Dynamic Variation And Clinical Significance Of The Cytokine’s Level Changes Of Children With Acute Leukemia During Chemotherapy

BackgroundStudies have shown that cytokines and the mechanism of tumor how it occurred and developed are very closely related in recent years, acute leukemia is a hematological malignancy. Studies have shown that the expression and regulation of cytokine abnormalities may be involved in the occurrence and development of leukemia in recent years, such as cytokines play anti-tumor effect. But the tumor cells inhibit the function of immune cells through the secretion of inhibitory cytokines, which lead to immunocompromised state or immunosuppressive state. It can inhibit the host’s anti-tumor immune response in the immune response induced effect on multiple links. Cellular immunity plays an important role in the anti-tumor immune response, which has been confirmed in the incidence and the development process of leukemia. But leukemia etiology and pathogenesis is not yet fully elucidated, it is not only the heavy financial and emotional burden of the disease on the family and society, but also seriously affect the future quality of life of the children, in order to further improve quality of life of children leukemia, exports research focus in the field of pathogenesis and specific treatment measures of children with acute leukemia, which is not only needed to improve the cure rate of acute leukemia, but also to protect our limited medical resources. Once the body occurs inflammation response, there will produce cytokines such as interleukin which promote the neutrophil oozing inflammation localized theoretically speaking. Cytokines and infection are closely related. Clinical work also has been more studies confirm the close contact between cytokines and infection at home and abroad. Now the hot spot is the indexes which can early diagnosis and prediction of prognosis of febrile neutropenia. We must increase the cure rate of febrile neutropenia of clinical work in order to further improve the cure of children with acute leukemia rate, because febrile neutropenia after transplantation or chemotherapy are the main cause of death of patient with leukemia. Patients with febrile neutropenia are lack of specific symptoms,blood cultures has the limited clinical significance due to the limitations of the various technical conditions, serum CRP play an important role in early diagnosis, differential diagnosis, condition assessment and efficacy determine, however, because there is no good specificity, clinical applications have a certain restriction. Now the early judgment of the type of febrile neutropenia in patients with acute leukemia is hard to determine in clinical working. Therefore, there is the need to find to get up early to judge granulocyte lack predictor of febrile neutropenia, cytokines are likely to get up early predictor of infection. The antibiotic treatment is difficult to have a clear purpose; this study provides a theoretical basis that cytokines can be the indicators of early diagnosis and prevention of serious infection. We who have a clear purpose can use antibiotic treatment, to avoid the waste of antibiotics and other medical resources.Aim1.1To evaluate the changes of the immune microenvironment before and after high-dose chemotherapy children with acute leukemia, explore the clinical significance of variation of the level of C-reactive protein （CRP）, interleukin （IL）-6, interleukin-8and interferon （IFN）-γ and examine the correlation between them, in order to provide a theoretical basis for its monitoring indicators can be used as the response to treatment of children with acute leukemia.1.2We investigate the variation of CRP, IL-6, IL-8and IFN-γ of children with acute leukemia before and after febrile neutropenia, and explore the clinical significance of variation of the level of CRP, IL-6, IL-8and IFN-γ, and examine the correlation between CRP and cytokine （including IL-6, IL-8, IFN-γ）。 