BREEDING AND GENETICS Antibody Responses to Combinations of Antigens in White Leghorn Chickens of Different Background Genomes and Major Histocompatibility Complex Genotypes E. A. DUNNINGTON, C. T. LARSEN, W. B. GROSS, and P. B. SIEGEL Poultry Science Department, Virginia Polytechnic Institute and State University, Blacksburg, Virginia, 24061 ABSTRACT Antibody responses in chickens to SRBC, Newcastle disease (NDV), and Brucella abortus (BA) were compared when given singly or in combination. Chickens used in the present experiment originated from a cross and then backcrossing of White Leghorn populations that had been selected for high (HA) or low (LA) antibody response to SRBC antigens. The populations used in the current study were segregating for MHC haplotypes B 13 and B21. The experiment had a 2 x 3 x 6 factorial arrangement of treatments (two background genomes: HA and LA; three MHC haplotypes: B13B13, B13B21, and B 21 ^ 21 ; and six antigen treatments: SRBC, NDV, or BA only, SRBC plus NDV, SRBC plus BA, and NDV plus BA). Antigens were administered either i.v. (SRBC) or i.m. (NDV and BA) when chicks were 42 days of age. Blood was obtained 27 days later (69 days of age) for antibody determinations. A significant background genome by MHC haplotype interaction for BA antibodies was due to relatively high titers in Line HA chickens of MHC genotypes B13B13 and B13B2i. Background genome by MHC genotype interactions were not significant for SRBC or NDV antibodies. Antibody titers to SRBC were higher in background genome HA than LA, and similar among MHC genotypes. Antibodies to NDV were lower in chickens of MHC genotype B21B^, but there were no differences due to background genome. For each of the three antigens, antibody responses were highest when administered singly rather than in combination. Antibody titers were lower for SRBC when given with BA, and for BA titers when given with NDV. For NDV titers, combination with either of the other two antigens caused a reduction in antibody response to NDV antigen. (Key words: major histocompatibility complex, chickens, antibody response, background genome, vaccination program) 1992 Poultry Science 71:1801-1806
INTRODUCTION Losses in commercial poultry production due to disease outbreaks and expenses involved in vaccination and biosecurity programs are considerable. Both genetic and nongenetic means of protecting poultry from disease may have great economic impact. Experimentally, divergent selection for immunoresponsiveness (in terms of anti-
Received for publication May 11, 1992. Accepted for publication July 6, 1992.
body response to injection of SRBC) has been successful in chickens, which demonstrates genetic influence on this trait (Siegel and Gross, 1980; van der Zijpp et al, 1983; Pinard et al, 1990). Such selection has also been accompanied by correlated changes in frequencies of haplotypes at the MHC (Dunnington et al, 1984; Pinard et al, 1990), making it difficult to ascertain whether the changes in immune response were due to background genome or to MHC haplotype. Particular haplotypes at the MHC have been shown to influence resistance to various diseases (Bacon, 1987; Loudovaris et al, 1989; Gavora, 1990;
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DUNNINGTON ET AL.
Dietert et al, 1991). Examples of such influence by MHC haplotypes or haplotype combinations include Marek's disease (Martin et al, 1989; Blankert et al, 1990), Salmonella pullorum (Pevzner et al, 1977), and various bacterial and viral challenges (Gross et al, 1980). Other conditions appear to be unchanged by MHC influence, including response to Escherichia coli and Eitneria tenella (Dunnington et al, 1991, 1992b). Results concerning effects of MHC haplotypes on body weights and production traits are not consistent, with some reports indicating differences (Abplanalp et al, 1992; Sato et al, 1992) and others finding no differences (Dunnington et al, 1992a). Although commercial poultry may be vaccinated for several diseases at the same time, little work has been reported on genetic aspects of multiple versus single immunizations on antibody titers (Gyles et al, 1986). The objectives of the present study were to measure antibody responses to SRBC (a foreign protein), Newcastle disease virus (NDV, a virus), and Brucella abortus (BA, a bacterium) when given singly or in combination in White Leghorn chickens of different MHC haplotype combinations and genetic backgrounds.
