Establishment and characterization of a squamous cell carcinoma of the vulva in tissue culture and immunologic evaluation of the host

Abstract

A continual epithelial cell culture has been established from a squamous cell carcinoma of the vulva metastatic to the right inguinal lymph node. Initial characterization of the cell line (designated LT2), including morphology, growth properties, karyotyping, and secretion of carcinoembryonic antigen (CEA) and the beta subunit of human chorionic gonadotropin (HCG-β) is reported. The LT2 cell line satisfies the following criteria of malignancy: (1) establishment in culture for more than one and one-half years exceeding more than 70 generations; (2) heteroploid karyotype; (3) abnormal cytology; (4) growth in soft agar suspension; and (5) tumorigenicity in nude mice. This cell line has also been found to be free from contamination by fibroblasts, mycoplasma, bacteria, and fungi. With the LT2 cell line, the specific tumor-host immunorelationship was assayed by autologous (host lymphocyte versus LT2 cells) cytotoxicity and lymphoblast transformation in the presence of tumor extracts or mitomycin C-treated LT2 cells. In vivo patient immunocompetence was evaluated specifically by skin testing with autologous tumor extracts and nonspecifically with bacterial recall skin test antigens and dinitrochlorobenzene. In vitro nonspecific immunocompetence was evaluated by total peripheral white blood cell and lymphocyte counts, T cell, active T cell and B cell determinations, lymphocyte response to the mitogen phytohemagglutinin, and spontaneous lymphocyte-mediated cytotoxicity versus cultured bronchogenic carcinoma cells. Correlation of immunocompetence and the status of the tumor-host relationship with the patient's clinical course, and the potential significance of LT2 CEA and HCG-β production are discussed. The patient's response to bacillus Calmette-Guerin immunotherapy is evaluated.