spontaneous malignomas in tupaia (tree shrew)

American Journal of Primatology 2:177-189 (1982)

Spontaneous Malignomas in Tupaia (Tree Shrew) G. DARAI ~ N I ) L. ZOLLER Institut fur Medizinische Virologie, University of Heidelberg W . IIOFMANN A N D P. MOLLER Pathologisches Institut, Uniuersit.y of Heidelberg A. SCHWAIER Battelle-Institut e. V.. Frankfurt urn Main R.M. FLUGEL Institute fur Virusforschung, Ueutsches Krebsforschu,lgsrentrum Heidelberg

Nine spontaneous malignomas of the tree shrew were detected and analysed during an observation period of nine years. The tumours were histopathologi- cally examined and classified. All malignomas developed in imported Tupaia only. From the tumour cells of two different animals new Tupaia herpesviruses were isolated. This is the first report on spontaneous malignomas of Tupaia in captivity.

Key words: Tupaia, tumours, histopathology, herpesvirus

INTRODUCTION

Tupaia (tree shrew), a member of the family Tupaiidae, is regarded as one of the most primitive prosimians, bridging the gap between insectivores and primates [Fiedler et al, 1956; Moore & Goodman, 19681. The phylogenetic placement of the Tupaia has led to much interesting discussion [Dene et al, 1978; Goodmann, 1966; Lehmann et al, 19741, especially after recently published reports on the first discoveries of fossil tree shrews of the eocene and miocene [Chopra and Vasishat, 1979; Chopra et al, 1979; Maier, 1977; Martin, 19791. Our aim was to test animals suitable for laboratory experiments in medicine and biology which are more closely related to primates than to rodents [Helmstaedter et al, 1977a, b; Sakai et al, 1979; Schwaier, 1979 a, b; Schwaier et al, 1979; Steinhausen et al, 1978; Taugner et al, 19761. From this point of view several laboratories are engaged in studying Tupaia in different fields of science. We have been testing Tupaias for about nine years and have concentrated on tumour induction by chemicals, radiation, and viruses. With respect to these investigations, knowledge and information on the spectra and incidence of spontaneous tumours in Tupaia is of special interest. Elliot et al[1966] described a breast cancer in a tree shrew. This breast tumour, which was classified as an adenocarcinoma, is the only tumour which Elliot and co- workers observed during the study of tree shrews on a Malayan peninsula over a period of four years, during which about 400 tree shrews were carefully observed for detection of tumours.

Received August 10, 1981; accepted October 14, 1981

Address reprint requests t o Priv.Doz.Dr.rned.Gholarnreza Dnrai, Institut fur Medizinische Virologie der Univer- sitat Heidelberg, Im Neuenheimer Feld 324, 6900 Heidelberg 1, FRG.

0275-2565/82/0202-0177$04.00 0 1982 Alan R. Liss, Inc.

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MATERIALS AND METHODS

During a study of tree shrew (Tupaia belangeri) in our colony [Schwaier, 1973,19751, we observed six malignomas. The breeding colony was started with 35 imported animals, which were used for breeding only. They were maintained until natural death occurred. Up to now 1650 Tupaias were bred in captivity. Among these, 1464 animals were used for short-term experiments, which were terminated before the animals were two years old. One-hundred-seventy animals reached an age of more than two years. Of these, 71 survived to three years and 16 reached four years of age. Life expectancy of Tupaias is 8 to 14 years. In addition, we received a sacrificed Tupaia belangeri with spontaneously developed malignoma from Dr. von Holst, Bayreuth, one from Dr. Raab, Miinchen, and histopathological material from Dr. Kuhn, Gottingen. We included these three malignomas in our investigation, which is the subject of this report.

RESULTS AND DISCUSSION

The biological data as well as the history of animals which developed spontaneously malignant tumours are given in Table I.

Description of case 1: A lymphoma was detected in an adult female 7-8-year-old Tupaia (animal No.26) (Table I). Histopathological investigation revealed a discordant hyperplasia of the lymphoreticular tissues with formation of nodal, follicle-like pro- liferates. Two types of cells were observed. The first cell type was small with polymorphic nuclei of clear appearance, the nucleoli were small; the predominant part of the other cell type was of medium to large size with large, rounded nuclei. The chromatin showed marginal aggregations with marginal nucleoli. Mitoses were relatively frequent. Several thick reticulin fibers were sparsely distributed after silver-staining. Tumour cell infiltration into the following areas were observed: The parenchyme of the pancreas, the peribronchial lymph-follicles, the connective tissue of the pelvis of the kidney, the subserous membrane of duodenum, and the cerebral meninges. Diagnosis of this tumour was established according to Kiel classification [Lennert, 19781 as a high-grade malignant lymphoma, centroblastic type without sclerosis (Fig. 1A) or follicular center cell lymphoma predominantly large,” [Lukas & Collins, 19751, infiltrating the pancreas, the kidney (peripelvin), the peribronchial lymph nodes, and the subserous membrane of the duodenum, periportal fields of the liver, and meninges (Fig. lB , C). Case 2: A generalized lymphoproliferative disorder resembling human Hodgkin’s

