Major Histocompatibility Complex Class IV Restriction

Fragment Length Polymorphism Markers in Replicated

Meat-Type Chicken Lines Divergently Selected for
High or Low Early Immune Response

ZEHAVA UNI,1-2 MICHAL GUTMAN,3 GABRIAL LEITNER,1 ESTHER

LANDESMAN,1 DAN HELLER,1 and AVIGDOR CAHANER3
Department of Animal Science and Department of Genetics,
Faculty of Agriculture, The Hebrew University of Jerusalem,
Rehovot 76100, Israel
ABSTRACT Information on MHC may improve the efficiency of selection for
immunological traits via the application of marker assisted selection or by
selecting directly for a specific restriction fragment length polymorphism
(RFLP) band or MHC haplotype. An experimental procedure is presented here
for identifying MHC genes that are related to early immune response. A Class
IV cDNA clone was used to probe Southern blots of erythrocyte genomic DNA
from chickens. Chickens were taken from the second (S2) and third (S3)
generations of replicated lines divergently selected for high antibody response
(HC1, HC2) or low antibody response (LCI, LC2) to Escherichia coli vaccination
at 10 days of age. These selection criteria have been found to be associated with
other immunological parameters. The hypothesis that these selected lines differ
in their MHC loci was evaluated by comparing the frequencies of MHC RFLP
markers (single RFLP bands) and haplotypes (patterns of RFLP bands). The
significant differences between LC and HC in the frequency of many MHC
RFLP bands and of five MHC haplotypes indicate that early antibody produc-
tion is influenced by MHC genes. The reliability of the association between the
selection and frequency differences was tested and proven in most cases by
analysis of the replicated lines. These differences in RFLP markers represent a
change in allelic frequencies in MHC genes, probably due to selection. The
results imply a connection between the Class IV genes and early antibody
production, and they show the potential of prospective breeding not only by
immunological phenotype but also by genotype (i.e., using RFLP markers of the
MHC).
(Key words: chicken, restriction fragment length polymorphism, major
histocompatibility complex, Class IV, immune response)
1993 Poultry Science 72:1823-1831

