Engraftment of human synovium into severe combined immune deficient mice. Migration of human peripheral blood T cells to engrafted human synovium and to mouse lymph nodes.

To determine the feasibility of using the C.B-17 scid/scid (severe combined immune deficient, SCID) mouse as a recipient of human synovial xenografts, we have engrafted human synovium under the renal capsule of SCID mice, and determined synovial graft survival and histologic characteristics 4 to 7 wk after tissue implantation. Both normal and inflammatory synovial tissue grew well in SCID mice and maintained histologic and phenotypic components of the fresh synovial tissue before implantation. However, the number of T cells in synovial grafts decreased after implantation. To determine whether leukocytes could migrate to human synovial xenografts, either allogenic or autologous PBMC were injected in the peritoneum of SCID mice bearing synovial xenografts. We found that 7 days after i.p. injection of autologous or allogeneic PBMC, injected T cells had selectively migrated to human synovial grafts and to SCID mouse lymph nodes. Our data demonstrate that normal and inflammatory human synovial tissues will grow in SCID mice and serve as recipients for autologous and allogenic peripheral blood human T cells injected i.p. into engrafted mice.

Experimental murine cytomegalovirus infection in severe combined immunodeficient mice.

Severe combined immunodeficient (scid) and BALB/c mice were experimentally infected with murine cytomegalovirus (MCMV). Scid mice infected by the intraperitoneal route died or were moribund at dose-dependent times ranging from 12 to 13 days after inoculation for > or = 3.00 x 10(4) plaque forming units (pfu) of virus to 25 days for 1.17 x 10(2) pfu. Histologic lesions included severe adrenal necrosis at low doses and splenic necrosis at high doses. Multinucleate hepatocytes with multiple inclusion bodies were observed at all doses. In visceral organs, the inflammatory response consisted of cell necrosis and neutrophil infiltration. Scid mice infected with 1.00 x 10(3) pfu by the intranasal route were moribund by 24 or 25 days after inoculation. Viral titers in tissues examined from these mice increased in all organs examined until they became moribund. BALB/c mice infected intranasally had detectable virus titers in the adrenal glands, salivary glands, lungs, and spleen by 7 or 14 days after inoculation, but decreased thereafter. These mice remained clinically normal through the infection. In BALB/c mice, histologic lesions were present only in the salivary glands.

Successful engraftment of human postnatal thymus in severe combined immune deficient (SCID) mice: differential engraftment of thymic components with irradiation versus anti-asialo GM-1 immunosuppressive regimens.

To develop a model of human thymus growth in vivo, we have implanted postnatal human thymus under the renal capsule of severe combined immune deficient (SCID) mice and assayed for graft survival and graft characteristics 1-3 mo after engraftment. Three groups of SCID mice were engrafted with postnatal human thymus: untreated SCID mice, SCID mice pretreated with 400 cGy of gamma irradiation 1-5 d before engraftment, and SCID mice treated with intraperitoneal anti-asialo GM-1 antiserum every 4-5 d during engraftment. In the untreated group of SCID mice, only 37% of grafts survived and consisted of human thymic microenvironment components and human immature thymocytes. Irradiation of SCID mice before engraftment improved survival of human thymic grafts to 83%, but these grafts were largely devoid of thymocytes and contained only thymic microenvironment components with large numbers of thymic macrophages. Treatment of SCID mice with anti-asialo GM-1 antiserum throughout the engraftment period also promoted human thymus engraftment (70%) and induced SCID B cell Ig production (SCID[Ig+]) in 38% of animals. In SCID(Ig-) anti-asialo GM-1-treated mice, the human thymic grafts were similar in content to those in untreated SCID mice. However, in anti-asialo GM-1-treated animals with grafts that became SCID(Ig+), all animals were found to have mouse-human chimeric grafts in that the human thymic microenvironment (human fibroblasts, thymic epithelium, vessels) was colonized by murine T cells. These data demonstrate that human postnatal thymus will grow as xenografts in SCID mice, and that the components of human thymus that engraft are dependent on the immunosuppressive regimen used in recipient mice. A striking finding in this study was the induction of T and B lymphopoiesis in SCID mice by abrogation of NK cell activity with in vivo anti-asialo GM-1 treatment. These data strongly suggest that asialo GM-1+ NK cells and/or macrophages play a role in mediation of suppression of lymphopoiesis in SCID mice.

