[Spontaneous cytotoxicity treated with anti-tumor drugs of splenocytes of rats on a model of hepatoma cell monolayer cultures].

A purpose of this research is the study of spontaneous cytotoxic activity of effector cells (EC) of the innate immunity of animals against target cells (TC) of cultured hepatoma. There are established differences in the cytotoxic potential of freshly non-activated mouse and rat splenocytes: TC exhibit resistance to splenocytes of mice C3HA and are exposed to active dose-dependent killing under the influence of splenocytes outbred rats. There were revealed two mechanisms of killing of clip-target by splenocytes of rats--secretory variant (Zajdel hepatoma) and the path of classical apoptosis (hepatomas HTC, MH-22a and BWTG3). Single intraperitoneal administration of anticancer drugs cyclophosphamide (CP) and 5-fluorouracil (5-FU) to animals inhibits secretory pathway, whereas the activity of the apoptotic mechanism is enhanced after administration of CF to animals and is unchanged under the action of 5-FU.

[Immunomodulating effect of cyclophosphamide on cytotoxic activity of rats and mice splenocytes].

This study examines the immunomodulating effect of cytostatic drug--cyclophosphamide (Cy)--on natural cytotoxic activity of rats and mice splenocytes. The cytotoxicity of the effector cells against the confluent monolayer cell lines of rat hepatoma Zajdela and HTC and mice hepatoma MH-22a was estimated by means the morphometric analysis. It was shown, that 48 h after single intraperitoneal injection of Cy produced a immunomodulating effect on the activity of splenocytes--suppressor action on cells of Zajdela hepatoma and immunopotentiating action on the target cells of HTC and MH-22a hepatomas. The possible mechanisms of immunopotentiating effect of Cy are discussed.

[The morphometric analysis of cytotoxical action of rats and mice splenocytes against confluent monolayer cell lines of hepatomas].

The present study was aimed to examine the possibility to use of the morphometric analysis for estimation of the total natural cytotoxic activity of rat and mice C3HA splenocytes against the cultured target cells which formed confluent monolayers. By means of this method we have revealed that two rat monolayer hepatoma cell lines -- HTC and Zajdela -- were sensitive to cytolysis mediated by rat effector cells, but not splenocytes of C3HA mice. The mild pretreatment with 0.5% paraformaldehyde of the rat splenocytes produced a significant decrease only in the cytotoxic activity against the target cells line Zajdela and don't affect the lysis of the other target HTC cells. These results suggest that the natural cytotoxic rat cells may mediate their lytic functions toward tested target lines by different ways. In the case of HTC cells as the targets the effectors induce death receptor-mediated apoptosis, as to target cells Zajdela they deliver lethal hit by perforin-granzyme exocytosis mechanism. The cultured cell monolayers of mice hepatoma -- MH-22a and BWTG3 -- cells showed under conditions of our experiments the resistance to cytolysis by homological effector cells; however, the hepatoma MH-22a cells were susceptible to killing mediated by rat cytotoxic splenocytes.

[The influence of tumor growth on natural cytotoxicity and activity of some lysosomal enzymes of human effector cells and the C3HA mouse splenocytes]. / Vliianie protsessa opukholevogo rosta na estestvennuiu tsitotoksichnost' i aktivnost' nekotorykh lizosomnykh fermentov éffektornykh kletok cheloveka i splenotsitov myshei linii C3HA.

In the course of malignant growth processes in patients with lung cancer, a decrease of natural cytotoxic activity of peripheral blood lymphocytes was observed. This process was accompanied by changes of activities of two lysosomal enzymes, arylsulfatase and acid phosphatase, suggesting participation of these enzymes in manifestation of effector functions of lymphocytes in cancer patients. The level of activity of granular enzyme, beta-glucuronidase, remained unchanged at all stages of disease. A study of natural killer activity of C3HA mice splenocytes after inoculation of transplantable hepatoma 22-a cells revealed a relative stability of the level of their cytotoxicity, and of the activities of lysosomal enzymes--arylsulfatase, acid phosphatase, alpha-mannosidase, acid lipase, N-acetyl-beta-D-glucosidase, and beta-galactosidase, beginning from the 3rd day after hepatoma implantation.

[Effect of immunosuppressors cyclophosphane and 5-fluorouracil on natural cytotoxicity and activity of lysosomal enzymes of splenocytes in C3HA mice]. / Vliianie immunosupressorov tsiklofosfana i 5-ftoruratsila na estestvennuiu tsitotoksichnost' i aktivnost' lizosomnykh fermentov splenotsitov myshei linii C3HA.

