Nude mouse model of the melanosis syndrome.

The occurrence of diffuse hyperpigmentation secondary to a xenografted human malignant melanoma was observed in nude mice. The patient from whom this cell line was established developed cutaneous hyperpigmentation after his disease became disseminated. Light and electron microscopy studies were performed on skin and organ biopsy specimens from the hyperpigmented mice. These studies indicated that this melanoma-associated melanosis was secondary to the release of pigment granules from the xenografted tumors and to the uptake of these granules by macrophages throughout the body, including those located in the dermis.

Establishment and characterization of two human pancreatic cancer cell lines tumorigenic in athymic mice.

Two human pancreatic cancer lines, RWP-1 and RWP-2, have been established from 2 patients with primary pancreatic cancer metastatic to the liver. The patients' tumors, the xenografted tumors, and tumors obtained by inoculation of nude mice with cultured RWP-1 and RWP-2 cells are all moderately-well-differentiated ductal cell adenocarcinomas. Ultrastructural analysis supports the tissue histopathology findings. Xenografts of RWP-1 tumors double every 10 days, whereas the doubling time of RWP-2 xenografts is 22 days. Both tumors contain mucin. RWP-1 and RWP-2 cells have a doubling time in culture of 45 hr and form colonies in soft agar. RWP-1 cultures appear to be morphologically heterogeneous; two distinct epithelial cell types can be identified. RWP-1 and RWP-2 have modal chromosome numbers of 64 and 62, respectively. Appreciable levels of glucose-6-phosphate dehydrogenase and lactic dehydrogenase were found in both cell lines and xenografts. RWP-1 and RWP-2 cells produce appreciable amounts of carcinoembryonic antigen, 1090 and 414 ng/10(6) cells, respectively.

Differential characteristics of two newly established human breast carcinoma cell lines.

Two human breast carcinoma cell lines, EP and MW, were established in culture from malignant pleural effusions. In addition to producing tumors in antithymocyte serum-immunosuppressed mice, both cell lines showed epithelial characteristics and anchorage-independent growth in soft agar. EP and MW differed in morphology (spindle-shaped versus round), chromosomal mode (hyperdiploid versus near triploid), estrogen receptor content (43.8 versus 5.1 fmol/mg protein), cloning efficiency (0.24 versus 15%), and activities (milliunits/10(6) cells) of creatine phosphokinase (25.7 versus 62.6) and lactate dehydrogenase (346.7 versus 778.5). Electron microscopy revealed that MW cells had more perinuclear filamentous material and more frequent intracytoplasmic vacuole formation than did EP cells. While having no effect on MW cells at the concentrations studied (10(-5) to 10(-11) M), beta-estradiol (10(-7) M) stimulated the growth of EP cells by 106% over the hormone-depleted control. In a variety of systems, EP was consistently the more drug sensitive of the two lines. In vitro, EP was significantly (p less than 0.001) more sensitive to methotrexate, vincristine, and 5-fluorouracil, respectively. In antithymocyte serum-mouse xenografts, EP displayed a greater response to three different dosages of a combination of cyclophosphamide, methotrexate, and 5-fluorouracil. One such dosage (cyclophosphamide, 32.0 mg/kg/day; methotrexate, 13.0 mg/kg/day; 5-fluorouracil, 190.0 mg/kg/day; for 1 day) reduced EP and MW tumor weights to 5.9 and 41% of controls, respectively. These results correlated well with the clinical responses.

"COLO 357," a human pancreatic adenosquamous carcinoma: growth in artificial capillary culture and in nude mice.

The human pancreatic cancer cell line COLO 357 has been xenografted s.c. in athymic Swiss mice. The xenografts grew well to form typical adenosquamous carcinomas. The cells were placed in a perfused artificial capillary system where they formed a solid tumor mass which survived for 7 weeks. In this system, the cells consumed glucose and released enzymes and carcinoembryonic antigen into the extracapillary space.

Immune complex deposits in Graves' disease and Hashimoto's thyroiditis.