INTRODUCTION order to avoid economic loss. It is clear

The incidence of disease in commercial
chicken flocks is of major economic con-
cern. Vaccines, antibiotics, and other drugs
are being used in the poultry industry in
Received for publication January 4, 1993.
Accepted for publication June 10, 1993.
iDepartment of Animal Science.
2
To whom correspondence should be addressed.
3
Department of Genetics.
1823
that immunocompetence and resistance to
many infectious diseases is in part geneti-
cally determined (Gavora and Spencer,
1983). Consequently, there has been in-
creasing interest, in the identification of
genetic and immunological markers that
could be used in breeding programs to
improve immunocompetence.
The usefulness and limitations of
marker-assisted selection (MAS) have
been discussed by Soller and Backmann
1824 UNI ET AL.
(1986) and Smith and Simpson (1986). The
more markers identified and the closer
they are to quantitative trait loci, the more
efficient MAS is likely to be. When a
certain marker or group of markers is
associated with a quantitative trait that is
important to the breeder, MAS is more
advantageous than conventional selection.
Developments in molecular biology
techniques have enabled more efficient
evaluation of potential markers in the
genome. In some species of farm animals,
strong evidence has been found for the
linkage between restriction fragment
length polymorphism (RFLP) and prolac-
tin (Cowan et al, 1989), production traits
(Jung et al, 1989), and disease resistance
(Lunden et al, 1990). However, in chick-
ens, polymorphisms identified by the use
of DNA technology and associations be-
tween RFLP markers and quantitative
traits have hardly been described. High
polymorphism of RFLP markers was re-
vealed in the MHC Class IV (B-G) region
following digestion of DNA from layer
chickens (Miller et al, 1988) and meat-type
chickens (Uni et al, 1992). These RFLP are
potential markers for MHC genes.
The MHC genes play an important role
in immune system functions (reviewed by
Bacon, 1987). In chickens, the MHC (B
complex) is located on an 8 million bp
microchromosome, of which 6 million bp
code for ribosomal RNA in the nuclear
organizer (Bloom and Bacon, 1985). The
MHC contains tightly linked genes B-L, B-
V, and B-G (Hala et al, 1988; Kaufman et
al, 1989) that code, respectively, for Class
I, Class II, and Class IV glycoproteins on
the cell surface. The B-F and B-L antigens
are similar to their mammalian homo-
logues in structure, function, and tissue
distribution (Guillemont and Auffray,
1989). The B-G antigen, however, is unique
to avian species and was originally
described as limited to erythrocytes and
their precursor cells (Pink et al, 1977).
Recent publications, however (Miller et al,
1990; Salomonsen et al, 1991a) have
shown that members of this superfamily
of immunoglobulins are also found on
thrombocytes, lymphocytes, and certain
epithelial cells in the bursa of Fabricius,
thymus, and intestine.
The grouping of these genes on one
chromosome is termed a B-haplotype.
Different B-G haplotypes have been num-
bered from Ba to Bn (Briles and Briles,
1982) and used for genetic studies. Most of
those studies have been conducted in
White Leghorn (WL) populations, using
serological tools for B-G haplotype iden-
tification. Several studies have demon-
strated the association between the MHC
and oncogenic diseases (Bacon et al, 1981)
or bacterial disease challenge survival
(Lamont et al, 1987). Nordskog (1983) and
Pevzner et al. (1975) reported different
levels of antibodies in different B haplo-
types. Based on all of the above, the MHC
region is a natural candidate for MAS,
especially for immunological traits.
In the present study, cDNA clone bg
32.1 (Miller et al, 1988) was used to probe
Southern blots of erythrocyte genomic
DNA from chickens. The examined chick-
ens were taken from the second (S2) and
third (S3) generations of replicated lines
divergently selected for high antibody
response (HC1, HC2) or low antibody
response (LCI, LC2) to Escherichia coli
vaccination at 10 days of age (Leitner et al,
1992). These selection criteria have been
found to be associated with other im-
munological parameters, such as antibody
response to Newcastle disease virus
(NDV) inactivated vaccine, SRBC, in-
creased phagocytic activity, and prolifera-
tive response to antigens or mitogens
(Pitcovski et al, 1987; Heller et al, 1992).
The selection therefore most probably
affects a general component of early
immiinocompetence, possibly by changing
allelic frequencies in MHC genes. The
hypothesis that these selected lines differ
in their MHC loci was evaluated by
comparing the frequencies of MHC RFLP
markers (single RFLP bands) and haplo-
types (patterns of RFLP bands).
MATERIALS AND METHODS
Birds
The analysis included 102 chickens from
the S2 generation and 102 chickens from the
S3 generation of Lines HC1, HC2, LCI, or
LC2. The immunological parameters of
these selection lines are presented in Heller
et al. (1992).
 CLASS IV MARKERS AND CHICKEN IMMUNE RESPONSE 1825
Restriction Fragment Length verify the consistency of the selection effect
Polymorphism of Class IV Major in those replicated lines.
Histocompatibility Complex
The DNA was extracted from samples of RESULTS
heparinized whole blood collected from
each chicken (Hillel et d., 1989). Genomic Frequencies of Restriction Fragment
DNA (10 /xg) was digested with the restric- Length Polymorphism of Class IV
tion endonucleases Pvull (83 chickens from Major Histocompatibility Complex
S2 generation and 100 chickens from S3
generation) or Bglll (102 chickens from S2 Digestion of DNA with Restriction
generation and 100 chickens from S3 genera- Enzyme Pvull. An RFLP analysis following
tion) prior to electrophoresis on a 1% digestion with the restriction enzyme Pvull
agarose gel. The DNA was transferred to revealed high levels of polymorphism and
nylon filters according to the method of complexity at the Class IV region. A total of
Southern (1975), hybridized (Hillel et ah, 31 polymorphic bands (1.6 to 8.0 kb) were
1989) with the 32P-labeled chicken Class IV observed in 183 chickens, taken at random
cDNA probe bg 32.1 (Miller et al., 1988), and from S2 and S3 generation birds of lines LCI,
subjected to autoradiography. LC2, HC1, and HC2. Twelve of these RFLP
bands were analyzed statistically.
Statistical Analysis In S2, five RFLP bands (P2, P4, P6, P24,
P25) had significantly different frequencies
Differences of band and haplotype fre- in the HC versus LC lines (Table 1). In S3, six
quencies between the pooled, divergently RFLP bands (P3, P4, P22, P23, P24, P25)
selected lines (LC and HC) were analyzed were found to have significantly different
by chi-square test using PROC FREQUEN- frequencies in the divergent lines (Table 2).
CIES (SAS Institute, 1987). In those cases in For Band P22, the significant difference
which LC differed significantly from HC, between HC1 and HC2, and the similarity
the differences between replicated lines between HC2 and LCI, suggests that the
were also analyzed by chi-square test to difference between pooled LC and HC was

