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.

New approach to vital analysis of functional activity in ABC transporters (markers for multidrug resistance) in solid tumors by the method of flow cytofluorometry.

We developed and described a new approach to vital analysis of functional activity of multidrug resistance markers (ABC transporters) in intact biopsy specimens from human solid tumors by the method of flow cytofluorometry. The algorithm of the study underwent revision, and the cell suspension was obtained in the final stage. Intensification of intracellular doxorubicin accumulation (fluorescence) and increase in the number of fluorescent cells and total fluorescence of doxorubicin-accumulating cells produced by ABC transporter inhibitor sodium azide served as the criteria of expression of these transporters in tumor tissue. Informative value of changes in various parameters of doxorubicin fluorescence is discussed. The increase in the count of fluorescent cells in the suspension of tumors cells after treatment with the inhibitor indicates the presence of tumor cells absolutely resistant to this preparation. The proposed method is technically simple, suitable for structurally different tumors, requires small amounts of the biopsy material and, therefore, can be used for routine analysis. The results of our analysis and spectrofluorometric assay of ABC transporters agree very closely, which suggest that this method is adequate for the purpose.

[Results of a 5-year screening for prostatic cancer]. / Itogi 5-letnego skrininga raka predstatel'noi zhelezy.

The results of 5-year screening (1996-2000) for prostatic cancer in 1129 males 40 to 80 years of age are presented. The examination included: measurement of blood levels of prostate-specific antigen (PSA), finger rectal examination, transrectal ultrasonic examination (TRUE) and, on demand, biopsy of the prostatic gland. Prostatic cancer was diagnosed in 1.5, 2.2 and 16% patients having PSA levels of 0-4.0, 4.0-10.0 and 10.0-30.0% ng/ml, respectively. At finger rectal examination prostatic cancer was suspected in 8% examinees, only in 33% of them the diagnosis was verified morphologically. By TRUE evidence 7% examinees were suspected and in 44.3% of them prostatic cancer was confirmed. Thus, biopsy proved necessary in 172 cases of 1129 examinees. In 64 (5.7%) males prostatic cancer was diagnosed and confirmed. Early prostatic cancer in the screened men and those consulted in the outpatient department of the National Cancer Research Center was detected in 77.7 and 22% men, respectively. The conclusion is made that men over 50 years of age should undergo prophylactic examination of the prostatic gland once a year.

[Inhibition of ABC-transporter(s)' function in non-small cell lung cancer cells by platinum drugs]. / Ingibirovanie funktsii ABC-transporterov v kletkakh nemelkokletochnogo raka legkogo pri vozdeistvii preparatov platiny.

With an account of the literature data that platinum drugs react with many cellular targets, including ATP and proteins, the authors suggested that disturbance of the function of energy-dependent ABC-transporters (markers of multidrug resistance, MDR) under the effect of platinum drugs could be a cause of increased efficacy of MDR agents (agents, MDR to which is developed by the classical mechanism) when used in combination with platinum drugs even in the treatment of multidrug resistant lung cancer. The cisplatin and carboplatin effect on accumulation of MDR doxorubicin in cells of non-small cell cancer was studied by flow cytometry with the use of biopsy specimens. The MDR phenotype of the tumors was determined by a change in doxorubicin intracellular accumulation under the action of the ABC-transporter(s)' inhibitors: verapamil and genistein (specific inhibitors of Pgp and MRP respectively) and sodium azide (an inhibitor of all energy-dependent ABC-transporters). The MDR phenotypes, i.e. Pgp-MRP+ or Pgp+MRP+, were detected in all the tumors investigated. Two types of changes in doxorubicin intracellular accumulation under the action of the inhibitors and the platinum drugs were shown: (a) an increase in doxorubicin cytoplasmic accumulation and (b) a change in subcellular distribution of the anthracycline (increased accumulation of doxorubicin in the cell nucleus and its higher binding to DNA). Cisplatin and carboplatin had an inhibitory effect on ABC-transporter(s) in all the tumors investigated but the effect of carboplatin was less pronounced. It was concluded that cisplatin and carboplatin stimulation of doxorubicin intracellular accumulation, as well as a change in subcellular distribution of the anthracycline under the action of the platinum drugs (increased doxorubicin accumulation in the cell nucleus) in multidrug resistant lung tumors could be at least partly explained by inhibition of the MDR transporter(s)' function. The results could provide a basis for the use of the sequential combination cisplatin (or carboplatin)-->doxorubicin in the treatment of multidrug resistant lung cancer.