From the perspective of cytokines, we can explore the relationship between the level of cytokine and the occurrence of febrileneutropenia of children with hematologic malignancies, we also can explore the relationship between the level of cytokine and the developing of febrile neutropenia of children with hematologic malignancies. The level of cytokine provides the basis for early clinical diagnosis, and prevention of serious infections.Methods1.1We selected40cases of children with hematologic malignancies from Department of Pediatrics of Nan fang Hospital of Southern Medical University, who met the standards of this study. The spirit was informed and voluntary principles. We detected levels of cytokines, which include CRP, IL-6, IL-8and IFN-γ. Serum samples were collected before chemotherapy （To）, after chemotherapy3days （T1）, after confirmation of febrile neutropenia,1days（T2）,3days（Ts）,7days after febrile neutropenia （i.e. the use of antibiotics7days, T4）, remission after chemotherapy （T5）.1.2Blood samples following detection:（1） In Nan fang Hospital laboratory, we use SYSMEX XE5000hematology analyzers and ancillary reagents to count of blood cells, specially absolute neutrophil count of the different time points（Unit is109/L）;（2） In Nan fang Hospital laboratory,we use immunonephelometry, which was by Roche automatic electrochemiluminescence immunoassay analyzer（ROCHE COBAS6000） and ROCHE supporting reagents for testing in strict accordance with the conventional steps,to detect40cases of children with acute leukemia with the levels of CRP of the different time points, normal value of CRP is less than5mg/L;（3） In the Kidney transplant immunology laboratory in Zhu Jiang Hospital,we used Luminex immunoassay （Bio-Plex suspension protein chip system, our test used Bio-Plex human cytokinemulti-plex kit and Bio-Plex cytokine reagent kit of Bio-Rad in USA）,which can detect simultaneously in a variety of cytokines in the same specimen,to detect40cases of children with hematologic malignancies of the different time points with levels of cytokines, which include IL-6, IL-8and IFN-y. The reaction occurred on the encoded microspheres of different fluorescent, which included antigen and antibody, ligend and receptor, enzyme and substrate, nucleic acid hybridization reaction. The instrument respectively detected coding and reporter fluorescent microspheres to achieve the purpose of the qualitative and quantitative through red and green laser beams. The Luminex detection cytokines is divided into three steps:step1:Probe molecules fixed; Step2:detection reaction; Third step:laser analysis. The data analysis application was Bio-Plex Manager software （version4.0）, its sensitivity is less than lOpg/ml.It can accurately detect the cytokine concentration; its range is1-32000pg/ml.1.3Statistics analysis:All data were analyzed using the SPSS13.0.All data were expressed as mean±standard deviation （X±S）; each set of data normality test. Between the two groups of the same indicator including IL-6, IL-8and IFN-γ signed2related-samples rank test, date of CRP uses2related-samples T test during group at each time point. Between the two indicators of IL-6, IL-8, IFN-γ and CRP used the Bivariate, correlation between CRP and IL-6used Pearson correlation, and the other two indicators used Spearman correlation. P<0.05presents statistics different.Results1The levels of CRP, IL-6, IL-8, IFN-γ changes of children with acute leukemia before and after high-dose chemotherapy1.1All data of CRP、IL-6、IL-8and IFN-γ were expressed as mean±standard deviation （X±S）, Value of CRP respectively was （4.51±8.42） mg/L、（4.82±8.54） mg/L and （19.82±36.86） mg/L in each time phase To、T1and T5; Value of IL-6respectively was （5.39±7.42） pg/ml、（6.45±9.69） pg/ml and （6.45±9.69）pg/ml in each time phase To、T1and T5; Value of IL-8respectively was （72.38±135.04） pg/ml、（20.76±48.37） pg/ml and （20.76±48.37） pg/ml in each time phase To、T1andT5; Value of IFN-y respectively was （71.24±18.48） pg/ml、（56.47±27.08） pg/ml and （56.47±27.08） pg/ml in each time phase To、T1and T5.1.2Compared T1、T5to T0,the levels of CRP, IL-6were no statistically significant （P>0.05）. Compared T1to To, the levels of IFN-γ and IL-8were statistically significant （P=0.005and P=0.002）; Compared T5to To. Compared T5to T1, the levels of IFN-y and IL-8were no statistically significant （P>0.05）1.3To, T1and T5phase of the levels of IL-6, IL-8, IFN-γ and CRP were not correlated （P>0.05）.2The levels of CRP, IL-6, IL-8, IFN-γ changes of children with acute leukemia before and after febrile neutropenia2.1All data of CRP. IL-6. IL-8and IFN-y were expressed as mean±standard deviation （X±S）.Value of CRP respectively was （36.0±39.73） mg/L、（24.86±39.88） mg/L and （24.19±45.08） mg/L in each time phase T2、T3and T4; Value of IL-6respectively was （30.18±64.6） pg/ml、（17.65±33.93） pg/ml and （15.49±25.08） pg/ml in each time phase T2、T3and T4; Value of IL-8respectively was （14.94±14.8） pg/ml.