[Melanoma cell lines as the basis for antitumor vaccine preparation].

The aim of the study was to obtain cell lines from tumor samples, and to determine phenotypic cell characteristics in order to choose the optimal line for vaccine preparation. 15 cell lines with stable growth, varying in cultural growth character and cytomorphology, were obtained from samples taken from patients with metastatic skin melanoma. Immunofluorescense method was used to determine the expression of T- and B-lymphocyte markers, antigens of major histocompatibility complex (MHC) class I and II, and CD86 co-stimulating molecule in the cell lines. The expression of melanocyte differentiation antigens and cancer/testicular antigens was evaluated using immunocytochemical assay. The results allowed the authors to distinguish three types of melanoma cell lines according to the expression of MHC molecules: MHC-negative; MHC class I positive; MHC classes I and II positive.

[Her-2/neu, Ki-67 expression and ploidy in breast carcinoma]. / Ekspressiia Her-2/neu, Ki-67 i ploidnost' pri rake molochnoi zhelezy.

Expression of the markers was studied immunocytochemically on cytological preparations of breast tumor, ploidy was assessed by flow cytofluorimetry. The material was divided into 2 groups: diploid--41.4% cases and aneuploid--58.6%. Hyperexpression of Her-2/neu in the first group was observed in 54.5%, in the 2nd group in 48.6% cases. Expression of Ki-67 in 63.4% and in 68.9%, respectively. Tumours with high proliferative activity were numerous in aneuploid tissues, and in diploid tumours moderate activity prevailed (p = 0.006). Direct correlation between the markers was observed, high expression of Ki-67 more frequently was associated with positive expression of Her-2/neu. Thus, aneuploid tumours with high proliferative activity and hyperexpression of Her-2/neu are more aggressive tumours of a large size.

[Comparison of immunocytochemical and immunohistochemical methods for biomarker detection in breast cancer]. / Sravnenie immunotsitokhimicheskogo i immunogistokhimicheskogo metodov opredeleniia biomarkerov pri rake molochnoi zhelezy.

The detection of the biological parameters of the tumor before the treatment beginning becomes of more importance. The present study aimed to carry out the comparative analysis of the molecular markers expression (P53, Ki-67, Her-2/neu, Bcl-2, Bax, ER, FasL and CD95) at the cytologic and the correspondent histologic samples. The 18 tissue samples of the breast cancer were investigated. The immunocytochemical and the immunohistochemical methods of the molecular markers determination were used. Our study showed the correlation between two methods and the possibility of the use of the immunocytochemical staining as routine method of the molecular markers expression determination.

[Apoptosis and proliferative activity of bone marrow cells in patients with aplastic syndromes as evidenced by trephine biopsy]. / Apoptoz i proliferativnaia aktivnost' kletok kostnogo mozga u bol'nykh s aplasticheskimi sindromami po dannym trepanobiopsii.

AIM: To clarify the effect of cyclosporin A (CSA) on the apoptosis and proliferative activity of bone marrow cells in patients with aplastic syndromes by trepanobiopsy evidence. MATERIALS AND METHODS: The TUNEL immunohistochemical assay was used to study apoptosis of bone marrow cells in the histological specimens from 24 patients: 8 with refractory anemia (RA), 8 with acute small-proportion leukemias [RA with excessive blasts (RAEB) + RAEB in transformation (RAEBt)], 3 with acute non-lymphoblastic leukemia (ANLL), 5 with lymphogranulomatosis (LGM). Control apoptosis examination was made in 10 patients treated with CSA. The proliferative activity of bone marrow cell was evaluated by bone marrow histological specimens from 10 patients (8 patients with RA and 2 with RAEB + RAEBt) at the onset of disease and during CSA therapy in the immunohistochemical test with primary nuclear antigen Ki-67 antigen antibodies. Changes in the proliferative activity and degree of apoptosis of bone marrow cells were assessed in relation to the cellularity detectable by the histological specimens. RESULTS: Patients with RA showed an increase in bone marrow cell apoptosis to 25.375 +/- 6.874 (-10.8 +/- 5.122 and 8.333 +/- 5.84 in controls and ANLL patients, respectively). The cells of hemopoiesis and stromal microenvironment are in the process of cell death. Higher bone marrow cellularity is observed in CSA-treated patients at clinical and hematological remission, which is followed by rises in the index of proliferation and the degree of apoptosis. CONCLUSION: The clinical effect of CSA in patients with aplastic syndromes is induced by its direct or mediated stimulating effect on pluripotent stem cells of hemopoiesis, by increasing bone marrow cellularity at the expense of clonal and/or normal hemopoiesis.

Fas (APO-1/CD95) Antigen: New Activation Marker for Evaluation of the Immune Status.

Using mAb ICO-160, we have studied Fas (APO-1/CD95) antigen expression on blood lymphocytes from healthy women, pregnant women and patients with chronic adnexitis, uterin myoma, ovarian cyst, ovarian and uterin cancer. In peripheral blood from healthy women Fas antigen was detected on 23.42 +/- 2.9% of lymphocytes. The healthy donors were divided in two different subgroups with low and high Fas expression. Expression of Fas antigen on lymphocytes was elevated (high expression subgroup) in all the groups of investigated patients, excepting the ovarian cancer ones with Fas expression similar to that in healthy donors. Comparison of expression of other differentiation antigens between healthy donors and the above groups of patients elucidated in the patients a kind of pronounced imbalance of the immune status and activation of the immune system. Additionally, in patients with ovarian cancer the imbalance of the immune status was reflected as altered ratio between helper and suppressor cells (the ratio was 0.73 in contrast to that 1.08 in group of healthy donors). As follows from the summarized results of all the trials, Fas antigen expression correlated with expression of other activation antigens as CD71 and CD25 (r = 0.05 and r = 0.52, respectively). Consequently, Fas (APO-1/CD95) antigen may be considered as the activation antigen and its expression may be used for assessing the immune status.