A. Tipold et al., RESTRICTION-FRAGMENT-LENGTH-POLYMORPHISM OF THE T-CELL RECEPTOR BETA-CHAIN GENE IN DOGS, Veterinary immunology and immunopathology, 51(3-4), 1996, pp. 225-233
Since T-cells and the T-cell receptor (TCR) play a pivotal role in the
response of the immune system, they are a target for pathogenesis stu
dies in immune mediated diseases and have been used to generate marker
s for T-cell dependent diseases in humans and dogs, TCR rearrangement
is generated at the genomic DNA level and can be analyzed by Southern
blotting techniques, In the present study this method to detect rearra
ngement of the TCR beta chain in the dog was critically examined, To s
earch for restriction fragment length differences due to either inheri
ted polymorphism or in diseases with suspected superantigen influence
(X-linked severe combined immune deficiency and canine juvenile polyar
teriitis syndrome) 13 dog families of three different breeds were exam
ined. In addition primary spleen cell cultures, stimulated with either
phytohemagglutinin A (PHA) or staphylococcus enterotoxin A (SEA) and
B (SEB) were studied. The germline digest pattern of the enzymes Pst I
, Sst I, Bgl II, Eco RI and Eco RV were identical in all dogs examined
with the exception of one dog with canine juvenile polyarteriitis syn
drome, In this dog an additional band was found in the Bgl II and Eco
RV digestion suggestive of specific TCR rearrangement. Bam HI digestio
n revealed restriction fragment length polymorphisms (RFLPs) showing M
endelian inheritance. After digestion of the genomic DNA extracted fro
m PHA, SEA or SEB stimulated spleen cells and Southern blot analysis,
no differences in fragment patterns between the unstimulated cells and
the stimulated cells could be detected, An important point to conside
r before a specific pattern variation between dogs is classified to be
a marker for a specific disease or is used in pathogenesis studies, i
s the possibility of an inherited RFLP, especially after Barn HI diges
tion. In such studies the combined examination of the parents and the
offspring must be recommended.