Enteric pathology and Salmonella-induced cell death in healthy and SIV-infected rhesus macaques

The goal of this study was to morphologically characterize a ligated ileal loop model of Salmonella enterica serotype Typhimurium infection in rhesus macaques (Macaca mulatta) and to verify the occurrence of Salmonella-induced cell death in vivo. Eight adult healthy male rhesus macaques were used for ligated ileal loop surgery. Four macaques had been intravenously inoculated with simian immunodeficiency virus (SIV) mac251. Ileal ligated loops were inoculated with wild-type (WT) S. Typhimurium strain IR715 (ATCC14028 nal (r)), an isogenic noninvasive mutant strain (ATCC14028 nal (r) ΔsipAΔsopABDE2), or sterile Luria Bertani broth. Loops were surgically removed at 2, 5, and 8 hours post-inoculation (hpi). Intestinal samples were processed for histopathology, immunohistochemistry for detecting Salmonella, terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick end labeling (TUNEL), and transmission electron microscopy. Combined histopathology scores were similar between SIV-infected and control macaques. As expected, the invasion-deficient mutant was less pathogenic than WT S. Typhimurium. Neutrophil infiltrate in the intestinal mucosa correlated with bacterial loads (r = 0.7148; P < .0001) and fluid accumulation (r = 0.6019; P < .0001) in the lumen of the intestinal loops. Immunolabeled WT S. Typhimurium was observed in the epithelium and lamina propria at the tip of the villi at 2 hpi, progressing toward deeper lamina propria at 5-8 hpi. Most TUNEL-positive cells localized to the lamina propria, and some had morphological features of macrophages. Ultrastructurally, bacteria were observed intracellularly in the lamina propria as well as within apoptotic bodies. This study provides morphological evidence of Salmonella-induced cell death in vivo in a relevant nonhuman primate model.     

Cellular immunologic studies of malignant lymphoma in rhesus macaques.

Cells from malignant lymphoma in 10 rhesus macaques were examined for lymphocyte surface markers. Three had features of T cells, 5 had features of B cells, and 2 lacked evidence of either B- or T-cell differentiation. Correlation between the histologic classification of cell type and the B- or T-cell nature of the neoplasms was not evident. Evaluation of serum electrophoresis, mitogen responses tests, and previous histologic studies suggest that the development of the neoplastic lymphocyte proliferation occurred following or during an abnormal immunologic response.     

Nodular Pneumocystis carinii pneumonia in SIV-infected macaques.

Pneumocystis carinii (PC) pneumonia is a frequent manifestation of the acquired immunodeficiency syndrome (AIDS) in humans and macaques. An unusual nodular type of PC pneumonia was observed in two simian immunodeficiency virus (SIV)-inoculated rhesus macaques (Macaca mulatta). These animals developed clinical signs of simian AIDS, including anorexia, weight loss, dyspnea, and collapse. Grossly, both animals had multifocal tan-white nodules 1-10 mm in diameter scattered throughout the lungs. One animal had similar nodules involving the diaphragm and thoracic wall. The lungs were characterized by severe PC pneumonia with numerous large nodules consisting of foamy material that compressed adjacent tissue. The nodules had central areas of necrosis and lysis of alveolar septa. Varying degrees of necrotizing vasculitis were observed in areas of nodular PC pneumonia. The presence of PC in intra-alveolar spaces and nodular lesions was confirmed by immunohistochemistry. No evidence of other agents, including viral inclusions, bacteria, fungi, and lung mites, was detected. The animal with the most severe nodular PC pneumonia had vascular involvement with extrapulmonary spread to the diaphragm, thoracic wall, and regional lymph nodes. This unusual type of nodular PC pneumonia has been rarely seen in human AIDS patients.     

CYP1B1 is polymorphic in cynomolgus and rhesus macaques

Cytochrome P450 (CYP) 1B1 is involved in the metabolic activation of various procarcinogens, and some CYP1B1 genetic variants alter CYP1B1-dependent procarcinogen metabolism. Cynomolgus and rhesus macaques are frequently used in toxicity tests due to their evolutionary closeness to humans. In this study, we attempted to identify CYP1B1 genetic variants in 13 cynomolgus and 4 rhesus macaques. A total of 17 genetic variants were identified, including 8 non-synonymous genetic variants, indicating that, similar to humans, CYP1B1 is polymorphic in macaques. These CYP1B1 genetic variants could be the basis for understanding potential inter-animal differences in macaque CYP1B1-dependent metabolism of promutagens.     

