Histopathology and distribution of cells harboring bovine leukemia virus (BLV) proviral sequences in ovine lymphosarcoma induced by BLV inoculation.

Six sheep with lymphosarcoma induced by hypodermic inoculation of bovine leukemia virus (BLV) materials were examined to elucidate the relation between pathologic lesions and integration of BLV provirus in cellular DNAs. Antibodies to BLV gp-antigens had been detected since the 3rd week after the inoculation, and BLV was positive when checked 3 months later. Lymphosarcomas followed the leukemic phase in 4 sheep. The other 2 sheep showed initial lesions of lymphosarcoma and were aleukemic clinically. Five animals were killed by enthanasia and autopsied at 2.5 to 3.5 years postinoculation (pi) because of their diseased condition. One animal died 10 years pi following the 4th leukemic episode. Sarcomatous lesions were confirmed grossly and histologically, and the proliferating neoplastic cells were classified into lymphocytic, prolymphocytic, lymphoblastic and histiocytic types. Integration of BLV provirus in cellular DNAs of the peripheral blood lymphocytes (PBL) and neoplastic cells of sarcomatous lesions was examined by Southern blotting technique. BLV provirus was demonstrated in the PBL of all infected animals and in most of the sarcomatous lesions of the spleen, kidney and lymph nodes except 4 lymph nodes showing slight neoplastic infiltration. The results indicated that ovine lymphosarcoma could be caused by BLV and the cells carrying proviral information seemed to be disseminated and proliferate in the lesions.     

Thymic lymphosarcoma of T cell lineage in a koala (Phascolarctos cinereus)

Objective To diagnose and characterise thymic lymphosarcoma in a koala.
Design A pathological case.  

Animal

Seven-year-old female koala.
Procedure The neoplastic process was investigated macroscopically, haematologically, histologically and immunohistologically.
Results The koala had difficulty swallowing because of a medial swelling in the lower neck. Biopsy of this mass and blood examination revealed lymphosarcoma with a leukaemic manifestation; necropsy and histopathological examination showed the mass to be thymus. Palatine tonsils, cervical, axillary and mesenteric lymph nodes, spleen, liver, gut, bronchi, genitalia and bone marrow were infiltrated by neoplastic cells. Immunohistological staining of the thymic mass, cervical and mesenteric lymph nodes, bone marrow, spleen and gut revealed the neoplastic cells to be of T lymphocyte origin (positive for both anti-human CD3 and CD5).  

Conclusions

It is speculated that the neoplastic process originated in the thymus and was disseminated by bloodborne neoplastic cells: This first report of thymic lymphosarcoma in a marsupial confirms that antibodies raised originally to investigate human lymphoid neoplasia can cross-react with neoplastic lymphocytes in koalas.     

Further phenotypic characterization of target cells for bovine leukemia virus experimental infection in sheep

To determine the phenotype of target cells for bovine leukemia virus (BLV) infection in sheep, we analyzed blood lymphocytes from BLV-infected clinically healthy and leukemic sheep by use of monoclonal antibodies. In clinically healthy and leukemic sheep that were BLV-infected, the blood concentration of T lymphocytes was within normal values, but the number of B lymphocytes was increased in several cases. In addition, the number of blood lymphocytes expressing the BLV antigen correlated well with that of B lymphocytes. Double immunofluorescence staining demonstrated that lymphocytes expressing BLV antigens bore B-cell but not T-cell surface markers. Moreover, neoplastic cells in the lymph nodes of leukemic sheep were stained immunohistochemically with an anti-B monoclonal antibody but not with any of anti-T monoclonal antibody tested, indicating that tumor cells are of B-lymphocyte origin. Collectively, these results show that BLV antigen-positive cells obtained from BLV-infected sheep that have no clinical signs and BLV-induced lymphosarcoma cells belong to the B-lymphocyte lineage.     

Hypereosinophilic paraneoplastic syndrome in a cat with intestinal T cell lymphosarcoma

A 10-year-old, neutered female, domestic shorthair cat was presented with a recent history of weight loss, polydipsia, diarrhoea and vomiting. On physical examination, intestinal thickening and mesenteric lymph node enlargement were apparent. Clinical investigations revealed peripheral blood eosinophilia, eosinophilic abdominal effusion and eosinophilic mesenteric lymphadenitis. There was a temporary response to treatment with glucocorticoids but signs progressed and the cat was euthanased. On histology, there was eosinophilic infiltration and fibroplasia of intestine and mesenteric lymph nodes. Large aggregates of neoplastic round cells in the intestine and lymph nodes were identified as T lymphocytes using immunohistochemistry. A diagnosis of intestinal T cell lymphosarcoma was made. This case demonstrates that hypereosinophilic paraneoplastic syndrome may occur in cats with lymphosarcoma. Eosinophil chemotaxis may be a response to the production of interleukin-5 by neoplastic lymphocytes.     

