Xenotransplantation of cryopreserved equine squamous cell carcinoma to athymic nude and SCID mice

Cryopreserved equine ocular squamous cell carcinoma (SCC) was inoculated subcutaneously into 15 athymic nude and 15 SCID mice. Xenotransplantation resulted in tumor growth in two athymic nude mice and 1 SCID mouse. Histological appearance and immunohistochemical characterization using cytokeratin 5/6 markers and p53 markers of the tumor grown in mice was in full accord with the original equine tumors. No evidence of metastasis was noted in any mouse. This model may serve as a relevant in vivo model for studying the biology of equine ocular SCC and for the testing of new therapeutic modalities.     

Heterotransplantation of equine carcinoma cells in athymic (nude) mice

Nondifferentiated equine carcinoma cells from a primary lesion were implanted subcutaneously in athymic (nude) mice. The cells were implanted at 2 sites each in 3 mice. At 1 of the 6 inoculation sites, a tumor developed, which invaded surrounding tissues, as shown by histopathologic examination. Karyotype analysis verified that the tumor was of equine origin. Cells from this tumor were serially heterotransplanted 20 times without change in growth rate. Once established in nude mice, this equine carcinoma cell line was stored cryogenically and then was successfully reimplanted into nude mice. All of the implants developed into tumors, over 20 generations. Preliminary screening of antineoplastic drugs indicated that this tumor line is sensitive to cyclophosphamide. Because of its ease of handling and high reimplantation efficiency, this tumor line should prove useful in equine cancer research.     

A review of equine mucocutaneous squamous cell carcinoma

Squamous cell carcinoma (SCC) accounts for approximately 20% of all equine mucocutaneous (MC) tumours and continues to present a therapeutic challenge to practitioners. Most MC‐SCC are locally invasive and slow to metastasise, but metastasis to local lymph nodes is not uncommon. The most common location for MC‐SCC is the periorbital region, with the eyelid most commonly affected. Although only 13% of MC‐SCC involves the external genitalia, MC‐SCC is the most common neoplasm of male genitalia. Equine caballus papillomavirus‐2 has recently been linked to MC‐SCC and may prove to be necessary for tumour development. Risk factors may include chronic exposure to ultraviolet light and chronic skin irritation. Horses developing genital MC‐SCC tend to be older compared to those with periorbital MC‐SCC. Histopathology is required for definitive diagnosis of MC‐SCC, although horse phenotype and lesion location may suggest MC‐SCC. Several treatment modalities have been successful in eliminating or managing MC‐SCC, with surgical excision and intratumoural chemotherapy yielding the best results. Other treatment options including cryotherapy, hyperthermia, radiotherapy and photodynamic therapy are often used as adjunctive therapies. Early recognition of tumours and prompt intervention are associated with a positive outcome.     

Naturally-occurring Sendai virus infection of athymic nude mice.

Nude (nu/nu) mice, Balb/c-derived, responded to a naturally-occurring Sendai virus infection in a different manner than conventional mice. They developed a chronic debilitating disease and a persistent viral infection of the respiratory tract with intranuclear inclusion bodies in tracheal, bronchial and bronchiolar epithelial cells, laryngeal and tracheal glandular epithelium and in type I and II alveolar cells. The infection was identified by serologic and tissue culture studies, the mouse antibody production test and ultrastructural examination of pulmonary lesions. Phlebitis of pulmonary veins, suppurative rhinitis and otitis media accompanied the viral infection while some mice developed a secondary bronchopneumonia.     

Factors influencing morbidity and outcome of equine ocular squamous cell carcinoma

A retrospective review of 41 cases of equine ocular squamous cell carcinoma with follow up information for 33 horses is presented. An apparent prevalence in heavy horse breeds was identified. Age and sex distribution were similar to those previously reported. High recurrence (42.4 per cent) and low metastasis (6 per cent) rates were seen. Initial tumour location, primary treatment used, or patient case history had no apparent influence on final outcome. The willingness of owners to pursue with continuous treatment correlated with a high degree of success.     

Challenges in the treatment of equine periocular squamous cell carcinoma

Management of periocular squamous cell carcinoma is challenging because of the need for adjunctive therapy, the adverse effects of therapies and the frequent recurrence of SCC. Appropriate treatment of equine ocular SCC usually involves surgical excision combined with adjunctive therapy selected as appropriate for the anatomic site of the lesion. Metastasis of periocular SCC has been reported to occur in approximately 6–15% of cases. The horse owner should be carefully educated to understand that for best long‐term results from the treatment of periocular SCC, they must be diligent in observing signs of recurrence or metastasis and be willing to have the horse examined as soon as adverse signs are observed. Furthermore, external beam radiation may be effective in the treatment of metastatic SCC and further clinical studies of this treatment modality are needed.     

