期刊
CLINICAL & TRANSLATIONAL IMMUNOLOGY
卷 8, 期 11, 页码 -出版社
WILEY
DOI: 10.1002/cti2.1090
关键词
B cell; differentiation; memory; monocyte; naive; TLR
类别
资金
- Australian National Health and Medical Research Council (NHMRC)
- NHMRC [1071916, APP1102792, 1103367]
- Melbourne International Research Scholarship (MIRS)
- Melbourne International Fee Remission Scholarship (MIFRS)
- Australian Government Department of Health
- National Health and Medical Research Council of Australia [1103367, 1071916] Funding Source: NHMRC
Objectives A fundamental question in influenza research is whether antibody titre decline upon successive exposure to variant strains is consequent to recall of cross-reactive memory B cells that competitively inhibit naive B-cell responses. In connection, it is not clear whether naive and memory B cells remain phenotypically distinct acutely after activation such that they may be distinguished ex vivo. Methods Here, we first compared the capacity of anti-Ig and Toll-like-receptor (TLR) 7/8 and TLR9 agonists (R848 and CpG) to augment human B-cell differentiation induced by IL-21 and sCD40L. The conditions that induced optimal differentiation were then used to compare the post-activation phenotype of sort-purified naive and memory B-cell subsets by FACS and antibody-secreting cell (ASC) ELISPOT. Results Sort-purified naive and memory B cells underwent robust plasmablast and ASC formation when stimulated with R848, but not CpG, and co-cultured with monocytes. This coincided with increased IL-1 beta and IL-6 production when B cells were co-cultured with monocytes and stimulated with R848, but not CpG. Naive B cells underwent equivalent ASC generation, but exhibited less class-switch and modulation of CD27, CD38 and CD20 expression than memory B cells after stimulation with R848 and monocytes for 6 days. Conclusion Stimulation with R848, IL-21 and sCD40L in the presence of monocytes induces robust differentiation and ASC generation from both naive and memory B-cells. However, naive and memory B cells retain key phenotypic differences after activation that may facilitate ex vivo discrimination and better characterisation of acute responses to variant antigens.
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