MICA is a stress-induced protein that, as a rule, is not expressed in healthy tissues, but appears in large amounts on the surface of cells undergoing malignant transformation. In humans, this protein can either initiate antitumor immune response, or facilitate tumor cells for their escape of destruction. After shedding from tumor cell surface, soluble MICA enters blood circulation and contributes to decreased activity of effector cells, due to inactivation of NKG2D receptor. The aim of our study was to determine differences in circulating sMICA concentrations in sera of patients with different types of malignant lymphomas, and to evaluate the impact of sMICA upon NKG2D-positive cytotoxic lymphocytes. In experimental models with C1R-MICA cells, the MICA shedding was shown to occur from the surface of cultured tumor cells into the extracellular space. A reduced NKG2D expression dependent on sMICA concentration in the culture medium was demonstrated by flow cytometry in peripheral mononuclear cells, thus suggesting a role of serum sMICA in suppression of antitumor immune response. The sMICA detection was performed in patients with various types of B- or T-cell non-Hodgkin’s lymphomas by means of enzyme-linked immunosorbent assay. The groups of patients with increased sMICA content were identified and compared with the control group. Minimal amounts of serum sMICA were registered in the control group, with the median of 20 pg/ml. In a combined group of patients with various B-cell lymphomas, an increase in circulating sMICA amounts was shown, at the levels of more than 6.5 times exceeding the control values. The highest values of sMICA were recorded among the patients with chronic lymphocytic leukemia, diffuse large cell lymphoma, and multiple myeloma. Maximal sMICA levels among the investigated groups of patients were observed in the group of patients with T-cell anaplastic lymphoma (median of 574 pg/ml). The study provides preliminary evidence for a suppressive effect of different (immuno)chemotherapy components, in particular, rituximab and radiation therapy, upon serum sMICA contents in the lymphoma patients. Thus, elevated serum sMICA levels in patients with hematological malignancies may be considered as an additional criterion for application of the antitumor therapy. sMICA monitoring at different stages of cytostatic treatment may be useful in order to evaluate its efficiency.

MICA is a stress-induced protein that, as a rule, is not expressed in healthy tissues, but appears in large amounts on the surface of cells undergoing malignant transformation. In humans, this protein can either initiate antitumor immune response, or facilitate tumor cells for their escape of destruction. After shedding from tumor cell surface, soluble MICA enters blood circulation and contributes to decreased activity of effector cells, due to inactivation of NKG2D receptor. The aim of our study was to determine differences in circulating sMICA concentrations in sera of patients with different types of malignant lymphomas, and to evaluate the impact of sMICA upon NKG2D-positive cytotoxic lymphocytes. In experimental models with C1R-MICA cells, the MICA shedding was shown to occur from the surface of cultured tumor cells into the extracellular space. A reduced NKG2D expression dependent on sMICA concentration in the culture medium was demonstrated by flow cytometry in peripheral mononuclear cells, thus suggesting a role of serum sMICA in suppression of antitumor immune response. The sMICA detection was performed in patients with various types of B- or T-cell non-Hodgkin’s lymphomas by means of enzyme-linked immunosorbent assay. The groups of patients with increased sMICA content were identified and compared with the control group. Minimal amounts of serum sMICA were registered in the control group, with the median of 20 pg/ml. In a combined group of patients with various B-cell lymphomas, an increase in circulating sMICA amounts was shown, at the levels of more than 6.5 times exceeding the control values. The highest values of sMICA were recorded among the patients with chronic lymphocytic leukemia, diffuse large cell lymphoma, and multiple myeloma. Maximal sMICA levels among the investigated groups of patients were observed in the group of patients with T-cell anaplastic lymphoma (median of 574 pg/ml). The study provides preliminary evidence for a suppressive effect of different (immuno)chemotherapy components, in particular, rituximab and radiation therapy, upon serum sMICA contents in the lymphoma patients. Thus, elevated serum sMICA levels in patients with hematological malignancies may be considered as an additional criterion for application of the antitumor therapy. sMICA monitoring at different stages of cytostatic treatment may be useful in order to evaluate its efficiency.

