Vet Pathol 37:184–186 (2000)

BRIEF COMMUNICATIONS and CASE REPORTS

Primary Epitheliotropic T-cell Lymphoma of the Urinary Bladder in a Dog

P. MAIOLINO AND G. DEVICO

Abstract. A 7-year-old, intact female mixed-breed dog was presented for evaluation of hematuria. Physical
examination revealed a suprapubic mass. Ultrasonographic examination showed a large lobular mass occupying
the urinary bladder. At the owners’ request, the dog was euthanatized and a postmortem examination was
performed. Necropsy confirmed the presence of a lobular mass of about 5- to 6-cm diameter protruding into
the lumen of the bladder. Histologically, the mass was composed of a large number of atypical lymphoid cells
in the lamina propria and mucosal epithelium. Immunohistochemically, the neoplastic cells expressed CD3 but
not CD79␣ or keratin and vimentin, supporting a diagnosis of T-cell lymphoma.

Key words: Dogs; epitheliotropism; urinary bladder; T-cell lymphoma.

Primary malignant lymphoma of the urinary bladder is
extremely rare in humans.5,8,10 In veterinary medicine, only
eight cases have been reported: two in dogs, two in cats, and
four in cows.9 In this paper we describe the histologic and
immunohistochemical features of a primary malignant lym-
phoma of the urinary bladder with epitheliotropism in a dog,
a finding not previously reported in this species.
A 7-year-old, intact female, mixed-breed dog was pre-
sented to the Veterinary Medicine Faculty, University of Na-
ples, for examination after the owners complained that their
pet had constant hematuria. The dog had no other clinical
history. Physical examination revealed a suprapubic mass.
Plain abdominal radiographs and ultrasonographic exami-
nation showed the presence of a large lobular mass of uni-
form density in the area of the urinary bladder. No other
abnormalities could be seen. A tentative diagnosis of a tumor
was made. In view of the poor prognosis, the owners re-
quested euthanasia rather than an exploratory laparotomy.
At postmortem examination, abnormalities were confined
to the urinary bladder. The bladder was grossly abnormal
and the whole organ was removed. The gross pathologic
examination confirmed the presence of a large, lobular, 5- to
6-cm-diameter, light tan mass protruding from the ventral
mucosal surface, with no extension to the trigone evident.
On the cut surface, the bladder mucosa was markedly thick-
ened with prominent rugae and multiple foci of hemorrhage
and ulcers. No evidence of metastasis or other gross abnor-
malities were found.
Representative areas of the tumor were fixed in 10% for-
malin for 48 hours and then processed for paraffin embedding.
Four-micrometer sections were mounted on poly-L-lysine
coated glass slides and incubated at 37 C overnight to opti-
mize tissue adhesion to the slide. The sections were dewaxed
in xylene, dehydrated in graded alcohols, and stained with
hematoxylin and eosin. For immunohistochemical analysis
some sections were washed three times for 5 minutes each
time in 0.01 M phosphate-buffered saline (PBS), pH 7.2–7.4.
Endogenous peroxidase activity was blocked with hydrogen
peroxide 0.3% in absolute methanol for 30 minutes. Before
184
the immunohistochemical procedure (a streptavidin–biotin
peroxidase method) with a commercial kit (LSAB Kit, Dako,
Milan, Italy) some sections were treated with 0.1% trypsin
(Sigma, Milan, Italy) for 1 hour at 37 C for detection of CD3
and some were incubated twice for 5 minutes at 700 W in
citrate buffer (pH 6.0) in a microwave oven for detection of
CD79␣ and keratin. Then the sections were incubated over-
night at 4 C with one of the following antibodies diluted in
PBS containing 0.1% bovine serum albumin (BSA, Sigma)
and sodium azide 0.05%: CD3 (T-cell marker, diluted 1:50,
Dako), CD79␣ c␥ (clone HM57, diluted 1:50, Dako), keratin
(Keratin cocktail CK22, prediluted, Biomeda, Milan, Italy),
vimentin (clone V9, diluted 1:50, Dako). Biotinylated anti-
mouse and rabbit immunoglobulins (LSAB Kit, Dako), dilut-
ed in PBS, were used as secondary antibody, and were applied
for 30 minutes. After washing in PBS, the sections were in-
cubated in streptavidin conjugated to horseradish peroxidase
in Tris-HCl buffer containing sodium azide 0.015% (LSAB
Kit, Dako) for 30 minutes and finally exposed to the 3,3⬘-
diaminobenzidine tetrahydrochloride chromogen (DAB, Sig-
ma) containing hydrogen peroxide 0.03%, and counterstained
with Harris’ hematoxylin, dehydrated, cleared, and cover-
slipped with Eukitt (Bio-Optica, Milan, Italy). Sections of a
normal dog lymph node were used as positive controls and
to ensure specificity of the antibodies. Negative controls con-
sisted of sections incubated with PBS.
Histologically, the mucosa and submucosa of the urinary
bladder were massively infiltrated by a large number of me-
dium to large atypical lymphoid cells. The cells were usually
round with scant pale eosinophilic or large optically clear
cytoplasm. Nuclei were small, round to irregularly convo-
luted with finely aggregated and uniformly distributed chro-
matin, and contained rare nucleoli. Mitotic figures were fre-
quent (two to four per high-power field). The neoplastic cells
invaded the mucosal epithelium, with effacement of the stro-
mal–epithelial junction, which was often eroded and re-
placed by the atypical lymphoid cell population expressing
CD3 (Fig. 1). The intraepithelial lymphocytes were often
surrounded by a halo of clear cytoplasm. A great number of
Vet Pathol 37:2, 2000 Brief Communications and Case Reports 185