Infectious diseases of the upper respiratory tract: implications for toxicology studies.

The consequences of adventitious infectious agents upon the interpretation of toxicology studies performed in rats and mice are incompletely understood. Several prevalent murine pathogens cause alterations of the respiratory system that can confuse the assessment of chemically induced airway injury. In some instances the pathogenesis of infection with these agents has been relatively well studied in the lower respiratory tract. However, there are few well-controlled studies that have examined the upper respiratory region, which result in interpretive problems for toxicologic pathologists. The conduct and interpretation of both short-term and chronic rodent bioassays can be compromised by both the clinical and subclinical manifestations of infectious diseases. This paper reviews several important infectious diseases of the upper airway of rats and mice and discusses the potential influence of these conditions on the results of toxicology studies.

Whole-cell protein patterns of nonhemolytic group B, type Ib, streptococci isolated from humans, mice, cattle, frogs, and fish.

Whole-cell protein and physiological patterns of nonhemolytic group B, type Ib, streptococci isolated from humans, cattle, frogs, fish, and mice were compared. Isolates from humans, fish, and mice were identical. Only minor differences were seen in the isolates from human, bovine, and frog sources. Nonhemolytic group B streptococci from humans, fish, and mice and, to a lesser extent, from cattle and frogs share several characteristics, including a high similarity in proteins (on the basis of molecular weight); this suggests that they may have a common ancestry.

Fatal pneumonia with terminal emaciation in nude mice caused by pneumonia virus of mice.

Athymic nude mice used as sentinel animals in a mouse holding room died of pneumonia 17 to 32 weeks after being placed in the room. Lesions in the pulmonary parenchyma consisted of monocytic exudate, epithelial cell necrosis, hemorrhage, fibrin deposition and interstitial fibrosis. Septal edema, septal cell necrosis and septal capillary stasis were common, but there was limited sloughing of bronchial lining epithelium. Indirect fluorescence microscopy (IFA) of lung sections using pneumonia virus of mice (PVM) antibody was positive. The pneumonia and IFA results were reproduced in euthymic mice inoculated experimentally with lung suspension from naturally infected mice or with tissue culture fluid from cultures infected with American Type Culture Collection PVM. The lungs of a naturally infected nude mouse were studied by transmission electron microscopy. Virus growth was found on Type II alveolar epithelium and on poorly differentiated replacement alveolar epithelium. Virus particles appeared as long exophytic filaments containing one to six linearly arranged nucleocapsids. Inclusion bodies and intracellular virus structures were not observed.

The mouse bioassay for the detection of estrogenic activity in rodent diets: I. A standardized method for conducting the mouse bioassay.

A standardized procedure was developed for conducting the mouse bioassay for detecting estrogenic activity in rodent diets. Studies were conducted with CD-1 mice to determine the appropriate weaning age and length of bioassay period. Uterine growth curves were generated from mice weaned at 15 days of age and fed a negative control diet until 28 days of age. These mice showed slow regular increases in uterine weights from 15 22 days of age followed by rapid uterine growth in some mice from 24 to 28 days of age. Estrogenic bioassays using female mice weaned at 15 days of age and fed the positive control diets containing 4 or 6 ppb diethylstilbestrol (DES) demonstrated significant (P less than 0.05) increases in uterine weight and in uterus to body weight (U:BW) ratios over those of mice fed the negative control diet without DES for 5, 7 or 9 days after weaning. In contrast, mice weaned at 17 days of age showed significant (P less than 0.05) increases in uterine weight and in U:BW ratios only at 5 days after weaning. Six ppb DES was required in the positive control diet to produce a 1.5 fold increase in the U:BW ratio over those of mice fed the negative control diet. It was concluded that mice should be weaned at 15 days of age and that the bioassay period should be terminated at 7 days, when the mice are 22 days old, for best reproducible results. The criteria for a valid bioassay should include the demonstration of a significant statistical increase in the U:BW ratios of mice fed the DES positive diet over those of mice fed the negative control diet.

