The Effect Of Antiviral Therapy With Nucleoside (Acid) Analogues On The Capacity Of Dendritic Cell From Patients With Chronic Hepatitis B

Objective To establish the method of culturing DCs from human peripheral blood and study the difference in the phenotype and the capacity of stimulating leukocyte between patients with chronic hepatitis B and healthy subjects.Furthermore,we explored the change of the phenotype and the capacity of the DCs from patients receiving effective nucleotide antiviral therapy.Methods Selecting CHB without receiving antiviral treatment,PBMC were separated from peripheral blood,and then cultured in vitro with medium containing GM-CSF IL-4 TNF-αinto DCs,using light microscopy to monitor the morphologic features.The phenotype of DCs was identified by flow cytometry.Using mixed leukocytes reaction(MLR) detected the stimulating index(SI) of DCs to stimulate T cell proliferate.We separated patients' peripheral blood to culture DCs and tested their function again after receiving lamivudine and adefovir alternant therapy for 52 weeks,and comparing with ten healthy subjects and ten CHB patients with resisting lamivudine.Results1.PBMC from CHB patients which were cultured in vitro with medium containing GM-CSF 1000u/ml,rhIL-4 1000u/ml and TNF-α1000u/ml could be induced into typical DCs with typical morphology,phenotype and biological activity.However,comparing with DCs from healthy people,DCs from CHB patients had less dendritic protuberances.The figure of DCs from patients after antiviral treatment improved.2.The survival rates of DCs from healthy subjects,CHB before antiviral treatment and after receiving effective nucleotide antiviral therapy,and CHB patients with resisting lamivudine were (88.51±5.793)%,(82.84±7.588)%,(84.73±9.106)%,(83.68±6.752)%,respectively. There is no obvious statistical difference by analysis of variance.3.The expressions of costimulating molecules CD80,CD86,HLA-DR in the surface of DCs from CHB patients before antiviral treatment were(32.16±6.340)%,(36.65±6.867)%,(44.77±8.127)%,respectively.Compareing with healthy subjects they were obvious low,the difference had statistical meaning(P＜0.01).The expressions of CD80,HLA-DR in DCs from CHB patients after receiving effective nucleotide antiviral therapy were(44.39±5.391)%and (65.02±8.801)%,respectively,and they were both increasing comparing with that before antiviral treatment,and the difference had statistical meaning(P＜0.05),however,they were still low compareing with healthy subjects.The expression of CD86 was increasing,but it had no obvious statistical meaning(P＞0.05).The expressions of costimulating molecules CD80,CD86,HLA-DR in the surface of DCs of CHB patients with resisting lamivudine were (32.85±8.068)%,(38.34±4.665)%and(45.41±6.752)%,and there was no obvious statistical difference from that of CHB patients before receiving effective nucleotide antiviral therapy,and compareing with that of healthy subjects they were obvious low.4.The stimulation index(SI) that DCs from CHB patients before antiviral treatment stimulated lymphocytes to proliferate was 1.39±0.375,and obvious lower than the SI of DCs from healthy subjects,the difference had statistical meaning(P＜0.01).The SI of DCs from CHB patients after receiving effective nucleotide antiviral therapy was 1.65±0.590,and it was higher than that before antiviral treatment(P＜0.05).CHB patients with resisting lamivudine were not different from CHB patients before receiving effective nucleotide antiviral therapy,compareing with that of healthy subjects it was obvious low,Conelusion The PBMC from CHB patients under using GM-CSF,rhIL-4 and TNF-αcould be induced into typical DC with biological activity,however their costimulating molecules and the ability of stimulating lymphocytes to proliferate were decreased.These showed that HBV may decrease the function of DCs of CHB patients.It would result that the special Th and CTL aiming at HBV were not activated efficiently,and then HBVs could not be cleard efficiently.The costimulating molecules and the ability of stimulating lymphocytes to proliferate of DCs from CHB patients after receiving effective nucleotide antiviral therapy could be recovered.It show that nucleotide antiviral drugs not only decreased HBV load by restraining reverse-transcriptase enzyme,but could improve the ability of DCs of patients,and then improved the immunity function of patients.