The thyroid glands of four patients with Graves' disease and five patients with Hashimoto's thyroiditis were investigated to demonstrate in vivo immune complex deposition. By electron microscopy, electron-dense deposits were observed in the follicular basal lamina--basement membrane--(FBL) often associated with lymphocytic and plasma cell infiltration. A positive correlation was obtained with all cases by immunofluorescent studies using anti-IgG, IgA, IgM, C3 and antithyroglobulin conjugated serums. The staining was of a granular pattern and coincided to the FBL region. No discrepancies were noted in electron microscopic and immunofluorescent observations between patients with Graves' disease and Hashimoto's thyroiditis, and the occasional observation of immune complexes in areas devoid of infiltrate in some patients with Graves' disease. Morphologically, the deposits were found to be similar to those described in the Obese Strain chickens with spontaneous autoimmune thyroiditis.

Antitumor effects of polyvalent and monovalent vaccines coupled with interleukin-2 in a metastatic melanoma model.

We have previously reported that preimmunization of mice with formalinized extracellular antigens (fECA) derived from melanoma cells, in combination with interleukin 2 (IL-2) treatment and surgical resection, decreased subsequent tumor growth and increased survival of mice in a new model for spontaneous metastasis of melanoma. In this study, we have modified the sequence of tumor growth and therapy to more closely mimic the clinical situation. Mice were challenged subcutaneously in the tail with 5 x 10(5) B16 F10 melanoma cells and, by day 21, all of them had developed localized melanoma tumors. The primary tumor-bearing tails of control and experimental animals were then resected distal to the base of the tail, and therapy of the mice was initiated the following day. Groups of mice received different polyvalent and monovalent murine melanoma vaccines (including native or formalin treated extracellular antigens, intact melanoma cells, or purified B700 antigen), with or without concomitant low doses of IL-2. The results demonstrate that the vaccine therapies elicited significant increases in survival of the mice, accompanied by reductions in the size of lymph nodes and in the number of pulmonary metastases. These effects, particularly with the intact melanoma cell vaccine, could be improved even further with concomitant IL-2 treatment.

A new mouse model of experimental melanoma for vaccine and lymphokine therapy.

The annual incidence of malignant melanoma is estimated at 10-12 per 100,000 inhabitants in countries of central Europe and the United States, and alarmingly there has been a dramatic upward trend in that estimate. The B16 murine melanoma is a rapidly growing metastatic tumor of spontaneous origin, as are human malignant melanomas. Melanoma cells produce specific antigens which are uniquely different from normal cellular antigens, and the expression of such antigens is the cornerstone for preparation of anti-melanoma vaccines. One major problem in evaluating the effectiveness of vaccination and other biologic therapies is the variability of experimental tumor models. A new metastatic model of experimental melanoma which was developed in our laboratory imitates the major clinical stages of malignant metastatic melanoma: stage I, primary (local) tumor growth and bone marrow invasion; stage II, regional lymph node involvement; and stage III, metastasis to distant organs, such as the lungs. This model has been used successfully for screening vaccines constructed in our laboratory. Immunization with formalinized vaccines (of extracellular antigens, intact melanoma cells, or B700 antigen) or irradiated vaccines (of intact melanoma cells) partially inhibit primary melanoma tumor growth, reduce metastasis to regional lymph nodes and lungs, and significantly increase mean survival time. These anti-tumor effects were improved when polyvalent and monovalent vaccines were combined with IL-2 therapy. We also compared the immunogenic activity of vaccines made from B16 melanoma cells transfected with genes encoding murine IL-2 or GM-CSF, and effects on tumor bearing mice were compared with or without therapy using the corresponding lymphokines. In sum, comparison of antibody production, growth of primary melanoma tumors, number of surviving mice, mean survival time, and percent of mice with lung metastases, showed that the best course of immunotherapy involves vaccination of mice with irradiated B16 melanoma cells transfected to secrete GM-CSF, coupled with GM-CSF therapy.

Intrasplenic vaccination against experimental melanoma.

