Progress 10/01/03 to 09/30/08
Outputs
OUTPUTS: Genes of the chicken major histocompatibility complex (MHC) include B-F, MHC class I; B-L, MHC class II; and B-G, MHC class IV. The genes are closely linked on chromosome 16. Recombination between the B-F and B-G regions of the MHC has been valuable to define genetic control of immune responses more precisely. Congenic lines, each containing a single unique MHC recombinant, were produced on the genetic background of highly inbred Line UCD 003 (B17B17). After ten backcross generations to the background line, these congenic lines have estimated 99.9% genetic uniformity. The susceptible-resistant pigeon (Columba livia) model has been employed to understand genetic components of atherosclerotic cardiovascular disease. The White Carneau (WC) pigeon develops naturally-occurring (non-induced, spontaneous) atherosclerosis without elevated plasma cholesterol levels or other known risk factors. These non-induced atherosclerotic lesions are similar to the human lesions in morphology and ultrastructure, even occurring at similar anatomical sites along the arterial tree. The Show Racer (SR) pigeon is resistant to the atherosclerosis development under identical diet and housing conditions, and with similar blood cholesterol levels. Breeding experiments demonstrated aortic atherosclerosis susceptibility to be inherited as an autosomal recessive Mendelian trait. Results were presented at the annual project meeting, the Poultry Science Association meeting and in peer-reviewed publications. PARTICIPANTS: Participants C. M. Ashwell N. C. State University W. E. Briles Northern Illinois University J. E. Fulton Hy-Line International M. M. Miller Beckman Research Institute P. Wakenell University of California, Davis TARGET AUDIENCES: Poultry breeding and health companies, poultry producers and other poultry scientists. Other scientists interested in animal health issues. PROJECT MODIFICATIONS: New collaborations have begun with N. C. State University and the USDA Avian Disease and Oncology Laboratory, East Lansing, MI.
Impacts
Major histocompatibility (B) complex haplotype affects the incidence of lymphoma tumors following infection of chickens with oncogenic strains of Marek's disease virus (MDV). MHC-B genes contributing to resistance to Marek's disease (MD) mapped to the MHC-B subregion marked by MHC class I antigens. Following infection with the highly virulent RB-1B strain of MDV, two serologically identical recombinant haplotypes, BR2 and BR4 carried in semi-congenic lines, differed in the MD incidence. A larger challenge trial found that these two recombinant 003.R2 and 003.R4 lines, now fully congenic, also had a highly significant difference in MD. The MD incidence was 19% (27/140) in the 003.R2 line and 47% (61/130) in the 003.R4 line (P<0.0001) indicating that the recombinant haplotypes must differ over some portion of the region between the serological BG and BF markers used to identify them. The BR2 and BR4 crossover breakpoints were are separated by less than 1934 bp as resolved by sequencing and SNP mapping. The breakpoints identify the single BG1 gene as the locus affecting the difference in MD incidence observed in the 003.R2 and 003.R4 lines. The BG1 alleles in the two lines are identical in coding region, but differ in 3'-untranslated region (3'UTR). The receptor-like molecule encoded by the BG1 gene contains an immunoreceptor tyrosine-based inhibition motif (ITIM). Experiments are underway to learn how the 3'UTR difference influences BG1 gene activity. Further work is attempting to distinguish whether BG1 contributes in initial responses to infection, in responses to emergence of MDV from its brief latency or in later immune responses to the emerging tumors. Replicate primary aortic smooth muscle cell cultures were prepared from 5 one to three-day-old White Carneau (WC, suseptible) or Show Racer (SR) squabs of mixed genders. Cultures were allowed to grow for 7-8 days until a monolayer was obtained. Proteins were extracted sequentially from 1 x 108 cells. Proteins from each extraction fraction on two-dimensional sodium dodecyl sulfate polyacrylamide gels. Spots on digital gel photographs were analyzed with Phoretix software to identify relative pI and molecular weight of differentially-expressed proteins. There were 170 differentially-expressed proteins observed. Certain zones corresponding to pI and MW ranges contain proteins exclusively associated with either atherosclerosis susceptibility (WC) or resistance (SR). In the WC cells, 88 differentially-expressed proteins were found. Nearly half (41) of these were located in unique susceptibility zones. In SR cells, 82 differentially-expressed proteins were observed with approximately one-third (29) in unique resistance zones. Some of the proteins (such as smooth muscle myosin phosphatase, myosin heavy chain, fatty acid binding protein, ribophorin, heat shock protein, and TNFalpha-inducing factor), corresponded to current hypotheses to explain atherogenesis. Genetic stocks developed and maintained at the University of New Hampshire were the foundation of this research. The work could not have been done without those stocks.