Lymphosarcoma development in sheep experimentally infected with bovine leukaemia virus

Twelve sheep were experimentally infected with a phytohemagglutinin (PHA) treated short term culture of lymphocytes from a cow naturally infected with BLV at the PL stage. Five of 12 (42%) BLV infected sheep had histologically confirmed lymphosarcoma 10-16 months after infection. The PBL's were increased to leukemic levels 3-21 weeks before death due to lymphoblastic leukemia. Lymphocyte proliferation and appearance of immature lymphocytes and lymphoblastic cells in the blood were a characteristic feature of tumour development following inoculation with an Australian strain of BLV. In contrast to a number of previous studies the peripheral lymph nodes of all infected sheep were clinically normal throughout the experimental period but at death gross tumours were evident in the mesentric lymph nodes and the heart in all cases. All the other lymph nodes, liver, spleen, kidney and lung were histologically infiltrated with lymphoid tumour cells. Gross tumours were present in the abomasum (1 out of 5) in the urinary tract (2 out of 5) and in the uterus (1 out of 2). The majority of the tumour cells isolated from the various tissues were centroblastic demonstrating that the malignant leukemia in experimentally infected sheep was of a multicentric centroblastic type. The central nervous system was not involved in any case.     

Evidence of decreased concanavalin A induced suppressor cell activity in the peripheral blood and pulmonary lymph nodes of sheep with ovine progressive pneumonia

Concanavalin A (Con A) induced suppressor cell activity was evaluated in a group of ovine progressive pneumonia virus-antibody positive sheep (OPPV+). Decreased levels of suppressor activity were observed in the peripheral blood and regional pulmonary lymph nodes of animals with clinical and pathological evidence of ovine progressive pneumonia (OPP) when compared to animals with no lesions and animals with caseous lymphadenitis involving the lungs and pulmonary lymph nodes. Decreased levels of Con A stimulated lymphocyte proliferation was also observed in the peripheral blood and iliac lymph nodes of sheep with OPP. Sheep with OPP were found to be hypogammaglobulinemic. Sera from OPPV+ sheep had no effect on T and B cell mitogen stimulated responses of lymphocytes from sheep seronegative for antibodies to ovine progressive pneumonia virus (OPPV-) when compared to normal sheep serum.     

Extravasation of lymphocytes via paracortical venules in sheep lymph nodes: visualization using an intracellular fluorescent label

In rodents and humans, lymphocytes extravasate into lymph nodes via specialized paracortical venules lined with high endothelium (HEV). Sheep and other ruminants do not have morphologically defined HEV in their lymph nodes. It has been assumed that lymphocyte extravasation in these species proceeds via analogous structures; i.e., paracortical venules lined with low to medium endothelium. In this study, lymphocyte suspensions were prepared from surgically excised lymph nodes of sheep and labeled with an intracellular fluorescent dye, H33342. Labeled cells were infused intravenously back into donors, and sheep were killed at various intervals after infusion. Frozen sections of lymph nodes were examined microscopically for the location of labeled cells. Ten minutes after infusion, labeled cells were seen in the lumen of venules located in the paracortical region of the nodes. At later time points, cells were seen apparently migrating through the venule walls and in the adjacent paracortical tissue. Similar experiments were performed in which H33342-labeled murine lymphocytes were infused into syngeneic mice. When equivalent cell numbers (based on animal size) were infused, no obvious differences were seen between location and kinetics of appearance of labeled cells in lymph nodes of sheep compared to those of mice. These results indicate that lymphocyte extravasation in sheep proceeds via paracortical venules in lymph nodes. The function of these venules appears to be analogous to HEV in nonruminant species.     

