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cDNA sequence and Fab crystal structure of HL4E10, a hamster IgG lambda light chain antibody stimulatory for γδ T cells.

Verdino P, Witherden DA, Podshivalova K, Rieder SE, Havran WL, Wilson IA - PLoS ONE (2011)

Bottom Line: We generated antibody HL4E10 as a probe to identify novel costimulatory molecules on the surface of γδ T cells which lack the traditional αβ T cell co-receptors CD4, CD8, and the costimulatory molecule CD28.The crystal structure of the HL4E10 Fab at 2.95 Å resolution reveals a rigid combining site with pockets faceted by solvent-exposed tyrosine residues, which are structurally optimized for JAML binding.As the HL4E10 antibody is uniquely costimulatory for γδ T cells, humanized versions thereof may be of clinical relevance in treating γδ T cell dysfunction-associated diseases, such as chronic non-healing wounds and cancer.

View Article: PubMed Central - PubMed

Affiliation: Department of Molecular Biology, The Scripps Research Institute, La Jolla, California, United States of America.

ABSTRACT
Hamsters are widely used to generate monoclonal antibodies against mouse, rat, and human antigens, but sequence and structural information for hamster immunoglobulins is sparse. To our knowledge, only three hamster IgG sequences have been published, all of which use kappa light chains, and no three-dimensional structure of a hamster antibody has been reported. We generated antibody HL4E10 as a probe to identify novel costimulatory molecules on the surface of γδ T cells which lack the traditional αβ T cell co-receptors CD4, CD8, and the costimulatory molecule CD28. HL4E10 binding to γδ T cell, surface-expressed, Junctional Adhesion Molecule-Like (JAML) protein leads to potent costimulation via activation of MAP kinase pathways and cytokine production, resulting in cell proliferation. The cDNA sequence of HL4E10 is the first example of a hamster lambda light chain and only the second known complete hamster heavy chain sequence. The crystal structure of the HL4E10 Fab at 2.95 Å resolution reveals a rigid combining site with pockets faceted by solvent-exposed tyrosine residues, which are structurally optimized for JAML binding. The characterization of HL4E10 thus comprises a valuable addition to the spartan database of hamster immunoglobulin genes and structures. As the HL4E10 antibody is uniquely costimulatory for γδ T cells, humanized versions thereof may be of clinical relevance in treating γδ T cell dysfunction-associated diseases, such as chronic non-healing wounds and cancer.

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Related in: MedlinePlus

Amino-acid sequence alignment of the HL4E10 hamster IgG light chain with those of other hamster antibodies.The HL4E10 lambda light chain is aligned with H28.710 (Genbank accession no. U17165 [32]), 145.2c11 (U17870 [31]), and 1F4 (S80615 [30]). Identical residues are in red and homologous exchanges are in green. Signal peptides are underlined, CDR loops are shaded: CDR L1 is in yellow, CDR L2 is in cyan, CDR L3 is in orange-red, and residues which are rarely observed in antibodies at particular locations are shaded gray. Kabat numbering is used throughout, as well as the definition of CDRs.
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pone-0019828-g001: Amino-acid sequence alignment of the HL4E10 hamster IgG light chain with those of other hamster antibodies.The HL4E10 lambda light chain is aligned with H28.710 (Genbank accession no. U17165 [32]), 145.2c11 (U17870 [31]), and 1F4 (S80615 [30]). Identical residues are in red and homologous exchanges are in green. Signal peptides are underlined, CDR loops are shaded: CDR L1 is in yellow, CDR L2 is in cyan, CDR L3 is in orange-red, and residues which are rarely observed in antibodies at particular locations are shaded gray. Kabat numbering is used throughout, as well as the definition of CDRs.

Mentions: Using degenerate primers derived from N-terminal protein sequences, we determined the complete cDNA sequence for the hamster IgG HL4E10. The light chain cDNA comprises 699 base pairs (Genbank accession no. HM369134) encoding a 19-residue signal peptide and a 213 amino acid mature protein chain (Fig. 1). The HL4E10 heavy chain (HM369133) comprises 1389 base pairs encoding a 19-residue signal peptide and a 443-residue mature protein chain (Fig. 2).


cDNA sequence and Fab crystal structure of HL4E10, a hamster IgG lambda light chain antibody stimulatory for γδ T cells.

