Tumor-Infiltrating Lymphocytes in Breast Cancer. Association with Clinical and Pathological Parameters.

In patients with primary resectable breast cancer, a positive correlation between the age and the count of CD16+ lymphocytes and a negative correlation of this parameter with the number of regulatory CD4+CD25+CD127- cells and proliferative activity of Ki-67 tumor cells were revealed. Higher level of Ki-67 was associated with reduced number of effector lymphocytes (CD8+ and CD16+) and elevated content of regulatory CD8+CD11b-CD28- T cells. The absence of expression of estrogen receptors was associated with reduced cytotoxic potential of CD8+ T cell in comparison with ER+ breast cancer. The percentage of CD8+ lymphocytes (CD3+CD8+ and CD8+CD11b+CD28+) among lymphocytes infiltrating the tumor was higher in PR+ breast cancer than in PR- tumors. With increasing the tumor load, the number of lymphocytes expressing CD16 marker and their cytotoxic potential decreased.

[Lymphocyte receptors that regulate the immune-response--the key to the management of antitumor immunity].

Tumor uses various mechanisms to "escape" from immune surveillance including the important role of dysregulation of interaction of signals from corresponding co-stimulatory and co-inhibitory receptors (so-called "control points immunity"-- immune "checkpoints") modulating T-cell activation process. A creation of targeted drugs impacting on immune "checkpoints"" is a new promising trend in modern oncoimmunology. This review is devoted to a brief characterization of some receptors of immune competent cells (such as activation and inhibitor), which play a crucial role in the interaction of the immune system and tumors as well as targeted drugs acting on them for therapeutic purpose.

[Phenotype of immunocompetent cells and its role in antitumour immune response].

The paper contains an overview of the role of certain minor subpopulations of lymphocytes in tumour growth and of the main results obtained in the studies of the phenotype of immunocompetent cells from cancer patients. Special attention is given to selected indicators of immunity as predictors of the disease and efficacy of immunotherapy.

Suppressor cells in melanoma and lung cancer: correlation with the clinical stage.

Mononuclear blood cells from 40 healthy donors, 25 patients with melanoma and 35 patients with lung cancer were tested for suppressor cell activity and response to phytohemagglutinin (PHA). The suppressor cell activity was measured by determining the ability of Mitomycin C (MMC)-treated mononuclear cells from patients and controls to suppress the in vitro response of responder cells to PHA. Mononuclear cells from one healthy donor were stored in liquid nitrogen for use as responder cells. Indomethacin-sensitive suppression was also measured. In both groups of patients the mean values of suppressor cell activity in co-culture were higher than those in control group. When patients were subdivided in groups according to the clinical stage, the level of suppressor cell activity increased with the stage. Indomethacin-sensitive suppression was detected in some patients but the differences between patient and control groups were not statistically proved. An inverse relationship was observed between indomethacin-sensitive suppression and the PHA response.

Clinical significance of standard CD assessment in acute leukemia.

The data of detailed studies of immunophenotype of blast cells in 426 patients with acute leukemias are presented. Diagnostic and prognostic significance of different marker expression has been evaluated in groups of patients with ALL and AML. Frequency distribution of T1, T2, T3, pre-B, B, common, Ia and null subvariants identified according to immunoclassification of Baryshinkov et al. was studied in 250 children with ALL. These subvariants differed both in duration of disease (p = 0.0015) and in duration of first complete remission (p = 0.0031). The use of monoclonal antibodies of VI-series in 90 patients with ALL allowed to describe an immunophenotype of the subvariants in detail. The mosaic expression of myeloid antigens CD11, CD14, CD15, CDw65 identified by MoAbs VIM-12, VIM-13, VIM-D5 and VIM-2, respectively, on blast cells of patients with AML was shown. The expression of CD11 (ICO-GM1) or CD15 (ICO-G2) was prognostically unfavorable in children with AML (p = 0.0028). The expression of T-cell markers (E-receptor, CD7, reactivity with anti-T-cell serum) on blasts was prognostically favorable in children with AML (p = 0.003). So the data of immunophenotyping are of great value for accurate diagnosis and prognosis of acute leukemias.

CD95 (FAS/APO-1) antigen is a new prognostic marker of blast cells of acute lymphoblastic leukaemia patients.