Pathogenesis of simian AIDS in rhesus macaques inoculated with the SRV-1 strain of type D retrovirus

Type D retrovirus was isolated from rhesus macaques with simian acquired immunodeficiency syndrome (SAIDS) and transmitted to healthy rhesus macaques with tissue culture medium containing the virus. The clinical, immunologic, and lymph node morphologic changes were observed in 9 rhesus macaques for 52 weeks after inoculation. A spectrum of clinical signs developed including early death, persistent SAIDS, and apparent remission. Animals that died or developed persistent SAIDS had characteristic lymphoid depletion, persistently depressed peripheral blood mononuclear cell (PBMC) mitogenic response, and decreased serum immunoglobulins. The SAIDS retrovirus (SRV) was recovered from PBMC of 8 of the animals after inoculation. Virus could not be recovered from PBMC of one animal in remission, but this animal developed serum-neutralizing antibodies to SRV after inoculation. Seven of the animals seroconverted to SRV after inoculation, all 9 were seronegative for human T-lymphotropic virus-III, and 5 animals tested were seronegative to human T-lymphotropic virus-I. These findings support the etiologic role of the type D retrovirus in SAIDS and further define the pathogenesis of this disease.     

Pulmonary cryptosporidiosis in simian immunodeficiency virus-infected rhesus macaques

Cryptosporidiosis is a common opportunistic infection in the gastrointestinal tract of human and nonhuman primates with AIDS. Pulmonary infection associated with Cryptosporidium spp. has not been previously reported in monkeys. Two macaques experimentally infected with simian immunodeficiency virus (SIV) had lesions containing cryptosporidial organisms involving the trachea, lungs, bile ducts, pancreas, and intestine. The pulmonary sections revealed moderate to severe bronchopneumonia associated with cryptosporidiosis. Numerous 2-4 microm oval Cryptosporidium spp. organisms were present in the cytoplasm of alveolar macrophages, multinucleated giant cells, and intestinal epithelial cells. Giant cells were positive for SIV by in situ hybridization. These are the first reported cases of cryptosporidiosis with involvement of pulmonary parenchyma in SIV-infected macaques.     

A case of intestinal Mycobacterium simiae infection in an SIV-infected immunosuppressed rhesus monkey.

Although Mycobacterium simiae was identified and classified more than three decades ago, only a few cases are mentioned in the current literature. After experimental simian immunodeficiency virus infection, a 9-year-old female rhesus monkey (Macaca mulatta) developed progressive immunosuppression and gastrointestinal disease very similar to the clinical and pathomorphologic features of Johne's disease, which is caused by M. paratuberculosis. Acid-fast-positive bacteria reacted immunohistochemically with antibodies against M. paratuberculosis and M. bovis but were not useful for differentiation because of a high degree of cross-reactivity. In contrast to immunohistochemistry and histopathology, biochemical methods and cycle sequencing analysis of the 16S ribosomal RNA identified M. simiae as the disease-causing pathogen. This case demonstrates the importance of molecular biological methods for the diagnosis of M. simiae infection in monkeys.     

Feline herpesvirus 1-associated facial and nasal dermatitis and stomatitis in domestic cats.

Feline herpesvirus-associated dermatitis has rarely been reported. Recently we documented a unique ulcerative and often persistent facial dermatitis or stomatitis syndrome associated with feline herpesvirus 1. We believe this syndrome is relatively common, with the 10 cases in our series diagnosed between 1996 and 1997. The syndrome is associated with epithelial cell necrosis, eosinophilic inflammation, and intraepithelial herpesvirus inclusion bodies. The prevalence of eosinophilic inflammation and low number of inclusion bodies may lead to the misdiagnosis of allergic dermatitis or a lesion within the eosinophilic granuloma complex group of disorders. Feline herpesvirus 1 can be identified in lesional tissue by PCR methodology. Most of our cases developed under circumstances suggesting reactivation of latent herpesvirus infection, and previous glucocorticoid therapy or stress from overcrowding may have played a role in lesion development. Cats with ulcerative dermatitis, especially of the face and nose, and cats with stomatitis should be evaluated for the presence of feline herpesvirus. Treatment options include surgical excision, topical or systemic antibiotic therapy to treat secondary bacterial infection, and oral alpha interferon.     

Lymphocyte subpopulations in spontaneous disease of rhesus macaques.

The immunologic status of rhesus macaques (Macaca mulatta) with naturally occurring disease was evaluated by determining the percentages of B and T lymphocytes and mitogen responsiveness of lymphocytes in peripheral blood and lymph nodes. The B lymphocytes were identified by the presence of cell surface immunoglobulin and receptors for complement. The T lymphocytes were identified by their ability to form spontaneous rosettes with sheep red blood cells. Rhesus macaques with idiopathic or primary amyloidosis had normal lymphocyte characteristics. Peripheral blood lymphocytes from rhesus macaques with atypical tuberculosis had decreased percentages of spontaneous rosette-forming cells and depressed responses to concanavalin A, and those with chronic diarrhea or chronic arthritis were also found to have abnormal peripheral blood lymphocyte characteristics. The percentage of B and T lymphocytes in normal lymph nodes was variable, making simultaneous histologic examination necessary for evaluation of diseased animals.     