The effect of tamoxifen on tumours induced by cells from a canine mammary carcinoma cell line in athymic nude mice

Tumours were induced in CBA athymic nude mice by subcutaneous injection of REM 134 cells. These cells were from a continuous line derived from a canine mammary carcinoma and have no detectable oestrogen receptors. In vitro, the anti-oestrogen tamoxifen was growth inhibitory at a concentration of 10(-6) M and adding 10(-8) M oestradiol-17 beta did not reverse this effect. The relative rate of growth of the tumours induced by the cultured cells was the same in male and female mice. Oral tamoxifen at a dose of 1 mg mouse-1 week-1 suppressed the early growth rate significantly and to the same extent in male and female mice; conversely subcutaneous tamoxifen at 2 mg mouse-1 week-1 had only a transient early effect. These results argue against tamoxifen acting solely as an antagonist for oestrogen.     

裸鼠过度角化症牛棒状杆菌的分离鉴定

2010年8月一批进口入境的裸鼠在隔离期间发生了疑似国外文献报道的过度角化性皮炎,为了确诊该病及鉴定该病的病原,本研究从患病裸鼠皮肤组织分离到一株革兰氏阳性杆菌。细菌纯化后经Biolog自动生物鉴定仪系统初步鉴定为牛棒状杆菌(C.bovis);提取细菌基因组,PCR扩增16S rRNA基因,测序结果经比对与GenBank中的C.bovis同源性在99%以上;培养物感染裸鼠没有出现临床症状,但组织学检查显示,感染裸鼠表皮棘皮层增厚,表现出轻微的C.bovis感染特有的棘皮症。由此可以确认这次入境裸鼠暴发的疫病为过度角化症,病原为C.bovis。本研究首次报道了在我国实验动物设施暴发的裸鼠过度角化症,提示裸鼠感染C.bovis导致的过度角化症是实验动物质量监测中不可忽视的传染病。     

Topical 5-fluorouracil as an adjunct treatment in equine corneolimbal squamous cell carcinoma

An 11-year-old crossbreed mare was presented with a nodular, pink mass at the temporal corneolimbal junction of the right eye. Histopathology confirmed the diagnosis of invasive squamous cell carcinoma. The mare underwent lamellar keratectomy and conjunctivectomy under general anaesthesia, followed by adjunctive topical chemotherapy with 1% 5-fluorouracil solution for 10 days post-operatively. Adverse effects included moderate conjunctival inflammation and exuberant granulation tissue formation at the surgical site, responding to surgical debulking and topical corticosteroid application. No mass regrowth was present at 12 months following discharge. This report describes the successful use of topical 1% 5-FU as an adjunctive treatment for equine ocular squamous cell carcinoma.     

Induction of virus-producing tumours in athymic nude mice by bovine papillomavirus type 4

Bovine papillomavirus type 4 (BPV-4), the causative agent of alimentary papillomatosis, has been used to infect, in vitro, fragments of palatine mucosa from late term bovine fetuses. These small explants were placed beneath the renal capsule of athymic nude mice where they grew to produce, at first, squamous epithelial cysts containing BPV-4 genomic DNA and, later, papillomas which were morphologically identical to those of cattle and which contained large amounts of replicating virus. The possible utility of this technique in assessing neutralising antibodies in vaccine development is discussed.     

The use of mitomycin C as an adjunctive treatment for equine ocular squamous cell carcinoma

OBJECTIVE: To assess the effectiveness of topical mitomycin C application as an alternative adjunctive therapy to CO2 laser ablation in the treatment of equine ocular squamous cell carcinoma. DESIGN: A retrospective clinical study of eight client owned horses in which 10 affected eyes were treated for ocular squamous cell carcinoma over a 17 month period. (March 2003 to August 2004). PROCEDURE: Each horse was given a general anaesthetic to allow CO2 laser ablation of the lesion(s). Mitomycin C at a concentration of 0.4 mg/mL was then applied intraoperatively to the affected areas for 1 or 5 minutes. Postoperatively a triple antibiotic eye ointment was applied to the eye twice daily and each horse was treated with systemic non-steroidal anti-inflammatory drugs for 7 days. RESULTS: Recurrence following treatment was noted in three eyes. Two of these were retreated and no further signs developed. The remaining eye was enucleated at the request of the owner. Overall 90% of treated eyes appeared free of tumour a minimum of 11 months post treatment. CONCLUSION: In this case series mitomycin C appeared to offer a valid alternative to other adjunctive therapies currently employed in the treatment of equine ocular squamous cell carcinoma.     