disease was observed in a female tree shrew of 7-8 years of age (animal No. 30, Table I). Histological examination revealed hyperplasia of the lymphoreticular tissue and a polymorphic cellular population consisting of immunoblasts, numerous Hodgkin-like cells and Reed-Sternberg-like cells, eosinophilic granulocytes and a marked lymphocytic depletion (Fig. 2A). The spleen showed a diffuse infiltration pattern, liver and peripelvin tissue of kidneys were likewise infiltrated (Fig. 2B). Case 3: A malignant lymphoma, immunocytoma polymorphic subtype [Lennert et al,

19781 (Fig. 3A) was detected in a female, 8-year-old tree shrew (animal No. T,, Table I). Metastasis of tumour was also found in liver, spleen, and kidneys. A second independent malignoma-carcinoid tumour of pancreas was also detected (Fig. 3B). Cases 4 and 5: Two primary, hepatocellular carcinomas with metastases in the lungs

were found in two male animals (animals No. 29 and 31, Table I). Histopathologicd examination of the tumour of Tupaia No. 29 showed multiple tumour foci in liver with large polymorphic cells, very large nucleoli with multiple inclusion bodies (Fig. 4Ah and numerous mitoses and aggregated tumour cells in some interalveolar septula of the lung. This animal had also developed a second malignoma- an epidermoid carcinoma of skin (Fig. 4B, C, Table I)-independent of the hepatocellular carcinoma.

Malignant Tumors in Tupaia 179

Table I. Biological Data of Tree Shrews Which Developed Neoplasias

Weight/Ageh of animals Beginning of tumour development/

Code number when sacrificed Time of sacrifice of Tupaiu* (p lyear ) Sex (months)

26 15017-8 female 1 30 10517-8 female 1 TGd 19418-9 female unknown 2gC 23018-9 male 6 31 13117-8 male unknown 32 19819-10 female unknown T,e 165/5-6 female unknown TMf 185110-11 female unknown 8 17515-6 female 3

"Animal Nos. 8,26,29,20.31, and 32 were imported from Bangkok in 1971 (colony of Rattelle Institute). bAverage weigh1 of adult tupaia 230 * 20gm (male and 190 + 20gm [female). 'In September 1977 Tupuia No.29 developed several small tumours on the cheek close to the right eye region. The tumours were removed for histopathological examination. which revealed a precancerous keralosis (Fig. 2C). During the following 14 months a rapid growth of one walnut-sized tumour close to thc eyelid occurred. Histopathological examination revealed an epidermoid carcinoma of the skin (Fig. 2). dThe histopathological materials, biological data, and history of tumour induction was provided by Dr. Kuhn, Gottingen. eThe sacrificed animal was provided by Dr. von Holst, Bayreuth. fThe sacrificed animal was provided by Dr. Raab. Munchen.

Histopathological examination of Tupaia No. 31 revealed essentially the same results as described above for tree shrew No. 29, except that there was no second malignoma. In addition, this carcinoma possessed numerous necrotic foci. In contrast to the above mentioned liver carcinoma, this tumour showed partially trabecularly shaped aggregations of tumour cells surrounded by septa of the connective tissue of liver. The tumour cells showed bright and acidophilic cytoplasm. The nuclei were swollen, as were two or three nucleoli (Fig. 5A). Few mitoses were visible.