The mouse bioassay for the detection of estrogenic activity in rodent diets: II. Comparative estrogenic activity of purified, certified and standard open and closed formula rodent diets.

A major source of exogenous estrogenic substances, which may affect laboratory animals, comes from the diet. To test the possibility that commercially available rodent diets may significantly influence uterine weights and uterine:body weight (U:BW) ratios, estrogen bioassays were performed using female CD-1 mice weaned at 15 days of age and assigned randomly to a variety of commercial test diets or to a control diet (Purina #5002) containing 0 or 6 ppb added diethylstilbestrol (DES) for comparison. Mice were housed five per cage and given deionized water and feed ad libitum. Uterine:BW ratios from 15 mice per diet were determined after 3, 5 and 7 days of feeding. Mice fed The American Institute of Nutrition purified diet (AIN-76A) or the Purina #5015 natural ingredient breeder diet had significantly (P less than 0.05) increased U:BW ratios at 3, 5 and 7 days post weaning when compared to the control diet without added DES. This increase in U:BW ratios was similar to the U:BW ratios observed in a natural ingredient maintenance diet (Purina #5002), containing 6 ppb of DES. These results show that significant differences exist in the level of substances which can cause increase in uterine weight in some commercial diets. The diet may be important when performing or comparing certain types of studies, especially those relating to estrogenic substances. A standardized diet with minimal estrogenic activity may be desirable for such studies. It is unclear from the present studies what substances might be responsible for the uterine growth promoting activity in the diets examined.

The mouse bioassay for the detection of estrogenic activity in rodent diets: III. Stimulation of uterine weight by dextrose, sucrose and corn starch.

We have shown previously that mice fed the American Institute of Nutrition (AIN-76A) purified diet experience a significant increase in uterine:body weight (U:BW) ratios when compared to the U:BW ratios of mice fed a closed formula natural ingredient diet (Certified Rodent Chow #5002) for 7 days. The AIN-76A purified diet contains 5% corn oil and 65% carbohydrates with 50% of the carbohydrates coming from sucrose or dextrose and 15% from corn starch. The objective of this study was to determine whether the fat and carbohydrate content contributed to the unexpected uterine growth promoting activity observed in mice fed the AIN-76A diet. Estrogen bioassays were performed using CD-1 mice weaned at 15 days of age and assigned randomly to the negative control diet (Certified Rodent Chow #5002) or to the positive control diet (#5002) containing 4 or 6 ppb DES for comparison or to the test diets. The test diets were prepared by adding sucrose, dextrose, corn starch, corn oil or soybean oil to the #5002 negative control diet at 10% w/w concentration. Uterine:BW ratios were determined at 7 days post-feeding. The uterine weights and the U:BW ratios of mice fed the test diets containing dextrose, corn starch, or corn oil, were increased significantly (P less than 0.05) over those of mice fed the negative control diet. The uterine weights and U:BW ratios of mice fed the test diets containing sucrose or soybean oil also were increased over those of mice fed the negative control diet. These increases in uterine weights and U:BW ratios were similar to the increases in uterine weights and U:BW ratios of mice fed the positive control diet containing 4 ppb DES. It was concluded that the fats and carbohydrates caused preferential increases in uterine weights and in U:BW ratios and may account for the estrogen-like uterine growth promoting activity observed in mice fed the AIN-76A purified diet.

Reproductive performance of three species of Callitrichidae.

The reproductive performance of three species of Callitrichidae housed under identical conditions, including relatively spacious caging and visual screening, was compared. The three species were Callithrix jacchus, Saguinus fuscicollis, and Saguinus oedipus. Callithrix jacchus was the most reliable breeder, with the lowest percentage of stillborn and the highest post-weaning survival. While the reproductive performance of S oedipus was better than that generally reported for other colonies, the reproductive performance of the Saguinus species was still poorer than that of C jacchus. The two Saguinus species did not differ significantly in either percentage of stillborn or post-weaning survival. None of the species showed infertility due to lactation, but the mean inter-birth interval was significantly longer for S oedipus than for either C jacchus or S fuscicollis.