（15.67±13.8） pg/ml and （18.35±16.25） pg/ml in each time phase T2、T3and T4; Value of IFN-y respectively was （74.45±16.78） pg/ml、（66.19±16.43） pg/ml、（61.83±23.59） pg/ml in each time phase T2、T3and T4.2.2Compared T2、T3to To, the level of CRP significantly increased after infection early in the fever, and the level of CRP increased in24hours. Compared T2. T3to To,the levels of CRP were statistically significant （P<0.05）.Compared T4to To, Compared T4to T2, Compared T4to T3, the levels of CRP were not statistically significant （P>0.05）. There was bivariate correlation analysis for IL-6in each phase T2, T3and T4:here were significant positive correlation between T2and T3（r=0.773, P=0.00）, but T4and T3were not correlated （P>0.05）。2.3Compared T2. T3to To、the levels of IL-6, IL-8were statistically significant （P<0.05）, the level of IL-8decreased after infection, but the level of IL-6level increased in24hours.Compared T4to To, Compared T4to T2, Compared T4to T3, the levels of IL-6, IL-8were not statistically significant （P>0.05）2.4There was bivariate correlation analysis for IL-6in each phase T2, T3and T4: There were significant positive correlation between T2and T3（r=0.453, P=0.023）, There were also significant positive correlation between T2and T4（r=0.720, P=0.006）, butT4and T3were not correlated （P=0.54）。2.5There was bivariate correlation analysis for IL-8in each phase T2, T3and T4: T2and T3, T4and T2, T4and T3were not correlated （P>0.05）.2.6The median level of IFN-y had no significant changed in each time phase To、T2、T3and T4,the difference was not statistically significant （P>0.05）.2.7Correlation between CRP and IL-6:There were significant positive correlation between CRP and IL-6in each time phase T2（CRP-T2/IL-6-T2）、T3（CRP-T3/IL-6-T3） and T4（CRP-T4/IL-6-T4） except the underlying value before chemotherapy （To）（P<0.05）. Correlation coefficients respectively were rcRp-T2/IL-6-T2=0.621, r CRP-T3/IL-6-T3=0.805, r CRP-T4/IL-6-T4-0.955.2.8Correlation between IL-6and IL-8:Correlation between IL-8and IL-6: There were significant positive correlation between CRP and IL-6in each time phase T2（IL-8-T2/IL-6-T2）、T3（IL-8-T3/IL-6-T3） and T4（IL-8-T4/IL-6-T4） except the underlying value before chemotherapy （To）（P<0.05）. Correlation coefficients respectively were r IL₈-T2/IL-6-T2=0.670, r IL.8-T3/IL-6-T3=0.463, r IL-8-T4/IL-6-T4=0.724.2.9T2,T3and T4phase of the levels of CRP and IL-8, CRP and IFN-y, IL-8and IFN-γ, IL-6and IFN-γ were not correlated （P>0.05）.Conclusions1.1The level of IFN-y may reflect the immune status of children with acute leukemia before and after chemotherapy. IFN-y participates in the immune reconstitution after chemotherapy in children with acute leukemia. IFN-y can be used as an indicator of response to therapy monitoring of leukemia in children. The levels of IL-8is involved in leukemia pathogenesis needs further study。1.2CRP、IL-8and IL-6can respectively be used as a laboratory indicator of febrile neutropenia of early diagnosis of children with acute leukemia, the levels of CRP、IL-8and IL-6can be useful markers to identify febrile neutropenia of patients acute leukemia. Compared T2to To, obvious difference between them prompts more serious infection, the longer course of treatment, poor prognosis.1.3The highest value of CRP, IL-6occurred in24hours after fever. There was positive correlation between the level of the highest value of CRP and IL-6and the level of CRP and IL-6in each time phase T3、T4. The higher value of CRP and IL-6in24hours, the slower will them decline.This can determine whether the effectiveness of antibiotic treatment。1.4United CRP with IL-6or united IL-8with IL-6as a diagnosis, determine the efficacy endpoint of the model is worthy of application in clinical work.1.5IFN-y may play the slight role in the patient with febrile neutropenia of children with acute leukemia.