background genomes (Dunnington et al, 1989; Martin et al, 1990). Individuals used in the current experiment were from one of the two background genomes (HA or LA) and had one of the following genotypes at the MHC: B13B13, B13B21, or B%B21. In each case, the subpopulations were segregating for B13 and B21. No selection for antibody titer was conducted in these populations after the two selected lines were crossed. At hatch all chicks were wing-banded, vaccinated for Marek's disease, and placed in floor pens with wood shavings litter. They were provided ad libitum access to a diet containing 20% CP and 2,685 kcal of ME/kg. All chickens were weighed at 8 wk of age (BW8). Antigen Administration
Within each of the six background genome by MHC genotype subpopulations, chicks were assigned at random to one of seven treatments: uninoculated control, SRBC only, NDV only, BA only, SRBC plus NDV, SRBC plus BA, or NDV plus BA. There were from 9 to 11 chicks in each subgroup. A combination of all three antigens was not given because there was not a sufficient number of chicks for this subclass. At 42 days of age, chicks from the control group were bled and tested for base line MATERIALS AND METHODS antibody levels of the antigens to be administered. Also at 42 days of age, chicks Genetic Stocks and Husbandry from each subclass were inoculated with the appropriate antigen or antigens. The Chickens for the current experiment SRBC were administered i.v. (.1 mL of .25% were White Leghorns from lines that had saline suspension). The NDV1 was adbeen selected for high (HA) or low (LA) ministered i.m. (.01 mL killed Bl Newcastle antibody response to SRBC (Siegel and vaccine of chick embryo origin). The BA2 Gross, 1980). A correlated response to this was administered i.m. (.05 mL of BA selection was that line HA was 96% B21 and bacterin). Twenty-seven days after inoculaLine LA was 96% B 13 by the ninth genera- tion, a blood sample was obtained from tion of selection (Dunnington et al, 1984). each chick via the brachial vein for antibody Previous to the current experiment, chick- determinations for SRBC, NDV, and BA. ens from these two lines were crossed and then subpopulations were backcrossed to each of the original selected lines for three Laboratory Procedures generations. This mating procedure alAntibody response to SRBC antigen was lowed development of subpopulations that measured by the microhemagglutination were a rrunimum of 93% from the respective method of Wegmann and Smithies (1966) and expressed as the logarithm of the reciprocal of the last dilution in which there ^alsbury Laboratories, Inc., Charles City, IA 50616. was agglutination. Hemagglutination inhi2 bition (HI) antibody titers in response to National Vet Service Lab, Ames, IA 50011.
COMBINATIONS OF ANTIGEN CHALLENGES
NDV were determined by the HI test (Beard, 1989). The Brucella Standard Tube agglutination (STA) test for detection of BA antibody was carried out in 3-mL tubes according to Alton et al. (1975), starting with a serum dilution of 1:25. Statistical Analyses
1803
response had changed with selection (Table 2). There were no differences in antibody response to SRBC antigens of chickens having different MHC genotypes. Antibody response to NDV antigens was not different due to the background genome, but was lower in B21B21 individuals than in those of the other MHC genotypes.
Analyses of variance were conducted for BW8 and antibody titers to SRBC, NDV, and BA with the main effects of the analyses being line (background genome), MHC genotype, and antigen treatment. The control (uninoculated) group was used only to ascertain base line levels of antibodies and was not included in any analyses. Influences of all first-order interactions were also tested. Where first order interactions were significant, analyses were conducted within each of the main effects. When separation of means was necessary, Duncan's multiple range tests were conducted.
For all three of the antigens administered, the treatment (whether an antigen was given singly or in combination with another) did not interact with background genome or MHC genotype. For SRBC, higher response was obtained when SRBC was given alone or with NDV (Table 3). For NDV, combination with either of the other antigens reduced the antibody response. For BA, higher response was obtained when given alone or with SRBC.
RESULTS
DISCUSSION
Differences Due to Administration of Antigens
Inheritance of the immune system may be viewed in the context of 1) the genetics Base line (preinoculation) antibody titers of antibody formation and the immune in response to antigens for SRBC, NDV, and response; 2) the MHC complex; and 3) BA were all negative. At 27 days postinocu- blood group antigens (Stevens, 1991). Specificity and complexity of imlation controls were also negative. munoresponsiveness make selection difficult, especially in terms of different Differences Due to Background types of defense to classes of antigen Genome and Major (Gyles etal, 1986). The current experiment Histocompatibility Complex Genotypecompared the effects of background geThere were significant background ge- nome of populations selected for high and nome by MHC genotype interactions for low response to a foreign protein (SRBC) BW8 and for BA antibody titers (Table 1). and MHC genotypes on response to viral For BW8, this interaction was due to low (NDV) and a bacterial (BA) inoculation mean BW of B21B21 in Line HA. The singly and in combination. The kinetics of interaction for BA titers resulted from response to different antigens may vary higher values in Line HA chickens of and response to SRBC is quite rapid. An genotypes B13B13 and B13B21 compared interval of 27 days postimmunization was with all other groups. chosen because previous work with these For SRBC and NDV responses, back- lines has shown that the difference in lines ground genome by MHC genotype interac- for SRBC antibody response remains tions were not significant. Chickens of the (Ubosi et al, 1985) after 27 days and background genome that had been selected responses to NDV and BA should be for high 5-day postinoculation antibody apparent. Also, the immunodominant anresponse to SRBC injection (Line HA) had tigen of BA is the 0-chain of the greater SRBC antibody titers even at 27 lipopolysaccharide and is located on the days after injection, indicating that persis- surface of the organism (Diaz et al, 1968). tence of immune response as well as peak The STA test detects antibodies to the Titers of Uninoculated Controls
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DUNNINGTON ET AL.