Fig. 1. Urinary bladder, dog. Neoplastic lymphocytes, with a positive immunoreaction to CD3 antigen (arrowhead)
infiltrate the mucosal epithelium with effacement of the epithelial–laminal propria junction. Streptavidin biotin peroxidase
method, diaminobenzidine (DAB) chromogen, Mayer’s hematoxylin counterstain. Bar ⫽ 30 ␮m.

histiocytic cells were also scattered throughout the neoplastic
tissue imparting a ‘‘starry sky’’ appearance. The neoplastic
cells were positive for the T-cell marker anti-human CD3
(Fig. 2) but not for vimentin, keratin, and CD79␣, supporting
a diagnosis of T-cell lymphoma. No evidence was found of
bacterial infection.
Malignant lymphoma of the urinary bladder is known to
occur in several domestic species as a part of a multicentric
disease or as an extension from other sites. However, pri-
mary malignant lymphoma of the urinary bladder is known
to be a rare pathologic event both in humans and animals.
In humans, like most extranodal lymphomas, the neoplasms
are usually considered to be of B-cell origin, and according
to recent studies1 the mucosa-associated lymphoid tissue
lymphomas constitute a large proportion of these cases.
However, to our knowledge, epitheliotropism in bladder
lymphomas has never been described. The epitheliotropism
refers to the affinity of the malignant lymphocyte to epithe-
lial structures, mainly epidermis and cutaneous adnexae and
usually occurs in cutaneous T-cell lymphomas (e.g., mycosis
fungoides) in humans and animals3,4 and in canine gastro-
intestinal T-cell lymphomas.2,11 In canine cutaneous lympho-
ma, the epitheliotropism of the neoplastic elements has been
suggested to result from the expression on their cell surface

Fig. 2. Urinary bladder, dog. Neoplastic lymphocytes, with a positive immunoreaction to CD3 antigen (small arrow-
heads). Note the presence of histiocytic cell CD3 negative (large arrowheads). Streptavidin–biotin peroxidase method,
diaminobenzidine (DAB) chromogen, Mayer’s hematoxylin counterstain. Bar ⫽ 25 ␮m.
186 Brief Communications and Case Reports
of specific adhesion molecules (e.g., integrins), which me-
diate a direct interation with keratinocytes. This surface pat-
tern has also suggested that the neoplastic lymphoid T-cell
could originate from a memory subpopulation.6,7 In conclu-
sion, the anatomical localization and the histologic and the
immunohistochemical findings of present case, are consistent
with the diagnosis of primary malignant T-cell lymphoma of
the urinary bladder with epitheliotropism in a dog.
Acknowledgement
We wish to thank Mr. R. Ilsami for his technical assis-
tance.
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Request reprints from Dr. P. Maiolino, Dipartimento di Pa-
tologia e Sanità Animale, Settore Anatomia Patologica, Fa-
coltà di Medicina Veterinaria, Università degli Studi di Na-
poli Federico II, Via Delpino, 1-80137 Napoli (Italy) and
from Prof. G. DeVico, Instituto di Patologia Generale ed
Anatomia Patologia, Facoltà di Medicina Veterinaria, Univ-
ersità degli Studi di Messina, Via S. Cecilia, 30-98123 Mes-
sina (Italy).