We analyzed CD95(Fas/APO-1) antigen expression on bone marrow blasts in 38 children with acute lymphoblastic leukemia (ALL) receiving a treatment in the Department of Leukaemias at the Cancer Research Center in 1987-1989 years (n = 22) and in 1994-1997 years (n = 16). CD95 antigen expression was studied by monoclonal antibodies (MoAbs) IPO-4 in indirect immunofluorescence analysis. CD95 antigen was expressed on 35.8 +/- 7.5% bone marrow blasts, most frequently (63.6%) in the clinically favourable Pre-B ALL. Only in this group CD95 antigen expression was correlated with CD10 antigen expression that has a positive influence to the time of complete remission in ALL patients. Our data showed that CD95 expression on blast cells is a favourable prognostic sign, associated with increased relapse-free and total survival. On the contrary, the absence of CD95 antigen on blasts is an unfavourable sign for disease evolution.

[Isolation from the human peripheral blood of a fraction of mononuclear cells with high cytotoxicity against K-562 and HeLa target cells]. / Vydelenie iz perifericheskoi krovi cheloveka fraktsii mononuklearov s vysokoi tsitotoksichnost'iu protiv kletok-mishenei linii K-562 i HeLa.

Adherent cells were released from human peripheral blood mononuclears and the remaining mononuclears were separated by the density gradient centrifugation on percoll. Five fractions obtained were tested for cytotoxicity against K-562 and HeLa tumour target cells. Cell morphology and reactivity to some monoclonal antibodies were also studied. Fractions 2 and 3 were enriched by large granular lymphocytes (LGL) and BMA-070 positive cells and showed increased cytotoxicity only to K-562 target cells, while fraction 1 contained 9.5% of monocytes, but no LGL, and showed increased cytotoxicity against both K-562 and HeLa target cells.

[Tumor-induced immunosuppression in breast cancer patients]. / Izuchenie opukholeindutsirovannoi immunosupressii u bol'nykh rakom molochnoi zhelezy.

A comparative study of immunological reactivity of healthy volunteers and cancer patients with different stages of the disease was carried out. The study of 8 types of suppressor cells, chemoreceptor sensitivity and blast transformation of lymphocytes, natural killer cells, the level of immunoglobulins, regulatory subpopulation of lymphocytes, etc., has revealed several alterations in regulatory and effector links of immunity in the patients with late stages of the disease. The detailed immunological monitoring in the course of varying schemes of treatment is emphasized.

[The sensitivity of the blood lymphocytes in stomach cancer patients to met-enkephalin, dopamine and serotonin]. / Chuvstvitel'nost' limfotsitov krovi bol'nykh rakom zheludka k met-énkefalinu, dofaminu i serotoninu.

Sensitivity of blood lymphocytes to the stimulating effect of met-enkephalin and dopamine was shown to decrease at late stages of the gastric cancer (IIIB-IV) or in the case of metastases to other organs, while to the inhibitory effect of serotonin it increased as compared to early stages (I-IIA). Using specific blockers of opioid, dopamine and serotonin receptors it was shown that these effects were mediated through stimulation of respective type receptors.

[Expression of surface markers of lymphocytes and their relation to suppressor function in lymphogranulomatosis]. / Ekspressiia poverkhnostnykh markerov limfotsitov i ikh sviaz' s supressornoi funktsiei pri limfogranulematoze.

The patients with Hodgkins disease possessed an increased number of the T gamma-cells, a decreased number of T mu cells and higher endogenic suppressor activity as compared with healthy donors. Subpopulation of ferritin-carrying lymphocytes was found only in patients, whereas in donors these cells were not revealed at all. It was shown that ferritin receptors expressed only on T gamma-cells. For determination of the functional role of T-cell subpopulation before and after Con-A stimulation were studied. The suppression activity correlated with the T gamma-cell levels.

[Suppressor function of the blood lymphocytes in patients with stomach cancer undergoing gastrectomy combined with splenectomy]. / Supressornaia funktsiia limfotsitov krovi bol'nykh rakom zheludka pri provedenii gastréktomii v kombinatsii so splenéktomiei.

Suppressor status of immunity in 34 patients with gastric cancer of the III-IV stages was studied before and after (15 days and 3 months) gastrectomy and gastrosplenectomy. The control group consisted of 42 healthy donors. It was shown that gastrectomy caused inhibition of the endogenous activity, but stimulated the function of the prostaglandin-producing suppressor cells while combined gastrosplenectomy induced inhibition of the function of both endogenous and prostaglandin-producing suppressor cells.

