Molecular cloning and characterization of human thyroid peroxidase autoantibodies of lambda light chain type.
IgG class thyroid peroxidase (TPO) autoantibodies with kappa light (L) chains predominate in serum and the genes for a large repertoire of such autoantibodies have been characterized. The present study was performed to clone and characterize TPO autoantibodies with lambda L chains which comprise approximately 20% of serum TPO autoantibodies. From a combinatorial IgG H/lambda L chain cDNA library in the phage display vector pComb3, 24 TPO-binding clones with lambda L chains were isolated, comprising three different heavy (H) and light (L) chain combinations. These combinations utilized two genes from the Vlambda II and IIIb families (closest germline genes DPL11 and hsigg11150) and three genes from the VH1, VH3 and VH4 families (VH26, 4.34 and hv1L1). The deduced amino acid sequences of these H chains were quite different from those of kappa F(ab) isolated using the same H chain library. We expressed the proteins for these three lambda F(ab), as well as for a lambda F(ab) (Humlv318 L chain/DP10-like H chain) previously isolated from another patient. The affinities for TPO of the lambda F(ab) (Kd 8 x 10(-10) M to 10(-7) M) were lower than those of the kappa F(ab) (Kd approximately 10(-10) M). For two lambda F(ab), both H and L chain genes were close to germline configuration, but there was no straightforward relationship between the extent of somatic mutation from germline configuration and affinity for TPO. All four lambda F(ab) bound less well to denatured TPO as to native TPO. The three F(ab) for which sufficient protein could be expressed for competition studies all recognized domain B within the immunodominant region on TPO previously identified using F(ab) with kappa L chains. Aside from these TPO-specific F(ab), only a few other human IgG class, organ-specific autoantibodies with lambda L chains have been characterized at the molecular level. Our study significantly augments the small database on this category of autoantibodies in general.
Portolano
S
S
Thyroid Molecular Biology Unit, Veterans' Administration Medical Center, San Francisco, California 94121, USA.
Prummel
M F
MF
Rapoport
B
B
McLachlan
S M
SM
D000595:Amino Acid Sequence; D000915:Antibody Affinity; D000918:Antibody Specificity; D001323:Autoantibodies / Q000737:chemistry / Q000235:genetics* / Q000302:isolation & purification; D001483:Base Sequence; D003001:Cloning, Molecular; D005803:Genes, Immunoglobulin / Q000276:immunology; D006801:Humans; D016056:Immunodominant Epitopes / Q000737:chemistry; D007140:Immunoglobulin Fab Fragments / Q000235:genetics; D007145:Immunoglobulin kappa-Chains / Q000235:genetics; D007146:Immunoglobulin lambda-Chains / Q000737:chemistry / Q000235:genetics* / Q000302:isolation & purification; D007453:Iodide Peroxidase / Q000276:immunology*; D008969:Molecular Sequence Data; D011489:Protein Denaturation / Q000276:immunology
D001323:Autoantibodies; D016056:Immunodominant Epitopes; D007140:Immunoglobulin Fab Fragments; D007145:Immunoglobulin kappa-Chains; D007146:Immunoglobulin lambda-Chains; D007453:Iodide Peroxidase