TABLE 1. Frequency of 12 RFLP1 bands obtained using restriction enzyme PPWII

in the second generation of chicken lines selected for high (HC) or low (LC)
antibody response to Escherichia coli

Replicated lines Pooled lines

RFLP band 2 LCI LC2 HC1 HC2 LC HC P(X2)
(30)3 (30) (48) (58) (60) (106)
(kb) C°'1
(—)
8.0 PI 23 16 14 12 20 13 NS
7.5 P2 26 20 4 12 23 9 .004
5.2 P3 0 0 8 3 0 6 .056
4.7 P4 16 20 2 4 16 2 <.0001
4.5 P5 100 94 100 90 97 94 NS
4.3 P6 47 37 33 28 41 27 .020
2.3 P20 33 13 23 24 23 24 NS
2.0 P21 20 33 12 20 27 17 .085
1.9 P22 18 13 23 24 27 23 .067
1.8 P23 16 10 16 5 13 10 NS
1.7 P24 17 27 12 4 22 7 .004
1.6 P25 13 16 0 7 15 2 .001
Restriction fragment length polymorphism.
2P1... P25 designate the different RFLP bands.
32n.
1826 UNI ET AL.

TABLE 2. Frequency of 12 RFLP1 bands obtained using restriction enzyme PPMII

in the third generation of chicken lines selected for high (HC) or low (LC)
antibody response to Escherichia coli

Replicated lines Pooled lines

RFLP band 2 LCI LC2 HCl HC2 LC HC P(x2)
(52)3 (58) (32) (58) (110) (90)
(kb) (°/,^
8.0 PI 19 14 22 13 16 17 NS
7.5 P2 26 12 6 20 14 16 NS
5.2 P3 4 2 9 22 3 18 <.0001
4.7 P4 15 12 0 5 14 4 .008
4.5 P5 100 82 100 86 89 91 NS
4.3 P6 37 44 25 38 40 33 .082
2.3 P20 27 7 25 33 17 33 NS
2.0 P21 30 37 50 36 34 41 NS
1.9 P22 19 10 37* 19 14 25 .025
1.8 P23 2 7 13 19 4 17 .003
1.7 P24 21* 36 9* 0 29 3 <.0001
1.6 P25 13 7 0 0 20 0 .001

'Restriction fragment length polymorphism.

2
P 1 . . . P25 designate the different RFLP bands.
32n.
'Indicates a significant difference (P < .05) between a pair of replicated lines.

random and not due to selection. Although
the replicated lines differed significantly in
the frequency of Band F24, its frequency in
LCI and LC2 was much higher than that in
HCl and HC2 (Table 2).
Digestion of DNA with Restriction
Enzyme BgMI. The use of a second six-
cutter restriction enzyme revealed a similar
degree of polymorphism and complexity at
the Class IV region. A total of 25 poly-
morphic bands (2.0 to 9.5 kb) were observed
in an experimental population of 202 chick-
ens taken at random from the S2 and S3
generations of the replicated divergent
lines. Seven of the 25 RFLP bands were
analyzed statistically. In S2, four RFLP
bands (B19, B20, B21, and B24) had signifi-
cantly different frequencies in LC and HC
birds (Table 3). For Band B19, the significant
difference between HCl and HC2, and the
similarity between HCl, LCI, and LC2
suggest that the difference between pooled
LC and HC was random and not due to
selection. Although the HC replicated lines
differed significantly in Band B20, its fre-
quency in LCI and LC2 was much higher
than that in HCl and HC2. Statistical
analysis of pairs of replicated lines showed
that the difference between the pooled LC
and HC in RFLP Band B20 could be random
and not due to selection. In S3 generation
birds, the frequency of these RFLP bands
and of another band (B25) were found to be
significantly different in the divergent lines
(Table 4).
Frequencies of Major
Histocompatibility Complex Class IV
Restriction Fragment Length
Polymorphism Haplotypes
Haplotypes were determined according
to their "restriction pattern" and by family
group analysis, as described by Uni et al.
(1992) and Landesman et al. (1993). A total
of 14 different haplotypes (A, C, D, E, F, G,
H, K, L, R, T, U, V, and X) were identified
and designated by letters in 102 chickens
from the replicated divergent lines (LCI,
LC2; HCl, HC2) of S3 generation birds.
Among the chickens examined, a large
variety of B-G haplotypes in all kinds of
combinations was found. Only 14% of the
chickens were homozygous. Figure 1 ex-
hibits a representative Southern blot depict-
ing the RFLP pattern of 19 chickens from
two selection lines (LCI; HCl), after diges-
tion with Pvull (Figure la) or BglU (Figure
lb) and hybridization with Class IV probe
 CLASS IV MARKERS AND CHICKEN IMMUNE RESPONSE 1827
TABLE 3. Frequency of seven RFLP1 bands obtained using restriction enzyme BglU
in the second generation of chicken lines selected for high (HC) or low (LC)
antibody response to Escherichia coli