Effects of two immunosuppressors, cyclophosphane abd 5-fluorouracyl, used in clinical practice for treatment of oncological diseases, were assessed in respect to cytotoxicity and activity of several lysosomal enzymes located in splenocyte granules of C3HA mice. 48 h after a single intraperitoneal injection, both preparations produced a marked decrease in their cytotoxic activity, which was accompanied by a pronounced splenopathy. Both preparations were shown to decrease activity of arylsulfatase. Administration of cyclophosphane brought about the rise of activity of acid lipase as compared to control. Activities of acid phosphatase, alpha-mannosidase, beta-galactosidase, and N-acetyl-beta-D-glucosidase did not change after administration of the used immunosuppressors. It may be suggested that only arylsulfatase and acid lipase are involved in performance and(or) manifestation of the natural killer activity in splenocytes of the C3HA mice after their administration with cyclophosphane or 5-fluorouracyl.

[The response of peripheral blood lymphocytes from lung cancer patients to in-vitro treatment with recombinant interleukin-2]. / Izuchenie otveta limfotsitov perifericheskoi krovi bol'nykh rakom legkogo na obrabotku in vitro rekombinantnym interleikinom-2.

The cytotoxic activity of peripheral blood lymphocytes of 135 lung cancer patients was studied. A wide range of cytotoxicity values was shown--from 0 to 88%. During the randomized investigation of these patients no strong correlation was found between the levels of cytotoxicity and the cancer stages. A certain tendency of increasing the number of patients with weak cytotoxic activity upon progressing disease was followed. The lymphocytes with a strong natural killer activity did not react on the in vitro incubation with interleukin-2. The lymphocytes with a strong was obtained when the lymphocytes with the lowest cytotoxicity levels were incubated with this immunomodulator.

[The comparative characteristics of the immunogenic and immunomodulating activity of different antigenic preparations of the causative agent of Q fever Coxiella burnetii]. / Sravnitel'naia kharakteristika immunogennoi i immunomoduliruiushchei aktivnosti raznykh preparatov antigenov vozbuditelia Q-likhoradki Coxiella burnetii.

The immunogenicity of the Coxiella burnetii corpuscle antigen and two its components, obtained after the treatment with chloroform-methanol or extraction by trichloroacetic acid, was investigated. All the studied antigens were highly immunogenic. A strong correlation was discovered between the immunogenic and immunomodulating properties of the C. burnetii antigens. Cells of the mononuclear phagocytic system are the targets for stimulating action of the studied antigens. A cyclic increase of antibacterial activity and oxygen-dependent metabolism of the peritoneal cells was noted, in addition to the increase of the natural splenocyte cytotoxicity caused by the action of these antigens. The role of cytokines in rising the activity of peritoneal cells and splenocytes, caused by immunization of mice with the above antigenic substances, is discussed.

Prognostic value of non-MHC-restricted killer cell activity in lung cancer.

The prognostic value of peripheral blood non-MHC-restricted cytotoxicity against the myeloid leukaemic line K562 in lung cancer patients was studied. At the time of diagnosis and before operation, 57 patients with lung cancer were tested for cytotoxicity and subsequently followed for up to 4 years. In addition, 145 lung cancer patients, 30 patients with non-neoplastic lung diseases and 76 healthy donors were tested for cytotoxicity without the follow-up, in order to correlate the stage of lung cancer and the growth rate of tumours to the level of non-MHC-restricted cytotoxicity. On average, lung cancer patients had similar non-MHC-restricted cytotoxicity to the controls. However, patients with stage II-IV diseases showed an impaired activity, stages III and IV differing significantly from the controls. This result shows that the decline in natural killer (NK) activity is associated with tumour burden. Patients with slowly growing neoplasms had stronger cytotoxic activity than patients with fast or moderately progressing disease. In the follow-up study, the whole material of 57 patients showed only a slight correlation between cytotoxicity and survival: 42% of the patients with strong activity survived for more than 2.5 years, whereas 6% of the patients with weak activity did so. In stage I patients there was no correlation between cytotoxicity and survival, nor was there a correlation in patients with stages II-IV of the disease. Hence, in our group of patients the determination of cytotoxicity preoperatively yielded no prognostic information beyond that already available from staging. However, those stage II-IV patients that survived for 1 year or more after the diagnosis and cytotoxicity tests, showed a significant correlation between cytotoxicity and survival.

