| In vitro Assessment of Immunological Synapse Formation by Flow Cytometry
|
|
Author:
Date:
2016-03-20
[Abstract] In adaptive immune system, formation of immunological synapse between T cells and antigen presenting cells (dendritic cells, B cells, and macrophages) or target cells (tumor cells and viral-infected cells) is critical for the execution of T cell immune responses via cytokine secretion or direct killing activity. Here, we describe the practical methods that directly measure the number of conjugates as a result of immunological synapse formation between T cells and superantigen-loaded B cells or between cytotoxic T cells and antigen-loaded target cells by dual-color flow cytometry.
[摘要] 在适应性免疫系统中，T细胞和抗原呈递细胞（树突细胞，B细胞和巨噬细胞）或靶细胞（肿瘤细胞和病毒感染细胞）之间的免疫突触的形成对通过细胞因子执行T细胞免疫应答至关重要 分泌或直接杀死活动。 在这里，我们描述了通过双色流式细胞术直接测量T细胞和加载超抗原的B细胞之间或细胞毒性T细胞和抗原负载的靶细胞之间的免疫突触形成的结合物数量的实际方法。
|
|
|
| Adoptive Transfer of Memory B Cells
|
|
Author:
Date:
2015-08-20
[Abstract] The adoptive transfer of antigen-specific B cells into mice that cannot recognize that specific antigen has two main advantages. The first is determining exactly when the B cells were transferred and exposed to antigen. The second is that all B cells that can bind that antigen are the ones that were transferred; no new antigen-specific B cells will emerge from the bone marrow. Thus all B cells that were exposed to the antigen and still alive after at least 4 weeks (8 weeks or more is ideal), are memory B cells.
Splenic B cells from B1-8 mice were prepared with an EasySep Mouse B ...
[摘要] 将抗原特异性B细胞过继转移到不能识别该特异性抗原的小鼠中具有两个主要优点。 第一种是确切地确定B细胞何时转移并暴露于抗原。 第二是可以结合该抗原的所有B细胞是转移的; 从骨髓中不会出现新的抗原特异性B细胞。 因此，暴露于抗原并在至少4周（8周或更长时间是理想的）后仍存活的所有B细胞是记忆B细胞。
根据制造商的方案用EasySep小鼠B细胞富集试剂盒制备来自B1-8小鼠的脾B细胞。 将单细胞悬浮液静脉内转移到受体小鼠的尾静脉中。 每只小鼠转移约100万个NP + B细胞。 转移后约12-24小时，用在明矾中沉淀的50μgNP-CGG腹膜内免疫小鼠。
|
|
|