TABLE 1. Mean body weight at 8 wk of age (BW8) and antibody titers for Brucella abortus antigen1 for White Leghorn chickens differing in background genome and major histocompatibility complex genotype Background genome2 Genotype
HA
LA flWR
B13B13 B13fl21 B21B21
556 554 478
B13fll3 B13B21 B21B21
i~\
,„
± 11" ± lla ± 10"
3.14 ± 2.83 ± 2.03 ±
NS NS ** Brucella abortus antibody .29* *» .25" *» .23b NS
574 ±15 a 570 ± 10» 575 ± l l a titer 1.88 ± .26" 1.67 ± .29a 2.09 ± .28"
a b
- Means ± SE within a column and subheading with no common superscripts differ significantly (P < .05). iBackground genome by MHC genotype interactions were significant (P < .05). 2 HA = background genome selected for high antibody response to SRBC; LA = background genome selected for low response to SRBC. "Indicates highly significant difference (P < .01) between means within a row.
TABLE 2. Mean antibody titers to sheep erythrocytes and Newcastle disease (NDV) for White Leghorn chickens differing in background genomes and in major histocompatibility complex genotypes1 Genome and genotype Background genome HA LA MHC genotype
SRBC
NDV
4.23 ± .20" 3.56 ± .19b
5.08 ± .19" 4.73 ± .28"
3.58 ± .30* 3.96 ± .23* 4.10 ± .21*
5.37 ± .30» 5.16 ± .31' 4.28 ± .24b
2
B13B13 B13B21 B21B21 ab
' Means ± SE within a column and subheading with no common superscripts differ significantly (P < .05). background genome by MHC genotype interactions were not significant (P > .05). 2 HA = background genome selected for high antibody response to SRBC; LA = background genome selected for low response to SRBC.
TABLE 3. Mean titers in response to sheep erythrocytes, Newcastle disease (NDV), and Brucella abortus (BA) depending on whether each antigen was administered alone or with another antigen Antigen inoculated
Body weight at 8 wk
Antibody titer SRBC
NDV
(g) SRBC 559 ± l l a 4.35 ± .22a NDV 559 ± 13a 5.69 ± BA 545 ± 14a 3.70 ± .23* 4.92 ± SRBC + NDV 560 ± 13a 3.57 ± .28b SRBC + BA 529 ± 11* a 4.24 ± NDV + BA 539 ± 12 ab ' Means ± SE within a column with no common superscripts differ
BA
.29" .28b .26b
2.70 ± .20*
2.19 ± .19ab 2.02 ± .19b
significantly (P < .05).
COMBINATIONS OF ANTIGEN CHALLENGES
0-chain antigen. The 0-chain of BA is a homopolymer of perosamine (Caroff and Perry, 1984; Caroff et al., 1984), and therefore, antibodies against the 0-chain are not directed against protein epitopes (NDV antibodies). Antibody titers for each antigen were higher when given singly than in combination. These results suggest that genetic background and vaccine combinations should be considered in immunization programs.