Influence of epidermal growth factor on growth of bovine mammary tissue in athymic nude mice

Bovine mammary tissue obtained from midpregnant Holstein heifers by surgical biopsy was transplanted subcutaneously to ovariectomized athymic nude mice (n = 5 heifers). After 3 weeks recovery, mice were either sham operated or sialoadenectomized (submandibular salivary glands removed). After an additional week, mice were injected with saline or 17 beta-estradiol + progesterone (1 microgram + 1 mg/day) for 2 days. In addition, half of the sialoadenectomized mice were injected with epidermal growth factor (5 micrograms/day). Grafted tissue was removed and rate of deoxyribonucleic acid (DNA) synthesis estimated by incorporation of 3H thymidine. Estradiol + progesterone increased the incorporation of 3H thymidine from 77 +/- 20 dpm/micrograms DNA to 472 +/- 53 dpm/micrograms DNA. In sialoadenectomized mice, DNA synthesis was increased from 88 +/- 16 dpm/micrograms DNA (saline treated) to 360 +/- 29 dpm/micrograms DNA (estradiol + progesterone treated). In sialoadenectomized mice treated with epidermal growth factor, DNA synthesis in estradiol + progesterone treated mice was 529 +/- 36 dpm/micrograms DNA, compared to 112 +/- 30 dpm/micrograms DNA in sialoadenectomized mice treated with epidermal growth factor. These data indicate that sialoadenectomy of athymic nude mice decreased the ability of transplanted bovine mammary tissue to increase DNA synthesis in response to estradiol and progesterone. This inhibition was removed by epidermal growth factor treatment. These data suggest a physiological role of epidermal growth factor in regulating development and hormone responsiveness of bovine mammary tissue.     

Heterotransplantation of canine sarcoma cells (BLU284) in athymic (nude) mice

Undifferentiated canine sarcoma cells from a primary lesion were implanted subcutaneously in athymic (nude) mice. Tumors were implanted at 2 sites in each of 4 mice, and 6 of the 8 inoculation sites developed into tumors. Tumors grew and invaded surrounding tissues, as shown by histologic examination. Karyotype analysis verified that tumors were of canine origin. Cells from this tumor were serially heterotransplanted 26 times without consistent change in growth rate. Once established in nude mice, samples from 2 generations of this canine sarcoma cell line were stored cryogenically and were implanted into nude mice. Over 26 generations, 93.7% of fresh implants developed into tumors. Preliminary screening of antineoplastic drugs indicated that this tumor line was sensitive to cyclophosphamide and vincristine. The difference in sensitivity of this heterotransplanted tumor in nude mice toward the various antineoplastic agents provides a useful model for the elucidation of biochemical bases of antineoplastic drug resistance in tumors.     

Host response to bovine ocular squamous cell carcinoma

Specific humoral and cellular immunologic responses to autologous and heterologous tumors were evaluated in 35 Hereford cows with ocular squamous cell tumors and in 6 healthy cows. Sera from 5 healthy cows and 23 ocular tumor-bearing cows were evaluated for antibody to tumor, using radioimmunoassay, passive hemagglutination, agglutination, and microagglutination assays with various soluble tumor antigen and whole tumor cell preparations. Antibody to tumor was detected in only 2 cows. Using the microagglutination assay, antibody to autologous and heterologous tumor cells was found in the sera of 2 cows inoculated intraocularly with purified, viable autologous tumor cells. Twenty-eight tumor-bearing cows and 3 healthy cows were evaluated for delayed-type hypersensitivity responses, using 5 tumor preparations; positive skin test responses were not observed.     

Squamous cell carcinoma of the equine stomach

Summary

Both clinical and pathological features of seven horses suffering from a squamous cell carcinoma of the stomach are presented. The main complaints in the horses, aged six years or more, were loss both of weight and of condition. Metastases had often developed.

The diagnostic difficulties and possibilities are discussed.     

Establishment of Theileria parva-infected bovine tissue culture in Swiss and athymic (nude) mice

Theileria parva-infected bovine lymphoid cells, grown in culture, were inoculated by different routes into neonatal and adult Swiss mice immunosuppressed by irradiation, thymectomy or inoculation of anti-lymphocyte serum. Tumour-like masses, composed of parasitized bovine lymphoid cells, formed at the site of subcutaneous inoculation in immunosuppressed neonatal and adult mice, but consistent establishment of cells following intra-peritoneal inoculation occurred only in neonatal mice. In all cases the degree of cellular establishment was proportional to the degree of immunosuppression. The best “take” was in irradiated neonatally thymectomized mice.Cells underwent short-term multiplication in mice but, as immune competence returned, the cells were rejected. There was no evidence that cells, on passage, became more adapted to grow in mice, nor that mouse cells became parasitized.Culture-derived cells were also inoculated subcutaneously into irradiated and non-irradiated nu/nu, nu/+ and Swiss mice. Tumour-like masses, composed of parasitized bovine lymphoid cells, developed at the site of inoculation in all irradiated mice. In nu/+ and Swiss mice these masses regressed after 2–3 weeks, but in the athymic nu/nu mice there was generally no rejection or cellular degeneration and parasitized cells became widely disseminated in the host's tissues and organs, in some cases causing death.T. parva-infected cells could not be established in non-irradiated nu/nu mice, nor when irradiated nu/nu mice were inoculated by the intra-peritoneal route. “Take” in irradiated neonatal nu/nu mice was also poor.Cells were passaged three times in irradiated nu/nu mice inoculated subcutaneously and it seems probable that indefinite passage of T. parva in mice can now be achieved.