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Molecular and functional characterization of genes encoding horse MHC class I antigens.
Abstract: Sequence and functional analyses were undertaken on two cDNAs and a genomic clone encoding horse major histocompatibility complex (MHC) class I molecules. All of the clones were isolated from a single horse that is homozygous for all known horse MHC class I and class II antigens. The two cDNAs (clones 8-9 and 1-29) were isolated from a lymphocyte library and encode polymorphic MHC antigens from two loci. The genomic cosmid clone, isolated from a sperm library, contains the 8-9 gene. All three genes were expressed in mouse L-cells and were recognized by alloantisera and, for the cDNAs, by alloreactive cytotoxic T lymphocytes. A total of 3815 bp of the genomic clone were sequenced, extending from 429 bp upstream (5') of the leader peptide through the 3' untranslated region. Promoter region motifs and an intron-exon structure characteristic of MHC class I genes of other species were found. A subclone containing 407 bp of the promoter region was inserted into a chloramphenicol acetyl transferase reporter plasmid, tested in transient transfection assays, and found to have promoter activity in heterologous cells. This genomic clone will enable detailed studies of MHC class I gene regulation in horse trophoblasts, and in horse retroviral infections.
Publication Date: 2001-11-17 PubMed ID: 11862413DOI: 10.1007/s00251-001-0384-xGoogle Scholar: Lookup The Equine Research Bank provides access to a large database of publicly available scientific literature. Inclusion in the Research Bank does not imply endorsement of study methods or findings by Mad Barn.
- Journal Article
- Research Support
- Non-U.S. Gov't
- Research Support
- U.S. Gov't
- Non-P.H.S.
- Research Support
- U.S. Gov't
- P.H.S.
Summary
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The research article discusses the molecular and functional studies done on genes responsible for encoding horse Major Histocompatibility Complex (MHC) class I antigens. These analyses were performed on two cDNAs and one genomic clone, all originating from the same horse, which is homozygous for known MHC class I and II antigens.
Horse MHC class I molecules
- The research aims to understand the structure of horse MHC I antigens, which play a key role in immune response.
- The cDNAs and genomic clone were examined, the former two originating from a lymphocyte library while the genomic clone was isolated from a sperm library.
- The cDNAs and genomic clone encode the MHC class I antigens characteristic of two separate loci.
Expression in Mouse L-Cells
- All three genes were expressed in mouse L-cells to further their characterization.
- The expression in mouse L-cells showed that the isolated genes were recognized by alloantisera, which confirms the validity of the genomic clone and the two cDNAs.
- Moreover, the cDNAs were recognized by alloreactive cytotoxic T lymphocytes, which provides further evidence for the correct isolation and expression of the genes in question.
Genomic Clone Analysis
- Focusing on the genomic clone, a total of 3815 base pairs (bp) were sequenced, extending from 429 bp upstream of the leader peptide through the 3′ untranslated region.
- Key motifs and an intron-exon structure similar to other species’ MHC class I genes were observed.
Subcloning and Promoter Activity Analysis
- An additional insight was gleaned by inserting a subclone with 407 bp of the promoter region into a chloramphenicol acetyl transferase reporter plasmid.
- This construct was then tested in transient transfection assays to determine the promoter activity in non-native (heterologous) cells.
Future Research
- The report ends by explaining the potential utility of the genomic clone in future research.
- It will enable a closer look at MHC class I gene regulation in horse trophoblasts, which are involved in placenta formation, and during retroviral infections in horses.
Cite This Article
APA
Carpenter S, Baker JM, Bacon SJ, Hopman T, Maher J, Ellis SA, Antczak DF.
(2001).
Molecular and functional characterization of genes encoding horse MHC class I antigens.
Immunogenetics, 53(9), 802-809.
https://doi.org/10.1007/s00251-001-0384-x Publication
Researcher Affiliations
- Department of Veterinary Microbiology and Preventive Medicine, Iowa State University, Ames, IA 50011-1250, USA. scarp@iastate.edu
MeSH Terms
- Amino Acid Sequence
- Animals
- Base Sequence
- Conserved Sequence
- DNA, Complementary / genetics
- Exons
- Genes, MHC Class I
- Histocompatibility Antigens Class I / genetics
- Horses / genetics
- Horses / immunology
- In Vitro Techniques
- Introns
- L Cells
- Male
- Mice
- Molecular Sequence Data
- Promoter Regions, Genetic
- Sequence Homology, Nucleic Acid
- T-Lymphocytes, Cytotoxic / immunology
Grant Funding
- HD-15799 / NICHD NIH HHS
- HD-80436 / NICHD NIH HHS
Citations
This article has been cited 5 times.- Miller D, Tallmadge RL, Binns M, Zhu B, Mohamoud YA, Ahmed A, Brooks SA, Antczak DF. Polymorphism at expressed DQ and DR loci in five common equine MHC haplotypes. Immunogenetics 2017 Mar;69(3):145-156.
- Ramsay JD, Leib SR, Orfe L, Call DR, Tallmadge RL, Fraser DG, Mealey RH. Development of a DNA microarray for detection of expressed equine classical MHC class I sequences in a defined population. Immunogenetics 2010 Sep;62(9):633-9.
- Tallmadge RL, Campbell JA, Miller DC, Antczak DF. Analysis of MHC class I genes across horse MHC haplotypes. Immunogenetics 2010 Mar;62(3):159-72.
- Mealey RH, Lee JH, Leib SR, Littke MH, McGuire TC. A single amino acid difference within the alpha-2 domain of two naturally occurring equine MHC class I molecules alters the recognition of Gag and Rev epitopes by equine infectious anemia virus-specific CTL. J Immunol 2006 Nov 15;177(10):7377-90.
- Tallmadge RL, Lear TL, Antczak DF. Genomic characterization of MHC class I genes of the horse. Immunogenetics 2005 Nov;57(10):763-74.
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