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Gross, and P. B. Siegel, 1984. Allelic frequencies in eight alloantigen systems of chickens selected for high and low antibody response to sheep red blood cells. Poultry Sci. 63:1470-1472. Dunnington, E. A., W. B. Gross, A. Martin, and P. B. Siegel, 1992b. Response to Eimeria tenella of chickens selected for high or low antibody response and differing in haplotypes at the major histocompatibility complex. Avian Dis. 36:49-53. Dunnington, E. A., A. S. Larsen, N. P. O'SuUivan, and P. B. Siegel, 1992a. Growth and egg production traits in chickens as influenced by major histocompatibility types and background genomes. J. Anim. Breed. Genet. 109:188-196. Dunnington, E. A., A. Martin, R. W. Briles, W. E. ACKNOWLEDGMENTS Briles, W. B. Gross, and P. B. Siegel, 1989. Antibody responses to sheep erythrocytes for Thanks are expressed to G. Schurig for White Leghorn chickens differing in haplotypes supplying BA antigen and the procedure of the major histocompatibility complex (B). for testing BA results. Anim. Genet. 20:213-216. Dunnington, E. A., P. B. Siegel, and W. B. Gross, 1991. Escherichia coli challenge in chickens REFERENCES selected for high or low antibody response and differing in haplotypes at the major histocomAbplanalp, H., K. Sato, D. Napolitano, and J. Reid, patibility complex. Avian Dis. 35:937-940. 1992. Reproductive performance of inbred congenic Leghorns carrying different haplotypes for Gavora, J. S., 1990. Disease genetics. Pages 805-846 in: Poultry Breeding and Genetics. R. D. Crawford, the major histocompatibility complex (B-blood ed. Elsevier Publishing Co., New York, NY. groups). Poultry Sci. 71:9-17. Alton, G. G., L. M. Jones, and D. E. Pietz, 1975. Gross, W. B„ P. B. Siegel, R. W. HaU, C. H. Domermuth, and R. T. Dubose, 1980. ProducLaboratory techniques in Brucellosis. Pages tion and persistence of antibodies in chickens. 2. 64-124 in: World Health Organization MonoResistance to infectious diseases. Poultry Sci. 59: graph Series. No. 55, World Health Organiza205-210. tion, Geneva, Switzerland. Bacon, L. D., 1987. Influence of the major histocom- Gyles, N. R., H. Fallah-Moghaddam, L. T. Patterson, J. K. Skeeles, C. E. WhitfiU, and L. W. Johnson, patibility complex on disease resistance and 1986. Genetic aspects of antibody responses in productivity. Poultry Sci. 66:802-811. chickens to different classes of antigens. Poultry Beard, C. W., 1989. Pages 192-194 in: A Laboratory Sci. 65:223-232. Manual for the Isolation and Identification of Avian Pathogens. H. G. Purchase, L. H. Arp, C. Loudovaris, T., B. H. Yoo, B. L. Sheldon, and K. J. Fahey, 1989. The chicken major histocompatibilH. Domermuth, and E. P. Pearson, ed. 3rd ed. ity complex, its associations with immune The American Association of Avian Patholoresponsiveness and disease resistance. Pages gists, Kennett Square, PA. 83-86 in: Proceedings Australian Poultry Science Blankert, J. J., M.G.J. Tilanus, B. G. Hepkema, G.A.A. Symposium, University of Sydney, Sydney, Albers, E. Egberts, and A. J. van der Zijpp, 1990. Australia. MHC-associated resistance against Marek's disease in White Leghorn chickens: Refined typing Martin, A., E. A. Dunnington, W. E. Briles, R. W. Briles, and P. B. Siegel, 1989. Marek's disease of B-G and B-F alleles using protein and DNA and major histocompatibility complex haploanalysis. Pages 457-460 in: Proceedings IV types in chickens selected for high or low World Congress Genetics Applied to Livestock antibody response. Anim. Genet. 20:407-414. Production, Edinburgh, Scotland. Caroff, M., D. R. Bundle, M. B. Perry, J. W. Martin, A., E. A. Dunnington, W. B. Gross, W. E. Briles, R. W. Briles, and P. B. Siegel, 1990. Chenwonogrodsky, and J. R. Duncan, 1984. Production traits and alloantigen systems in Antigen S-type lipopolysaccharide of Brucella lines of chickens selected for high or low abortus 1119-3. Infect. Immun. 16:384-388. antibody responses to sheep erythrocytes. Caroff, M., and M. B. Perry, 1984. Structure of the Poultry Sci. 69:871-878. 0-chain of the phenol phase soluble cellular lipopolysaccharide of Versinia enterocolitica sero- Pevzner, I. Y., C. Trowbridge, and A. W. Nordskog, type 0:9. Eur. J. Biochem. 139:195-200. 1977. Selection for high and low antibody response to Salmonella pullorum in chickens. Diaz, R., L. M. Jones, D. Leons, and J. B. Wilson, 1968. Genetics 86:s48-49.(Abstr.) Surface antigens of smooth brucellae. J. Bacteriol. 96:893-901. Pinard, M. H., M. A. van der Meulen, M. B. Dietert, R. R., R. L. Taylor, Jr., and M. F. Dietert, 1991. Kreukniet, M.G.B. Nieuwland, and A. J. van der Biological function of the chicken major Zijpp, 1990. Divergent selection for antibody histocompatibility complex. Crit. Rev. Poult. production in chickens: Differences in major Biol. 3:111-129. histocompatibility complex (MHC) haplotype distribution. Pages 477-480 in: Proceedings IV Dunnington, E. A., R. W. Briles, W. E. Briles, W. B.
