IL-17 A,ST2,TLR4 in aGVHD group were significantly higher than those in non aGVHD group(P<0.05);Fourteenth days after transplantation,the levels of IL-2,IL-4,IL-17 A,IL-21,IL-23,TNFR1 in aGVHD group were significantly higher than those in non aGVHD group(P<0.05),while the level of IFN-γ was significantly lower than that of non aGVHD group(P<0.05);Twenty-eighth days after transplantation,the levels of IL-2,IL-7,IL-17 A,TNFR1,aGVHD group was significantly higher than that in non aGVHD group(P<0.05),the levels of IFN-γ was significantly lower than those in non aGVHD group(P<0.05);Fifty-sixth days after transplantation,the levels of IL-1β,IL-2,IL-4,IL-7,IL-17 A,IL-21,IL-23,ST2,TF in aGVHD group were significantly higher than those in non aGVHD group(P<0.05);100th days after transplantation,the levels of IL-7,IL-23 and TF in aGVHD group were significantly higher than those in non aGVHD group(P<0.05).2.Comparison between Ⅲ-Ⅳ class aGVHD and II class aGVHD: Seventh days before transplantation,the serum level of IL-8 in the former was significantly higher thanthat in the latter(P<0.05);Seventh days after transplantation,the serum levels of IL-1 and IL-22 were significantly higher in the former than the latter(P<0.05);There was no significant difference in serum marker levels between the two groups(P>0.05)at fourteenth days after transplantation;Twenty-eighth days after transplantation,the serum level of TLR4 was significantly higher in the former than that of the latter(P<0.05);Fifty-sixth days after transplantation,the serum levels of TNFR1 were significantly higher in the former than that of the latter(P<0.05);At 100 th days after transplantation,the serum levels of IL-21 were significantly lower than the latter(P<0.05).3.Comparison of 22 serum biomarkers in patients with aGVHD before and after treatment,it was found that there was a significant decrease in the level of TLR4 in the patients with aGVHD after treatment(P<0.05).4.Comparison of aGVHD patients with poor effect of hormone therapy and good curative effect: Seventh days before transplantation,the serum level of IL-7 in the former was significantly lower than that in the latter(P<0.05);Seventh days after transplantation,the serum levels of IL-17 were significantly lower in the former than the latter(P<0.05);There was no significant difference in serum marker levels between the two groups(P>0.05)at fourteenth days after transplantation;Twenty-eighth days after transplantation,the serum level of IL-23 was significantly lower in the former than that of the latter(P<0.05);Fifty-sixth days after transplantation,the serum levels of IL-6 and IL-12p40 were significantly higher in the former than that of the latter(P<0.05);At 100 th days after transplantation,the serum levels of IL-8,IL-22,ST2 and TNFR1 were significantly higher than the latter(P<0.05).Conclusions:1.Protein chip detection aGVHD group compared with non GVHD group,in before and after transplantation at the different time point expression have significant differences in 12 serum biological markers,it is closely related with the occurrence and development of aGVHD.They can be used as auxiliary diagnostic indexes of aGVHD,and is helpful for the early diagnosis and intervention of aGVHD.2.Compared with II grade aGVHD and Ⅲ-Ⅳ grade aGVHD has five serum markers with high expression and a low expression,indicating that the expression level of the sixserum markers may be related to the severity of aGVHD,which can be used as the basis for the diagnosis of Ⅲ-Ⅳ grade aGVHD,and provide reference for clinical treatment.3.TLR4 serum levels after treatment significantly decreased,indicating a better therapeutic effect for aGVHD patients,and provide a new means of pre judgment for the immune targeted therapy of aGVHD.4.3 serum markers(IL-7,IL-17 A,IL-23)low expression and 6 serum markers(IL-6,IL-8,IL-12p40,IL-22,ST2,TNFR1)high expression may be related to the effect of hormone therapy in aGVHD,and it is a reference index to predict whether the patient is resistant to the hormone.