Replicated lines Pooled lines

RFLP band* LCI LC2 HC1 HC2 LC HC P(x2)
(52)3 (52) (42) (58) (104) (100)
(kb) (°',\
2.9 B18 40 24 43 21 33 30 NS
2.8 B19 24 24 24* 7 24 14 .037
2.5 B20 25 17 8* 0 21 4 .004
2.4 B21 42 35 48 44 39 46 .012
2.2 B23 0 7 4 5 4 5 NS
2.1 B24 6 7 30 35 7 32 <.O01
2.0 B25 26 24 31 20 25 25 NS
Restriction fragment length polymorphism.
2
B18...B25 designate the different RFLP bands.
32n.
'Indicates a significant difference (P < .05) between a pair of replicated lines.

bg 32.1. This figure also demonstrates the
designation of the B-G haplotypes and some
of the restriction fragment bands that were
analyzed statistically. Among the seven
chickens from the LC line, five chickens
were carrying Haplotype K in different
combinations (with Haplotype R, A, or G).
Among the 12 chickens from the HC line,
six were carrying Haplotype D in different
combinations (with Haplotype V or R). Of
these 19 chickens, 5 were homozygous. The
interpretation of these RFLP patterns as
haplotypes of the MHC region were fol-
lowed by evaluation of their frequencies
within the selection lines. Only haplotypes
that were found in at least 8 birds out of the
102 examined chickens were analyzed
statistically. Several haplotypes (A, C, F, K,
U, and X) appeared only or mainly among
either LC or HC birds (Table 5). With the
exception of Haplotype K (statistical analy-
sis between pairs of replicated lines showed
that the difference between the pooled LC
and HC could be random and not due to the

TABLE 4. Frequency of seven RFLP1 bands obtained using restriction enzyme BglU
in the third generation of chicken lines selected for high (HC) or low (LC)
antibody response to Escherichia coli

Replicated lines Pooled lines

RFLP band* LCI LC2 HC1 HC2 LC HC P(x2)
(54)3 (58) (32) (56) (112) (88)
(kb) (o/\

2.9 B18 43 25 38 44 34 42 .074

2.8 B19 22 30 11 7 23 8 <.001
2.5 B20 7 18 4 0 13 1 <.001
2.4 B21 33 26 37 50 29 47 <.001
2.2 B23 6 6 6 3 6 4 NS
2.1 B24 9 7 40 38 8 38 <.O01
2.0 B25 22 31 15 16 26 16 .046
Restriction fragment length polymorphism.
2
B18 . . . B25 designate the different RFLP bands.
32n.
1828 UNI ET AL.

