B-cell repertoire dynamics after sequential hepatitis B vaccination and evidence for cross-reactive B-cell activation.
B-cell repertoire dynamics after sequential hepatitis B vaccination and evidence for cross-reactive B-cell activation.
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DOI：
10.1186/s13073-016-0322-z
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发表时间：
2016-06-16
期刊：
影响因子：
12.3
通讯作者：
Kelly DF
中科院分区：
文献类型：
--
作者：
Galson JD;Trück J;Clutterbuck EA;Fowler A;Cerundolo V;Pollard AJ;Lunter G;Kelly DF
不同的B细胞库对于识别和应答感染性抗原和疫苗抗原至关重要。B细胞受体（BCR）基因的高通量测序现在可以用于深入研究B细胞库，并可能比传统的免疫学方法更清楚地了解B细胞反应。在这里，我们使用高通量BCR测序提供新的见解B细胞动力学的主要过程后，B型肝炎疫苗接种。9名未接种过疫苗的参与者接种了3剂B型肝炎疫苗（0、1和2或7个月）。将总BCR库的高通量Illumina测序与分选的疫苗抗原富集的B细胞的靶向测序相结合，以分析每种疫苗接种后总库和疫苗特异性库的纵向应答。使用ELISpot测定每种疫苗接种后的疫苗特异性细胞数。从总BCR库中解卷积疫苗特异性序列簇显示疫苗特异性序列簇占总序列簇的<0.1%，并且具有某些定型特征。尽管在第一次疫苗给药后通过ELISpot未检测到疫苗特异性B细胞，但在三次疫苗给药中的每一次后扩增了疫苗特异性BCR序列簇。与后续接种后检测到的BCR簇相比，首次接种疫苗后检测到的这些疫苗特异性BCR簇具有不同的特性;它们突变更多，即使在接种前也以低频率存在，并且似乎来源于更成熟的B细胞。这些结果证明了我们的疫苗特异性BCR分析方法的高灵敏度，并提出了B细胞对新抗原反应的另一种观点。在对第一剂疫苗的反应中，许多疫苗特异性BCR簇似乎主要来源于先前活化的交叉反应性B细胞，这些细胞对疫苗抗原具有低亲和力，需要后续剂量以产生更高亲和力的B细胞。本文的在线版本（doi：10.1186/s13073-016-0322-z）包含补充材料，可供授权用户使用。
A diverse B-cell repertoire is essential for recognition and response to infectious and vaccine antigens. High-throughput sequencing of B-cell receptor (BCR) genes can now be used to study the B-cell repertoire at great depth and may shed more light on B-cell responses than conventional immunological methods. Here, we use high-throughput BCR sequencing to provide novel insight into B-cell dynamics following a primary course of hepatitis B vaccination. Nine vaccine-naïve participants were administered three doses of hepatitis B vaccine (months 0, 1, and 2 or 7). High-throughput Illumina sequencing of the total BCR repertoire was combined with targeted sequencing of sorted vaccine antigen-enriched B cells to analyze the longitudinal response of both the total and vaccine-specific repertoire after each vaccine. ELISpot was used to determine vaccine-specific cell numbers following each vaccine. Deconvoluting the vaccine-specific from total BCR repertoire showed that vaccine-specific sequence clusters comprised <0.1 % of total sequence clusters, and had certain stereotypic features. The vaccine-specific BCR sequence clusters were expanded after each of the three vaccine doses, despite no vaccine-specific B cells being detected by ELISpot after the first vaccine dose. These vaccine-specific BCR clusters detected after the first vaccine dose had distinct properties compared to those detected after subsequent doses; they were more mutated, present at low frequency even prior to vaccination, and appeared to be derived from more mature B cells. These results demonstrate the high-sensitivity of our vaccine-specific BCR analysis approach and suggest an alternative view of the B-cell response to novel antigens. In the response to the first vaccine dose, many vaccine-specific BCR clusters appeared to largely derive from previously activated cross-reactive B cells that have low affinity for the vaccine antigen, and subsequent doses were required to yield higher affinity B cells. The online version of this article (doi:10.1186/s13073-016-0322-z) contains supplementary material, which is available to authorized users.