TY - JOUR

T1 - Polymorphisms of the T cell receptor CD3delta and CD3varepsilon chains affect anti-CD3 antibody binding and T cell activation

AU - Boding, Lasse

AU - Nielsen, Martin Weiss

AU - Bonefeld, Charlotte Menné

AU - von Essen, Marina Rode

AU - Nielsen, Bodil Lisbeth

AU - Lauritsen, Jens Peter H

AU - Hansen, Ann Kathrine

AU - Nielsen, Morten Milek

AU - Kongsbak, Martin

AU - Rubin, Maria

AU - Vennegaard, Marie Torp

AU - Ødum, Niels

AU - Geisler, Carsten

N1 - Keywords: Amino Acid Sequence; Amino Acid Substitution; Animals; Antigens, CD3; Cell Line; Embryonic Stem Cells; Gene Knock-In Techniques; Lymphocyte Activation; Mice; Mice, Inbred C57BL; Mice, Inbred Strains; Mice, Knockout; Molecular Sequence Data; Muromonab-CD3; Polymorphism, Single Nucleotide; Protein Binding; Protein Structure, Tertiary; Sequence Alignment; Structure-Activity Relationship; T-Lymphocytes

PY - 2010

Y1 - 2010

N2 - T cell receptor (TCR) structure and function have been thoroughly studied for decades. Production and analyses of knock-out and knock-in mice with mutations in the CD3 chains have contributed significantly to these studies. The generation of such gene-modified mice relies on the availability of suitable embryonic stem (ES) cell lines. Traditionally, ES cell lines from the 129 mouse strains have been used followed by backcrossing to the C57BL/6 strain. In the present study, we demonstrate the existence of polymorphisms in the CD3 genes from mice of the 129 and C57BL/6 strains. These polymorphisms result in amino acid substitutions in the ectodomains of both the CD3delta and CD3varepsilon chains in 129 mice compared to C57BL/6 mice. The amino acid substitutions do not change the stoichiometry or surface expression level of the TCR complex in 129 T cells but cause reduced anti-CD3 antibody binding to 129 T cells. Further, when stimulated with mitogenic anti-CD3 antibodies, T cells from the 129 strains show reduced expression of the activation marker CD69, Ca(2+) flux, IL-2 production and proliferative responses compared to C57BL/6 T cells. These findings demonstrate that polymorphisms of the CD3delta and varepsilon ectodomains exist in mice, and that some of these polymorphisms lead to amino acid substitutions which cause structural changes and affect anti-CD3 antibody binding. Thus, functional T cell studies should be interpreted with caution when anti-CD3 antibodies are used for stimulation of T cells derived from gene-modified mice originating from 129 ES cell lines.

AB - T cell receptor (TCR) structure and function have been thoroughly studied for decades. Production and analyses of knock-out and knock-in mice with mutations in the CD3 chains have contributed significantly to these studies. The generation of such gene-modified mice relies on the availability of suitable embryonic stem (ES) cell lines. Traditionally, ES cell lines from the 129 mouse strains have been used followed by backcrossing to the C57BL/6 strain. In the present study, we demonstrate the existence of polymorphisms in the CD3 genes from mice of the 129 and C57BL/6 strains. These polymorphisms result in amino acid substitutions in the ectodomains of both the CD3delta and CD3varepsilon chains in 129 mice compared to C57BL/6 mice. The amino acid substitutions do not change the stoichiometry or surface expression level of the TCR complex in 129 T cells but cause reduced anti-CD3 antibody binding to 129 T cells. Further, when stimulated with mitogenic anti-CD3 antibodies, T cells from the 129 strains show reduced expression of the activation marker CD69, Ca(2+) flux, IL-2 production and proliferative responses compared to C57BL/6 T cells. These findings demonstrate that polymorphisms of the CD3delta and varepsilon ectodomains exist in mice, and that some of these polymorphisms lead to amino acid substitutions which cause structural changes and affect anti-CD3 antibody binding. Thus, functional T cell studies should be interpreted with caution when anti-CD3 antibodies are used for stimulation of T cells derived from gene-modified mice originating from 129 ES cell lines.

U2 - 10.1016/j.molimm.2010.06.012

DO - 10.1016/j.molimm.2010.06.012

M3 - Journal article

C2 - 20638133

VL - 47

SP - 2450

EP - 2457

JO - Molecular Immunology

JF - Molecular Immunology

SN - 0161-5890

IS - 15

ER -