Germinal Center Reaction Blockade

To induce primary responses 2.5 × 10⁴ HEL-binding B cells from tamoxifen-treated SWHEL/Syk^fl/fl or SWHEL/Syk^fl/flRMCM mice were transferred into B6.SJL recipient mice together with 5 × 10⁸ HEL-conjugated sheep RBCs (SRBC). To generate a memory pool, splenocytes containing an equivalent of 5 × 10⁵ HEL-binding B cells from SWHEL/Syk^fl/+RMCM or SWHEL/Syk^fl/flRMCM mice were transferred into (B6 x B6.SJL)F1 recipient mice together with 1 × 10⁹ HEL^3×-conjugated SRBC. HEL (Sigma-Aldrich) or HEL^3× (a gift from R. Brink) (17 (link)) was conjugated to SRBC (Patricell) as described previously (18 ). The germinal center reaction was blocked by three injections of 300 μg anti-CD40L (MR1) or control Hamster IgG (BioXcell) every second day.

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Ackermann J.A., Nys J., Schweighoffer E., McCleary S., Smithers N, & Tybulewicz V.L. (2015). Syk Tyrosine Kinase Is Critical for B Cell Antibody Responses and Memory B Cell Survival. The Journal of Immunology Author Choice, 194(10), 4650-4656.

Publication 2015

anti-CD40L (MR1) control Hamster IgG

Anti cd40l B cells Germinal center Hamster Memory Mice Rbcs Sheep Tamoxifen Three

Corresponding Organization :

Other organizations : GlaxoSmithKline (United Kingdom)

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Variable analysis

independent variables

Genotype of mice (SWHEL/Syk^fl/fl or SWHEL/Syk^fl/flRMCM)
Tamoxifen treatment of mice
Dose of HEL-conjugated sheep RBCs (5 × 10^8 or 1 × 10^9)
Injection of anti-CD40L (MR1) or control Hamster IgG

dependent variables

Primary response of HEL-binding B cells (2.5 × 10^4)
Memory pool of HEL-binding B cells (5 × 10^5)

control variables

Mouse strains (B6.SJL recipient mice, (B6 x B6.SJL)F1 recipient mice)
HEL and HEL^3× conjugation to sheep RBCs

positive control

None specified

negative control

Injection of control Hamster IgG

Annotations

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