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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.
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| Label | Type | Description | Intervention Names |
|---|---|---|---|
| virologic breakthrough | CHB patients withdraw NAs with virologic breakthrough |
| |
| virologic response | CHB patients withdraw NAs with virologic response |
| |
| healthy group | The healthy (non-HBV infected) group completed a standard HBsAg vaccination scheduled before entering the study |
|
| Name | Type | Description | Arm Group Labels | Other Names |
|---|---|---|---|---|
| T-bet B cell | Diagnostic Test | The frequency, function, and phenotype of T-bet B cells was tested |
|
| 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 |
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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
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CHB patients on NAs therapy
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| Name | Affiliation | Role |
|---|---|---|
| Xuesong Liang, DR | Chaihai hospital | Principal Investigator |
| Facility | Status | City | State | ZIP | Country | Contacts |
|---|---|---|---|---|---|---|
| Changhai hospital | Shanghai | 200433 | China |
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| ID | Term |
|---|---|
| D013375 | Substance Withdrawal Syndrome |
| D019694 | Hepatitis B, Chronic |
| ID | Term |
|---|---|
| D019966 | Substance-Related Disorders |
| D064419 | Chemically-Induced Disorders |
| D001523 | Mental Disorders |
| D006509 | Hepatitis B |
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pbmc
| D000086982 |
| Blood-Borne Infections |
| D003141 | Communicable Diseases |
| D007239 | Infections |
| D018347 | Hepadnaviridae Infections |
| D004266 | DNA Virus Infections |
| D014777 | Virus Diseases |
| D006525 | Hepatitis, Viral, Human |
| D006521 | Hepatitis, Chronic |
| D006505 | Hepatitis |
| D008107 | Liver Diseases |
| D004066 | Digestive System Diseases |
| D002908 | Chronic Disease |
| D020969 | Disease Attributes |
| D010335 | Pathologic Processes |
| D013568 | Pathological Conditions, Signs and Symptoms |