LC HC

Kt 1 4 5 10 11 12 13 14 15 16 17 18 19

5.1-

4.2-

3.5-

2-
Ml U p . ,

KR AK AC KK KG KG DO DV DV XV CC DD GX UG VX TU DD DR
1
LC HC
1 2 3 4 5 6 V •8 9 10 11 12 13 14 15 16 17 18 19

!liifflt*S|
6

4-

• . B ir • - W ^ ^ 5 •
820 B21
2.3-
• • - i " f I - £ •
W — # •B24 # • w w

KR AK AC KK KG KG DD DV DV XV CC DD GX UG VX TU OD DR
FIGURE 1. A representative Southern blot depicting the restriction fragment length polymorphism patterns
obtained following digestion with PvuU (a) and BglTl (b) of genomic DNA (from chicken lines selected to high
(HC) or low (LC) antibody response to Escherichia coli (LC = 7 and HC = 12 chickens) and hybridization with Class
IV probe bg 32.1. Restriction fragment sizes were estimated on the basis of Hmdm-digested 5-phage DNA
fragments. Some RFLP fragments are marked. P2, P3, P17, P20, P24, P25 on Figure la and B19, B20, B21, B24, B25
on Figure lb are Class rv RFLP fragment numbers (P for PuuII and B for Bg/II). Class IV Haplotype designation is
indicated at the bottom of each lane.
 CLASS IV MARKERS AND CHICKEN IMMUNE RESPONSE 1829
TABLE 5. Frequency of B-G haplotypes in the chicken lines selected for high (HC) or low
(LC) antibody response to Escherichia coli

Replicated lines Pooled lines

Haplotypes LCI LC2 HCl HC2 LC HC P(x2)
(54)1 (58) (34) (58) (112) (92)

\'u)
A 16 7 0 2 11 1 .005
C 12 15 0 0 13 0 <.001
D 30 18 6 28 24 20 NS
E 16 15 20 8 15 12 NS
F 6 0 10 14 3 12 .025
G 8 9 16 18 8 17 NS
K 12* 30 10* 0 21 4 <.001
R 0 7 23 4 4 11 NS
U 0 0 6 14 0 11 <.001
X 0 0 6 12 0 9 .005
Ln titer 2 .9 1.3 3.5 3.8 1.2 3.6
i2n.
2
Line means of antibody response to E. coli vaccination at 10 days of age.
'Indicates a significant difference (P < .05) between a pair of replicated lines.