Publications
- Connors, C., Anderson, J. L., Smith, S. C. and Taylor, Jr. R. L., 2008. Genetic Factors of Atherosclerosis in White Carneau Pigeons (Columba livia). 17th UNH COLSA Undergraduate Research Conference 2008 p. 12
- Smith, S. C., Smith, E. C., Gilman, M. L., Anderson, J. L. and Taylor, Jr. R. L. 2008. Differentially expressed soluble proteins in aortic cells from atherosclerosis-susceptible and resistant pigeons. Poult. Sci. 87(Suppl. 1):68-69
- Wang, Y., Goto, R. M., Wakenell, P. S., Taylor, Jr., R. L. and Miller, M. M. 2008. Genetic resistance to GAHV-2 induced lymphoma in the chicken model. 11th Int. Conf. in Malignancies in AIDS and Other Acquired Immunodeficiencies, Bethesda, MD in press
- Chapman, M. A., Taylor, Jr., R. L. and Wideman, Jr., R. F. 2007. Analysis of plasma serotonin levels and hemodynamic responses following chronic serotonin infusion in broilers challenged with bacterial lipopolysaccharide and microparticles. Poult. Sci. 87:116-124
- Smith, S. C., Smith, E. C., Gilman, M. L., Anderson, J. L. and Taylor, Jr. R. L. 2008. Differentially expressed soluble proteins in aortic cells from atherosclerosis-susceptible and resistant pigeons. Poult. Sci. 87:1328-1334
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Progress 10/01/06 to 09/30/07
Outputs
OUTPUTS: Chicken major histocompatibility complex (MHC) genes that are closely linked on chromosome 16 include B-F, MHC class I; B-L, MHC class II; and B-G, MHC class IV. Recombination within the chicken major histocompatibility complex B-F and B-G regions has been useful to define genetic control of immune responses more precisely. Six congenic lines, each containing a single unique MHC recombinant, were produced on the genetic background of Inbred Line UCD 003 (B17B17). These congenic lines have undergone ten backcrosses to the background line and thus have 99.9% genetic uniformity. A new recombinant, designated R13 (BF17-BG23), was found in a single male from the tenth BC generation for R1 (BF24-BG23). An additional backcross to Line UCD 003 increased the number of R13 birds. Results were presented at the annual project meeting, the Poultry Science Association meeting and in peer-reviewed publications. 1. Presentations - to whom, when, title, where. University of Maryland,
Department of Animal & Avian Sciences Seminar, College Park, MD 2007 Major histocompatibility (B) complex control of responses against Rous sarcomas. North Carolina State University Department of Poultry Science Seminar, Raleigh, NC, 2007 Immune response control by the chicken major histocompatibility (B) complex Texas A&M University, Department of Poultry Science Seminar, College Station, TX 2007 Genetics of immune response in chickens University of Wisconsin, Research Animal Resource Center Seminar, Madison, WI 2007 Genetics of immune response in chickens and animal care issues University of New Hampshire, Genetics Seminar, Durham, NH, 2007 Genetics control of responses against Rous sarcoma virus tumors. Chapman, M. A., R. L. Taylor, Jr., and R. F. Wideman, Jr., 2007. 5-HT osmotic minipump study in ascites susceptible and resistant lines. Poult. Sci. 86(Suppl. 1):221 Poultry Science Association Annual meeting, San Antonio, TX Wilkinson, N. G., L. M. Yates, R. T. Kopulos, W. E.