[The expression of lymphocytic function-associated antigen determined by IKO-11 monoclonal antibodies in patients with nephroblastoma and neuroblastoma]. / Ekspressiia limfotsitarnogo funktsiia-assotsiirovannogo antigena, opredeliaemogo monoklonal'nymi antitelami IKO-11, u bol'nykh nefroblastomoi i neiroblastomoi.

The importance of NK-cells was shown in 12 patients with nephroblastoma and 10 patients with neuroblastoma using ICO-11 monoclonal antibodies. The numbers of circulating LFA-1+ cells were shown to be different in the untreated and treated patients. A certain regular influence of antitumour therapy on the amount of circulating LFA-1+ cells was established.

[Standardization of ICO-1 monoclonal antibodies against monomorphic Ia-like (Dr) antigens]. / Standartizatsiia monoklonal'nykh antitel IKO-1 protiv monomorfnykh Ia-podobnykh (Dr) antigenov.

ICO-1 Mab were obtained following BALB/c mouse immunization with 24-week human fetal thymocytes. Cloning for two times by the method of limited dilutions led to a hybridoma with a stable production of G3 isotype Mab. ICO-1 Mab have immunoprecipitated an antigen consisting of two polypeptide chains with molecular weight 29 and 34 kDal. The antigen expression was enhanced following PHA or alloantigen activation of blood mononuclear cells. ICO-1 Mab inhibited the alloantigenic response of blood mononuclear cells. Mab detected 29% of antigen-positive cells in the peripheral blood of healthy adults. When the reaction of ICO-1 Mab was compared with that of Mab against monomorphic Ia-like antigens OKLa, anti-Ia-BRL, BMA-021 and HLA-Dr on blood cells from healthy donors and patients with leukemia proved to be identical.

[ICO-63 monoclonal antibodies to the differentiation antigen of hemopoietic cells suitable for immunophenotyping of human solid tumors]. / Monoklonal'nye antitela IKO-63 k differentsirovochnomu antigenu gemopoéticheskikh kletok, prigodnye dlia immunofenotipirovaniia solidnykh opukholei cheloveka.

ICO-63, new monoclonal antibodies (MCAB), used for immunodiagnosis, were produced by the standard hybridoma technique ICO-63 MCAB reacted with 50% granulocytes, 20% thrombocytes and endothelial cells, whereas they did not react with peripheral blood mononuclear cells, thymocytes and erythrocytes. The molecular weight of ICO-63 MCAB-identified antigen was about 100 kD. ICO-63 MCAB reacted with 12 out of 15 neuroblastomas, with some solid tissue sarcomas and melanomas, but did not react with tumours of the epithelial origin.

[ICO-1 monoclonal antibodies against the constant region of Ia-like (Dr) antigens]. / Monoklonal'nye antitela IKO-1 protiv konstantnoi chasti Ia-podobnykh (Dr) antigenov.

Mouse monoclonal antibodies ICO-1 to constant part of Ia-Like (Dr) antigens were produced. Hybridoma continuously produced IgG3 antibodies during 22 passages in vivo, more than 7 months in vitro. Antibodies specifically bound to 29.1 +/- 2.3% of peripheral blood mononuclear cells of healthy people recognized the antigen on B lymphocytes and 44.2 +/- 3.4% of monocytes. This antigen was absent on granulocytes and T lymphocytes. Using monoclonal antibodies ICO-1 antigenically positive cells were detected in 40 patients with B-CLL, in 12 of 35 patients (34.3%) with chronic granulocytic leukemia at the stage of blastic crisis, in 43 of 65 patients (66.1%) with ALL, in 14 of 38 patients (36.8%) with lymphosarcoma and in 17 of 30 patients with acute myelomonocytic leukemia. The antibodies responded to surface antigens in the reaction of indirect surface immunofluorescence, complement-dependent cytotoxic reaction and radioimmune tests.

[ICO-35 monoclonal antibodies to the antigen of acute lymphoblastic leukemia]. / Monoklonal'nye antitela IKO-35 k antigenu ostrogo limfoblastnogo leikoza.