[The effect of antithymocyte serum on the natural cytotoxicity of C3HA mouse splenocytes in the early periods after 20-methylcholanthrene administration]. / Izuchenie vliianiia antitimotsitarnoi syvorotki na estestvennuiu tsitotoksichnost' splenotsitov myshei C3HA na rannikh srokakh posle vvedeniia 20-metilkholantrena.

The role of T-lymphocytes in natural cytotoxicity of splenocytes of C3HA mice after a single injection of 20-methylcholanthrene (20-MC) was investigated. A splenocyte suspension was treated with anti-T-cell serum and complement. This treatment was not shown to exert influence on the natural cytotoxicity of splenocytes within 1-13 days after injecting 20-MC.

Coxiella burnetii antigens may induce resistance to tumour growth.

Coxiella burnetii antigens stimulate the defence against growth of hepatoma 22a cells. The antigen-stimulated mice survived longer, they considerably later developed palpable tumours and showed a retarded tumour growth. The enhanced resistance to tumour growth may be explained by at least 2 interrelated phenomena; namely by the induction of interferon-like activity and an increased NK cell activity.

[The effect of macrophages on the natural killer activity of the splenocytes of C3HA mice in the early stages of 20-methylcholanthrene-induced carcinogenesis]. / Vliianie makrofagov na estestvennuiu killernuiu aktivnost' splenotsitov myshei C3HA na rannikh stadiiakh kantserogeneza, vyzvannogo 20-metilkholantrenom.

The influence of macrophages on NK cell activity of C3HA mice on 1, 7 and 13 days after single i. m. injection of 20-methylcholanthren was investigated. It is shown that macrophages significantly stimulate this activity on the 13th day after carcinogen application.

[Natural killer activity of the splenocytes in C3HA strain mice during 20-methylcholanthrene-induced carcinogenesis]. / Estestvennaia killernaia aktivnost' splenotsitov myshei linii C3HA v usloviiakh kantserogeneza, vyzvannogo 20-metilkholantrenom.

The induction of tumor in C3HA mice after intramuscular injection with 20-methylcholanthrene is accompanied by a decrease in natural antitumor resistance. This conclusion is based on the observation of the decreased natural killer activity per total number of splenocytes, from the time of carcinogen application till the appearance of tumor nodes.

[Characteristics of the morphofunctional changes in the liver of C3HA mice due to immunization with a homogenate of normal syngeneic liver]. / Kharakteristika morfofunktsional'nykh izmenenii pecheni myshi C3HA vsledstvie immunizatsii gomogenatom normal'noi singennoi pecheni.

Immunization of C3HA mice with homogenate of normal syngeneic liver at a dosage causing elevation of antitumor resistance damages the liver of recipients. The damage involves the formation of foci of necrosis and the alteration in activities of some tissue specific and embryonal enzymes. The damage is reversible; its degree and the rate of reverse depends on a dosage of homogenate injected.

[Mouse splenocyte sensitization to normal tissue antigens and natural killer activity. III. The effect of partial hepatectomy]. / Sensibilizatsiia splenotsitov myshei k normal'nym tkanevym antigenam i estestvennaia killernaia aktivnost'. III. Vliianie chastichnoi gepatéktomii.

Partial hepatectomy leads to both increasing of natural cell-mediated activity and sensibilization level (SL) of splenocytes of hepatectomized mice towards antigens of the syngeneic liver. The wave-like variability of SL was shown with sharp increase at 3, 6 and 9 days after operation. Natural killer activity was elevated on the 2nd and 10th days with a significant decrease on the 3-4th days after operation. It is assumed that the variability in the functional activity of splenocytes under study may characterized splenocytes of different populations.

[Sensitization of murine splenocytes to normal tissue antigens and natural killer activity. II. The effect of immunization with allogeneic and xenogeneic tissues]. / Sensibilizatsiia splenotsitov myshei k normal'nym tkanevym antigenam i estestvennaia killernaia aktivnost'. II. Vliianie immunizatsii allogennymi i ksenogennymi tkaniami.

Immunization of C3HA mice with allogeneic tissue (liver of DBA mice) leads to wave-like variability in the natural killer activity of their recipient splenocytes. This variability is similar to that of NK-activity due to syngeneic immunization and is characterized by a sharp increase on days 1, 3-4 and 6 after immunization. Xenogeneic immunization exerts no influence on NK-activity, if compared with the injection of 0.14 M NaCl.

