Feng Feng, Yuen Rachel, Wang Yumei, Hua Axin, Kepler Thomas B, Wetzler Lee M
Department of Microbiology, Boston University, Boston, MA 02118, USA.
Department of Mathematics and Statistics, Boston University, Boston, MA 02118, USA.
iScience. 2023 Dec 14;27(1):108749. doi: 10.1016/j.isci.2023.108749. eCollection 2024 Jan 19.
Generating large-scale, high-fidelity sequencing data is challenging and, furthermore, not much has been done to characterize adjuvants' effects at the repertoire level. Thus, we introduced an IgSeq pipeline that standardized library prep protocols and data analysis functions for accurate repertoire profiling. We then studied systemically effects of CpG and Alum on the Ig heavy chain repertoire using the ovalbumin (OVA) murine model. Ig repertoires of different tissues (spleen and bone marrow) and isotypes (IgG and IgM) were examined and compared in IGHV mutation, gene usage, CDR3 length, clonal diversity, and clonal selection. We found Ig repertoires of different compartments exhibited distinguishable profiles at the non-immunized steady state, and distinctions became more pronounced upon adjuvanted immunizations. Notably, Alum and CpG effects exhibited different tissue- and isotype-preferences. The former led to increased diversity of abundant clones in bone marrow, and the latter promoted the selection of IgG clones in both tissues.
生成大规模、高保真的测序数据具有挑战性，此外，在库水平上表征佐剂的作用方面所做的工作不多。因此，我们引入了一种IgSeq流程，该流程标准化了文库制备方案和数据分析功能，以进行准确的库分析。然后，我们使用卵清蛋白（OVA）小鼠模型系统地研究了CpG和明矾对Ig重链库的影响。在IGHV突变、基因使用、CDR3长度、克隆多样性和克隆选择方面，对不同组织（脾脏和骨髓）和同种型（IgG和IgM）的Ig库进行了检查和比较。我们发现，在未免疫的稳态下，不同区室的Ig库表现出可区分的特征，在佐剂免疫后，差异变得更加明显。值得注意的是，明矾和CpG的作用表现出不同的组织和同种型偏好。前者导致骨髓中丰富克隆的多样性增加，后者促进了两种组织中IgG克隆的选择。
