Analysis of the equine lymphocyte antigen system by Southern blot hybridization.
Abstract: Fourteen Standardbred horses homozygous for one of six equine lymphocyte antigen (ELA) specificities (A1, A3, A4, A5, A6, or A10) were analyzed by Southern blot hybridization using DNA probes derived from the mouse major histocompatibility complex (MHC). Total genomic DNA from peripheral lymphocytes was digested with the restriction enzymes Hind III, Pvu II, or Eco RI. Twenty-three to thirty-three bands were generated for individual horses with the class I cDNA probe. The resulting band patterns revealed 12-14 nonpolymorphic fragments, which is consistent with the highly conserved Qa/Tla genes seen in other species. The remaining 10-19 bands displayed significant polymorphism; no two animals had identical band patterns when studied with all three enzymes. The polymorphism was markedly decreased between animals of the same ELA serotypes. Unique bands were identified in both A1 horses and all four A6 animals. Pvu II digestions of lymphocyte DNA were hybridized with mouse MHC class II probes. A cDNA probe for the E alpha gene revealed only a single nonpolymorphic band. In contrast, a cDNA probe for the H-2 A alpha locus displayed three to five strong bands in each animal with polymorphism that was most pronounced between horses of different ELA serotypes. Genomic DNA probes for A beta and E beta genes both revealed multiple polymorphic bands. However, cross-hybridization between these two probes prevented distinction between A beta and E beta equivalent loci. The reduced polymorphism evident within ELA specificities is consistent with the concept that the equine lymphocyte antigen system includes two families of closely linked MHC genes.
Publication Date: 1987-01-01 PubMed ID: 2880799DOI: 10.1007/BF00768832Google 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
- U.S. Gov't
- P.H.S.
Summary
This research summary has been generated with artificial intelligence and may contain errors and omissions. Refer to the original study to confirm details provided. Submit correction.
This research studied the equine lymphocyte antigen (ELA) system in horses using Southern blot hybridization and DNA probes from the mouse major histocompatibility complex (MHC). The study showed significant variation in the band patterns, suggesting that the ELA system contains two closely linked MHC gene families.
Objective and Methodology
This study sought to analyze the Equine Lymphocyte Antigen (ELA) system by using Southern blot hybridization, a method used to check for presence of a specific DNA sequence within a DNA sample. The researchers carried out this analysis on fourteen standardbred horses. These horses were homozygous (possessing two identical forms of a particular gene) for one of the six ELA specificities: A1, A3, A4, A5, A6, or A10.
- The DNA samples used for this study were derived from lymphocytes, a type of white blood cell. The DNA was dissected via three restriction enzymes: Hind III, Pvu II, and Eco RI.
- The researchers applied a DNA probe from the mouse’s major histocompatibility complex (MHC), a set of cell-surface proteins essential for organism’s immune system, to conduct the Southern blot hybridization.
Results and Observations
- When the class I cDNA probe was applied, between 23 to 33 bands were produced for each horse. Out of these, 12-14 were nonpolymorphic fragments, signifying they did not vary in form. This trait is shared with their counter parts in other species, the highly conserved Qa/Tla genes.
- The remaining 10-19 bands exhibited prominent polymorphism (genetic variation). No similar band patterns were found in any two animals when studied under all the three enzymes.
- There was however, marked decrease in variation among animals of the same ELA serotypes. Unique banding patterns were identified for both A1 horses and all four A6 horses.
- There were more findings when lymphocyte DNA digested with Pvu II was studied with the mouse MHC class II probes. A cDNA probe for the E alpha gene only showed a singular, nonpolymorphic band. However, a significant polymorphism was observed within the bands revealed by a cDNA probe for the H-2 A alpha locus.
- The probes for A beta and E beta genes also unraveled various polymorphic bands. Cross-hybridization between these two probes hindered distinction between A beta and E beta equivalent loci though.
Interpretation and Conclusion
- The outcomes of this research indicate that the ELA system comprises of two interconnected families of MHC genes. This is inferred from the reduced polymorphism found within ELA specificities.
- A clear understanding of genetic variations within ELA could be of great significance for studying immune response in horses, potentially leading to improved vaccination strategies and disease resistance.
Cite This Article
APA
Alexander AJ, Bailey E, Woodward JG.
(1987).
Analysis of the equine lymphocyte antigen system by Southern blot hybridization.
Immunogenetics, 25(1), 47-54.
https://doi.org/10.1007/BF00768832 Publication
Researcher Affiliations
MeSH Terms
- Animals
- Antigens / analysis
- DNA / genetics
- Horses / genetics
- Horses / immunology
- Lymphocytes / immunology
- Nucleic Acid Hybridization
- Nucleotide Mapping
- Polymorphism, Genetic
- Polymorphism, Restriction Fragment Length
Grant Funding
- CA 39070 / NCI NIH HHS
References
This article includes 15 references
- Wettstein PJ, Faas S, Buck DA. Class I and class II restriction pattern polymorphisms associated with independently derived RT1 haplotypes in inbred rats.. Immunogenetics 1985;22(1):9-22.
- Steinmetz M, Frelinger JG, Fisher D, Hunkapiller T, Pereira D, Weissman SM, Uehara H, Nathenson S, Hood L. Three cDNA clones encoding mouse transplantation antigens: homology to immunoglobulin genes.. Cell 1981 Apr;24(1):125-34.