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World Congress Genetics Applied to Livestock Production, Edinburgh, Scotland. Sato, K., H. Abplanalp, D. Napolitano, and J. Reid, 1992. Effects of heterozygosity of major histocompatibility complex haplotypes on reproduction and viability of Leghorn hens sharing a common inbred population. Poultry Sci. 71:18-26. Siegel, P. B., and W. B. Gross, 1980. Production and persistence of antibodies to sheep erythrocytes. I. Directional selection. Poultry Sci. 59:1-5. Stevens, L., 1991. Genetics and Evolution of the Domestic Fowl. Cambridge University Press,
Port Chester, NY. van der Zijpp, A. J., K. Frankena, J. Boneschanscher, and M.G.B. Nieuwland, 1983. Genetic analysis of primary and secondary immune responses in the chicken. Poultry Sci. 62:565-572. Ubosi, C. O., E. A. Dunnington, W. B. Gross, and P. B. Siegel, 1985. Divergent selection of chickens for antibody response to sheep erythrocytes: Kinetics of primary and secondary immunizations. Avian Dis. 29:347-355. Wegmann, T. G., and O. Smithies, 1966. A simple hemagglutinin system requiring small amounts of red cells and antibodies. Transfusion 6:67-75.
INTRODUCTION Losses in commercial poultry production due to disease outbreaks and expenses involved in vaccination and biosecurity programs are considerable. Both genetic and nongenetic means of protecting poultry from disease may have great economic impact. Experimentally, divergent selection for immunoresponsiveness (in terms of anti-
Received for publication May 11, 1992. Accepted for publication July 6, 1992.
body response to injection of SRBC) has been successful in chickens, which demonstrates genetic influence on this trait (Siegel and Gross, 1980; van der Zijpp et al, 1983; Pinard et al, 1990). Such selection has also been accompanied by correlated changes in frequencies of haplotypes at the MHC (Dunnington et al, 1984; Pinard et al, 1990), making it difficult to ascertain whether the changes in immune response were due to background genome or to MHC haplotype. Particular haplotypes at the MHC have been shown to influence resistance to various diseases (Bacon, 1987; Loudovaris et al, 1989; Gavora, 1990;
1801
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DUNNINGTON ET AL.
Dietert et al, 1991). Examples of such influence by MHC haplotypes or haplotype combinations include Marek's disease (Martin et al, 1989; Blankert et al, 1990), Salmonella pullorum (Pevzner et al, 1977), and various bacterial and viral challenges (Gross et al, 1980). Other conditions appear to be unchanged by MHC influence, including response to Escherichia coli and Eitneria tenella (Dunnington et al, 1991, 1992b). Results concerning effects of MHC haplotypes on body weights and production traits are not consistent, with some reports indicating differences (Abplanalp et al, 1992; Sato et al, 1992) and others finding no differences (Dunnington et al, 1992a). Although commercial poultry may be vaccinated for several diseases at the same time, little work has been reported on genetic aspects of multiple versus single immunizations on antibody titers (Gyles et al, 1986). The objectives of the present study were to measure antibody responses to SRBC (a foreign protein), Newcastle disease virus (NDV, a virus), and Brucella abortus (BA, a bacterium) when given singly or in combination in White Leghorn chickens of different MHC haplotype combinations and genetic backgrounds.
background genomes (Dunnington et al, 1989; Martin et al, 1990). Individuals used in the current experiment were from one of the two background genomes (HA or LA) and had one of the following genotypes at the MHC: B13B13, B13B21, or B%B21. In each case, the subpopulations were segregating for B13 and B21. No selection for antibody titer was conducted in these populations after the two selected lines were crossed. At hatch all chicks were wing-banded, vaccinated for Marek's disease, and placed in floor pens with wood shavings litter. They were provided ad libitum access to a diet containing 20% CP and 2,685 kcal of ME/kg. All chickens were weighed at 8 wk of age (BW8). Antigen Administration
Within each of the six background genome by MHC genotype subpopulations, chicks were assigned at random to one of seven treatments: uninoculated control, SRBC only, NDV only, BA only, SRBC plus NDV, SRBC plus BA, or NDV plus BA. There were from 9 to 11 chicks in each subgroup. A combination of all three antigens was not given because there was not a sufficient number of chicks for this subclass. At 42 days of age, chicks from the control group were bled and tested for base line MATERIALS AND METHODS antibody levels of the antigens to be administered. Also at 42 days of age, chicks Genetic Stocks and Husbandry from each subclass were inoculated with the appropriate antigen or antigens. The Chickens for the current experiment SRBC were administered i.v. (.1 mL of .25% were White Leghorns from lines that had saline suspension). The NDV1 was adbeen selected for high (HA) or low (LA) ministered i.m. (.01 mL killed Bl Newcastle antibody response to SRBC (Siegel and vaccine of chick embryo origin). The BA2 Gross, 1980). A correlated response to this was administered i.m. (.05 mL of BA selection was that line HA was 96% B21 and bacterin). Twenty-seven days after inoculaLine LA was 96% B 13 by the ninth genera- tion, a blood sample was obtained from tion of selection (Dunnington et al, 1984). each chick via the brachial vein for antibody Previous to the current experiment, chick- determinations for SRBC, NDV, and BA. ens from these two lines were crossed and then subpopulations were backcrossed to each of the original selected lines for three Laboratory Procedures generations. This mating procedure alAntibody response to SRBC antigen was lowed development of subpopulations that measured by the microhemagglutination were a rrunimum of 93% from the respective method of Wegmann and Smithies (1966) and expressed as the logarithm of the reciprocal of the last dilution in which there ^alsbury Laboratories, Inc., Charles City, IA 50616. was agglutination. Hemagglutination inhi2 bition (HI) antibody titers in response to National Vet Service Lab, Ames, IA 50011.