[Sensitization of mouse splenocytes to normal tissue antigens and natural killer activity. I. The effect of immunization with normal syngeneic tissues]. / Sensibilizatsiia splenotsitov nyshei k normal'nym tkanevym antigenam i estestvennaia killernaia aktivnost'. I. Vliianie immunizatsii normal'-nymi singennymi tkaniami.

Immunization of C3HA mice with homogenized syngeneous liver led to sensibilization of splenocytes towards liver antigens and to the increase in natural killer cell activity towards K-562 cells. The dynamics of sensibilization and cytotoxicity in different time intervals after immunization has led to a conclusion about a correlation between the two above phenomena.

[Correlation of the mutagenic and carcinogenic activity of various aromatic amines]. / Korreliatsiia mutagennoi i kantserogennoi aktivnosti nekotorykh aromaticheskikh aminov.

Six aromatic amines were studied for their mutagenic activity using Salmonella typhimurium TA 100 and TA 98. 3,3'-dichlorobenzidine, ortho-tolidine, benzidine, 2-naphthylamine proved to be mutagenic, while aminotobias acid and 2,2',4,4'-tetraaminodiphenyl did not show mutagenic activity. Chronic experiments on mice and rats using these compounds revealed a close correlation between carcinogenicity and mutagenicity.

[Intercellular information transfer in the process of immunogenesis. IX. The induction of the synthesis of antiphage antibodies in lymphosarcoma cells of rats as affected by different fractions of immune nuclear RNA]. / O mezhkletochnoi peredache informatsii v protsesse immunogeneza. IX. Induktsiia sinteza antifagovykh antitel v kletkakh limfosarkomy krys pod vliianiem razlichnykh fraktsii immunoi iadernoi RNK.

Various fractions of the immune nuclear RNA were isolated from spleens of phage T2 immunized rats. The fractions were compared for their ability to induce anti-phage T2 antibody synthesis in transplantable lymphosarcoma cells. The most active proved to be the nuclear sap RNA and its subfraction with sedimentation constant of 10 S. The 4S and 26S subfractions RNA were less stable and in some experiments failed to induce antibody synthesis.

[Intercellular information transfer in the process of immunogenesis. VI. The induction of antiphage antibody synthesis in the cells of rat transplantable lymphosarcoma under the influence of splenic RNA from rats and mice immunized with phage T2]. / O mezhkletochnoi peredache informatsii v protsesse immunogeneza. VI. Induktsiia sinteza antifagovykh antitel v kletkakh perevivnoi limfosarkomy krys pod vliianiem RNK selezenki krys i myshei, immunizirovannykh fagom T2.

It has been proved that nuclear and cytoplasmic RNAs, isolated from spleens of T2 phage immunized rats and mice, can induce T2 phage antibodies in cells of the transplantable rat lymphosarcoma. With the nuclear RNA from rat spleens, the effect is persisting in a number of subsequent cell generations. The data presented are principally in accord with results of the authors' previous studies in which lymphosarcoma cells were treated with RNA extracted from spleens of rat immunized with sheep red cells. These results well compare with the authors' earlier advanced hypothesis suggesting a possible involvement of RNA in deblockation of genes responsible for the synthesis of the antibodies in question.

[Mutagenic action of nitroso compounds on Escherichia coli cells]. / Izuchenie mutagennogo deistviia nitrozosoedineniia na kletki Escherichia coli.

An attempt to induce some forward and back mutations in two Escherichia coli strains (his- and HfrH requiring thiamine) under the action of the carcinogenic nitrosamines--dimethylnitrosamine (DMN) and diethylnitrosamine (DEN)--is described. For this purpose the cells of E. coli were treated with 5% DMN or 1% DEN for 1 hour at 37 degrees C in 0.14 M NaCl. It was shown that the sensitivity of both strains to both nitrose compounds was not the same. DEN was 5-fold as toxic as DMN for the E. coli cells. DMN and DEN induced neither mutations of resistance to 10(-3) M valine, nor reversions in histidine-dependent strain. These mutations were obtained after the cells were treated with 0.1 M NaNO2. Lethal effects of DMN increased more than in 5 times and the toxicity of DEN did not change in hydroxylating mixture, in which nitrosamines derived to active compounds. Under these conditions both carcinogenes showed a mutagenic activity. DEN proved to be about twice as strong mutagenically as DMN. Thus, in our experiments we could see that DMN and DEN could induce both forward and back mutations in E. coli.