Functional in vitro studies of recombinant human immunoglobulin G and immunoglobulin A anti-D

Transfusion. 2007 Feb;47(2):306-15. doi: 10.1111/j.1537-2995.2007.01107.x.

Abstract

Background: The use of anti-D purified from human serum to prevent hemolytic disease of the fetus and newborn due to D is well established. Owing to supply and safety reasons, however, an unlimited and non-plasma-derived source of antibodies for Rhesus prophylaxis is needed.

Study design and methods: Recombinant human immunoglobulin G (IgG)1, IgG2, IgG3, IgG4, IgA1, and IgA2 anti-D with the same variable region were expressed in Chinese hamster ovary cells. The effector functions of these antibodies were assessed by an antibody-dependent cell-mediated cytotoxicity (ADCC) assay and a chemiluminescence (CL) method for detection of respiratory burst.

Results: In the ADCC assay, IgG1, IgG3, and IgA1 did the best and were as active as a currently used prophylactic polyclonal anti-D. IgG4 and IgA2 were moderately active, whereas IgG2 was not active. In the CL assay, IgG1 and IgG3 were active but much less so than a currently used prophylactic polyclonal anti-D. For some effector cell preparations, IgG4 was active in the CL assay, whereas IgG2, IgA1, and IgA2 were not. A mixture of IgG1 and IgG3 showed a synergistic effect in the CL assay and did as well as the prophylactic polyclonal anti-D in ADCC and CL. Mixtures of IgA1 and either IgG1 or IgG3 showed no synergistic effect.

Conclusion: A mixture of recombinant human IgG1 and IgG3 anti-D could be of value in future Rhesus prophylaxis.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Antibody Specificity
  • CHO Cells
  • Cricetinae
  • Cricetulus
  • Gene Expression / immunology
  • Humans
  • Immunoglobulin A / genetics*
  • Immunoglobulin A / immunology
  • Immunoglobulin G / genetics*
  • Immunoglobulin G / immunology
  • In Vitro Techniques
  • Recombinant Proteins / genetics*
  • Recombinant Proteins / immunology
  • Respiratory Burst
  • Rh-Hr Blood-Group System / immunology*
  • Transfection

Substances

  • Immunoglobulin A
  • Immunoglobulin G
  • Recombinant Proteins
  • Rh-Hr Blood-Group System
  • Rho(D) antigen