Research reportLimiting dilution analysis of virus-specific memory B cells by an ELISPOT assay
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Pre-existing immunity modulates responses to mRNA boosters
2023, Cell ReportsScalable production and immunogenicity of a cholera conjugate vaccine
2021, VaccineCitation Excerpt :Nitrocellulose bottom plates (MAHAS4510, Millipore) were coated with OSP:BSA (100 ng/well), or keyhole limpet hemocyanin (KLH; Pierce Biotechnology) (2.5 μg/ml, negative control) or goat anti-mouse IgG (Southern Biotech) [30,31,34]. We assessed memory B cell responses after the final round of immunization as previously described [30,31,34]. We used unstimulated samples as negative controls and subtracted responses to KLH as a control for the plate.
CD19-positive antibody-secreting cells provide immune memory
2018, Blood AdvancesCitation Excerpt :Therefore, the potential secreted immunoglobulin contribution to humoral immunity of CD19+ ASCs is greater than CD19− ASCs based on frequency and tissue distribution. Long-lived immunity and the presence of high-affinity serum IgG antibodies specific for previously encountered antigens, like vaccine antigens, was described as existing for decades independent of memory B cells3,17,40,41 and was thought to be contributed only by CD19− BM ASCs. Our work shows both CD19+ and negative ASCs also exist in the spleen and have similar capability as the BM ASCs to recognize vaccine antigens (Figure 3).
Safety Profile and Immunologic Responses of a Novel Vaccine Against Shigella sonnei Administered Intramuscularly, Intradermally and Intranasally: Results From Two Parallel Randomized Phase 1 Clinical Studies in Healthy Adult Volunteers in Europe
2017, EBioMedicineCitation Excerpt :Exploratory immunological outcomes, only for cohort 3 of study 2, included assessment of memory B cells (MBC), specific for the OAg and its GMMA carrier, and of antibody secreting cells (plasmablasts) on PBMCs obtained at baseline and at 28 days after the third immunization (MBC) and at 7 days after the first immunization (plasmablasts). Assays were performed at GSK Vaccines, Siena, Italy, by ELISpot, following standard procedures (Slifka and Ahmed, 1996). Serum anti-S.
An optimized method for enumerating CNS derived memory B cells during viral-induced inflammation
2017, Journal of Neuroscience MethodsCitation Excerpt :Culture conditions designed to quantify virus-specific Bmem have largely been optimized using cells derived from lymphoid tissue and rely on various in vitro stimulation strategies to convert Bmem into ASC, which are then measured by ELISPOT in a separate plate. In vitro this process requires proliferation and thus prolonged stimulation more than 24 h, the minimum time required to trigger division by Bmem (Slifka and Ahmed, 1996b). For instance, following LCMV infection maximal virus-specific Bmem conversion into ASC was initially achieved by stimulating splenocytes from LCMV infected mice at serial dilutions with irradiated LCMV infected carrier splenocytes as a source of viral antigen for 6 days p.i (Slifka and Ahmed, 1996b).