COMBINATIONS OF ANTIGEN CHALLENGES
NDV were determined by the HI test (Beard, 1989). The Brucella Standard Tube agglutination (STA) test for detection of BA antibody was carried out in 3-mL tubes according to Alton et al. (1975), starting with a serum dilution of 1:25. Statistical Analyses
1803
response had changed with selection (Table 2). There were no differences in antibody response to SRBC antigens of chickens having different MHC genotypes. Antibody response to NDV antigens was not different due to the background genome, but was lower in B21B21 individuals than in those of the other MHC genotypes.
Analyses of variance were conducted for BW8 and antibody titers to SRBC, NDV, and BA with the main effects of the analyses being line (background genome), MHC genotype, and antigen treatment. The control (uninoculated) group was used only to ascertain base line levels of antibodies and was not included in any analyses. Influences of all first-order interactions were also tested. Where first order interactions were significant, analyses were conducted within each of the main effects. When separation of means was necessary, Duncan's multiple range tests were conducted.
For all three of the antigens administered, the treatment (whether an antigen was given singly or in combination with another) did not interact with background genome or MHC genotype. For SRBC, higher response was obtained when SRBC was given alone or with NDV (Table 3). For NDV, combination with either of the other antigens reduced the antibody response. For BA, higher response was obtained when given alone or with SRBC.
RESULTS
DISCUSSION
Differences Due to Administration of Antigens
Inheritance of the immune system may be viewed in the context of 1) the genetics Base line (preinoculation) antibody titers of antibody formation and the immune in response to antigens for SRBC, NDV, and response; 2) the MHC complex; and 3) BA were all negative. At 27 days postinocu- blood group antigens (Stevens, 1991). Specificity and complexity of imlation controls were also negative. munoresponsiveness make selection difficult, especially in terms of different Differences Due to Background types of defense to classes of antigen Genome and Major (Gyles etal, 1986). The current experiment Histocompatibility Complex Genotypecompared the effects of background geThere were significant background ge- nome of populations selected for high and nome by MHC genotype interactions for low response to a foreign protein (SRBC) BW8 and for BA antibody titers (Table 1). and MHC genotypes on response to viral For BW8, this interaction was due to low (NDV) and a bacterial (BA) inoculation mean BW of B21B21 in Line HA. The singly and in combination. The kinetics of interaction for BA titers resulted from response to different antigens may vary higher values in Line HA chickens of and response to SRBC is quite rapid. An genotypes B13B13 and B13B21 compared interval of 27 days postimmunization was with all other groups. chosen because previous work with these For SRBC and NDV responses, back- lines has shown that the difference in lines ground genome by MHC genotype interac- for SRBC antibody response remains tions were not significant. Chickens of the (Ubosi et al, 1985) after 27 days and background genome that had been selected responses to NDV and BA should be for high 5-day postinoculation antibody apparent. Also, the immunodominant anresponse to SRBC injection (Line HA) had tigen of BA is the 0-chain of the greater SRBC antibody titers even at 27 lipopolysaccharide and is located on the days after injection, indicating that persis- surface of the organism (Diaz et al, 1968). tence of immune response as well as peak The STA test detects antibodies to the Titers of Uninoculated Controls
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DUNNINGTON ET AL.
TABLE 1. Mean body weight at 8 wk of age (BW8) and antibody titers for Brucella abortus antigen1 for White Leghorn chickens differing in background genome and major histocompatibility complex genotype Background genome2 Genotype
HA
LA flWR
B13B13 B13fl21 B21B21
556 554 478
B13fll3 B13B21 B21B21
i~\
,„
± 11" ± lla ± 10"
3.14 ± 2.83 ± 2.03 ±
NS NS ** Brucella abortus antibody .29* *» .25" *» .23b NS
574 ±15 a 570 ± 10» 575 ± l l a titer 1.88 ± .26" 1.67 ± .29a 2.09 ± .28"
a b
- Means ± SE within a column and subheading with no common superscripts differ significantly (P < .05). iBackground genome by MHC genotype interactions were significant (P < .05). 2 HA = background genome selected for high antibody response to SRBC; LA = background genome selected for low response to SRBC. "Indicates highly significant difference (P < .01) between means within a row.