Neutrophil chemotaxis in rhesus macaques (Macaca mulatta)

Chemotactic responses of isolated peripheral blood neutrophils from rhesus macaques (Macaca mulatta) were studied, using a micropore filter method. Cell migration toward zymosan-activated serum was similar to that of human cells, whereas the response to N-formyl methionyl leucyl phenylalanine (FMLP) was weaker than was that in human cells, requiring higher concentrations of FMLP for maximal migration. Optimal FMLP concentrations for attraction of rhesus neutrophils and human neutrophils were 5.0 X 10(-7)M and 1.0 X 10(-8)M, respectively. The chemotactic responses of the 2 neutrophils to complement (zymosan-activated serum) were similar. However, rhesus neutrophils required a higher concentration of the formyl peptide, FMLP, for maximal migratory response.     

Pulmonary functions in conscious and anesthetized rhesus macaques.

Techniques for measurements of selected pulmonary functions for a period of 6 hours in conscious and lightly anesthetized rhesus macaques are described. Among all variables measured, only higher PaCO2 (367 +/- 32 mm of Hg) occurred in macaques breathing O2 as compared with those breathing room air. Tidal volume, respiratory rate, minute volume, O2 consumption, and specific ventilation in conscious and anesthetized monkeys were not significantly different. Further, respiratory and metabolic values from lightly anesthetized macaques in the present study agree well with the reports of others. However, differences were the higher values for O2 consumption, tidal volume, respiratory rate, and minute volume obtained with the presently described techniques as compared with certain reported data.     

Progression of an orbital T-cell rich B-cell lymphoma to a B-cell lymphoma in a dog

An 11-year-old Shetland Sheepdog was presented for exophthalmos caused by a locally extensive, poorly defined mass located behind the right eye. The primary orbital mass was identified by light microscopy and immunohistochemistry as a T-cell rich B-cell lymphoma (TCRBCL) composed predominantly of BLA.36-positive large neoplastic lymphoid cells admixed with fewer CD3- and CD79a-positive small lymphocytes. The dog was treated for lymphoma, but 6 months after presentation it was euthanatized for suspected hepatic and gastrointestinal metastasis. Gross findings revealed an enlarged liver with multiple well-demarcated, randomly distributed 0.1-1.5-cm white nodules, five firm white submucosal jejunal nodules, and ileocecal, mediastinal, and hilar lymphadenopathy. Metastatic liver lesions consisted of sheets of monomorphic large neoplastic lymphoid cells that effaced and expanded portal and centrilobular zones. These cells were morphologically similar to the large neoplastic cells of the original orbital tumor and were CD3-negative and variably BLA.36-positive, consistent with B-cell lineage. Similar cells comprised the jejunal nodules and effaced the lymph nodes. The progression of TCRBCL to a diffuse B-cell lymphoma in this case is consistent with reported human cases and has not been previously reported in the dog.     

B-cell conjunctival lymphoma in a cat

A bilateral conjunctival neoplasm was surgically removed from a 13-year-old Domestic Short-haired female cat and examined by histopathology. The neoplasm was infiltrative, nonencapsulated and composed of large sheets of densely packed, round to polygonal cells. Neoplastic cells were variably sized with scant to modest amounts of cytoplasm and oval to round nuclei. On immunohistochemistry the neoplastic cells stained positively for BLA.36 and negative for CD-3 antibodies. On the basis of histopathology and immunohistochemical findings, a diagnosis of conjunctival B-cell lymphoma was made. This is the first case with immunohistochemical characterization of conjunctival lymphoma in a cat.     

实验性猕猴1型糖尿病模型的建立及评价

7～10岁健康猕猴15只,随机分为Ⅰ、Ⅱ、Ⅲ组,每组5只,分别按照100、125、150mg/kg的剂量一次性静脉注射链脲菌素,连续观察血糖、血浆C肽和胰岛素浓度的动态变化,观察期15周.结果:与注射链脲菌素前比较,注射后1周,Ⅰ组猕猴血糖浓度明显升高,血浆C肽和胰岛素浓度显著下降(P＜0.01),随后趋于正常水平,而Ⅱ、Ⅲ组猕猴血糖浓度明显升高并在随后的时间内持续维持在高水平,血浆C肽和胰岛素浓度持续维持在低水平(P＜0.001),Ⅲ组动物至15周全部死亡.结论:按照125mg/kg的剂量一次性给猕猴静脉注射链脲菌素,可成功复制1型糖尿病动物模型,其维持高血糖和低胰岛素及C肽浓度至少15周.     

