An isotype switched and somatically mutated rheumatoid factor clone isolated from a MRL-lpr/lpr mouse exhibits limited intraclonal affinity maturation

J Immunol. 1994 May 1;152(9):4489-99.

Abstract

Employing site-directed mutagenesis we have reconstructed and expressed the germ-line precursor of an expanded rheumatoid factor (RF) clone. This RF clone, designated clone F, was isolated from an autoimmune MRL/MpJ-lpr/lpr mouse. Most of the clone members were extensively mutated and isotyped-switched. The predominant isotype of clone F was gamma 3. The RF bound specifically to the MRL gamma 2 a allotype (Igh-1j) but not to the B6 gamma 2a allotype (Igh-1b). The germ-line antibody was also found to bind gamma 2a in an RF assay. The affinities of the germ-line RF and representative members of the clone were measured in an ELISA-based equilibrium binding assay. The dissociation constant (Kd) of the germ-line RF was 2.5 x 10(-6) M. All of the expressed clone members had affinities within a two- to sixfold range of the germ line, indicating that the mechanisms of somatic hypermutation and selection resulted in only limited affinity maturation of this autoantibody clone.

Publication types

  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Amino Acid Sequence
  • Animals
  • Antibody Affinity
  • Antibody Diversity / genetics
  • B-Lymphocytes / immunology
  • Base Sequence
  • Cloning, Molecular
  • DNA, Complementary / genetics
  • Hybridomas / immunology
  • Immunoglobulin Class Switching / genetics*
  • Kinetics
  • Mice
  • Mice, Mutant Strains
  • Molecular Sequence Data
  • Mutagenesis, Site-Directed
  • Rheumatoid Factor / genetics*
  • Rheumatoid Factor / immunology
  • Rheumatoid Factor / metabolism
  • Transfection

Substances

  • DNA, Complementary
  • Rheumatoid Factor