Metastatic thymic lymphosarcoma in a calf

Metastatic thymic lymphosarcoma was diagnosed in a 16-month-old mixed-breed heifer with a history of progressive weight loss. Physical examination revealed cachexia, pale mucous membranes, large peripheral lymph nodes, and a 15 X 40-cm mass in the ventral portion of the neck, extending cranially from the thoracic inlet. Neoplastic lymphocytes were identified in aspirates of pleural effusion and bone marrow. Histologic examination of necropsy specimens substantiated metastatic dispersal of the tumor into lymphoid tissue, liver, intestine, heart, and kidney. This case differs from other reported cases of thymic lymphosarcoma because of the involvement of organs other than the thymus and lymph nodes. Analytical flow cytometry was performed on mononuclear leukocytes obtained from blood by use of density gradient centrifugation. The majority of cells (65%) appeared to be of an immature, poorly differentiated phenotype, on the basis of the small numbers of cells stained with monoclonal antibodies specific for the following cell surface markers: 8% BoCD2 (IL-A26; sheep erythrocyte receptor on T-lymphocytes); less than 10% B-lymphocytes as determined by expression of MHC class II proteins and surface immunoglobulin; 12% monocytes (IL-A24); and 5% null cells (IL-A29). Although the leukemic cell population did not express traditional surface markers for T-lymphocytes, we hypothesize that the leukemic cell population represents an early stage of T-cell maturation that has failed to differentiate and express characteristic cell surface antigens.     

Comparison of core needle biopsy and fine-needle aspiration of enlarged peripheral lymph nodes for antemortem diagnosis of enzootic bovine lymphosarcoma in cattle

OBJECTIVE: To determine whether antemortem core needle biopsy and fine-needle aspiration of enlarged peripheral lymph nodes could be used to distinguish between inflammation and lymphosarcoma in cattle. DESIGN: Prospective study. ANIMALS: 25 cattle with enlarged peripheral lymph nodes. PROCEDURES: Antemortem biopsies of the selected lymph nodes were performed with an 18-gauge, 12-cm core needle biopsy instrument. Fine-needle aspirates were performed with a 20-gauge, 4-cm needle. Specimens were analyzed by pathologists who were unaware of clinical findings and final necropsy findings, and specimens were categorized as reactive, neoplastic, or nondiagnostic for comparison with necropsy results. RESULTS: Sensitivity and specificity of core needle biopsy ranged from 38% to 67% and from 80% to 25%, respectively. Sensitivity of fine-needle aspiration ranged from 41% to 53%, and specificity was 100%. Predictive values for positive test results ranged from 77% to 89% for core needle biopsy and were 100% for fine-needle aspiration. Predictive values for negative test results were low for both core needle biopsy and fine-needle aspiration. CONCLUSIONS AND CLINICAL RELEVANCE: Results indicated that core needle biopsy and fine-needle aspiration can aid in the antemortem diagnosis of bovine enzootic lymphosarcoma. Results of fine-needle aspiration of enlarged peripheral lymph nodes were more specific and more predictive for a positive test result than were results of core needle biopsy.     

Lymphosarcoma and associated immune-mediated hemolytic anemia and thrombocytopenia in horses

Three horses with equine lymphosarcoma were examined because of clinical signs including chronic weight loss, respiratory distress, peripheral edema, and chronic colic. Clinicopathologic findings included evidence of an immune-mediated hemolytic anemia. Immune-mediated thrombocytopenia also was diagnosed in 1 of the horses and suspected in another. One horse died in spite of treatment, 1 died 5 hours after surgical removal of a tumor encircling the jejunum, and 1 was euthanatized because of deteriorating condition. Necropsy of each horse revealed extensive neoplastic infiltration of peripheral lymph nodes and abdominal or thoracic viscera with neoplastic lymphocytes.     

Immunomorphologic and morphometric changes in pulmonary lymph nodes of sheep with progressive pneumonia

Morphologic, immunohistochemical, and morphometric studies were conducted on the posterior mediastinal lymph nodes of eleven sheep with naturally occurring ovine progressive pneumonia and four apparently healthy sheep with no pulmonary lesions (three seropositive, one seronegative for antibody to ovine progressive pneumonia virus). Compared with lesion-free sheep, sheep with ovine progressive pneumonia had a seven-fold increase in B lymphocyte areas and a 21/2-fold increase in T lymphocyte areas of these lymph nodes. Immunochemistry revealed cytoplasmic immunoglobulin G in scattered cells of germinal centers, medullary cords and interfollicular areas and membrane-associated immunoglobulin G in dendritic cells of germinal centers. Immunoglobulin M staining cells were widely scattered in germinal centers and medullary cords. Although B cell hyperplasia seemed to be the predominant process in lymph nodes of sheep with ovine progressive pneumonia, this was not accompanied by the expected degree of plasmacytosis, morphologically and immunohistochemically. These findings may represent an aberrancy of immunoregulation in ovine progressive pneumonia.     

Experimental induction of caseous lymphadenitis in sheep by intralymphatic inoculation of Corynebacterium ovis

Lesions histologically similar to those of natural caseous lymphadenitis developed in sheep popliteal lymph nodes following injection of viable Corynebacterium ovis cells into afferent lymphatic ducts. Histopathological examination of affected lymph nodes suggested that exotoxin might be involved in the establishment, progressive development and persistence of caseous lymphadenitis lesions through localised cytotoxicity.     

