Physiological level production of antigen-specific human immunoglobulin in cloned transchromosomic cattle

PLoS One. 2013 Oct 24;8(10):e78119. doi: 10.1371/journal.pone.0078119. eCollection 2013.

Abstract

Therapeutic human polyclonal antibodies (hpAbs) derived from pooled plasma from human donors are Food and Drug Administration approved biologics used in the treatment of a variety of human diseases. Powered by the natural diversity of immune response, hpAbs are effective in treating diseases caused by complex or quickly-evolving antigens such as viruses. We previously showed that transchromosomic (Tc) cattle carrying a human artificial chromosome (HAC) comprising the entire unrearranged human immunoglobulin heavy-chain (hIGH) and kappa-chain (hIGK) germline loci (named as κHAC) are capable of producing functional hpAbs when both of the bovine immunoglobulin mu heavy-chains, bIGHM and bIGHML1, are homozygously inactivated (double knockouts or DKO). However, B lymphocyte development in these Tc cattle is compromised, and the overall production of hpAbs is low. Here, we report the construction of an improved HAC, designated as cKSL-HACΔ, by incorporating all of the human immunoglobulin germline loci into the HAC. Furthermore, for avoiding the possible human-bovine interspecies incompatibility between the human immunoglobulin mu chain protein (hIgM) and bovine transmembrane α and β immunoglobulins (bIgα and bIgβ) in the pre-B cell receptor (pre-BCR) complex, we partially replaced (bovinized) the hIgM constant domain with the counterpart of bovine IgM (bIgM) that is involved in the interaction between bIgM and bIgα/Igβ; human IgM bovinization would also improve the functionality of hIgM in supporting B cell activation and proliferation. We also report the successful production of DKO Tc cattle carrying the cKSL-HACΔ (cKSL-HACΔ/DKO), the dramatic improvement of B cell development in these cattle and the high level production of hpAbs (as measured for the human IgG isotype) in the plasma. We further demonstrate that, upon immunization by tumor immunogens, high titer tumor immunogen-specific human IgG (hIgG) can be produced from such Tc cattle.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • B-Lymphocytes / metabolism
  • Cattle
  • Cell Proliferation
  • Chromosomes, Artificial, Human / genetics
  • Chromosomes, Artificial, Human / immunology
  • Chromosomes, Artificial, Human / metabolism
  • Humans
  • Immunoglobulin G / genetics
  • Immunoglobulin G / immunology
  • Immunoglobulin G / metabolism
  • Immunoglobulin Heavy Chains / genetics
  • Immunoglobulin Heavy Chains / immunology
  • Immunoglobulin Heavy Chains / metabolism
  • Immunoglobulin M / genetics
  • Immunoglobulin M / immunology
  • Immunoglobulin M / metabolism
  • Immunoglobulin mu-Chains / genetics
  • Immunoglobulin mu-Chains / immunology
  • Immunoglobulin mu-Chains / metabolism
  • Immunoglobulins / genetics
  • Immunoglobulins / immunology
  • Immunoglobulins / metabolism*

Substances

  • Immunoglobulin G
  • Immunoglobulin Heavy Chains
  • Immunoglobulin M
  • Immunoglobulin mu-Chains
  • Immunoglobulins

Grant support

This work was financed by internal funding of Hematech, Inc., which no longer exists and no longer has financial interest in the work. Hematech has transferred all of its business to Sanford Applied Biosciences, LLC. There are no current external funding sources for this study. Kyowa Hakko Kirin provided permission for the work to be published. Financial interest in this work is currently owned by Sanford Applied Biosciences who did have a role in the decision to publish as well as the preparation of the manuscript. While it is considered beneficial to the interests of Sanford Applied Biosciences to publish this work, this consideration played no role in the study design, data collection and analysis and preparation of the manuscript. This is evidenced by the inclusion of now independent authors with no affiliation to Sanford Applied Biosciences.