Case 6: Malignant mixed tumour, osteoid sarcoma, and squamous cell carcinoma of the uterus was found in a female Tupaia (animal No.32, Table I). Histopathological examination showed that the mucous coat of the uterus contained a glandular-cystic hyperplasia. Starting at the myometrium, star- and spindle-like cells were observed, which lay on small trabecula of osteoid substance (Fig. 6A, C). Nuclei of tumour cells showed multiple nucleoli and chromatin aggregation. I t is remarkable that numerous aggregations of squamous carcinoma cells (Fig. 6B) with central necrosis are situated within the sarcoma area. Case 7: A carcinoid tumour of pancreas with metastases in lung and the adrenal gland

was observed in a female Tupaia (animal No.8, Table I and Fig. 7A). In addition, two carcinomas of the apocrine sweat gland were detected in two tree shrews (Case 8), which were provided by Dr. von Holst, Bayreuth, Tupaia T,, and by Dr. Raab, Munchen, Tupaia T, in Table I (Case 9), Both tree shrews developed slowly growing tumours of bean-size at the epidermis of the left flank. Histopathological examination showed that Tupaia T, was a tubular carcinoma of the apocrine sweat gland (Fig. 8A) and Tupaia T, a papillary carcinoma of the apocrine sweat gland (Fig. 8B, C) [Weiss and Frese, 19741.

Parallel to the histopathological investigations of the Tupaia malignomas, cell cultures of the different tumours were established in order to study a possible viral etiology of

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Fig. 1. Microphotographs of malignoma of l'upaza no. 26. A) Section of high-grade malignant centro-blastic lymphoma. 13) Metastasis of tumour in meninges. C) Metastasis of tumour cells in liver (HQE, x 350).


Fig. 2. Microphotographs of malignoma of Tupaia no. 30. A) Section of tumour like “Hodgkin’s disease, lymphocytic depletion,” large arrows show Hodgkin’s cells and small arrow shows Reed-Sternberg giant cells (H&E, x 350). B) Metastasis of tumour in liver indicated by arrows (H&E, x 180)

182 Darai et a1

Fig. 3. Microphotographs of two different turnours of Tupaia no. TG. A) Section of malignant lymphoma. im- rnunocytoma. polymorphic subtype. B) Section of pancreas infiltrated with carcinoid tumour cells IH&E, x 1401

184 Daraietal

Fig. 5. Microphotographs of malignoma of Tupaia no. 31. A) Section of a primary hepatocellular carcinoma (H&E, x 3501. B) Large metastasis of tumour in the lung (H&E, x 40).

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Fig. 7. Microphotographs of carcinoid tumour of pancreas of Tupnia no. 8. A) Section of pancreas, large arrow shows carcinoid tumour cells and small arrow shows an Island of Langerhans. B) Metastasis of the tumour cells in the lung indicated with white arrow (H&E, x 350).


Fig. 8. Microphotographs of papillary and tubularly carcinoma of apocrine sweat gland of TUpQiQ no. TB and TM. A) Section of skin of Tupaia no. TB shows typical picture for tubular carcinoma of apocrine sweat gland, (H&E, x 70). B.C) Sections of skin of Tupaiu no. TM shows typical picture for papillary carcinoma of apocrine sweat gland (H&E, magnificaton: 8B x 120, 8C x 240).

188 Daraietal

these neoplasias. It is remarkable and of special interest that two types of herpesviruses were isolated and characterised from a malignant lymphoma (Tupaia No.26) and from Hodgkin’s disease [Darai et al, 1979a, b]. These findings complement studies on the isolaton of herpesviruses from a malignant lymphoma of owl monkeys [Schneider, 19751, from a spontaneous myelomonocytic leukemia of orangutan [Rasheed et al, 19771, from Burkitt’s lymphoma, and of nasopharyngeal carcinomas of man [Epstein et al, 1965; Zur Hausen et al, 19701. Although an endogenous retrovirus was isolated from Tupnia cell cultures after activation with iododeoxyuridine [Flugel et al, 19781, our attempts to demonstrate the presence of RNA tumour viruses by forming reverse transcriptase assays with the Tupaia tumour cell cultures were not successful.

CONCLUSION

Nine malignomas of tupaia were detected and analysed during an observation period of nine years. Our colony comprised 1650 animals, from the F1 to F8 generation; all malignomas developed in the later part of their lives. Neoplasias did not occur in animals bred in captivity. Whether the spontaneous induction of these malignomas is due to ad- vanced age or environmental factors can only be determined after detailed experimental investigation. There was a remarkably broad spectrum of malignomas, including even such a rare tumour as the apocrine sweat gland carcinoma. The isolation of two herpesviruses from the malignant non-Hodgkin-lymphoma (Tupaia No.26) and from Hodgkin’s disease (Tupaia No.30), is of great importance for virology and comparative pathology, as well as for the possible etiopathogenesis of malignant lymphoma and mor- bus Hodgkin of humans.

ACKNOWLEDGMENT

We dedicate this work to the 60th birthday of Professor Dr. Klaus Munk.

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spontaneous malignomas in tupaia (tree shrew)

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