The immunodominant component of a formalinized extracellular antigen (fECA) vaccine prepared from B16 F10 melanoma cells is the melanoma-associated antigen B700. We now demonstrate that a single prophylactic intrasplenic inoculation of B700 antigen (1-10 mu g) stimulates the production of antibodies which have antiproliferative effects on B16 F10 melanoma cells in vitro. In addition, potential cytotoxic effects of splenocytes from B700 antigen inoculated mice were evaluated for two cellular immune effector functions, natural killer (NK) cell activity and lymphokine activated killer (LAK) cell activity; both activities were increased following B700 antigen inoculation. Intrasplenic injection of B700 antigen elicited an increase in the expression of the CD25 surface antigen (IL-2 R alpha) by T lymphocytes and up-regulated the expression of IL-2 R alpha mRNA. Thus both humoral and cellular cytotoxic immune responses might play roles in the decreased growth of primary tumors in B700 antigen inoculated mice and in the higher survival rate in this group of animals.

Immunization of mice with irradiated melanoma tumor cells transfected to secrete lymphokines and coupled with IL-2 or GM-CSF therapy.

We compared the immunogenic activity of irradiated vaccines prepared from B16 F10 melanoma cells with one made from B16 F10 melanoma cells transfected with genes encoding murine IL-2 or GM-CSF. Vaccines were studied in the conditions of treatment of C57BL/6 mice with or without the corresponding lymphokines. Control and prevaccinated mice were challenged with parental B16 F10 murine melanoma cells (5 x 10(5)) subcutaneously in the midtail to examine growth of the primary (local) tumor in the middle of the tall and metastases to the lungs. This experimental model is very close to the clinical stages of metastatic melanoma. The effectiveness of preimmunization of mice was determined by the levels of antibody production to a melanoma-associated antigen termed B700. The comparison of antibody production, growth of primary melanoma tumors, number of mice surviving at the end of the observation period, mean survival time and per cent mice with metastases in the lungs showed that the best course of immunotherapy was prevaccination of mice with a vaccine of irradiated B16 F10 melanoma cells transfected to secrete GM-CSF, coupled with GM-CSF therapy.

Cutaneous lupus erythematosus associated with melanoma and BCG vaccine therapy.

Abnormalities in host immunologic defenses may be associated with melanoma and lupus erythematosus, In two patients, cutaneous lupus erythematosus developed in the course of melanoma. Immunoglobulin deposits and complement were seen on direct immunofluorescence of exposed uninvolved skin. One patient was undergoing immunotherapy with BCG vaccine when the eruption occurred.

Rhinophycomycosis entomophthorae.

Rhinophycomycosis entomophthorae is a variant type of subcutaneous phycomycosis of the face that is caused by Entomophthora coronata. The case we describe was the first patient reported in the world literature, to our knowledge. His infection reappeared after an extended dormant period. Although his condition exhibited an early partial response to surgical and medical management, the condition became refractory to treatment.

Pustular herpes gestationis.

Pustular eruptions during pregnancy and the puerperium are rare. A case of herpes gestationis (HG) in which the lesions were predominantly pustules and in which a vesicular or bullous component was not noted is reported. The differential diagnosis of pustular eruptions associated with pregnancy is discussed.

Nonmelanoma skin cancer mortality. A population-based study.

To estimate the magnitude of nonmelanoma skin cancer mortality and describe its parameters, we reviewed the medical records of all deaths certified as due to this cause among Rhode Island residents from 1979 through 1987. After excluding acquired immunodeficiency syndrome-associated Kaposi's sarcoma, we confirmed that nonmelanoma skin cancer was the cause of death for 51 individuals, a quarter of the number of melanoma deaths reported. The age-adjusted nonmelanoma skin cancer mortality rate was 0.44/10(5) per year. Fifty-nine percent were due to squamous cell carcinoma, and 20% were due to basal cell carcinoma. Most appeared actinically induced. Among deaths from squamous cell carcinoma, the mean age was 73 years. At least 80% of the squamous cell carcinomas metastasized, and 47% arose on the ear. None appeared due to refusal of treatment. Among deaths from basal cell carcinoma, the mean age was 85 years, and refusal of surgical intervention was documented in 40%. Study of nonmelanoma skin cancer mortality provides for estimation of the magnitude of this problem, complements other studies of prognosis, and helps guide prevention, early detection, and treatment.

