EASL2019 FRI-153 强效乙型肝炎核心特异性B细胞反应与之相关 未

StephenW

FRI-153
Potent hepatitis B core-specific B cell responses associate with
clinical parameters in untreated and virally suppressed chronic
HBV patients
Thomas Vanwolleghem1,2, Anthony Grooshuismink2, Kim Kreefft2,
Magdeleine Hung3, Nikolai Novikov3, Andre Boonstra2. 1Antwerp
University Hospital, Gastroenterology and Hepatology, Antwerp,
Belgium; 2Erasmus University Medical Center, Gastroenterology and
Hepatology, Rotterdam, Netherlands; 3Gilead Sciences, Biology, Foster
City, United States
Email: [email protected]
Background and aims: Exhaustion of virus-specific T cells is a
hallmark of chronic HBV (cHBV) infections, but the HBV-specific B cell
response is less well studied. Previously, we identified B cell-related
transcriptomic changes in blood and liver of cHBV patients in
different clinical phases. We now examined, the number, phenotype
and function of HBcAg-specific B cells during cHBV, in comparison to
HBsAg-specific B cells.
Method: Serum and PBMC were obtained from 137 cHBV patients,
both untreated (n = 114) belonging to different clinical phases as NUC
treated (n = 23), and 22 healthy HBsAg-vaccinated controls. The
phenotype of overall and HBV-specific B lymphocytes was studied by
FACS using DyLight650 and DyLight550 dual fluorescently labelled
HBsAg and HBcAg in combination with antibodies against CD3, CD10,
CD19, CD27, CD21, CD38, and FcRL5. In vitro anti-HBs and anti-HBc
antibody production was measured after polyclonal PBMC stimulation
by ELISPOT assays. Anti-HBs and anti-HBc antibodies were
measured in serum and in supernatant by ELISA.
Results: Serum levels of anti-HBc, but not anti-HBs antibodies
associate with the clinical phases of cHBV, characterized by
increasing titers in patients with ALT rise (p < 0.0001). In vitro, a
similar profile is seen for the number of spot-forming anti-HBcproducing
cells and their levels in the supernatant of these cultures
(p < 0.0001). Also the number of HBcAg-specific B cells in blood
followed this pattern (p = 0.0035). In contrast, HBsAg-specific B cells
showno typical numeric or functional changes in cHBV and are vastly
outnumbered by HBcAg-specific B cells in blood (92.8-fold, P <
0.0001). HBcAg-specific B-cells are enriched for a CD21- and CD21 +
CD27 + memory B cell profile compared to total B cells (3.6-fold and
2.0-fold, P < 0.0001 respectively), but demonstrate a less activated
phenotype (activation marker CD38: 0.8-fold, P = 0.0017; inhibitory
IgG co-receptor FcRL5: 2.3-fold, P < 0.0001). Interestingly, complete
viral suppression in the NUC cohort, led to reduced numbers of
HBcAg-binding B cells and in vitro production of anti-HBc antibodies
(all P < 0.05). For the total chronic HBV cohort, HBV DNA levels
positively correlated with in vitro anti-HBc production levels (r =
0.388, P = 0.0008) and with the number of HBcAg-binding B cells (r =
0.323 P < 0.05).
Conclusion: HBcAg-specific B cells vastly outnumber HBsAg-specific
B cells in blood of chronic HBV patients, are enriched for a classical
memory B cell phenotype and show no impairment with increasing
HBV DNA titers.

StephenW

FRI-153
强效乙型肝炎核心特异性B细胞反应与之相关
未经治疗和病毒性抑制慢性乙型肝炎的临床参数
HBV患者
Thomas Vanwolleghem1,2，Anthony Grooshuismink2，Kim Kreefft2，
Magdeleine Hung3，Nikolai Novikov3，Andre Boonstra2。 1Antwerp
安特卫普大学医院，胃肠病学和肝病学，
比利时; 2伊拉斯姆斯大学医学中心，消化内科和
荷兰鹿特丹的肝病学; 3Gilead Sciences，Biology，Foster
美国城市
电子邮件：[email protected]
背景和目的：用尽病毒特异性T细胞是一种
慢性HBV（cHBV）感染的标志，但HBV特异性B细胞
反应不太好。以前，我们确定了B细胞相关的
cHBV患者血液和肝脏转录组学的变化
不同的临床阶段。我们现在检查了数量，表型
与cBBV相比，HBcAg特异性B细胞的功能和作用
HBsAg特异性B细胞。
方法：从137例cHBV患者中获得血清和PBMC，
未处理（n = 114）属于不同临床阶段的NUC
治疗（n = 23）和22个健康的HBsAg疫苗对照。该
总体和HBV特异性B淋巴细胞的表型研究
FACS使用DyLight650和DyLight550双荧光标记
HBsAg和HBcAg联合抗CD3，CD10抗体，
CD19，CD27，CD21，CD38和FcRL5。体外抗HBs和抗HBc
在多克隆PBMC刺激后测量抗体产生
通过ELISPOT分析。抗-HBs和抗-HBc抗体
通过ELISA测量血清和上清液。
结果：血清抗-HBc水平，而非抗HBs抗体水平
与cHBV的临床阶段相关，其特征在于
ALT升高患者的滴度增加（p <0.0001）。体外，a
对于形成斑点的抗HBc产生的数量，可以看到类似的特征
细胞及其在这些培养物上清液中的水平
（p <0.0001）。还有血液中HBcAg特异性B细胞的数量
遵循这种模式（p = 0.0035）。相比之下，HBsAg特异性B细胞
显示cHBV的典型数字或功能变化，并且非常广泛
血液中HBcAg特异性B细胞数量超过92.8倍，P <
0.0001）。 HBcAg特异性B细胞富含CD21-和CD21 +
CD27 +记忆B细胞谱与总B细胞相比（3.6倍和3倍）
分别为2.0倍，P <0.0001），但表现出较少的活化
表型（活化标记物CD38：0.8倍，P = 0.0017;抑制性
IgG共受体FcRL5：2.3倍，P <0.0001）。有趣的是，完整
NUC队列中的病毒抑制导致数量减少
HBcAg结合B细胞和体外产生抗HBc抗体
（均P <0.05）。对于总HBV慢性HBV队列，HBV DNA水平
与体外抗HBc生成水平正相关（r =
0.388，P = 0.0008）和HBcAg结合B细胞的数量（r =
0.323 P <0.05）。
结论：HBcAg特异性B细胞数量远远超过HBsAg特异性
慢性HBV患者血液中的B细胞富含经典
记忆B细胞表型并且随着增加而没有显示出损伤
HBV DNA滴度。