Toll Like Receptor 2 And 9 Express In Peripheral Blood Monocyte Of Chronic Severe Hepatitis B

Objective To approach the role of Toll like receptor 2 (TLR2), Toll like receptor 9 (TLR9) and tumor necrosis factor-a (TNF-α) in the pathogenesis of chronic severe hepatitis B and their relationship with patients severe levels by studying the expression of TLR2 and TLR9 in peripheral blood mononuclear cells (PBMC) and serum TNF-αof chronic severe hepatitis B patients.Methods The heparin anticoagulant and procoagulant venous blood samples from 45 cases of chronic severe hepatitis B patients (patients group) and 23 cases of healthy control (control group) were collected. PBMC were separated from heparin anticoagulant venous blood. TLR2 and TLR9 protein in lymphocytes and monocytes were detected by flow cytometry.TLR2 and TLR9 mRNA in PBMC were detected by fluorescence quantitative PCR. The serum was separated from procoagulant venous blood and TNF-αin serum was detected by ELISA.Results 1. The results of flowcytometry were as follows. (1)The positive percentage of monocytes TLR2 protein in patient and control groups was 92.11±16.43 and 81.84±29.80 respectively. The difference had no statistical significance (P>0.05). The positive percentage of lymphocyte TLR2 protein in the two groups was 9.73±5.37 and 2.18±2.46 respectively. The difference had no statistical significance too (P>0.05). The MFI of monocyte TLR2 protein in patient and control groups was 149.82±43.93 and 170.25±76.43 respectively, having no statistical difference too (P>0.05). The MFI of lymphocyte TLR2 protein in the two groups was 6.04±5.21 and 4.26±2.46 respectively, having no statistical difference too (P>0.05). (2) The positive percentage of monocyte TLR9 protein in patient and control groups was 81.65±32.31 and 55.03±21.60 respectively. The difference of two groups had no statistical significance (P>0.05).The positive percentage of lymphocyte TLR9 protein in the two groups was 75.64±37.15 and 23.29±18.87 respectively, showing significant difference (P<0.05). The MFI of monocyte TLR9 protein in patient and control groups was 140.61±48.35 and 225.85±90.11 respectively, showing significant difference too (P<0.05). The MFI of lymphocyte TLR9 protein in the two groups was 37.39±18.36 and 19.51±11.13 respectively, showing significant difference too (P<0.05).2.The results of TLR2和TLR9 mRNA in PBMC were as follows. The relative expression amount of TLR2 mRNA in patients and control groups was 0.209±0.226 and 0.356±0.310 respectively. The difference of two groups had no statistical significance (P>0.05). The relative expression amount of TLR9 mRNA in patients and control groups was 0.014±0.028 and 0.253±0.159 respectively, showing significant difference (P<0.05).3.The serum TNF-α(pg/ml) of patients and control groups was 260.15±71.35 and 158.66±99.234 respectively, showing significant difference (P<0.05).4. The correlation analysis of above results and clinical data were as follows. The MFI of monocyte TLR9 protein had a positive correlation with serum HBeAg level(r=0.382, P<0.05). TLR9 mRNA relative expression amount had a positive correlation with prothrombin time activity(PTA) (r=0.313, P<0.05). TNF-a and PTA showed a negative correlation(r=-0.372, P<0.05). TLR9 mRNA relative expression amount had a positive correlation with the MFI of monocyte TLR9 protein (r=0.459, P<0.01).Conclusions (1) TLR2 is mainly expressed in monocyte and weakly expressed in lymphocyte. TLR9 is expressed in both monocyte and lymphocyte on a certain ratio. (2) The lower express of TLR9 is relevant with the pathogenesis of chronic severe hepatitis B. (3) TLR9 has a negative correlation and TNF-αhas a positive correlation with patients'severe levels.