Briles, and R. L. Taylor, Jr., 2007. Antibody response against bovine red blood cells in major histocompatibility (B) complex recombinant R13. Poult. Sci. 86(Suppl. 1):143 Poultry Science Association Annual meeting, San Antonio, TX 2. Publications - just the number = two (peer reviewed research publications, reviews, monographs, etc.) 3. Graduate, undergraduate, post docs completed study with you (Name, degree, date). None (3 M. S. and 2 Ph.D committee member) 4. Courses, labs, lectures where results from this research have been incorporated. ANSC 612 Genetics of Domestic Animals ANSC/GEN 706 Human Genetics 5. New varieties, seeds, etc. = None 6. New databases developed, web sites, web address, etc. = None 7. DNA and/or protein sequences completed = None 8. Advice/Testimony to local, state, federal committees, advisory groups, etc. = None 9. Herbarium specimens = None 10. Study collections, etc. = None
PARTICIPANTS: C. M. Ashwell (N. C. State University), W. E. Briles (Northern Illinois University), J. E. Fulton (Hy-Line International), M. M. Miller (Beckman Research Institute), P. Wakenell (University of California, Davis), R. F. Wideman (University of Arkansas), N. G. Wilkinson (University of New Hampshire graduate student training)
TARGET AUDIENCES: Poultry breeding and health companies, poultry producers and other poultry scientists. Other scientists interested in animal health issues. Efforts Results were presented at the Poultry Science Association meeting and in peer-reviewed publications. Other presentations occurred through classes and informal discussions with individuals or groups.
PROJECT MODIFICATIONS: All genetic stocks were eliminated by the University of New Hampshire (March 21, 2007). The lack of genetic stocks will cripple future research efforts.
Impacts
R13R13 chickens were compared to other MHC congenic recombinant lines for Rous sarcoma tumor outcome. Six-week-old chickens of each congenic line (R1, R2, R4, R5, R13) line were wing-web inoculated with 30 pock forming units of subgroup C Rous sarcoma virus (RSV). Tumors were scored for size six times over 10 weeks post-inoculation. A tumor profile index (TPI), was assigned from the tumor size scores [Collins et al., Immunogenetics 5:333, 1977]. The following TPI values were used: 1 = complete regression by 28 days, or earlier; 2 = complete regression by 42 or 56 days; 3 = complete regression by 70 days, or a decreasing slope, or complete regression by 56 days followed by recurrence; 4 = general upward trend, or plateau or slight regression after 56 days; and 5 = terminal tumor prior to 70 days. A repeated measures split-plot design in least squares ANOVA with hatch and B genotype as the main statistical effects was used to evaluate mean tumor sizes. The TPI values
were rank transformed and analyzed by ANOVA as described (Conover and Iman, Am. Statistician 35:124, 1981). Fisher's Protected LSD at P < 0.05 was used to separate significant means. Tumors grew in >90% of inoculated chickens. The tumor size increased in all genotypes through week 3 post-inoculation. Tumors in R2R2 and R4R4 chickens plateaued thereafter. R1R1 tumors continued to increase in size. R5R5 tumors also plateaued at a much lower size after week 3 post-inoculation. The mean TPI for R5R5 chickens was significantly lower than the TPI of R2R2, R4R4, and R13R13, but not R1R1. Gene differences generated through recombination impacted the growth of Rous sarcomas in MHC congenic recombinant lines. Genotype R5R5 with B-F/B-L21 had significantly lower mean TPI than the R2R2, R4R4, and R13R13 chickens. Genetic stocks developed and maintained until 2007 at the University of New Hampshire were the foundation of this research. The work could not have been done without those stocks.