Verdino P, Witherden DA, Podshivalova K, Rieder SE, Havran WL, Wilson IA - PLoS ONE (2011)

Amino-acid sequence alignment of the HL4E10 hamster IgG light chain with those of other hamster antibodies.The HL4E10 lambda light chain is aligned with H28.710 (Genbank accession no. U17165 [32]), 145.2c11 (U17870 [31]), and 1F4 (S80615 [30]). Identical residues are in red and homologous exchanges are in green. Signal peptides are underlined, CDR loops are shaded: CDR L1 is in yellow, CDR L2 is in cyan, CDR L3 is in orange-red, and residues which are rarely observed in antibodies at particular locations are shaded gray. Kabat numbering is used throughout, as well as the definition of CDRs.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC3101210&req=5

pone-0019828-g001: Amino-acid sequence alignment of the HL4E10 hamster IgG light chain with those of other hamster antibodies.The HL4E10 lambda light chain is aligned with H28.710 (Genbank accession no. U17165 [32]), 145.2c11 (U17870 [31]), and 1F4 (S80615 [30]). Identical residues are in red and homologous exchanges are in green. Signal peptides are underlined, CDR loops are shaded: CDR L1 is in yellow, CDR L2 is in cyan, CDR L3 is in orange-red, and residues which are rarely observed in antibodies at particular locations are shaded gray. Kabat numbering is used throughout, as well as the definition of CDRs.
Mentions: Using degenerate primers derived from N-terminal protein sequences, we determined the complete cDNA sequence for the hamster IgG HL4E10. The light chain cDNA comprises 699 base pairs (Genbank accession no. HM369134) encoding a 19-residue signal peptide and a 213 amino acid mature protein chain (Fig. 1). The HL4E10 heavy chain (HM369133) comprises 1389 base pairs encoding a 19-residue signal peptide and a 443-residue mature protein chain (Fig. 2).

Bottom Line: We generated antibody HL4E10 as a probe to identify novel costimulatory molecules on the surface of γδ T cells which lack the traditional αβ T cell co-receptors CD4, CD8, and the costimulatory molecule CD28.The crystal structure of the HL4E10 Fab at 2.95 Å resolution reveals a rigid combining site with pockets faceted by solvent-exposed tyrosine residues, which are structurally optimized for JAML binding.As the HL4E10 antibody is uniquely costimulatory for γδ T cells, humanized versions thereof may be of clinical relevance in treating γδ T cell dysfunction-associated diseases, such as chronic non-healing wounds and cancer.

View Article: PubMed Central - PubMed

Affiliation: Department of Molecular Biology, The Scripps Research Institute, La Jolla, California, United States of America.

ABSTRACT
Hamsters are widely used to generate monoclonal antibodies against mouse, rat, and human antigens, but sequence and structural information for hamster immunoglobulins is sparse. To our knowledge, only three hamster IgG sequences have been published, all of which use kappa light chains, and no three-dimensional structure of a hamster antibody has been reported. We generated antibody HL4E10 as a probe to identify novel costimulatory molecules on the surface of γδ T cells which lack the traditional αβ T cell co-receptors CD4, CD8, and the costimulatory molecule CD28. HL4E10 binding to γδ T cell, surface-expressed, Junctional Adhesion Molecule-Like (JAML) protein leads to potent costimulation via activation of MAP kinase pathways and cytokine production, resulting in cell proliferation. The cDNA sequence of HL4E10 is the first example of a hamster lambda light chain and only the second known complete hamster heavy chain sequence. The crystal structure of the HL4E10 Fab at 2.95 Å resolution reveals a rigid combining site with pockets faceted by solvent-exposed tyrosine residues, which are structurally optimized for JAML binding. The characterization of HL4E10 thus comprises a valuable addition to the spartan database of hamster immunoglobulin genes and structures. As the HL4E10 antibody is uniquely costimulatory for γδ T cells, humanized versions thereof may be of clinical relevance in treating γδ T cell dysfunction-associated diseases, such as chronic non-healing wounds and cancer.

Show MeSH
Related in: MedlinePlus