Analysis of the horse V(H) repertoire and comparison with the human IGHV germline genes, and sheep, cattle and pig V(H) sequences.
Abstract: We have constructed a chimeric antibody single-chain Fv (scFv) fragments phage-displayed library that combines an invariant human V(L) chain with the repertoire of V(H) domains amplified from a horse immunized against scorpion venom. To gain insight into the equine V(H) repertoire, the V(H) sequences of 46 unique clones randomly chosen from the library prior to antigenic selection were analyzed. Comparisons with previously reported equine V(H) sequences, as well as with the repertoire of human IGHV germline genes and known V(H) sequences of sheep, cattle and pig, suggest that the equine IGH locus harbors at least three IGHV gene families. Two families belong to clan II while the other was classified into clan I. The horse sequences were also found to encode a diverse repertoire of canonical structures. The most populated equine IGHV gene family, named IGHV1, and another family termed IGHV3, encode two out of the three canonical structures so far described for CDR1. The IGHV2 gene family has the third canonical structure at CDR1. In CDR2, nine loop lengths were found, with four of them matching the pattern of typical canonical structures. The remaining five CDR2 loop lengths are shorter or longer than those reported for human IGHV germline genes and known sequences of sheep, cattle and pig. The analysis of CDR3 loops indicates a length distribution broader than previous reports for horses; being similar to that of humans, sheep and pigs. Moreover, equine CDR3 loops were found to have a combination of lower content of cysteine and higher proportion of glycine not seen in the other species. This implies less constrained loops and therefore more apt for searching the conformational space of antigen-binding sites. Altogether, these findings reveal a more diverse perspective of the horse V(H) repertoire than previous estimations and lay foundations for future studies of the equine IGH locus.
Publication Date: 2005-12-07 PubMed ID: 16337682DOI: 10.1016/j.molimm.2005.10.017Google Scholar: Lookup
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- Comparative Study
- Journal Article
- Research Support
- Non-U.S. Gov't
Summary
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The research conducted an analysis of the horse V(H) repertoire and compared it with human, sheep, cattle, and pig V(H) sequences. The study created a phage-displayed library of antibody fragments from a horse immunized with scorpion venom and discovered a more diverse range of canonical structures in the equine V(H) repertoire than previously estimated.
Objective of the Study
- The main aim of the research was to gain insight into the equine V(H) repertoire, which pertains to the horse’s immune response, and their comparison with the repertoires of human, sheep, cattle, and pigs.
Methods and Materials
- The researchers constructed a chimeric antibody (scFv fragments phage-displayed library) using a horse that had been immunized against scorpion venom.
- The V(H) sequences from the library were then analyzed and compared with data from previously known equine V(H) sequences, sheep, cattle, pig, and human IGHV germline genes.
Findings
- The researchers found that the horse IGH locus harbors at least three IGHV gene families, which suggests a more diverse immune response than previously assumed.
- Two out of these three gene families belong to clan II, while the other comes under clan I.
- The gene families IGHV1 and IGHV3 were found to encode two out of the three known structures for complementarity determining regions (CDR1).
- The third canonical structure for CDR1 was found in the IGHV2 gene family.
- In the analyses of CDR2 loops, nine different loop lengths were found.
- The equine CDR3 loops displayed a broader length distribution, comparable to that of humans, sheep, and pigs and below the reported content of cysteine with a higher amount of glycine, implying its adaptability potential in the form of less constrained loops for antigen-binding sites.
Conclusion
- The findings provide a comprehensive view of the horse V(H) repertoire, suggesting it as more variable than assumed before.
- The study lays the foundation for future studies and exploration in the field of equine IGH locus.
Cite This Article
APA
Almagro JC, Martinez L, Smith SL, Alagon A, Estevez J, Paniagua J.
(2005).
Analysis of the horse V(H) repertoire and comparison with the human IGHV germline genes, and sheep, cattle and pig V(H) sequences.