Publications
- Schulten, E. S., Yates, L. M. and Taylor, Jr., R. L. 2007. Antibody response against sheep red blood cells in lines congenic for major histocompatibility (B) complex recombinant haplotypes. Int. J. Poult. Sci. 6:732-738
- Chapman, M. A., Taylor, Jr., R. L. and Wideman, Jr., R. F. 2007. Analysis of plasma serotonin levels and hemodynamic responses following chronic serotonin infusion in broilers challenged with bacterial lipopolysaccharide and microparticles. Poult. Sci. (in press)
- Chapman, M. A., Taylor, Jr., R. L. and Wideman, Jr., R. F. 2007. 5-HT osmotic minipump study in ascites susceptible and resistant lines. Poult. Sci. 86(Suppl. 1):221
- Wilkinson, N. G., Yates, L. M., Kopulos, R. T., Briles, W. E. and Taylor, Jr., R. L. 2007. Antibody response against bovine red blood cells in major histocompatibility (B) complex recombinant R13. Poult. Sci. 86(Suppl. 1):143
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Progress 10/01/05 to 09/30/06
Outputs
Chicken major histocompatibility complex (MHC) genes include B-F, MHC class I; B-L, MHC class II; and B-G, MHC class IV. The genes are closely linked on chromosome 16 in the chicken genome. Recombinants between the MHC B-F and B-G regions have been useful to define genetic control of immune responses more precisely. Inbred Line UCD 003, (B17B17) is the genetic base for six congenic lines each containing a single unique MHC recombinant. These congenic lines have undergone ten backcrosses to Line UCD 003 and thus have 99.9% genetic uniformity. A new recombinant, designated R13 (BF17-BG23), was found in a single male from the tenth BC generation for R1 (BF24-BG23). The number of R13 birds was increased through an additional backcross to Line UCD 003. This experiment tested the new recombinant for antibody production. A single R13B17 male mated to five R13B17 dams produced the experimental progeny segregating for R13R13, R13B17, and B17B17 genotypes. At 4 weeks of age,
all progeny were injected intravenously with 1 mL of 2.5% bovine RBC (BRBC) to induce a primary antibody response. Serum was collected from blood samples taken 7 days post-injection. Microtiter methods measured total and mercaptoethanol (ME)-resistant (IgG) antibodies against BRBC. Titers were expressed as the log2 of the reciprocal of the highest dilution giving visible agglutination. The same birds received another injection at 11 weeks of age to stimulate a secondary antibody response. All primary and secondary antibody titers against BRBC were evaluated by least squares ANOVA with hatch and B genotype as main effects. Fisher's Protected LSD at P < 0.05 separated significant means. Both primary and secondary total antibody titers ranked highest to lowest among R13B17, B17B17, and R13R13 genotypes, respectively. No primary antibody titers differed significantly. However, the secondary total antibody titer of genotype R13B17 was significantly higher than the titer of R13R13 but not
B17B17. ME-resistant antibodies did not differ significantly. The results suggest a complementary effect between R13 and B17 in the secondary antibody response.
Impacts
Poultry breeders and producers receive a substantial economic benefit through improved health. Greater understanding of the genes that affect avian immunity will improved poultry health.
Publications
- Fulton, J. E., Juul-Madsen, H., Ashwell. C.M., McCarron, A.M., Arthur, J.A., OSullivan, N. and R. L. Taylor, Jr. 2006. Molecular genotype identification of the Gallus gallus major histocompatibility complex. Immunogenetics 58:407-421
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Progress 10/01/04 to 09/30/05
Outputs
A new MHC recombinant, designated R13 (BF17-G23), was found in the tenth backcross generation for R1 (BF24-G23). A single R13B17 male mated to five R13B17 dams produced the experimental progeny segregating for R13R13, R13B17 and B17B17 genotypes. Four-week-old birds of each genotype were injected intravenously with 1 mL of 2.5% bovine RBC (BRBC) to induce a primary antibody response. Serum was collected from blood samples taken 7 days post-injection. Microtiter methods measured total and mercaptoethanol-resistant (IgG) antibodies against BRBC. Titers were expressed as the log2 of the reciprocal of the highest dilution giving visible agglutination. The same birds received another injection at 11 weeks of age to produce a secondary antibody response. The total and ME-resistant antibody titers for the primary and secondary responses were evaluated by least squares ANOVA. The primary total antibody titer to BRBC was highest in genotype B17B17, intermediate in R13B17 and
lowest in R13R13. No values differed significantly. The R13 recombination event did not involve genes affecting the antibody response. We reported previous data on subgroup A Rous sarcoma growth in chickens segregating for the B1 haplotype in combination with either B2 or B5. The B1B2 and B2B2 genotypes had significantly lower tumor profile index (TPI) than did the B1B1 genotype. Heterozygous B1B5 chickens had TPI significantly lower than either B1B1 or B5B5 chickens. This test used parents with 25% inbred Line 6.15-5 in a single sire mating with five dams per sire for each genotype, B1B2 and B1B5. Six-week-old chickens were wing-web inoculated with 15 pock forming units of subgroup C Rous sarcoma virus (RSV). A tumor profile index (TPI) was assigned based on the tumor size scores taken size six times over 10 weeks post-inoculation. Tumors grew differentially in progeny segregating for haplotype B1 in combination with either B2 or B5. B1B1 chickens had a TPI significantly higher than
either B1B2 or B2B2 chickens. The B5B5 chickens had significantly higher TPI than B1B5 or B1B1 chickens. B1 had a poorer response than B2 but was a better responder than B5 against subgroup C Rous sarcoma virus.
