ABSTRACT
Objective:
To determine the spectrum drift characteristics of CI4+CD25+Tregs TCR β chain CDR3 in
patients with different phases of acute
hepatitis B (AHB) and
chronic hepatitis B (CHB)
patients before and after the entecavir
treatment.
Methods:
Anticoagulation venous
blood was collected from 4 normal control subjects,3 AHB
patients with acute phase and convalescent phase,and 4 CHB
patients before and after the entecavir
treatment;and
peripheral blood mononuclear cells were isolated;CD4+ CD25+ Tregs were
separated by using the
magnetic beads,and total RNAs were extracted from CD4+ CD25+ Tregs and used for
reverse transcription.The TRBV CDR3 was amplified by
polymerase chain reaction (
PCR) with forward primers specific for 24 TRBV
families and one
fluorescence-labeled common reverse primer specific for the BC region.The
PCR products were sent out for Genescan,and results were analyzed for the TRBV
family CDR3 spectrum characteristics by using the Peak Scanner
Software vl.0.Data were analyzed with the comparative t-test to perform the
statistical analysis.
Results:
The CDR3 spectral types of the TRBV
family showed drift characteristics in 3 cases of AHB
patients with acute and convalescent phases;single/oligo peak spectral type
family was observed in most of
patients with acute phase;multiple peak spectral type was seen in
patients with convalescent phase;and the common spectrum shift of TRBV4,10,14,16,19
families seen in
patients with acute phase was changed to multiple peak spectral type.The clonal expansion of TRBV
family in the CD4+CD25+Tregs in PBMC from AHB
patients with convalescent phase was significantly lower than AHB
patients with acute phase (t =9.456,P =0.011).The clonal expansion of Tregs TRBV13.2,15,16,18,20
family seen in C HB
patients before
treatment may interfere the
virus removal through down-regulating the body's
immune response;and with the decline of
viral load in
serum after the
antiviral treatment,the clonal expansion of Tregs TRBV1,5.2,6,12,14,24
family may help body induce
immune tolerance and result in the HBV persistence.The clonal expansion of TRBV
family in the CI4+CD25+Tregs in PBMC from of CHB
patients after
antiviral treatment was increased (t =-0.666,P =0.553).
Conclusion:
TRBV4,10,14,16,19
family of spectrum shift seen in AHB
patients with acute phase was changed to multiple peak spectral type in
patients with convalescent phase,suggesting this transition may be associated with
HBsAg and
HBeAg turning to negative.The clonal expansion of Tregs TRBV13.2,15,16,18,20
family seen in CHB
patients before
treatment may interfere the
virus removal through down-regulating the body's
immune response;and with the decline of
viral load in
serum after the
antiviral treatment,the clonal expansion of Tregs TRBV1,5.2,6,12,14,24
family may help body induce
immune tolerance and result in the HBV persistence.