10x Genomics VDJ seqeuncing of IOMAgl germinal center B cells

This experiment was performed to identify somatic hypermutations in germinal center B cells of a mouse expressing the inferred germline of IOMA BCR after and a 5 step sequential immunization regimen. Overall design: This experiment was performed to identify somatic hypermutations in germinal center B cells of mouse HP3, homozyogous for the IghIOMAiGL and IgkIOMAiGL transgenes expressing the inferrred germline of IOMA after and a 5 step immunization regimen. The mice were immunized sequentially with mi3 nanoparticles. First primed with IGT2-mi3 and sequentially boosted with IGT1-mi3 (at week 5), 426c degly2 D279N-mi3 (week 10) , and twice with mosaic8-mi3 (week 15 and 20). HP3 was sacrified in week 23 and spleen and mesenteric lymph nodes were processed and stored in LN2. Then Germinal center B cells were sroted as live, singlet, lineage dump-, B220+, CD95+ CD38- lymphocytes.

本实验旨在鉴定经5步连续免疫方案免疫后、表达IOMA B细胞受体（BCR）推定种系序列的小鼠生发中心B细胞（germinal center B cells）中的体细胞超突变（somatic hypermutations）。
实验设计概述：本实验以携带Igh^IOMAiGL与Igk^IOMAiGL纯合转基因、表达IOMA BCR推定种系序列的小鼠HP3为研究对象，以mi3纳米颗粒进行序贯免疫：首次免疫使用IGT2-mi3，于第5周以IGT1-mi3加强免疫，第10周以426c degly2 D279N-mi3加强免疫，第15周及第20周两次以mosaic8-mi3加强免疫。小鼠HP3于第23周处死，获取脾脏与肠系膜淋巴结并置于液氮（LN2）中保存。随后将符合存活、单细胞、谱系dump阴性、B220+、CD95+CD38-表型的淋巴细胞分选为目标生发中心B细胞。