- Email: [email protected]
17. Genes encoding the transporters associated with antigen processing in the chicken are situated between the classical MHC class I genes and are highly polymorphic
Abstmcts.
6
Monduy
25th
B. 16.45
16.
- 17.00
Expression
of alternative
splice variants
WALDVOGEL,
AS.,“”
Institute
of bovine
ZAKHER,
of Veterinary
IL-4 in cultured
A., PERRON LEPAGE,
Pathology,
University
Four splice variants of bovine IL-4 (boIL-4) mRNA were detected by RT-PCR from ConA stimulated lymph node lymphocytes (LNCs): complete boIL-4 mRNA coding for all four exons, boIL-462 lacking exon 2, boTL-463 lacking exon 3, and boIL-46213 lacking exons 2 and 3. The influence of Thl and Th2 polarising culture conditions on the expression of these splice variants by LNCs was measured by real time RT-PCR. Stimulation with PHA and rhuIL-12 markedly enhanced expression of boIFN-y mRNA, and stimulation with PHA and rboIL-4 markedly increased boIL-4 mRNA. The expression of boIL-462 and boIL-463 mRNA was not influenced in a consistent way
Room
B. 17.15
17.
Genes
encoding
‘Institute
lymph
node cells and in viva
M.-F., HEUSSLER, Bern,
V.T.
Switzerland
by these culture conditions. The amount of boIL-4 exceeded the amount of boIL-462 always and boIL-463 was present in even lesser quantities. boIL-46213 mRNA was not detected in any of the samples. These results were the first strong evidence of an independent regulation of boIL-4 and boIL-4 splice variants. boIL-462 and boIL-463 mRNA were also detected in bovine tissues collected at necropsy. Interestingly, splice variant mRNA was occasionally detected in the absence of IL-4 mRNA. These results demonstrate that IL-4 mRNA is regulated independently from splice variant mRNA in vitro and in vivo.
the transporters the classical
associated with antigen processing in the chicken MHC class I genes and are highly polymorphic
for
Animal
Health,
B.A.,*:“:’ MILNE,
Compton, Nr. Newbury, Genome Campus,
S.,2 BECK, S.,z KAUFMAN,
Berks., Hinxton,
Transporters associated with antigen processing (TAP) pump peptides from the cytosol to the lumen of the endoplasmic reticulum where the peptides bind to MHC class I molecules. The genes encoding this transporter are found in the class II region of the MHC in mammals. In contrast, we have found that in the chicken the TAP genes are located in between the two classical class I genes (B-F1 and B-F2) in the chicken MHC. The TAP genes are in opposite transcriptional orientation, with the promoter region situated centrally. The promoter contains elements
B. 17.45
of Bern,
bovine
- 17.45
WALKER,
Room
2002
criteria for allele acceptance were applied to the remaining previously reported sequences, 11 further alleles were confirmed. This suggests that to date there is good evidence for 24 FLA-DRB alleles fulfilling nomenclature criteria. Preliminary results of Reference Strand Conformational Analysis on the same cat samples suggest individual cats have between one and three DRB loci.
(PCR) and clonal sequence analysis. Applying the strict criteria for assigning new alleles as used by the established mammalian MHC nomenclature committees, we have defined 13 FLADRB alleles, including four previously unreported alleles. We identified many sequences that were one or two base pairs different from these 13 defined alleles, and have shown that these are most likely artifacts of PCR amplification. When the same
Room
March
UK RG20 7NN; ‘The Cambs., CD10 ISA
are situated
between
J.’ Sanger
Centre,
Wellcome
Trust
that are commonly found in the promoters of MHC class I and class II genes; X/X2, S and Y boxes and an interferon response element. The TAP genes are highly polymorphic at both the nucleotide and protein level as seen in 14 inbred chicken lines studied. These polymorphisms are mostly found in the regions thought to be involved in peptide binding. We have reason to believe some of the polymorphisms may control the peptidebinding specificity for each haplotype, which result in the striking peptide-binding motifs found in chicken haplotypes studied.
~ 18.00
18.
Equine
MHC
polymorphism:
DNA
sequence
BROWN, J.J.,“” CARTER, S.D., OLLTER, W.E.R., THOMSON, Mammalian
Immunogenetics
Research
Group,
Faculty
of Veterinary
Normal immune surveillance depends on the ability of major histocompatibility complex class II molecules (MHC II) to bind peptide antigens and carry them to the cell surface for
Science,
analysis
at the DRA
locus
W., CLEGG, P.D., MATTHEWS, University
of Liverpool,
Liverpool
J.B L69 7ZJ
presentation to T-cells. Polymorphisms in DNA coding fork the MHC class IT antigen binding sites can significantly affect antigen binding. The MHC class II DRA locus is distinct from most other
6
Monduy
25th
B. 16.45
16.