Hindlimb lameness caused by systemic lymphosarcoma in a Warmblood mare

Equine lymphosarcoma is rare but has been reported in mediastinal lymph nodes, cutaneous nodules, the gastrointestinal system and peripheral lymph nodes. This report describes the clinical presentation of lymphosarcoma characterised by weight shifting, reluctance to ambulate or peripheral oedema. This case is extremely atypical in clinical presentation and justifies the need for consideration of lymphosarcoma when formulating a differential diagnosis of lameness with atypical presentation.     

乳胶凝集试验检测牛、羊淋巴结中的沙门氏菌

用沙门氏菌多价血清致敏乳胶并制成乳胶抗体,建立沙门氏菌乳胶凝集试验检测方法（LAT）。用该方法和常规分离培养检测法检测38份牛淋巴结、101份羊淋巴结,结果乳胶凝集试验牛淋巴结阳性21份,羊淋巴结阳性44份;常规分离培养检测法牛淋巴结阳性22份,羊淋巴结阳性48份,两种方法的阳性符合率牛淋巴结为81.8%,羊淋巴结为81.3%。试验表明乳胶凝集试验操作简便、快速、敏感性高、特异性强且可用于现场检测,是一种适合基层单位用来检测沙门氏菌的可靠方法。     

Clinical and anatomical features of lymphosarcoma in 118 cats

Objective To determine patients' characteristics and anatomical distribution of lesions in cats with lymphosarcoma. Design Prospective multi-institutional study of naturally occurring feline lymphosarcoma. Methods Veterinarians in Sydney were provided with free diagnostic laboratory services for suspect cases of feline lym-phosarcoma. Lymphosarcoma was diagnosed based on physical findings, radiographic and/or ultrasonographic images and results of cytological or histopathological examination. When owners were not interested in pursuing an antemortem diagnosis, suspect cases were collected for necropsy. Patients' characteristics and physical findings were recorded. A modified scheme for anatomical classification of lesions was devised including a ‘mixed’ category for cases which involved two or more anatomical forms. Results One hundred and eighteen cases were accrued over an 18 month period. The median age was 120 months and range 5 to 212 months. Age distribution was bimodal, with a small peak for cats less than 24 months, and a normal distribution centred on 97 to 120 months. Eighty cats were domestic crossbreds, 22 were Siamese or Oriental cats (including crosses), 6 were Burmese, 5 were purebred longhairs and the remaining 5 were one of a number of purebred shorthaired breeds. In comparison to 1017 consecutive cases admitted to our hospital for conditions other than lymphosarcoma, Siamese/Oriental cats were over-represented amongst lymphosarcoma cases (P = 0.0006). Male cats were also over-represented, accounting for 72 of 118 cases (P = 0.05). Abdominal lymphosarcoma was the most common anatomical form (43 cats), followed by mixed (39), nodal (20), mediastinal (9) and atypical (involving non-lymphoid organs, 7) forms. When analysed for specific organ involvement, 29 (25%) had mediastinal involvement, 71 (60%) had abdominal involvement including 60 (51%) with involvement of the intestinal tract and/or mesenteric lymph nodes and 36 (31%) with bilateral renal involvement, and 47 (40%) had peripheral lymph node involvement. No case of primary lymphoid leukaemia was identified. A noticeable subgroup of cats younger than 24 months had involvement of the anterior mediastinum with or without concurrent enlargement of cervical or axillary lymph nodes; Siamese/Oriental cats were over-represented in this subgroup. Among cases with nodal involvement, lymph nodes of the head and neck were frequently involved, mandibular nodes most commonly, followed by superficial cervical nodes. In seven cases a solitary node was affected. Conclusions Compared with similar surveys overseas, our cats were older and male cats were over-represented. There was a notable subgroup of young cats with mediastinal involvement. Siamese/Oriental cats were over-represented in this subgroup as well as in the larger population of cats with lymphosarcoma. Compared with overseas surveys, renal involvement, mixed cases and atypical cases (including nasal lymphosarcoma) were more common. A new subcategory of nodal lymphosarcoma, with involvement restricted to node(s) of head and neck, was identified.     

