Molecular characterization of single memory B cells

McHeyzer‐Williams, Michael G.; Nossal, G. J. V.; Lalor, Paul A.

doi:10.1038/350502a0

Cited by 150 publications

(111 citation statements)

References 26 publications

Supporting

Mentioning

103

Contrasting

Order By: Relevance

“…NP-specific ELISAs were done as previously described (25,26). Briefly, for IgM and IgG ELISAs, plates were coated overnight with 20 mg/ml NP(18)-BSA (Biosearch Technologies).…”

Section: Np-specific Elisasmentioning

confidence: 99%

IL-21 Receptor Is Critical for the Development of Memory B Cell Responses

Rankin

MacLeod

Keegan

et al. 2011

The Journal of Immunology

View full text Add to dashboard Cite

Development of long-term humoral immunity, characterized by the formation of long-lived plasma cells (PCs) in the bone marrow and memory B cells, is a critical component of protective immunity to pathogens, and as such it is the major goal of vaccination. However, the mechanisms involved in the generation of long-term humoral immunity remain poorly understood. In this study, we used IL-21R–deficient (IL-21R.KO) mice to examine the role of the IL-21 pathway in the development of the B cell memory response. Primary IgG serum Ab responses to the T cell-dependent Ag 4-hydroxy-3-nitrophenylacetyl (NP) hapten conjugated to chicken γ globulin were delayed in IL-21R.KO mice, but reached normal titers within 3 to 4 wk of immunization. IL-21R.KO mice formed germinal centers and generated normal numbers of PCs in their bone marrow. Additionally, memory B cell formation was similar in wild-type and IL-21R.KO mice. However, NP-specific memory B cells and PCs failed to expand following secondary immunization of IL-21R.KO mice, and consequently, secondary IgG Ab responses to NP hapten conjugated to chicken γ globulin were significantly impaired. These results identify the IL-21 pathway as a critical component of the memory B cell response.

show abstract

“…NP-specific ELISAs were done as previously described (25,26). Briefly, for IgM and IgG ELISAs, plates were coated overnight with 20 mg/ml NP(18)-BSA (Biosearch Technologies).…”

Section: Np-specific Elisasmentioning

confidence: 99%

IL-21 Receptor Is Critical for the Development of Memory B Cell Responses

Rankin

MacLeod

Keegan

et al. 2011

The Journal of Immunology

View full text Add to dashboard Cite

show abstract

“…Recently, evidence has come forward that B cells acting as precursors of memory cells enter a period of great sensitivity to tolerance induction shortly after their T-dependent activation (5). We have attempted to gain more precise knowledge of memory cell generation (6)(7)(8) and its abrogation by soluble antigen (7). Part of the strategy makes use of the fact that there are very few cells in the primary B-cell repertoire that can make antibody to a pure protein antigen when polyclonally stimulated, particularly when the enzymelinked immunosorbent assay (ELISA) readout is geared to detect only bivalent IgG1 antibody (6,7).…”

mentioning

confidence: 99%

Memory cell generation ablated by soluble protein antigen by means of effects on T- and B-lymphocyte compartments.

Karvelas

Nossal

1992

Proc. Natl. Acad. Sci. U.S.A.

View full text Add to dashboard Cite

Adult C57BL/6 mice were injected with 100 ,ug of soluble, freshly deaggregated human serum albumin (HSA) to produce partial immunologic tolerance. UnhEjected normal control (N) mice contain only 100 B cells in their spleens with the capacity to (i) be activated in vitro into clonal proliferation by Escherichia coli lipopolysaccharide plus interleukins 2, 4, and 5, (ii) form IgG1 as well as IgM antibody, and (iii) display specificity for HSA when only IgG1 is allowed to score in an enzyme-linked immunosorbent assay (ELISA). Such N mice generate ==50,000 clonable anti-HSA IgG1 antibody-forming cell precursors in their spleens after T-dependent immunization with HSA adsorbed onto alum and given with BordeteUa pertussis adjuvant. Mice prei'jected with soluble HSA (TOL)-generate far fewer anti-HSA IgG1 antibodyforming cell precursors, termed anti-HSA memory cells. Splenocytes were transferred from N or TOL mice into lethally irradiated syngeneic recipients together with syngeneic bone marrow. Whereas N splenocytes generated plentiful memory cells within 2 weeks in antigenically challenged recipients, TOL splenocytes did not. Work with Ly-5 congenic mice ruled out memory cell generation from either the host or the bone marrow inoculum within this limited time. N T cells plus TOL B cells showed consistently lowered memory cell generation. TOL T cells plus N B cells showed an even greater lowering of adoptive memory cell generation. Thus the lowered response capacity of TOL mice resided in the T-and B-cell compartments. Attempts to show a suppressor component within the T-cell population were inconclusive, but a profound defect in capacity to respond to HSA in vitro was exhibited by the CD4' T cells of TOL mice. B lymphocytes were harvested from T-dependently immunized mice 5 days after challenge, incubated with soluble HSA for 18 hr, and then adoptively transferred together with N T cells. The recently activated B cells were not rendered tolerant by this manipulation. The results argue for a major T-cell component in the process whereby soluble protein antigens ablate affinity maturation and memory cell generation.

show abstract

“…AID mice or GFP AID mice were sorted by FACS. The Ig heavy-chain V H 186.2 genes, which are preferentially used by high-affinity NP-specific B cells in C57BL/6 mice, were amplified by RT-PCR, and then PCR direct sequencing was performed (27)(28)(29)(30). As shown in Table S1, the use of the V H 186.2 gene segment in NP-binding B cells was more than 2.5-fold lower in LMP2A AID mice than in GFP AID mice.…”

Section: Lmp2a Expression In B Cells Results In B-cell Hyperactivatiomentioning

confidence: 99%