In order to know whether effector cells are capable of recognizing the epidermal targets in pemphigus vulgaris (PV) and bullous pemphigoid (BP), non-adherent cells (NAC) of peripheral blood from 27 primary PV, 19 BP patients and 12 healthy volunteers were used in cytotoxic tests. Autologous, allogenic and murine epidermal keratinocytes as well as autologous and allogenic fibroblasts were employed as targets. Total esterase activity of serine proteinases was measured in supernatants of samples and the results obtained were compared with the data received in 51Cr-release assay. These two techniques yielded similar results. The effector cells of PV and BP patients were found to lyse the epidermal target cells without presensitization in vitro. Epidermal cells from involved and uninvolved skin of PV patients, as well as from perilesional skin of BP patients, were highly sensitive to cytotoxic effect on the part of effector cells. The NAC activity in BP was higher than in PV. On the one hand, the obtained data suggest that PV and BP patients develop autoreactive cytotoxic T lymphocytes, sensitized to pemphigus and pemphigoid antigens, respectively, and on the other hand that epidermocytes in pemphigus and pemphigoid patients turned to be more fragile compared to normal human epidermal keratinocytes. It was supposed that cell-mediated cytotoxic reactions are probably related to enhanced proteolytic activity in the site of bullous eruption.

Hemocarboadsorption (HCA) with the following immunosuppressive therapy was used to treat 48 patients with pemphigus vulgaris (PV) and 31 patients with bullous pemphigoid (BP) in the acute phase. The levels of autoantibodies, eicosanoids, endoproteases and their inhibitors as well as interleukin (IL) 1 and IL 2 production and absorption of exogenous IL 2 were examined in the treatment dynamics. The therapeutic effect of HCA was displayed in prompt disease remission in 43 PV and 29 BP patients. Clinical improvement was accompanied by antibody elimination and interleukin cascade reaction normalization. HCA resulted in growth of patients' serum immunosuppressive activity. It was concluded that HCA has some immunoregulatory effect (Int J Artif Organs; 1990: 13: 181-88).

The fixation of pemphigus antibodies was revealed by indirect immunofluorescence in shedding grass snake epidermis. The antibodies of patients with pemphigus vulgaris (PV) were specifically binding to the antigenic substance which appears in the snake epidermis at the initial stage of integument change. At the peak of shedding, the fixation of PV antibodies was observed only in the upper layers of the ‘old’ epidermis although the intensity of fluorescent staining was considerably reduced. At the same stage of shedding, there was marked fixation of antibodies of patients suffering from pemphigus foliaceus (PF).The specific binding of PF antibodies was noted predominantly on the cell membranes of the ‘old’ keratinocytes, localized in the upper layers of the epidermis. In contrast to bullous pemphigoid antibodies, PV and PF antibodies failed to fix in the skin of the snake prior to or after physiologic shedding. The data obtained suggest that pemphigus may have an atavistic origin.

In order to clarify the role of natural cytotoxicity (NC) in the damage of epidermal keratinocytes (EK) in pemphigus vulgaris (PV) we used the direct 51Cr-release assay in studying the cytotoxic activity of large grandular lymphocytes (LGL), obtained from 34 acute PV patients and 19 healthy donors against EK of PV patients, donors, intact newborn BALB/c mice and mice with experimental PV, shedding and non-shedding grass snakes. We also investigated the effect of pemphigus antibodies and shedding snake serum protein (SSSP) upon EK sensitivity to cytotoxic effects of NC effectors. In other experiments, studied were the amount of serine proteinase secreted by donor LGL in the presence of membrane antigen of antibody- or SSSP-transformed EK, and direct effect of pemphigus antibodies and SSSP upon the release of radioactive label and endoproteinases from EK. It was found that EK, which had interacted with pemphigus antibodies or SSSP, became a sensitive target for NC effectors. Membrane antigen of antibody- and SSSP-transformed human, murine and snake EK induces the secretion of endproteinases by NC effector cells. The conclusion was made that the effect of both pemphigus antibodies and SSSP on EK resulted in the emergences of a new antigen on EK cellular membrane which is recognized by NC effectors and plays the role of the ligand in receptor/ligand interaction between LGL and their targets.

Development of novel methods of early diagnosis of lung cancer is one of the major tasks of contemporary clinical and experimental oncology. In this study, we utilized the tobacco nitrosamine 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanone (NNK)-induced lung cancer in A/J mice as an animal model for development of a new imaging technique for early diagnosis of lung cancer. Lung cancer cells in A/J mice overexpress nicotinic acetylcholine receptors. Longitudinal CT scans were carried out over a period of 8 months after NNK treatment, followed by PET/CT scans with 18F-Nifene that binds to α4-made nicotinic receptors with high affinity. PET/CT scans of lungs were also obtained ex vivo. CT revealed the presence of lung nodules in 8-month NNK-treated mice, while control mice had no tumors. Imaging of live animals prior to necropsy allowed correlation of results of tumor load via PET/CT and histopathological findings. Significant amount of 18F-Nifene was seen in the lungs of NNK-treated mice, whereas lungs of control mice showed only minor uptake of 18F-Nifene. Quantitative analysis of the extent and amount of 18F-Nifene binding in lung in vivo and ex vivo demonstrated a higher tumor/nontumor ratio due to selective labeling of tumor nodules expressing abundant α4 nicotinic receptor subunits. For comparison, we performed PET/CT studies with 18F-FDG, which is used for the imaging diagnosis of lung cancer. The tumor/nontumor ratios for 18F-FDG were lower than for 18F-Nifene. Thus, we have developed a novel diagnostic imaging approach to early diagnosis of lung cancer using 18F-Nifene PET/CT. This technique allows quantitative assessment of lung tumors in live mice, which is critical for establishing tumor size and location, and also has salient clinical implications.