Mol Immunol, 43(11), 1836-1845.
https://doi.org/10.1016/j.molimm.2005.10.017 Publication
Researcher Affiliations
- Florida International University, Department of Biological Sciences, FL 33199, USA. almagro@fiu.edu
MeSH Terms
- Amino Acid Sequence
- Animals
- Cattle
- Complementarity Determining Regions / genetics
- Complementarity Determining Regions / immunology
- Gene Library
- Germ Cells / metabolism
- Horses / genetics
- Horses / immunology
- Humans
- Immunoglobulin Heavy Chains / chemistry
- Immunoglobulin Heavy Chains / genetics
- Immunoglobulin Heavy Chains / immunology
- Immunoglobulin Variable Region / chemistry
- Immunoglobulin Variable Region / genetics
- Immunoglobulin Variable Region / immunology
- Molecular Sequence Data
- Nucleotides / genetics
- Peptide Library
- Phylogeny
- Sequence Alignment
- Sequence Analysis, Protein
- Sheep, Domestic / genetics
- Sheep, Domestic / immunology
- Swine / genetics
- Swine / immunology
Citations
This article has been cited 13 times.- Wibmer CK, Mashilo P. Exploiting V-Gene Bias for Rapid, High-Throughput Monoclonal Antibody Isolation from Horses. Viruses 2022 Sep 30;14(10).
- Rosenfeld R, Alcalay R, Zvi A, Ben-David A, Noy-Porat T, Chitlaru T, Epstein E, Israeli O, Lazar S, Caspi N, Barnea A, Dor E, Chomsky I, Pitel S, Makdasi E, Zichel R, Mazor O. Centaur antibodies: Engineered chimeric equine-human recombinant antibodies. Front Immunol 2022;13:942317.
- Stanfield RL, Wilson IA, Smider VV. Conservation and diversity in the ultralong third heavy-chain complementarity-determining region of bovine antibodies. Sci Immunol 2016 Jul;1(1).
- Al Qaraghuli MM, Palliyil S, Broadbent G, Cullen DC, Charlton KA, Porter AJ. Defining the complementarities between antibodies and haptens to refine our understanding and aid the prediction of a successful binding interaction. BMC Biotechnol 2015 Oct 24;15:99.
- Badial PR, Tallmadge RL, Miller S, Stokol T, Richards K, Borges AS, Felippe MJ. Applied Protein and Molecular Techniques for Characterization of B Cell Neoplasms in Horses. Clin Vaccine Immunol 2015 Nov;22(11):1133-45.
- Battista JM, Tallmadge RL, Stokol T, Felippe MJ. Hematopoiesis in the equine fetal liver suggests immune preparedness. Immunogenetics 2014 Nov;66(11):635-49.
- Walther S, Czerny CP, Diesterbeck US. Exceptionally long CDR3H are not isotype restricted in bovine immunoglobulins. PLoS One 2013;8(5):e64234.
- Tallmadge RL, Tseng CT, King RA, Felippe MJ. Developmental progression of equine immunoglobulin heavy chain variable region diversity. Dev Comp Immunol 2013 Sep;41(1):33-43.
- Sun Y, Liu Z, Ren L, Wei Z, Wang P, Li N, Zhao Y. Immunoglobulin genes and diversity: what we have learned from domestic animals. J Anim Sci Biotechnol 2012 Jun 20;3(1):18.
- Guo Y, Bao Y, Meng Q, Hu X, Meng Q, Ren L, Li N, Zhao Y. Immunoglobulin genomics in the guinea pig (Cavia porcellus). PLoS One 2012;7(6):e39298.
- Guo Y, Bao Y, Wang H, Hu X, Zhao Z, Li N, Zhao Y. A preliminary analysis of the immunoglobulin genes in the African elephant (Loxodonta africana). PLoS One 2011 Feb 25;6(2):e16889.
- Herzig CT, Lefranc MP, Baldwin CL. Annotation and classification of the bovine T cell receptor delta genes. BMC Genomics 2010 Feb 9;11:100.
- Keggan A, Freer H, Rollins A, Wagner B. Production of seven monoclonal equine immunoglobulins isotyped by multiplex analysis. Vet Immunol Immunopathol 2013 Jun 15;153(3-4):187-93.
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