TABLE 2. Mean antibody titers to sheep erythrocytes and Newcastle disease (NDV) for White Leghorn chickens differing in background genomes and in major histocompatibility complex genotypes1 Genome and genotype Background genome HA LA MHC genotype
SRBC
NDV
4.23 ± .20" 3.56 ± .19b
5.08 ± .19" 4.73 ± .28"
3.58 ± .30* 3.96 ± .23* 4.10 ± .21*
5.37 ± .30» 5.16 ± .31' 4.28 ± .24b
2
B13B13 B13B21 B21B21 ab
' Means ± SE within a column and subheading with no common superscripts differ significantly (P < .05). background genome by MHC genotype interactions were not significant (P > .05). 2 HA = background genome selected for high antibody response to SRBC; LA = background genome selected for low response to SRBC.
TABLE 3. Mean titers in response to sheep erythrocytes, Newcastle disease (NDV), and Brucella abortus (BA) depending on whether each antigen was administered alone or with another antigen Antigen inoculated
Body weight at 8 wk
Antibody titer SRBC
NDV
(g) SRBC 559 ± l l a 4.35 ± .22a NDV 559 ± 13a 5.69 ± BA 545 ± 14a 3.70 ± .23* 4.92 ± SRBC + NDV 560 ± 13a 3.57 ± .28b SRBC + BA 529 ± 11* a 4.24 ± NDV + BA 539 ± 12 ab ' Means ± SE within a column with no common superscripts differ
BA
.29" .28b .26b
2.70 ± .20*
2.19 ± .19ab 2.02 ± .19b
significantly (P < .05).
COMBINATIONS OF ANTIGEN CHALLENGES
0-chain antigen. The 0-chain of BA is a homopolymer of perosamine (Caroff and Perry, 1984; Caroff et al., 1984), and therefore, antibodies against the 0-chain are not directed against protein epitopes (NDV antibodies). Antibody titers for each antigen were higher when given singly than in combination. These results suggest that genetic background and vaccine combinations should be considered in immunization programs.
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Gross, and P. B. Siegel, 1984. Allelic frequencies in eight alloantigen systems of chickens selected for high and low antibody response to sheep red blood cells. Poultry Sci. 63:1470-1472. Dunnington, E. A., W. B. Gross, A. Martin, and P. B. Siegel, 1992b. Response to Eimeria tenella of chickens selected for high or low antibody response and differing in haplotypes at the major histocompatibility complex. Avian Dis. 36:49-53. Dunnington, E. A., A. S. Larsen, N. P. O'SuUivan, and P. B. Siegel, 1992a. Growth and egg production traits in chickens as influenced by major histocompatibility types and background genomes. J. Anim. Breed. Genet. 109:188-196. Dunnington, E. A., A. Martin, R. W. Briles, W. E. ACKNOWLEDGMENTS Briles, W. B. Gross, and P. B. Siegel, 1989. Antibody responses to sheep erythrocytes for Thanks are expressed to G. Schurig for White Leghorn chickens differing in haplotypes supplying BA antigen and the procedure of the major histocompatibility complex (B). for testing BA results. Anim. Genet. 20:213-216. Dunnington, E. A., P. B. Siegel, and W. B. Gross, 1991. Escherichia coli challenge in chickens REFERENCES selected for high or low antibody response and differing in haplotypes at the major histocomAbplanalp, H., K. Sato, D. Napolitano, and J. Reid, patibility complex. Avian Dis. 35:937-940. 1992. Reproductive performance of inbred congenic Leghorns carrying different haplotypes for Gavora, J. S., 1990. Disease genetics. Pages 805-846 in: Poultry Breeding and Genetics. R. D. Crawford, the major histocompatibility complex (B-blood ed. Elsevier Publishing Co., New York, NY. groups). Poultry Sci. 71:9-17. Alton, G. G., L. M. Jones, and D. E. Pietz, 1975. Gross, W. B„ P. B. Siegel, R. W. HaU, C. H. Domermuth, and R. T. Dubose, 1980. ProducLaboratory techniques in Brucellosis. Pages tion and persistence of antibodies in chickens. 2. 