Design of targeted B cell killing agents

Alexey V. Stepanov, Alexey A. Belogurov, Natalia A. Ponomarenko, Oleg A. Stremovskiy, Leonid V. Kozlov, Anna M. Bichucher, Sergey E. Dmitriev, Ivan V. Smirnov, Olga G. Shamborant, Dmitry S. Balabashin, Lidia P. Sashchenko, Alexander G. Tonevitsky, Alain Friboulet, Alexander G. Gabibov, Sergey M. Deyev

Research output: Contribution to journalArticle

23 Citations (Scopus)

Abstract

B cells play an important role in the pathogenesis of both systemic and organ-specific autoimmune diseases. Autoreactive B cells not only produce autoantibodies, but also are capable to efficiently present specific autoantigens to T cells. Furthermore, B cells can secrete proinflammatory cytokines and amplify the vicious process of self-destruction. B cell-directed therapy is a potentially important approach for treatment of various autoimmune diseases. The depletion of B cells by anti-CD20/19 monoclonal antibody Retuximab® used in autoimmune diseases therapy leads to systemic side effects and should be significantly improved. In this study we designed a repertoire of genetically engineered B cell killers that specifically affected one kind of cells carrying a respective B cell receptor. We constructed immunotoxins (ITs), fused with c-myc epitope as a model targeting sequence, based on barnase, Pseudomonas toxin, Shiga-like toxin E.coli and Fc domain of human antibody IgGγ1. C-MYC hybridoma cell line producing anti-c-myc IgG was chosen as a model for targeted cell depletion. C-myc sequence fused with toxins provided addressed delivery of the toxic agent to the target cells. We demonstrated functional activity of designed ITs in vitro and showed recognition of the fusion molecules by antibodies produced by targeted hybridoma. To study specificity of the proposed B cells killing molecules, we tested a set of created ITs ex vivo, using C-MYC and irrelevant hybridoma cell lines. Pseudomonas-containing IT showed one of the highest cytotoxic effects on the model cells, however, possessed promiscuous specificity. Shiga-like toxin construct demonstrated mild both cytotoxicity and specificity. Barnase and Fc-containing ITs revealed excellent balance between their legibility and toxic properties. Moreover, barnase and Fc molecules fused with c-myc epitope were able to selectively deplete c-myc-specific B cells and decrease production of anti-c-myc antibodies in culture of native splenocytes, suggesting their highest therapeutic potential as targeted B cell killing agents.

Original languageEnglish
Article numbere20991
JournalPLoS One
Volume6
Issue number6
DOIs
Publication statusPublished - 2011
Externally publishedYes

Fingerprint

B-lymphocytes
B-Lymphocytes
Cells
Immunotoxins
hybridomas
autoimmune diseases
Hybridomas
Shiga-like toxins
Shiga Toxins
Autoimmune Diseases
Poisons
Pseudomonas
therapeutics
epitopes
antibodies
cytotoxicity
Epitopes
toxins
Immunoglobulin G
Molecules

ASJC Scopus subject areas

  • Medicine(all)
  • Biochemistry, Genetics and Molecular Biology(all)
  • Agricultural and Biological Sciences(all)

Cite this

Stepanov, A. V., Belogurov, A. A., Ponomarenko, N. A., Stremovskiy, O. A., Kozlov, L. V., Bichucher, A. M., ... Deyev, S. M. (2011). Design of targeted B cell killing agents. PLoS One, 6(6), [e20991]. https://doi.org/10.1371/journal.pone.0020991

Design of targeted B cell killing agents. / Stepanov, Alexey V.; Belogurov, Alexey A.; Ponomarenko, Natalia A.; Stremovskiy, Oleg A.; Kozlov, Leonid V.; Bichucher, Anna M.; Dmitriev, Sergey E.; Smirnov, Ivan V.; Shamborant, Olga G.; Balabashin, Dmitry S.; Sashchenko, Lidia P.; Tonevitsky, Alexander G.; Friboulet, Alain; Gabibov, Alexander G.; Deyev, Sergey M.

In: PLoS One, Vol. 6, No. 6, e20991, 2011.

Research output: Contribution to journalArticle

Stepanov, AV, Belogurov, AA, Ponomarenko, NA, Stremovskiy, OA, Kozlov, LV, Bichucher, AM, Dmitriev, SE, Smirnov, IV, Shamborant, OG, Balabashin, DS, Sashchenko, LP, Tonevitsky, AG, Friboulet, A, Gabibov, AG & Deyev, SM 2011, 'Design of targeted B cell killing agents', PLoS One, vol. 6, no. 6, e20991. https://doi.org/10.1371/journal.pone.0020991
Stepanov AV, Belogurov AA, Ponomarenko NA, Stremovskiy OA, Kozlov LV, Bichucher AM et al. Design of targeted B cell killing agents. PLoS One. 2011;6(6). e20991. https://doi.org/10.1371/journal.pone.0020991
Stepanov, Alexey V. ; Belogurov, Alexey A. ; Ponomarenko, Natalia A. ; Stremovskiy, Oleg A. ; Kozlov, Leonid V. ; Bichucher, Anna M. ; Dmitriev, Sergey E. ; Smirnov, Ivan V. ; Shamborant, Olga G. ; Balabashin, Dmitry S. ; Sashchenko, Lidia P. ; Tonevitsky, Alexander G. ; Friboulet, Alain ; Gabibov, Alexander G. ; Deyev, Sergey M. / Design of targeted B cell killing agents. In: PLoS One. 2011 ; Vol. 6, No. 6.
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