- 17.00
Expression
of alternative
splice variants
WALDVOGEL,
AS.,“”
Institute
of bovine
ZAKHER,
of Veterinary
IL-4 in cultured
A., PERRON LEPAGE,
Pathology,
University
Four splice variants of bovine IL-4 (boIL-4) mRNA were detected by RT-PCR from ConA stimulated lymph node lymphocytes (LNCs): complete boIL-4 mRNA coding for all four exons, boIL-462 lacking exon 2, boTL-463 lacking exon 3, and boIL-46213 lacking exons 2 and 3. The influence of Thl and Th2 polarising culture conditions on the expression of these splice variants by LNCs was measured by real time RT-PCR. Stimulation with PHA and rhuIL-12 markedly enhanced expression of boIFN-y mRNA, and stimulation with PHA and rboIL-4 markedly increased boIL-4 mRNA. The expression of boIL-462 and boIL-463 mRNA was not influenced in a consistent way
Room
B. 17.15
17.
Genes
encoding
‘Institute
lymph
node cells and in viva
M.-F., HEUSSLER, Bern,
V.T.
Switzerland
by these culture conditions. The amount of boIL-4 exceeded the amount of boIL-462 always and boIL-463 was present in even lesser quantities. boIL-46213 mRNA was not detected in any of the samples. These results were the first strong evidence of an independent regulation of boIL-4 and boIL-4 splice variants. boIL-462 and boIL-463 mRNA were also detected in bovine tissues collected at necropsy. Interestingly, splice variant mRNA was occasionally detected in the absence of IL-4 mRNA. These results demonstrate that IL-4 mRNA is regulated independently from splice variant mRNA in vitro and in vivo.
the transporters the classical
associated with antigen processing in the chicken MHC class I genes and are highly polymorphic
for
Animal
Health,
B.A.,*:“:’ MILNE,
Compton, Nr. Newbury, Genome Campus,
S.,2 BECK, S.,z KAUFMAN,
Berks., Hinxton,
Transporters associated with antigen processing (TAP) pump peptides from the cytosol to the lumen of the endoplasmic reticulum where the peptides bind to MHC class I molecules. The genes encoding this transporter are found in the class II region of the MHC in mammals. In contrast, we have found that in the chicken the TAP genes are located in between the two classical class I genes (B-F1 and B-F2) in the chicken MHC. The TAP genes are in opposite transcriptional orientation, with the promoter region situated centrally. The promoter contains elements
B. 17.45
of Bern,
bovine
- 17.45
WALKER,
Room
2002
criteria for allele acceptance were applied to the remaining previously reported sequences, 11 further alleles were confirmed. This suggests that to date there is good evidence for 24 FLA-DRB alleles fulfilling nomenclature criteria. Preliminary results of Reference Strand Conformational Analysis on the same cat samples suggest individual cats have between one and three DRB loci.
(PCR) and clonal sequence analysis. Applying the strict criteria for assigning new alleles as used by the established mammalian MHC nomenclature committees, we have defined 13 FLADRB alleles, including four previously unreported alleles. We identified many sequences that were one or two base pairs different from these 13 defined alleles, and have shown that these are most likely artifacts of PCR amplification. When the same
Room
March
UK RG20 7NN; ‘The Cambs., CD10 ISA
are situated
between
J.’ Sanger
Centre,
Wellcome
Trust
that are commonly found in the promoters of MHC class I and class II genes; X/X2, S and Y boxes and an interferon response element. The TAP genes are highly polymorphic at both the nucleotide and protein level as seen in 14 inbred chicken lines studied. These polymorphisms are mostly found in the regions thought to be involved in peptide binding. We have reason to believe some of the polymorphisms may control the peptidebinding specificity for each haplotype, which result in the striking peptide-binding motifs found in chicken haplotypes studied.
~ 18.00
18.
Equine
MHC
polymorphism:
DNA
sequence
BROWN, J.J.,“” CARTER, S.D., OLLTER, W.E.R., THOMSON, Mammalian
Immunogenetics
Research
Group,
Faculty
of Veterinary
Normal immune surveillance depends on the ability of major histocompatibility complex class II molecules (MHC II) to bind peptide antigens and carry them to the cell surface for
Science,
analysis
at the DRA
locus
W., CLEGG, P.D., MATTHEWS, University
of Liverpool,
Liverpool
J.B L69 7ZJ
presentation to T-cells. Polymorphisms in DNA coding fork the MHC class IT antigen binding sites can significantly affect antigen binding. The MHC class II DRA locus is distinct from most other