Analysis of mutagen-induced chromosome damage in a primate species (Saguinus oedipus oedipus) at risk for spontaneous adenocarcinoma of the colon.

A high incidence of adenocarcinoma of the colon (greater than 16%) has been observed at necropsy in the South American primate, Saguinus oedipus oedipus (S. oedipus), while the disease has not been found in tamarins of the closely related species, Saguinus fuscicollis spp, housed in the same research colony. Cytogenetic analyses in cultured lymphocytes from 10 S. oedipus and 10 S. fuscicollis illigeri (S. fuscicollis) demonstrated no differences in the average frequencies of spontaneous or mitomycin C (MMC)-induced sister chromatid exchanges (SCEs) between animals of the two species. However, highly significant variability in MMC-induced chromosome lesions was observed between the individual S. oedipus, with one animal exhibiting increased sensitivity for both SCEs and chromosome breakages. At present we do not know the relationship, if any, between increased sensitivity to mutagen-induced cytogenetic lesions in specific S. oedipus tamarins and the increased risk for colon cancer that has been documented in this primate species. However, our cytogenetic findings in this one S. oedipus are similar to data obtained in evaluations of persons with several autosomal recessive conditions in which there is a genetic predisposition for developing malignancies.

Competition for a desired food in family groups of the common marmoset (Callithrix jacchus) and the cotton-top tamarin (Saguinus oedipus).

Four captive groups of common marmosets (Callithrix jacchus) and three groups of cotton-top tamarins (Saguinus oedipus) were each presented with a desired food and their consumption was monitored. Inequalities in consumption between individuals were found for each group of each species. Generally, adult females consumed most of the desired food, followed by juveniles, and finally adult males. High consumption levels by all individuals during isolated feedings indicated that consumption inequalities within groups were most likely due to competition rather than inter-individual preference differences. Aggressive defense of the food cup was performed most frequently by adult females in both species. In Callithrix jacchus groups, this aggression was most often directed against adult males, but in Saguinus oedipus groups, it was most often directed against juveniles. Modification of the method of food presentation, such that food was more dispersed, lessened, but did not eliminate, inequalities in consumption.

Mucous and argentaffin cells in colonic adenocarcinomas of tamarins and rats.

Primary and metastatic neoplasms of four naturally occurring colonic adenocarcinomas in the cotton-topped tamarin, Saguinus oedipus oedipus, and four chemically induced rat colonic adenocarcinomas were examined with light and electron microscopy. In each case, we observed mixtures of mucous and argentaffin cells in metastatic sites, proving that endocrine cells are an integral part of these tumors. The occurrence of mixed cell populations in metastases in both species suggests, but does not prove, that colonic tumor cells have the capacity for multi-directional differentiation and that neoplastic mucous and argentaffin cells in the colon have a common origin. We also observed mitotic figures in neoplastic argentaffin cells, confirming the observations of others that these cells can label with tritiated thymidine and divide.

Prevalence of ocular disease in a colony of tamarins and marmosets.

In a large tamarin and marmoset experimental colony, 526 animals were examined by biomicroscopy and ophthalmoscopy for the presence of ocular disease. In 109 animals, there were 147 abnormalities involving the eyelids, cornea, iris, lens, retina or optic nerve. Most abnormalities were unimportant in terms of ocular function, but a few did cause loss of vision and included diffuse, progressive retinal atrophy and a severely traumatized globe. The survey indicated that while usually minor, ocular disease in the tamarin and marmoset was widespread.

Therapy of filariasis in Tamarins.

A regimen of thiacetarsamide sodium (0.22 ml/kg twice daily for 2 days) plus levamisole phosphate (11 mg/kg/day for 10 days) was effective in eliminating unidentified microfilariae from the blood in seven of eight tamarins. No serious side effects resulted from the treatment. All of the animals were initially freed of circulating microfilariae after treatment, and five have remained microfilaria-negative for 1 year. Two of the tamarins died of causes unrelated to filariasis and were microfilaria-negative before death. One tamarin remained microfilaria-positive after two courses of this treatment.