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Rescue of HIV-1 broad neutralizing antibody-expressing B cells in 2F5 VH x VL knockin mice reveals multiple tolerance controls.

Publication ,  Journal Article
Verkoczy, L; Chen, Y; Bouton-Verville, H; Zhang, J; Diaz, M; Hutchinson, J; Ouyang, Y-B; Alam, SM; Holl, TM; Hwang, K-K; Kelsoe, G; Haynes, BF
Published in: J Immunol
October 1, 2011

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 V(H) 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 V(H) × V(L) KI mice, and find an even higher degree of tolerance control than observed in the 2F5 V(H) KI strain. Although B cell development was severely impaired in 2F5 V(H) × V(L) 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 V(H)/V(L) expression by immature/transitional B cells. Importantly, 7% of rescued B cells retained 2F5 V(H)/V(L) 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 V(H) × V(L) 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.

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Published In

J Immunol

DOI

EISSN

1550-6606

Publication Date

October 1, 2011

Volume

187

Issue

7

Start / End Page

3785 / 3797

Location

United States

Related Subject Headings

  • Mice, Transgenic
  • Mice, Inbred C57BL
  • Mice
  • Immunology
  • Immunoglobulin Variable Region
  • Immune Tolerance
  • HIV-1
  • HIV Infections
  • Genes, Immunoglobulin
  • Gene Knock-In Techniques
 

Citation

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ICMJE
MLA
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Verkoczy, L., Chen, Y., Bouton-Verville, H., Zhang, J., Diaz, M., Hutchinson, J., … Haynes, B. F. (2011). Rescue of HIV-1 broad neutralizing antibody-expressing B cells in 2F5 VH x VL knockin mice reveals multiple tolerance controls. J Immunol, 187(7), 3785–3797. https://doi.org/10.4049/jimmunol.1101633
Verkoczy, Laurent, Yao Chen, Hilary Bouton-Verville, Jinsong Zhang, Marilyn Diaz, Jennifer Hutchinson, Ying-Bin Ouyang, et al. “Rescue of HIV-1 broad neutralizing antibody-expressing B cells in 2F5 VH x VL knockin mice reveals multiple tolerance controls.J Immunol 187, no. 7 (October 1, 2011): 3785–97. https://doi.org/10.4049/jimmunol.1101633.
Verkoczy L, Chen Y, Bouton-Verville H, Zhang J, Diaz M, Hutchinson J, et al. Rescue of HIV-1 broad neutralizing antibody-expressing B cells in 2F5 VH x VL knockin mice reveals multiple tolerance controls. J Immunol. 2011 Oct 1;187(7):3785–97.
Verkoczy, Laurent, et al. “Rescue of HIV-1 broad neutralizing antibody-expressing B cells in 2F5 VH x VL knockin mice reveals multiple tolerance controls.J Immunol, vol. 187, no. 7, Oct. 2011, pp. 3785–97. Pubmed, doi:10.4049/jimmunol.1101633.
Verkoczy L, Chen Y, Bouton-Verville H, Zhang J, Diaz M, Hutchinson J, Ouyang Y-B, Alam SM, Holl TM, Hwang K-K, Kelsoe G, Haynes BF. Rescue of HIV-1 broad neutralizing antibody-expressing B cells in 2F5 VH x VL knockin mice reveals multiple tolerance controls. J Immunol. 2011 Oct 1;187(7):3785–3797.

Published In

J Immunol

DOI

EISSN

1550-6606

Publication Date

October 1, 2011

Volume

187

Issue

7

Start / End Page

3785 / 3797

Location

United States

Related Subject Headings

  • Mice, Transgenic
  • Mice, Inbred C57BL
  • Mice
  • Immunology
  • Immunoglobulin Variable Region
  • Immune Tolerance
  • HIV-1
  • HIV Infections
  • Genes, Immunoglobulin
  • Gene Knock-In Techniques