ICO-35 monoclonal antibodies (Mabs) were produced following BALB/c mouse immunization with peripheral blood cells from a patient with lymphoid type of chronic myeloid leukemia in blast crisis (SML BC) ICO-35. Mabs detect antigen on CD10-positive cells from patients with acute lymphoblastic leukemia, CML BC, CD10-positive cells of Reh line and are not bound to other cells. Comparative studies of reactivity of ICO-35 and K 503 Mabs to CD10 antigen revealed their similarity. However, in contrast to K503 ICO-35 Mabs do not react with granulocytes.

[The expression of leukocytic and related antigens on the cells of human breast cancer cells]. / Ekspressiia leikotsitarnykh i rodstvennykh im antigenov na kletkakh raka molochnoi zhelezy cheloveka.

Certain expression regularities of different markers in human breast cancer cells are demonstrated. Monomorphic determinants of HLA class II antigen are detected only in case of HLA class I expression and cancer-embryonal antigen (CEA); the latter is combined with HLA class I and ICO-84 antigens at a significantly higher rate. The expression effect of CEA and HLA-DR on the metastatic spreading rate of regional lymph nodes is marked. A heterogeneity of the single tumour and in patients with malignant breast cancer is determined by the presence of the "leucocytic" antigens and CEA. In parallel studies monoclonal antibodies (MAB) HMFG-1 and a common epithelial antigen are assayed. The antigen interacting with MAB HEA-125 is characterized by the monomorphic expression.

[The immunodiagnosis and immunotherapy of tumors]. / Immunodiagnostika i immunotherapiia opukholei.

Recent studies into the immunology of tumors have allowed the oncological use of immunological methods to be substantially extended. This increases the potentialities of immunodiagnosis, primarily to identify a number of tumor-associated antigens, which has promoted early differential diagnosis and monitoring of the course of disease. On the other hand, diagnosis of immunodeficiencies in cancer patients has become better: a panel of monoclonal antibodies to differentiated immunocompetent cell antigens has been developed, their subpopulations revealed, the time course of whose quantitative composition correlates with the clinical course and prognosis of disease. Evidence for immunological disorders serves as a basis for using different immunotherapies in the complex treatment of cancer patients. Present-day oncological care uses a great variety of immunomodulators; however, preferable treatment includes cytokines (interferons, interleukins) whose application substantially enhances the efficiency of treatment in patients having certain tumors. Progress in biotechnology and gene engineering has contributed to the development of new trends in the therapy of tumors, namely the design of anticancer vaccines obtained by inserting the genes of cytokines and their receptors, tumor-associated antigens and other agents into the genome of a tumor cell. So far single clinical observations are yielding promising results.

[Contemporary possibilities of clinical immunology in oncology]. / Sovremennye vozmozhnosti klinicheskoi immunologii v onkologii.

Clinical immunology involves studies of immunological diagnosis and monitoring of cancer patients, their immunity and treatment-induced changes, including those associated with immunotherapeutical exposures, immunodiagnosis of leukemias, estimation of solid tumor markers. Immunophenotyping of peripheral blood cells from patients with solid tumors treated with chemo- and immunotherapy has elucidated some relationships to evaluate the efficiency of treatment. Detailed characterization of the structure of leukemic clone has promoted development of individual treatment regimens. The authors' experimental studies will evaluate the effects of cytokines on the mechanisms of multidrug resistance caused by mdr-1 gene hyperexpression and on apoptosis processes diminished by bcl-2 gene hyperexpression.

[Change in the nonspecific cytotoxic activity of the lymphocytes depending on the method of their isolation]. / Izmenenie nespetsificheskoi tsitotoksicheskoi aktivnosti limfotsitov v zavisimosti ot metoda ikh vydeleniia.

The authors present the results of comparative study of nonspecific cytotoxicity of lymphocytes, obtained by different methods, on the cells of cultivated melanoma. The lymphocytes were obtained from defibrinated and heparinized blood of normal donors on a column made of cotton and of phycoll-verographine mixture. Lymphocytes obtained on the latter proved to possess greater cytotoxicity than those obtained on the former. Lymphocyte suspensions from the column, containing platelets were less toxic than lymphocyte suspensions without any platelets. Lymphocytes obtained on the column were more toxic against the melanoma cells of the 22nd--29th passage than against the cells of the 35th--41st passage. This regularity was more pronounced for lymphocyte-platelet suspensions.