Currently, cellular immunity can be estimated using the cluster of differentiation antigens or CD-markers. The panel of CD-markers including 6-8 parameters is usually used for assessment of cellular immunity. In our study we assessed not only subpopulations of T-, B-, NK-, NKT-lymphocytes, but also expression of activation markers (HLA-DR, CD25, CD38, CD69, CD314) using flow cytometry. The increase in the relative and absolute numbers of Treg-lymphocytes and NK-cells as well as the alpha chain of the IL-2 receptor on CD25 lymphocytes was observed in 20 patients with disseminated gastric and colorectal cancer. The percentage of CD25+ -cells and Treg-lymphocytes was also incresaed in 30 patients with skin melanoma. By comparing the expression of lymphocyte activation markers in the two groups of patients, it was shown that patients with gastric and colorectal cancer had the increased percentage of activated CD314+ , CD69+ NK-cells and patients with skin melanoma had the elevated level of activated CD95+ and CD38+ T-lymphocytes. This panel of markers can be used for monitoring immune response to immunotherapy for cancer.

Currently, cellular immunity can be estimated using the cluster of differentiation antigens or CD-markers. The panel of CD-markers including 6-8 parameters is usually used for assessment of cellular immunity. In our study we assessed not only subpopulations of T-, B-, NK-, NKT-lymphocytes, but also expression of activation markers (HLA-DR, CD25, CD38, CD69, CD314) using flow cytometry. The increase in the relative and absolute numbers of Treg-lymphocytes and NK-cells as well as the alpha chain of the IL-2 receptor on CD25 lymphocytes was observed in 20 patients with disseminated gastric and colorectal cancer. The percentage of CD25+ -cells and Treg-lymphocytes was also incresaed in 30 patients with skin melanoma. By comparing the expression of lymphocyte activation markers in the two groups of patients, it was shown that patients with gastric and colorectal cancer had the increased percentage of activated CD314+ , CD69+ NK-cells and patients with skin melanoma had the elevated level of activated CD95+ and CD38+ T-lymphocytes. This panel of markers can be used for monitoring immune response to immunotherapy for cancer.

0ne of the modern approaches for cancer treatment is based on the application of immunotherapy using activated cytotoxic lymphocytes. Search of methodological approaches for the preparation of activated lymphocytes in vitro is relevant. As a result of this study, the method for activation and culturing of lymphocytes for cancer patients have been perfected using cytokine IL-2 and IL-15. Peripheral mononuclear cells of cancer patients were culture using two different mediums based on RPMi-1640 with IL-2 for 10 days and X-vivo20 supplemented with IL-2 and IL-15 for 14 days. The expression of activation markers (CD38, CD69, CD25, HLA-DR and CD314) and subpopulations of lymphocytes were evaluated by the method of flow cytometry every 2 days. The expression of activation markers of lymphocytes increased after 3 days of culture in the first medium and after 5 days in the second one. We revealed that the activation of lymphocytes was faster in medium based on RPMI with IL-2, but the proliferation and viability of lymphocytes were lower than in the second medium. The culture medium based on RPMI with IL-2 can be recommended for more quickly obtaining of lymphokine-activated killer cells. The medium based on X-vivo20 with a combination of IL-2 and IL-15 can be recommended for a longer cultivation of lymphocytes and for escalating of lymphokine-activated killer cells. it has been shown that the combination of cytokines IL-2 and IL-15 not only has a positive influence on the proliferation activity of the lymphocytes and the expression of activation markers, but also on their viability.

The major medical problem in the treatment of skin melanoma is improvement methods of treatment, increasing their effectiveness and safety. In this study, adoptive immunotherapy, using lymphocytes activated in vitro, was performed in 15 patients with metastatic melanoma. Evaluated the phenotype of peripheral blood lymphocytes and activation markers (HLA-DR, CD25, CD314, CD38, CD69) before and 3-4 weeks after immunotherapy. It is shown that for these patients is characterized by increasing the number of CD25+ and Treg lymphocytes in the bloodstream, which has not changed after immunotherapy. Adoptive immunotherapy in combination with chemotherapy resulted in a decrease of absolute number of lymphocyte, B- and T-lymphocytes, T helper cells, NKT-cells, CD314+ lymphocytes, CD38+ lymphocytes and immature T-lymphocytes (CD3+CD38+) (р < 0,05). However, there was a positive dynamic to increase the percentage of NK-cells to 32% and CD69+NK-cells to 21% and significant increase in expression of HLA-DR on all lymphocytes (p < 0.05). Adoptive immunotherapy characterized by the absence of side effects and can be recommended as accompanying to basic radiation and chemotherapy.

