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 - 2010/9
Y1 - 2010/9
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 CD3δ and CD3e{open} 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, Ca2+ flux, IL-2 production and proliferative responses compared to C57BL/6 T cells. These findings demonstrate that polymorphisms of the CD3δ and e{open} 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 CD3δ and CD3e{open} 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, Ca2+ flux, IL-2 production and proliferative responses compared to C57BL/6 T cells. These findings demonstrate that polymorphisms of the CD3δ and e{open} 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
SN - 0161-5890
VL - 47
SP - 2450
EP - 2457
JO - Molecular Immunology
JF - Molecular Immunology
IS - 15
ER -