Distribution of tissue water and electrolytes in normal rhesus macaques.

Techniques for the determination of water and electrolytes in individual tissues of normal rhesus macaques (Macaca mulatta) are described. Base-line values are presented for intracellular and extracellular distributions of water and electrolytes in 14 tissues, including skin, skeletal muscle (gastrocnemius), cardiac muscle (left ventricle), lung, diaphragm, liver, renal cortex, outer medulla and inner medulla, cerebral cortex, cerebellum, thalamus-hypothalamus complex, medulla oblongata, and spinal cord.     

Cutaneous T-cell-rich B-cell lymphoma in a horse

Cutaneous malignant lymphomas are rare in horses and comprise predominantly T-cell-rich B-cell lymphomas. They are characterized by multiple tumour nodules affecting predominantly female horses with a survival rate of months to years. At the final stage, metastases to regional lymph nodes occur, whereas widespread organ involvement is rarely reported. In this case report, a cutaneous T-cell-rich B-cell lymphoma in a 7-year-old standardbred gelding with metastases is described. Clinically, multiple cutaneous and subcutaneous nodules, enlarged superficial lymph nodes, rapid weight loss, and ventral oedema were observed. In addition to the clinical findings, necropsy revealed tumour infiltration in multiple body lymph nodes, a solitary pleural mass, and few pulmonary and intestinal tumour nodules. Microscopically, all neoplasms were composed of a densely packed cell population consisting of large lymphoblastic cells expressing CD79a, and numerous small, round, CD3-positive T lymphocytes. With respect to these findings the diagnosis of a cutaneous T-cell-rich B-cell lymphoma with metastases was made.     

Neurolymphomatosis in a dog with B-cell lymphoma

Lymphoma in the left femoral nerve of a 10-year-old English Cocker Spaniel caused complete paralysis of the affected limb. Neoplastic cells were immunopositive for CD79a and Pax5 and negative for CD3. Neoplastic cells were in multiple lymph nodes and one kidney but spared bone marrow. The clinical and histologic features in this case resemble those of the rare human condition of neurolymphomatosis.     

Gastric stromal tumors in two rhesus macaques (Macaca mulatta)

Two female rhesus monkeys (Macaca mulatta, 10 and 24 years old) developed microcytic anemia and became terminally ill. At necropsy, large gastric masses were present, and, in one case, there were widespread abdominal metastases. Except for slightly atypical patterns, at the light microscopic level, the lesions resembled smooth muscle tumors. Ultrastructurally, however, cells in both tumors resembled primitive mesenchyme, while in one of the tumors, there were some characteristics of Schwann cells. No ultrastructural features of smooth muscle were present in either tumor. Vimentin and S-100 were detected immunohistochemically in both tumors. S-100 staining was more intense in the tumor with ultrastructural features of Schwann cells. Actin and desmin were not expressed in either gastric tumor, but diffusely stained a uterine tumor that was concomitantly present in one of the rhesus monkeys. The uterine tumor also exhibited typical ultrastructural features of smooth muscle. In the past, gastrointestinal stromal tumors in all species were thought to be of smooth muscle origin. Recently in human pathology, this conventional viewpoint has given way to the realization that there is a spectrum of neural crest and mesenchymal tumors. We report two gastric stromal tumors in two rhesus monkeys that histologically resembled smooth muscle tumors but were of neuroectodermal and primitive mesenchymal origin.     

Effect of staphylococcal enterotoxin B on cardiorenal functions in rhesus macaques.

Selected cardiovascular and renal functions were measured for 5 hours in conscious, chair-restrained, female rhesus macaques (Macaca mulatta) after IV (0.05 and 1.0 mg/kg) or oral (1.0 mg/kg) administration of staphylococcal enterotoxin B (SEB). Cardiovascular functions, renal hemodynamics, and renal metabolism were also studied between 6 and 11 hours after IV SEB inoculation. Oral SEB produced few changes in cardiorenal functions. In contrast, IV SEB produced hypotension, tachycardia, increased total peripheral and renal vascular resistance, and decreased cardiac and renal functions. The early significant (P less than 0.05) renal depression was not associated with hypotension (mean arterial blood pressure greater than 100 mm of Hg). However, all measured renal functions except extraction ratio of PAH were positively correlated with decreased blood pressure (r = 0.52 - 0.71) in the later phase of SEB toxemia. It is concluded that the kidney is one of the organs affected by IV SEB. Renal impairment may partially contribute to death during SEB enterotoxemia in macaques.