Prognostic evaluation of childhood Acute Lymphoblastic Leukaemia, using different immunological markers (CD3, CD19, CD10, Ki-67,P-gp) and serum IL-10, IL-6, IL-4, IL-8 and INF-γ.

number: 
2345
إنجليزية
Degree: 
Author: 
Rasha Raheem Abdulhamza
Supervisor: 
Dr.Abdul-RazakH.Ahmad
Dr.Salma AL- Hadad
year: 
2009

Abstract:

Acute lymphoblastic leukemia (ALL) is the most common malignancy occurring in childhood and accounts for 77% of all leukemia cases. In Iraq leukemia is in the first level among the cancer diseases in children in the year 2004 and is the most common malignancy, accounting for 34.97% of all cancer in 2004. Long-term survival is greater than 80% with appropriate treatment.The prognosis of children with acute lymphoblastic leukemia (ALL) has improved remarkably during the past four decades. The main reasons for this development are intensive multiagent chemotherapy and effective prophylactic treatment for central nervous system leukemia. Riskadjusted therapy according to recognized prognostic factors has also played
an important role.Hence this study tries to investigate the expression of prognostic markers on peripheral blood lymphocyte (PBL), like the expression of CD3,CD19, Ki67,P-gp and CD 10 in ALL patients, estimation of IL-4, IL-6, IL-8, IL10, INF-gamma serum levels in ALL patients, Correlate the results of PBL-phenotypic, proliferation ,drug resistant & maturations markers expression and serological estimation of Th1&Th2 cytokines & chemokines with different disease patterns, explore the possible role of these markers and serum cytokines &chemokines levels in prediction of the disease outcome. Initially Forty patients with ALL admitted to Oncology Unit from 10th of May 2008 to 20th of October 2008 in (Children Welfare Teaching
Hospital at Medical city in Baghdad) were included as patients group and thirty, age and sex matched apparently healthy children, were included in this study as a control group .The diagnosis in each case was established by clinical examination and confirmed by haematological examination (bone marrow and complete blood picture) Blood sample was taken from each patient and divided into 2 parts, one kept in heparinized blood tube to be used for lymphocyte separation, smear preparation and fixation on charged slide, wrapped, and kept at -20°C until assayed, While, the other kept in plain tube & allowed to clot, serum
was separated and collected, and dispensed in aliquots and kept at -20°C until assayed. CD3,CD19, CD10, Ki67, and P-gp expression detected using Immunocytochemistry staining technique, whereas, the serum concentration of IL-4, IL-6, IL-8, IL10, INF-gamma were determined using Sandwich Enzyme-Linked Immunosorbent Assay (ELISA). The studied patients consist of 20 females and 20 males; there was no statistical difference in the frequency of patient among age group. Mean age of the patients was (7.47 year) with a range from 5 month to12 year. Most of the patients 32/40(80 %) were L2 According to the French-American-British (FAB) criteria, and about 36/40 (90%) were originated from B cell, 30/40(75%) of patients were alive at the end of 2008. Regarding to the (WBC count) risk factor, the patient were classified into two groups, standard risk group (which the WBC count less than 50×109 cell/liter) 70% ,and high risk group (WBC count more than 50 ×109 cell/liter) 30% . Regarding to the age risk factor ,there were three groups ,
less than 1 years old ( 5% Percentage), second group between 1-10 years old (72.5 %, percentage),3 rd group more than 10 years the (22.5 % percentage ). Immunocytochemistry staining of CD10, Ki67, and P-gp revealed a highly statistical significant differences with higher mean percentage of expression in ALL patients when compared with the healthy control group. Percentage of expression for Ki-67 and P-gp were increased among most unfavorable groups like dead group, less than 1 years old group and Tcell patient group with a significant difference in the mean of Ki-67 expression on PBL among age risk group and significant difference in the mean of P-gp expression on PBL among WBC risk group ,and between age
risk group . CD10 expression increased among most favorable groups like B-cell group, alive group and standard risk group (WBC count less than 50×109 cell/liter) with no significant difference. The sera of ALL patients group revealed statistically significant difference with concentration of IL-10 and IL-6 when compared with that of the control group. And there is statistically significant difference between age group, type of leukaemia, WBC risk group and IL-10. And showed statistically significant difference between type of leukaemia, WBC risk group and IL-6.Serum IL-10 have correlations with age group and type of leukaemia while serum IL-6 have correlations with age group and Ki-67. The sera of ALL patients group revealed no statistically significant difference with concentration of IL-4, IL-8 and INF-gamma when compared with that of the control group. And there is no significant difference among these cytokines and study groups.