64-124 in: World Health Organization MonoResistance to infectious diseases. Poultry Sci. 59: graph Series. No. 55, World Health Organiza205-210. tion, Geneva, Switzerland. Bacon, L. D., 1987. Influence of the major histocom- Gyles, N. R., H. Fallah-Moghaddam, L. T. Patterson, J. K. Skeeles, C. E. WhitfiU, and L. W. Johnson, patibility complex on disease resistance and 1986. Genetic aspects of antibody responses in productivity. Poultry Sci. 66:802-811. chickens to different classes of antigens. Poultry Beard, C. W., 1989. Pages 192-194 in: A Laboratory Sci. 65:223-232. Manual for the Isolation and Identification of Avian Pathogens. H. G. Purchase, L. H. Arp, C. Loudovaris, T., B. H. Yoo, B. L. Sheldon, and K. J. Fahey, 1989. The chicken major histocompatibilH. Domermuth, and E. P. Pearson, ed. 3rd ed. ity complex, its associations with immune The American Association of Avian Patholoresponsiveness and disease resistance. Pages gists, Kennett Square, PA. 83-86 in: Proceedings Australian Poultry Science Blankert, J. J., M.G.J. Tilanus, B. G. Hepkema, G.A.A. Symposium, University of Sydney, Sydney, Albers, E. Egberts, and A. J. van der Zijpp, 1990. Australia. MHC-associated resistance against Marek's disease in White Leghorn chickens: Refined typing Martin, A., E. A. Dunnington, W. E. Briles, R. W. Briles, and P. B. Siegel, 1989. Marek's disease of B-G and B-F alleles using protein and DNA and major histocompatibility complex haploanalysis. Pages 457-460 in: Proceedings IV types in chickens selected for high or low World Congress Genetics Applied to Livestock antibody response. Anim. Genet. 20:407-414. Production, Edinburgh, Scotland. Caroff, M., D. R. Bundle, M. B. Perry, J. W. Martin, A., E. A. Dunnington, W. B. Gross, W. E. Briles, R. W. Briles, and P. B. Siegel, 1990. Chenwonogrodsky, and J. R. Duncan, 1984. Production traits and alloantigen systems in Antigen S-type lipopolysaccharide of Brucella lines of chickens selected for high or low abortus 1119-3. Infect. Immun. 16:384-388. antibody responses to sheep erythrocytes. Caroff, M., and M. B. Perry, 1984. Structure of the Poultry Sci. 69:871-878. 0-chain of the phenol phase soluble cellular lipopolysaccharide of Versinia enterocolitica sero- Pevzner, I. Y., C. Trowbridge, and A. W. Nordskog, type 0:9. Eur. J. Biochem. 139:195-200. 1977. Selection for high and low antibody response to Salmonella pullorum in chickens. Diaz, R., L. M. Jones, D. Leons, and J. B. Wilson, 1968. Genetics 86:s48-49.(Abstr.) Surface antigens of smooth brucellae. J. Bacteriol. 96:893-901. Pinard, M. H., M. A. van der Meulen, M. B. Dietert, R. R., R. L. Taylor, Jr., and M. F. Dietert, 1991. Kreukniet, M.G.B. Nieuwland, and A. J. van der Biological function of the chicken major Zijpp, 1990. Divergent selection for antibody histocompatibility complex. Crit. Rev. Poult. production in chickens: Differences in major Biol. 3:111-129. histocompatibility complex (MHC) haplotype distribution. Pages 477-480 in: Proceedings IV Dunnington, E. A., R. W. Briles, W. E. Briles, W. B.
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DUNNINGTON ET AL.
World Congress Genetics Applied to Livestock Production, Edinburgh, Scotland. Sato, K., H. Abplanalp, D. Napolitano, and J. Reid, 1992. Effects of heterozygosity of major histocompatibility complex haplotypes on reproduction and viability of Leghorn hens sharing a common inbred population. Poultry Sci. 71:18-26. Siegel, P. B., and W. B. Gross, 1980. Production and persistence of antibodies to sheep erythrocytes. I. Directional selection. Poultry Sci. 59:1-5. Stevens, L., 1991. Genetics and Evolution of the Domestic Fowl. Cambridge University Press,
Port Chester, NY. van der Zijpp, A. J., K. Frankena, J. Boneschanscher, and M.G.B. Nieuwland, 1983. Genetic analysis of primary and secondary immune responses in the chicken. Poultry Sci. 62:565-572. Ubosi, C. O., E. A. Dunnington, W. B. Gross, and P. B. Siegel, 1985. Divergent selection of chickens for antibody response to sheep erythrocytes: Kinetics of primary and secondary immunizations. Avian Dis. 29:347-355. Wegmann, T. G., and O. Smithies, 1966. A simple hemagglutinin system requiring small amounts of red cells and antibodies. Transfusion 6:67-75.