selection), all those haplotypes were signifi-
cantly different in their frequency in the
selection lines.
DISCUSSION
The field of molecular biology has
provided new tools and approaches to
animal breeding, including identification
of genes influencing quantitative traits. An
important condition for finding a marker
for a given trait is to use a probe that
reveals a high level of polymorphism in
the gene region examined. The bg 32.1
probe, which binds to the B-G genes,
reveals a very high degree of polymor-
phism (Miller et al, 1988; Uni et al, 1992)
and is therefore a suitable marker.
Based on serological and biochemical
information, B-G genes appear to exist
within a subregion that is physically
separated from B-F/B-L genes (Briles and
Briles, 1982; Miller et al, 1988). However,
Kaufman et al (1989) raised the question
of whether all B-G genes are separated
from all B-F/B-L genes. His evidence
indicates that some B-G genes are located
within the B-F/B-L subregion. His model
introduced the possibility that RFLP frag-
ments include Class I or Class II and Class
IV genes and can therefore be identified
by a Class IV probe. '
Miller et al. (1991) recently explained
the nature of the high degree of polymor-
phism in B-G antigens. They found that
the B-G polypeptides are composed of
single extracellular domains and single
membrane-spanning domains (all of them
with sequence diversity) and long
cytoplasmic tails, which are made up
entirely of differing numbers of units, each
consisting of seven amino acid residues
(heptads), which may themselves vary in
sequence. The large number of RFLP
bands obtained in the current study agrees
with their findings. Moreover, A PvuU
restriction site was located in the sequence
of multiple short (42 bp) repeats that had
been reported by Kaufman et al. (1989).
These repeats have been suggested to
have a function in the size heterogeneity
of B-G molecules, due to variation of their
cytoplasmic tail length.
The most polymorphic genes known in
eukaryotic organisms are associated with
the immune system functions of antigen
presentation and recognition. The findings
of Miller et al. (1991) revealed that the B-G
genes are similar to immunoglobulin,
having extracellular domains with varia-
ble sequences. Examination of the immune
function of B-G antigens (Salomonsen et
al, 1991b) showed an "adjuvant effect" of
chicken MHC polymorphic B-G antigens
and suggested rules for the polymorphic
1830 UNI ET AL.
B-G molecules, particularly for the genera-
tion of B cell diversity, in the immune
systems of birds. The recent information
about the function of B-G antigens empha-
sizes the potential usefulness of B-G RFLP
markers in selection for improved im-
munocompetency.
The significant differences between LC
and HC in the frequency of many MHC
RFLP bands and of five MHC haplotypes
indicate that early antibody production is
influenced by MHC genes. The reliability
of the association between the selection
and frequency differences was tested and
proven in most cases by analysis of the
replicated lines. These differences in RFLP
markers represent a change in allelic
frequencies in MHC genes, probably due
to tie selection. The increase in frequency
differences between LC and HC from S2 to
S3 observed for most RFLP bands provides
an additional indication of the association
between the selection and the change in
band frequencies.
Information on MHC may improve the
efficiency of selection for immunological
traits, via the application of MAS or by
selecting directly for a specific RFLP band
or MHC haplotype. The use of two
restriction enzymes increased the number
of RFLP markers, improved the definition
of MHC haplotypes (unpublished data),
and increased the chances of identifying
an association with immune traits. If a
certain fragment or group of fragments
(i.e., a haplotype) were to be associated
with a trait and if its effect were to be
large enough, MAS would be more advan-
tageous than conventional selection.
Analyses of the duplicated selection
lines, which differ significantly in several
immunological parameters at young age
(Heller et ah, 1992), enabled us to compare
the frequencies of RFLP markers. For the
first time in meat-type chickens, a signifi-
cant association between frequency of
Class r / MHC RFLP markers and immune
response was found. Differences between
LC and HC lines suggest some linkage
between the RFLP markers and the im-
mune characteristics of the lines. The
results imply a connection between the
Class IV genes and early antibody produc-
tion, and they show the potential of
prospective breeding not only by im-
munological phenotype but also by geno-
type (i.e., using RFLP markers of the
MHC).
ACKNOWLEDGMENTS
The authors thank M. M. Miller (Beck-
man Research Institute, Duarte, CA 91010)
for the bg 32.1 probe and J. Pitcovski
(Megal Technology Center, Kiriat-Shmona,
Israel) for his contribution. This work was
funded by Bundesministerium fur For-
schung und Technologie, Germany; Minis-
try of Science and Technology, Israel; and
Joint German Israel Research Projects.
REFERENCES
Bacon, L. D., 1987. Influence of the major histocom-
patibility complex on disease resistance and
productivity. Poultry Sci. 66:802-811.
Bacon, L. D., R. L. Witter, L. B. Crittenden, A. Fadly,
and J. Motta, 1981. B-haplotype influence on
Marek's disease, Rous sarcoma and lymphoid
