MUrine tools to catch high-affinity plasma cells (MUTCHAP)

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Price, Michael John (2021). MUrine tools to catch high-affinity plasma cells (MUTCHAP). University of Birmingham. Ph.D.

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Abstract

The generation of high-affinity antibodies is critical for natural and induced protective immune responses and production of efficacious therapeutic monoclonal antibodies. Germinal centre matured plasma cells (GCmat PCs) produce these long-term high-quality responses. The signals and mechanisms by which GCmat PCs are induced are not fully elucidated. We developed parallel strategies to generate novel gene manipulated mice where GCmat PCs are easily identifiable by fluorescent markers controlled by GC-specific S1pr2 and PC-specific Prdm1 expression.

Combination of validated transgenic alleles generated the High-Affinity LOw-affinity (HALO) PC mouse, able to distinguish between GCmat PCs and extrafollicular PCs. Kinetic assessment of early GC output confirmed previous results from our lab that GCmat PC output peaks early then declines. Furthermore, we detected transient re-expression of CD38 in GCmat PC precursors and GCmat transitioning PCs, an intriguing result with a yet undetermined function.

In tandem, we developed and characterised a novel split fluorescent reporter, diSplit670. CRISPR/Cas9 enhanced targeted insertion placed each part of diSplit670 under control of endogenous Prdm1 and S1pr2 in separate mouse embryonic stem cell (ESC) lines. To-date, germline transmission has been achieved for one targeted ESC line.

These novel tools would be uniquely suited to interrogate the biology of GCmat PCs.

Type of Work:

Thesis (Doctorates > Ph.D.)

Award Type:

Doctorates > Ph.D.

Supervisor(s):