Impacts
Poultry breeders and producers receive a substantial economic benefit through improved health. Greater understanding of the genes that affect avian immunity will improved poultry health.
Publications
- Tupick, T. A., Bloom, S. E., and Taylor, Jr., R. L. 2005. Major histocompatibility (B) complex gene dose effects on Rous sarcoma virus tumor growth. Int. J. Poult. Sci. 4:286-291
- Taylor, Jr., R. L., 2005. Chicken genetic resources at the University of New Hampshire. Proc. Chicken Genomics and Development Workshop. Cold Spring Harbor Lab. Press, p. 45
- Taylor, R. L., Jr., Briles, W. E., and Fulton, J. E. 2005. Characterizing Rous sarcoma growth for major histocompatibility (B) complex haplotype B61. Poult. Sci. 84(Suppl. 1): 40
- Miller, M. M., Goto, R. M., Wang, Y., Wakenell, P. S., and Taylor, Jr., R. L. 2005. Genetics of tumor suppression in the avian Mareks disease model. National Cancer Institute meeting p. 25
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Progress 10/01/03 to 09/30/04
Outputs
The chicken major histocompatibility complex (MHC) is defined by serologic reactions between erythrocytes and antibodies specific to the polymorphic MHC class I (BF) and class IV (BG) antigens. The microsatellite marker LEI0258 is located between the BG and BF MHC regions. PCR primers specific for LEI0258 were used to amplify DNA from various serologically defined MHC haplotypes. Twenty-six distinctive allele sizes were identified. Fourteen LEI0258 alleles had a unique association with a serologically defined MHC haplotype whereas 12 LEI0258 alleles were each found in 2 to 10 different MHC haplotypes. Some serologically well-defined MHC haplotypes shared a common LEI0258 allele size. The LEI0258 allele size and MHC haplotype association was very consistent for the same haplotype from multiple sources. Sequences were determined for fourteen different LEI0258 alleles. This two internal repeats having 13 bp and 12 bp lengths were found. Alleles size ranged from 185 to
563 bp and is due to changes in the number of both the 12 and 13 bp repeats, plus the presence or absence of an 8 bp deletion in the unique sequence region between the repeat and the primer. This marker represents a useful tool to identify MHC haplotype in chickens particularly for the initial development of serological reagents. Last year, we reported subgroup A Rous sarcoma growth in chickens segregating for the B1 haplotype in combination with either B2 or B5. The B1B2 and B2B2 genotypes had significantly lower tumor profile index (TPI) than did the B1B1 genotype. Heterozygous B1B5 chickens had TPI significantly lower than either B1B1 or B5B5 chickens. This test used B1B2 and B1B5 parents with 25% Line 6.15-5. A single sire mating with five dams was made for the genotypes B1B2 and B1B5. Six-week-old chickens were wing-web inoculated with 15 pock forming units of subgroup C Rous sarcoma virus (RSV). Tumors were scored for size six times over 10 weeks followed by assignment of a
tumor profile index (TPI) based on the tumor size scores. The overall tumor growth for the 70 B1B2 and 78 B1B5 chickens was generally regressive. MHC analysis of the progeny and statistical evaluation of tumor size and TPI are continuing.
Impacts
Poultry health will be improved by greater understanding of the genes that affect avian immunity. Improved health represents a substantial economic benefit to poultry breeders and producers.
Publications
- Erf, G. F. and Taylor, Jr., R. L. 2004. Ancillary Scientists Symposium - The avian immune system: Function and modulation: Introduction. Poult. Sci. 83:550-551
- Taylor, Jr., R. L. 2004. Major histocompatibility (B) complex control of responses against Rous sarcomas. Poult. Sci. 83:638-649
- Schulten, E. S., Briles, W. E. and Taylor, Jr., R. L. 2004. Rous sarcoma growth in lines congenic for major histocompatibility (B) complex recombinants. Poult. Sci. 83(Suppl. 1):147
- Fulton, J. E., Juul-Madsen, H., Ashwell, C. M., McCarron, A. M., and Taylor, Jr., R. L. 2004. Molecular genotype identification of the chicken major histocompatibility complex. Proc. 2004 Int. Soc. Anim. Genet. p25
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