leukosis virus-induced tumors in chickens.
Poultry Sci. 60:132-139.
Bloom, S. E., and L. D. Bacon, 1985. Linkage of the
major histocompatibility complex and the
nucleolar organizer in the chicken. J. Hered. 76:
146-154.
Briles, W. E., and R. W. Briles, 1982. Identification of
haplotypes of the chicken major histocompati-
bility complex (B). Immunogenetics 15:449-459.
Cowan, C. M., M R. Dentine, R. L. Ax, and L. A.
Schuler, 1989. Restricted fragment length poly-
morphism associated with growth hormone and
prolactin genes in Holstein bulls: evidence for a
novel growth hormone allele. Anim. Genet. 20:
157-165.
Gavora, J. S., and J. L. Spencer, 1983. Breeding for
genetic resistance: specific or general. World's
Poult. Sci. J. 43:137-148.
Guillemont, F., and C. Auffray, 1989. Molecular
biology of the chicken major histocompatibility
complex. Crit. Rev. Poult. Biol. 2:255-275.
Hala, K., A.-M. Chausse, O. Bourlet, O. Lassila, V.
Hasler, and C. Auffray, 1988. Attempt to detect
recombination between B-F and B-L genes
within the chicken B complex by serological
typing, in vitro MLR and RFLP analyses.
Immunogenetics 28:433-438.
Heller, E. D., G. Leitner, A. Friedman, Z. Uni, M.
Gutman, and A. Cahaner, 1992. Immunological
characteristics of meat-type chicken lines diver-
gently selected by antibody response to Es-
cherichia coli vaccination at 10 days of age. Vet.
Immunol. Immunopathol. 34:159-172.
Hillel, J., Y. Plozky, A. Haberfeld, U. Lavi, A.
Cahaner, and A. J. Jeffreys, 1989. DNA finger-
prints of poultry. Anim. Genet. 20:25-35.
Jung, Y. C, M. F. Rothschild, M. P. Flanagan, L. L.
Christian, and C. M. Warner, 1989. Association
of RFLP of swine leucocyte antigen class I genes
 CLASS IV MARKERS AND CHICKEN IMMUNE RESPONSE 1831
with production traits of Duroc and Hampshire Nordskog, A. W., 1983. Immunogenetics as an aid to
boars. Anim. Genet. 20:79-91. selection for disease resistance in the fowl.
Kaufman, J., J. Salomonsen, and K. Skjodt, 1989. B-G World's Poult. Sci. J. 39:199-209.
cDNA clones have multiple small repeats and Pevzner, I., A. W. Nordskog, and M. L. Kaeberle,
hybridize to both chicken MHC regions. Im- 1975. Immune response and the B blood group
munogenetics 30:440-451. locus in chickens. Genetics 80:753-759.
Lamont, S. J., C. Bohn, and N. Cheville, 1987. Genetic Pink, J.R.L., W. Droege, K. Hala, V. C. Miggiano, and
resistance to fowl cholera is linked to the major A. Ziegler, 1977. A three-locus model for the
histocompatibility complex. Immunogenetics 25: chicken major histocompatibility complex. Im-
284-289. munogenetics 5:203-216.
Landesman, E., Z. Uni, and E. D. Heller, 1993. Pitcovski, J., E. D. Heller, A. Cahaner, and B. A.
Classification of Major Histocompatibility Com- Peleg, 1987. Selection for early responsiveness of
plex class IV haplotypes by RFLP, in meat-type chickens to Escherichia coli and Newcastle dis-
chickens. Anim. Genet, (in press). ease virus. Poultry Sci. 66:1276-1282.
Leitner, G., Z. Uni, A. Cahaner, and E. D. Heller, Salomonsen, J., D. Dunon, K. Skjodt, D. Thorpe, O.
1992. Replicated divergent selection of meat- Vainio, and J. Kaufman, 1991a. Chicken major
type chickens for high or low early antibody histocompatibility complex-encoded B-G anti-
response to E. coli vaccination. Poultry Sci. 71: gens are found on many cell types that are
27-37. important for the immune system. Proc. Natl.
Acad. Sci. USA 88:1359-1363.
Lunden, A., S. Sigurdardottir, I. Edfors-Lilja, B.
Danell, J. Redel, and L. Andersson, 1990. The Salomonsen, J., H. Eriksson, K. Skjodt, T. Lundgreen,
effect of bovine major histocompatibility com- M. Simonsen, and J. Kaufman, 1991b. The
plex on bull breeding values for clinical mastitis "adjuvant effect" of the polymorphic B-G anti-
gens of the chicken major histocompatibility
and somatic cell counts in milk. Pages 497-500 complex analyzed using purified molecules
in: Proceedings of the 4th World Congress on incorporated in liposomes. Eur. J. Immunol. 21:
Genetics Applied to Livestock Production. XVI, 649-658.
Edinburgh, UK, July 23-27.
SAS Institute, 1987. SAS/STAT® Guide for Personal
Miller, M. M., H. Abplanalp, and R. Goto, 1988. Computers. 6th Edition. SAS Institute Inc., Cary,
Genotyping chickens for the B-G subregion of NC.
the major histocompatibility complex. Im- Smith, C, and S. P. Simpson, 1986. The use of genetic
munogenetics 28:374-379. polymorphism in livestock improvement. J.
Miller, M. M., R. Goto, S. Young, J. Chirivella, D. Anim. Breed. Gen. 103:205-217.
Hawke, and C. C. Miyada, 1991. Immunoglobu- Soller, M., and J. S. Backmann, 1986. Restriction
lin variable-region-like domains of diverse se- fragment length polymorphism in poultry
quence within the major histocompatibility breeding. Poultry Sci. 65:1474-1488.
complex of chicken. Proc. Natl. Acad. Sci. USA Southern, E., 1975. Detection of specific sequences
88:4377-4381. among DNA fragments separated by gel elec-
Miller, M. M., R. Goto, S. Young, J. Liu, and J. Hardy, trophoresis. J. Mol. Biol. 98:503-517.
1990. Antigens similar to major histocompatibil- Uni, Z., J. Hillel, R. Vaiman, A. Cahaner, and E. D.
ity complex B-G are expressed in the intestinal Heller, 1992. Restriction fragment length poly-
epithelium in the chicken. Immunogenetics 32: morphism of MHC class-rV (B-G) genotypes in
45-50. meat-type chickens. Anim. Genet. 23:319-324.