Effects of T-bet B Cells in Persistent Virus Control After Discontinuance of NAs in CHB Patients

Study Overview
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Brief Summary
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The immune mechanism of the nucleos(t)ide analogs (NAs) in inhibiting HBV replication effectively while having a low sustained virus control rate after drug withdrawal is unclear. B cell immunity and antibody response are the keys to prevent HBV reinfection and keep the virus under control. T-bet+ B, which can be regulated by IL-21, is a newly discovered major effector B cell in protection of pathogens and it is a main subtype of HBsAg-specific B cells. Thus, we suspect that T-bet+ B may play a role in ongoing controlling of the virus after withdraw of NAs in CHB patient. Based on our previous studies on CHB immunity, we use the RNAseq analyse, flow cytometry, and Elispot assay to analyze the frequency, function, and phenotype of B cells in CHB patients with different profiles after withdraw of NAs.

Official Title
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Effects of T-bet B Cells in Persistent Virus Control After Discontinuance of NAs in CHB Patients

Conditions
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Chronic HBV Infection, Drug Withdrawal

Intervention / Treatment
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* Diagnostic Test: T-bet B cell


Participation Criteria
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Eligibility Criteria
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Inclusion Criteria: 18 to 70 years old, no gender restriction, serum HBsAg positive than 6 months, can understand and sign informed consent, good compliance. Exclusion Criteria: coinfected with other hepatotropic virus such as hepatitis C virus,hepatitis D -virus,hepatitis E and hepatitis A etc; coinfected with HIV, markers such as ceruloplasmin, anti-nuclear antibodies and anti-mitochondrial antibodies for co-existent autoimmune and metabolic liver diseases were positive, with hepatocellular carcinoma(HCC) with uncontrollable extrehepatic disease, received glucocorticoid or other immune inhibitor therapy, pregnancy

Ages Eligible for Study
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Minimum Age: 18 Years
Maximum Age: 70 Years

Sexes Eligible for Study
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All

Accepts Healthy Volunteers
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Accepts Healthy Volunteers

Study Plan
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How is the study designed?
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Arms and Interventions

| Participant Group/Arm | Intervention/Treatment |
| --- | --- |
| virologic breakthrough<br>CHB patients withdraw NAs with virologic breakthrough | Diagnostic Test: T-bet B cell<br>* The frequency, function, and phenotype of T-bet B cells was tested<br>|
| virologic response<br>CHB patients withdraw NAs with virologic response | Diagnostic Test: T-bet B cell<br>* The frequency, function, and phenotype of T-bet B cells was tested<br>|
| healthy group<br>The healthy (non-HBV infected) group completed a standard HBsAg vaccination scheduled before entering the study | Diagnostic Test: T-bet B cell<br>* The frequency, function, and phenotype of T-bet B cells was tested<br>|

What is the study measuring?
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Primary Outcome Measures

| Outcome Measure | Measure Description | Time Frame |
| --- | --- | --- |
| phenotype of B cells | CD19+cells were sorted from different samples, the frequency, function, and phenotype of B cells were analyzed. | PBMC were collected 0、4、12、24 week after withdraw the NAs | 
| B cell ELISPOT assay | Sorted B cell subsets were stimulated for 5 days following which B cell ELISPOT was conducted. | Cells were stimulated with R-848 (1 μg/ml) and IL-2 (10 ng/ml) for 5 days at 37 °C to aid memory B cell differentiation | 

 Terms related to the study
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Keywords Provided by Centre Hospitalier Valida
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B cell, transcription factor bet, chronic hepatitis B, nucleotide analogs, function cure