- Mathis DJ, Benoist CO, Williams VE 2nd, Kanter MR, McDevitt HO. The murine E alpha immune response gene.. Cell 1983 Mar;32(3):745-54.
- Sood AK, Pereira D, Weissman SM. Isolation and partial nucleotide sequence of a cDNA clone for human histocompatibility antigen HLA-B by use of an oligodeoxynucleotide primer.. Proc Natl Acad Sci U S A 1981 Jan;78(1):616-20.
- Chardon P, Kirszenbaum M, Cullen PR, Geffrotin C, Auffray C, Strominger JL, Cohen D, Vaiman M. Analysis of the sheep MHC using HLA class I, II, and C4 cDNA probes.. Immunogenetics 1985;22(4):349-58.
- Bailey E. Population studies on the ELA system in American standardbred and thoroughbred mares.. Anim Blood Groups Biochem Genet 1983;14(3):201-11.
- Flavell RA, Allen H, Huber B, Wake C, Widera G. Organization and expression of the MHC of the C57 black/10 mouse.. Immunol Rev 1985 Jul;84:29-50.
- Chardon P, Vaiman M, Kirszenbaum M, Geffrotin C, Renard C, Cohen D. Restriction fragment length polymorphism of the major histocompatibility complex of the pig.. Immunogenetics 1985;21(2):161-71.
- Lew AM, Bailey E, Valas RB, Coligan J. The gene encoding the equine soluble class I molecule is linked to the horse MHC.. Immunogenetics 1986;24(2):128-30.
- Southern EM. Detection of specific sequences among DNA fragments separated by gel electrophoresis.. J Mol Biol 1975 Nov 5;98(3):503-17.
- Bailey E, Stormont C, Suzuki Y, Trommershausen Smith A. Linkage of loci controlling alloantigens on red blood cells and lymphocytes in the horse.. Science 1979 Jun 22;204(4399):1317-9.
- Pease LR, Nathenson SG, Leinwand LA. Mapping class I gene sequences in the major histocompatibility complex.. Nature 1982 Jul 22;298(5872):382-5.
- Steinmetz M, Hood L. Genes of the major histocompatibility complex in mouse and man.. Science 1983 Nov 18;222(4625):727-33.
- Stetler D, Das H, Nunberg JH, Saiki R, Sheng-Dong R, Mullis KB, Weissman SM, Erlich HA. Isolation of a cDNA clone for the human HLA-DR antigen alpha chain by using a synthetic oligonucleotide as a hybridization probe.. Proc Natl Acad Sci U S A 1982 Oct;79(19):5966-70.
- Lazary S, Bullen S, Müller J, Kovacs G, Bodo I, Hockenjos P, De Weck AL. Equine leukocyte antigen system. II. Serological and mixed lymphocyte reactivity studies in families.. Transplantation 1980 Sep;30(3):210-5.
Citations
This article has been cited 10 times.- Sasaki M, Hasebe R, Makino Y, Suzuki T, Fukushi H, Okamoto M, Matsuda K, Taniyama H, Sawa H, Kimura T. Equine major histocompatibility complex class I molecules act as entry receptors that bind to equine herpesvirus-1 glycoprotein D.. Genes Cells 2011 Apr;16(4):343-57.
- Tallmadge RL, Lear TL, Antczak DF. Genomic characterization of MHC class I genes of the horse.. Immunogenetics 2005 Nov;57(10):763-74.
- Kindt TJ, Singer DS. Class I major histocompatibility complex genes in vertebrate species: what is the common denominator?. Immunol Res 1987;6(1-2):57-66.
- Jouvenne P, Talbot PJ. Rapid identification of overlapping cDNA clones by Southern cross-hybridization.. Nucleic Acids Res 1988 Nov 11;16(21):10390.
- Ansari HA, Hediger R, Fries R, Stranzinger G. Chromosomal localization of the major histocompatibility complex of the horse (ELA) by in situ hybridization.. Immunogenetics 1988;28(5):362-4.
- Sachs DH, Germana S, el-Gamil M, Gustafsson K, Hirsch F, Pratt K. Class II genes of miniature swine. I. Class II gene characterization by RFLP and by isolation from a genomic library.. Immunogenetics 1988;28(1):22-9.
- Andersson L, Rask L. Characterization of the MHC class II region in cattle. The number of DQ genes varies between haplotypes.. Immunogenetics 1988;27(2):110-20.
- Ryder OA, Chemnick LG. Chromosomal and molecular evolution in Asiatic wild asses.. Genetica 1990;83(1):67-72.
- Albright D, Bailey E, Woodward JG. Nucleotide sequence of a cDNA clone of the horse (Equus caballus) DRA gene.. Immunogenetics 1991;34(2):136-8.
- Miller-Edge MA, Worley MB. In vitro responses of cheetah mononuclear cells to feline herpesvirus-1 and Cryptococcus neoformans.. Vet Immunol Immunopathol 1992 Jan 15;30(2-3):261-74.
Use Nutrition Calculator
Check if your horse's diet meets their nutrition requirements with our easy-to-use tool Check your horse's diet with our easy-to-use tool
Talk to a Nutritionist
Discuss your horse's feeding plan with our experts over a free phone consultation Discuss your horse's diet over a phone consultation
Submit Diet Evaluation
Get a customized feeding plan for your horse formulated by our equine nutritionists Get a custom feeding plan formulated by our nutritionists