Phenotyping of peripheral blood lymphocytes and immune activation markers of activation (HLADR, CD38, CD69, CD314, CD25) was performed in thirteen patients with disseminated forms of stomach and rectal cancer before cell treatment, and following adoptive immunotherapy with activated lymphocytes. It wasshown that the lymphocytes are well activated and are able to proliferate in vitro. It was revealed that the relative content of Tregs and NK cells is increased in these patients. After the courses of immunotherapy, initially low levels of B-cells (average 5%) remained in all the patients, whereas concentration of Тregs didn’t change. Increased expression of activation markers was revealed for all lymphocyte subsets (CD25, CD314, CD38, HLA-DR) and, especially, for T-lymphocytes (CD3+HLA-DR, CD3+CD38+). Significant decrease of T helpers, activated NK-cells (CD16+CD314+) and CD4+/CD8+ was noted in peripheral blood. A non-significant elevation in mature lymphocytes (CD45RO+) and reduced content of young lymphocytes (CD45RA+) were revealed. Adoptive immunotherapy is safe and well tolerated, being characterized by lack of side effects, and it may be recommended as a complement to conventional radio- and chemotherapy.

The authors consider general immunological effects of intravesical BCG vaccine as an independent method of immunotherapy of superficial cancer of the bladder (CB). 27 patients (15 males and 12 females, mean age 57 years) with a recurrence of superficial CB (T1-2N0M0) were treated. The patients had undergone transurethral or transvesical bladder resection and combined therapy. Intravesical immunotherapy of CB recurrence with BCG vaccine induced persistent changes in the immune system: stimulate lymphocyte activity and phagocytic activity of neutrophils, T-cell function, normalizes function of endogenic suppressors, increases the amount of IgM and CIC. Repeat courses of the vaccine maintain the above immunological effects for a long time.

The trial covered 58 patients with prostatic cancer aged 50-79. Before treatment these patients exhibited weak proliferative response of T-cells to PHA, increased number of T-suppressors/killers, enhanced suppressive influence of regulatory cells. Hormone chemotherapy produced immunodepressive and immunomodulating effects (normalization of the count of T-suppressors/killers and their function). Subsequent radiotherapy on the prostatic region (total dose 60 Gy) worsened immunodepression. Efficacy of placental suspension and levamisol depends on the treatment stage at which the latter were used: during hormone chemotherapy the addition of the suspension and levamisol corrects its immunodepressive effects, in subsequent radiotherapy immunodeficiency greatly increased.

A beneficial effect of bestatin used as adjuvant in the course of chemo- and radiotherapy of local cancer of the bladder manifested itself as a reduction of postradiation immunosuppression: increased count of T mu-helper/inductors and T gamma-suppressors/killers, enhancement of neutrophil phagocytic activity. Bestatin recovers radiation-impaired immune system by inhibition of T gamma-suppressor killers and their precursors generation, of endogenic regulators suppressive activity, normalization of T-lymphocyte and natural killer activity, stimulation of B-lymphocyte recovery.

During an open controlled study of 20 patients with rheumatoid arthritis (resistant forms) their lymph nodes were irradiated at a dose of 7.5 Gy. Clinical improvement, including reduced morning rigidity, the number of inflamed joints, and Riccis index, was noted shortly after therapy and 6 mos. after irradiation in 50% of the patients. Immunosuppression and moderate lymphopenia were noted in all of them. Of the side-effects there was nausea that disappeared without additional treatment. A conclusion has been made that irradiation of the lymph nodes at a total dose of 7.5 Gy is equally effective but less toxic than irradiation of 20 Gy.

Initial immunologic status in children suffering osteogenic sarcoma was retrospectively evaluated versus such parameters as site and extent of tumor, and clinical course. A complex of disorders in the immunologic system was established in osteogenic sarcoma patients. Such characteristics as levels of active, theophylline--sensitive and levamisole--reactivated T-cells were found clinically valuable and prognostically significant. These parameters may betray response of high- and low-affinity E-receptors of lymphocytes to serum factors of tumor growth.

The paper is concerned with immunological evaluation of different stages of combined therapy with local UHF hyperthermia in children with osteogenic sarcoma. Combined therapy (polychemo- and radiotherapy) was shown to cause a decrease in the number of immunocompetent cells, to enhance imbalance of immunoregulatory T-lymphocytes, to weaken T-lymphocyte function on PHA; immunosuppressive action of combined therapy did not depend on a tumor site. The incorporation of UHF-hyperthermia in the therapeutic scheme weakened the manifestations of secondary immunodeficiency, got back to normal the structure of T-lymphocyte population. A favorable immunomodulating effect of hyperthermia was more frequently observed in patients with crural bone tumors. The effect of hyperthermia was revealed after direct influence of thermotherapy but it was absent in continuation of combined treatment.