Transfer of immunity to Ostertagia circumcincta and IgA memory between identical sheep by lymphocytes collected from gastric lymph

Sheep rendered immune to Ostertagia circumcincta were challenged with 50,000 larvae and lymphocytes were collected from the gastric lymph up to eight days after challenge. The cells were transferred intravenously to genetically identical worm-free sheep which, together with controls, were challenged with 50,000 larvae and killed nine days later. Cells obtained during the donors lymphoblast response to challenge transferred partial immunity, measured either as stunting or loss of worms. Significantly less immunity was transferred by cells collected either before or after this response. Thus the responding cells can mediate protective immunity to O circumcincta. On the other hand the donor sheep remained immune to their challenge infection despite being depleted of these functional cells, showing that their presence was not essential for immunity to be maintained. Comparison of the immunoglobulin A (IgA) concentrations in the gastric lymph of recipient and control sheep showed that a local IgA response had also been transferred. Enumeration of mucosal mast cells suggested that a mastocytosis had been transferred to the two recipients which were most immune to challenge.     

Spontaneous regression of bovine cutaneous leukosis

Biopsies of skin from a 2-year-old heifer with spontaneously regressing dermal leukosis were examined. The heifer was not infected with bovine leukemia virus and was negative for tumor-associated antigens of enzootic bovine lymphosarcoma. By hematological standards for bovine leukemia, the heifer was positive at about 60 days post-occurrence of the disease (POD). At 53 days POD, lymphoblastic neoplastic cells in the dermis reacted with anti-T lymphocyte monoclonal antibody by the avidin biotin peroxidase complex method. The cells had intracytoplasmic clustered dense bodies under electron microscopy. From 53 to 83 days POD, figures of the transepithelial elimination (TE) against neoplastic cells and perivascular infiltration of small lymphocytes were in the dermis. Small lymphocytes reacted with anti-T lymphocyte monoclonal antibody. At necropsy there were no neoplastic lesions; there were flat lymph node-like tissues in the subcutis. Many germinal centers were seen in the lymphatic organs. Blood lymphocytes at 46 days POD were stimulated by phytohemagglutinin-P and concanavalin A. Sera, taken until 75 days POD and at necropsy, showed an inhibitory effect on mitogen-induced blastogenesis of normal bovine lymphocytes. These results suggested the existence of a spontaneous regressive mechanism against neoplastic lesions by TE and tumor immunity.     

PATHOLOGIC CORRELATION OF RESISTIVE AND PULSATILITY INDICES IN CANINE ABDOMINAL LYMPH NODES

We assessed the ability of the resistive index (RI) and pulsatility index (PI) to allow differentiation between normal, reactive, and neoplastic lymph nodes. Forty-seven medial iliac and 54 mesenteric lymph nodes from 83 dogs were evaluated sonographically. A cytologic sample was obtained in each dog that allowed categorization into one of the categories defined above. We found a significant difference in the RI and PI in nonneoplastic vs. neoplastic medial iliac and mesenteric lymph nodes. Values higher than 0.67 for the RI and 1.02 for the PI in medial iliac lymph nodes and higher than 0.76 for the RI and 1.23 for the PI in mesenteric lymph nodes had a high sensitivity and specificity for differentiating benign from neoplastic lymph nodes.     

T-cell lymphoma in a savanna monkey (Cercopithecus aethiops) probably related to simian T-cell leukemia virus infection

Lymphoma was seen in an 11-year-old female savanna monkey (Ceropithecus aethiops). The superficial inguinal and visceral lymph nodes were markedly enlarged, and their architecture was completely effaced by neoplastic cells. The neoplastic cells, which were highly pleomorphic, resembled those in adult T-cell lymphoma-leukemia in humans. Ultrastructurally the neoplastic cells were characterized by nuclear irregularity and clustered dense bodies, and almost all cells showed positivity for CD3. The animal had been reared with her family, and her mother and 2 brothers had antibodies reactive to human T-cell leukemia virus. This virus serologically cross-reacts with simian T-cell leukemia virus, which may be the causative agent of the present neoplasm.     

Influence of thymectomy on the susceptibility of cats to feline leukemia virus and lymphosarcoma.

Twelve cats were thymectomized at 5 weeks of age. Six of these cats were inoculated at 8 weeks of age and 6 at 4 months of age with the Rickard (R) strain of feline leukemia virus (FeLV), which produces a high incidence of thymic lymphosarcoma. Two groups of age-matched nonthymectomized cats were inoculated with the same FeLV-R stock. Thymectomy prior to FeLV infection had no influence on the induction of viremia or the incidence of lymphosarcoma. In the FeLV-inoculated nonthymectomized cats, lymphosarcoma developed in the thymus. In the thymectomized cats, lymphosarcoma developed in the intestine, mesenteric lymph nodes, and bone marrow, but the malignant lymphoblasts had surface markers characteristic of feline T lymphocytes. It was concluded that the presence of the thymus per se is not required for infection and oncogenesis by FeLV and that feline T lymphocytes may be transformed after peripheralization to other tissues.