[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.

[The role of epithelial antigens in diagnosis and staging of breast cancer]. / Rol' épitelial'nykh antigenov v diagnostike i stadirovanii raka molochnoi zhelezy.

The study of different epithelial antigen expression has been performed in 183 breast cancers. In 73 patients regional lymph nodes were studied as well. Panepithelial antigen Egp-34 (Mab HEA-125) was expressed in 100% of primary tumors and metastases. Antigen MUC-1 (Mab ICO-25) was identified in 93% of breast cancers. Monomorphic type of expression in tumor cells was typical for Egp-34, MUC-1 being in certain cases expressed as a proportion of cells. Additional immunohistochemical study of regional lymph nodes with Mab to Egp34 and MUC-1 provides a 10% increase in the rate of breast cancer metastases detection compared to histological examination alone. The carcinoembryonic antigen (CEA) was useful in identification of metastases only in CEA-positive breast cancers.

[Nitrullin -- a new original Russian drug of the nitrosomethylurea group]. / Nitrullin -- novyi original'nyi otechestvennyi preparat iz gruppy nitrozomocheviny.

Hematologic thrombopenia and leukopenia formation limits use of nitrullin as a toxic hazard. The drug showed moderate effect in treating inoperable non-small cell cancer of the lung and satisfactory end results. The treatment had marked symptomatic effect in patients with this cancer and, as a consequence, improved the quality of life. Nutrullin had immuno-modulating effect. Its application alone or in combination with VPN showed good results in the management of small-cell cancer of the lung.

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.

[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.

[An evaluation of the prognostic significance of antigen CD95(Fas/APO-1) expression on the cells of patients with a myelodysplastic syndrome, acute myeloid leukemia and chronic myeloleukemia]. / Otsenka prognosticheskoi znachimosti ékspressii antigena CD95(Fas/APO-1) na kletkakh bol'nykh s mielodisplasticheskim sindromom, ostrym mieloidnym leikozom i khronicheskim mieloleikozom.

AIM: The expression of CD95(Fas/APO-1) antigen was studied on bone marrow cells of 19 MDS patients, peripheral blood blast cells of 15 acute myeloid leukemia (AML) patients, blast cells and granulocytes of 68 patients with chronic myeloid leukemia (CML)--24 in chronic, 9 in accelerated phase and 35 in blastic crisis (BC)--by indirect surface immunofluorescence assay using flow cytometry (FACScan, Becton Dickinson, USA). RESULTS: CD95(Fas/APO-1) antigen was revealed on bone marrow cells of 8 out of 19 (36.8%) MDS patients; the percentage of antigen-positive cells was 38.1 +/- 19.2%; on 45.5 +/- 22.8% of cells in 6(45%) of 15 AML patients. Fas/APO-1 antigen was totally absent in CML chronic stage; its expression was found in 34% (12 of 35) of our patients with CML BC on peripheral blood blasts and in 56% (5 of 9) on peripheral blast cells of CML patients in acceleration phase. CONCLUSION: The data on overall survival of CD95-positive MDS patients suggest that the presence of Fas antigen is a favorable prognostic sign for patients with MDS. The patients from CD95-negative group represent a risk group both for survival and AML transformation. In CML BC group the survival does not depend upon Fas-antigen expression.

[An analysis of preliminary results in screening for prostatic cancer]. / Analiz predvaritel'nykh rezul'tatov skrininga raka predstatel'noi zhelezy.

450 males aged over 50 years free of urological symptoms were screened for prostatic cancer using three techniques; finger rectal examination (FRE), transrectal ultrasound investigation (TUI), assay for prostatic specific antigen in the serum (SPSA). SPSA quantities under 4 ng/ml, 4-10 ng/ml, 10-20 ng/ml, over 20 ng/ml were registered in 206(45.8%), 135(30%), 69(15.4%) and 40(8.8%) patients, respectively. Detectability of prostatic cancer increases by 33,37.9, 45.5, 69.2% due to TUI, FRE, TUI + FRE, all the three methods, respectively. Prostatic biopsy was needed in 102 (22.7%) cases. From the 450 examinees, prostatic cancer was diagnosed in 25 (5.6%). SPSA was high in all of them, higher than 10 ng/ml in 92%. 20 (80%) of 25 patients with cancer had early stages of the disease (TI-2). The study is going on.

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.