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Rescue of HIV-1 Broad Neutralizing Antibody-Expressing B Cells in 2F5 VH × VL Knockin Mice Reveals Multiple Tolerance Controls
- Source :
- The Journal of Immunology. 187:3785-3797
- Publication Year :
- 2011
- Publisher :
- The American Association of Immunologists, 2011.
-
Abstract
- The HIV-1 broadly neutralizing Ab (bnAb) 2F5 has been shown to be poly-/self-reactive in vitro, and we previously demonstrated that targeted expression of its VDJ rearrangement alone was sufficient to trigger a profound B cell developmental blockade in 2F5 VH knockin (KI) mice, consistent with central deletion of 2F5 H chain-expressing B cells. In this study, we generate a strain expressing the entire 2F5 bnAb specificity, 2F5 VH × VL KI mice, and find an even higher degree of tolerance control than observed in the 2F5 VH KI strain. Although B cell development was severely impaired in 2F5 VH × VL KI animals, we demonstrate rescue of their B cells when cultured in IL-7/BAFF. Intriguingly, even under these conditions, most rescued B cell hybridomas produced mAbs that lacked HIV-1 Envelope (Env) reactivity due to editing of the 2F5 L chain, and the majority of rescued B cells retained an anergic phenotype. Thus, when clonal deletion is circumvented, κ editing and anergy are additional safeguards preventing 2F5 VH/VL expression by immature/transitional B cells. Importantly, 7% of rescued B cells retained 2F5 VH/VL expression and secreted Env-specific mAbs with HIV-1–neutralizing activity. This partial rescue was further corroborated in vivo, as reflected by the anergic phenotype of most rescued B cells in 2F5 VH × VL KI × Eμ-Bcl-2 transgenic mice and significant (yet modest) enrichment of Env-specific B cells and serum Igs. The rescued 2F5 mAb-producing B cell clones in this study are the first examples, to our knowledge, of in vivo-derived bone marrow precursors specifying HIV-1 bnAbs and provide a starting point for design of strategies aimed at rescuing such B cells.
- Subjects :
- Immunology
Immunoglobulin Variable Region
Enzyme-Linked Immunosorbent Assay
HIV Infections
Mice, Transgenic
Cell Separation
Biology
Article
Clonal deletion
Immune tolerance
Flow cytometry
Mice
Immune Tolerance
medicine
Animals
Immunology and Allergy
Gene Knock-In Techniques
B-cell activating factor
Neutralizing antibody
B cell
B-Lymphocytes
Genes, Immunoglobulin
medicine.diagnostic_test
Flow Cytometry
Antibodies, Neutralizing
Virology
In vitro
Mice, Inbred C57BL
medicine.anatomical_structure
HIV-1
biology.protein
Bone marrow
Subjects
Details
- ISSN :
- 15506606 and 00221767
- Volume :
- 187
- Database :
- OpenAIRE
- Journal :
- The Journal of Immunology
- Accession number :
- edsair.doi.dedup.....f35db5161581bfee51a89fdd53016548