The clinical significance of periodic acid-Schiff-positive deposits in cuticle-proximal nail fold biopsy specimens.

In vivo capillary microscopic findings and proximal nail fold biopsy specimens from 19 patients with various connective tissue diseases, idiopathic Raynaud's phenomenon, and non-connective tissue diseases were studied. Periodic acid-Schiff-positive serous cuticular deposits were not specific to the group of patients with connective tissue disease. Generally, the severity of deposits correlated with the severity of the in vivo capillary microscopic pattern but not with disease severity or duration. The finding of these cuticular deposits may help to identify those patients with idiopathic Raynaud's phenomenon who are at risk to develop a connective tissue disease.

Human papillomavirus and widespread cutaneous carcinoma after PUVA photochemotherapy.

BACKGROUND: Oral psoralen with UV-A (PUVA) photochemotherapy is known to cause cutaneous malignancies and has been associated with cutaneous immunosuppression. Human papillomavirus infection has also been associated with cutaneous malignancies and with immunosuppressed individuals. We therefore sought evidence of human papillomavirus infection in a patient with a long history of PUVA therapy and multiple cutaneous malignancies. OBSERVATIONS: During a 15-year period, an otherwise healthy patient with psoriasis who had undergone a 10-year course of PUVA photochemotherapy developed 13 squamous cell carcinomas, eight lesions diagnosed as "squamous cell carcinoma vs keratoacanthoma," 14 other keratoacanthomas, six basal cell carcinomas, one melanoma in situ, and 18 other keratinocytic dysplasias. Twenty-two of the 30 lesions tested for human papillomavirus DNA by polymerase chain reaction were positive for type 16/18, including six of the seven basal or squamous cell carcinomas tested. CONCLUSION: We hypothesize that PUVA therapy-induced immunosuppression may play an important role in PUVA-related carcinogenesis by affecting the extent and pathogenicity of human papillomavirus infection.

Analysis of human serum proteins by molecular weight dependent acrylamide gel electrophoresis.

The technique of acrylamide gel electrophoresis of sodium dodecyl sulfate treated protein mixtures has been applied to the analysis of human serum proteins in the 70,000 to 250,000 molecular weight range. After staining, the bands are well defined, the molecular weight is defined and, hence, the identity of each can be estimated from the migration distance. In ambiguous cases, the identification of a band is confirmed by an independent method. The procedure is particularly valuable in the study of the gammopathies. The immunoglobulins migrate as expected, in contrast to the problems often encountered in the Ornstein-Davis gel method. Furthermore, light chains migrate more rapidly, are well separated from other serum proteins and, therefore, are readily detected. Paraproteins from three patients with documented gammopathies have been studied and characterized using this method.

Virilizing granulosa cell tumor responsive to human chorionic gonadotropin and oral contraceptive with 8-year followup.

Urinary and serum steroid measurements were obtained in a 48-year-old female before and after the removal of a virilizing granulosa cell tumor (GCT). Preoperatively, serum testosterone was markedly elevated while 17-ketosteroid (17-KS) and estrogen excretion were normal. Chorionic gonadotropin administration effected a marked rise in urinary 17-KS excretion and serum testosterone while oral contraceptive therapy for 21 days decreased serum testosterone to normal. With tumor resection, serum testosterone fell to normal and subsequent administration of HCG had no stimulatory effect on serum testosterone or urinary 17-KS excretion. The patient's clinical response and 8-year followup attest to the tumor's unusual virilizing role.

Inflammatory linear epidermal nevus caused by branchial cleft sinuses in a woman with numerous congenital anomalies.

A patient was seen with several congenital anomalies and bilateral branchial cleft sinuses that clinically resembled inflammatory linear verrucous epidermal nevi. This unusual condition most likely represented a cutaneous reaction to residual branchial remnants and prior surgical therapy.