[Are the properties of peripheral blood lymphocytes of patients with prostate cancer connected with the efficiency of radiation therapy?].

The genome damage (frequency of cells with micronuclei and chromosome aberrations), concentration of reactive oxygen forms (ROS), markers of lymphocytes activation, expression of proliferation (CD69, Ki67) and proapoptotic antigen (CD95), as well as the ability to adaptive response have been investigated in blood lymphocytes of healthy donors and patients with prostate gland cancer. The influence of hormone-therapy on lymphocytes properties and connection between the parameters studied with the effectiveness of treatment, which was estimated by the level of prostate specific antigen (PSA), have been investigated. It was discovered that the genome damage to the patients with prostate gland cancer lymphocytes does not differ from control. The increase of the ROS level and decrease of radiosensitivity (irradiation of isolated lymphocytes in vitro at a dose of 1 Gy) are observed but they are insignificant. The content of the cells expressing CD69 and CD95 markers doesn't change but the expression of proliferative activity marker Ki67 in cells decreases. Radiosensitivity of lymphocytes in patients with prostate gland cancer correlates with the CD95 markers expression--a higher radio sensitivity points to their predisposition to apoptotic death. The expression of the markers studied depends on the oxidative status--a high ROS level suppresses their expression. The hormone therapy applied before radiotherapy leads to the increase in radiosensitivity and decrease in ROS. As the MN test shows, the ability to adaptive response of the lymphocytes in patients with prostate gland cancer is increased as compared with lymphocytes of healthy donors but it is insignificant; moreover, hormones do not influence the ability to the adaptive response. The high oxidative status further the formation of the adaptive response. We suppose that the discovered correlation between the initial, before treatment, frequency of lymphocytes with micronuclei and treatment effectiveness, namely, the decreased number of damaged cells associated with the treatment efficiency, is very important for the treatment prognosis. The results obtained can be very important for the experimental justification and understanding a possible use of blood lymphocytes for the additional diagnostics of prostate gland cancer and prognosis for its successful treatment.

[Analysis of cellular bases of secondary thymic atrophy in irradiated mice by flow cytometry].

Using flow cytometry we looked for a thymocyte subpopulation responsible for the development of thymus urgent recovery and secondary atrophy in sublethally irradiated mice (4 Gy). It was expected that the number of these cells would grow before the urgent recovery peak and would drop during secondary atrophy. It was found out that DN3 thymocytes were the best for these criteria. The DN3 stage of thymocytes development is characterized by the rearrangement of the major portion of T-cell receptor genes. On the basis of this finding we have discussed the possibility of secondary atrophy correction using IL-7.

[Dynamics of T-Cell receptor gene rearrangement and T-lymphocytes migration from thymus during post-radiation regeneration].

Recovery and migration of T-cells from the thymus to the secondary lymphoid organs in mice after sublethal gamma irradiation were investigated by measuring T-cell receptor excision circles (TRECs). The TRECs level practically represents the cellularity of thymus, in particular it correlates with the quantity of T-cells which have rearranged TCR genes and express the receptor complex CD3-TCR. So, TRECs can be considered as one of the markers of these cells. TREC-containing cells form a subset of recent thymic emigrants in the secondary lymphoid organs. After a significant TREC decrease in the lymph nodes within the early phase (4 days) after irradiation, we registered the increase of their number during urgent organ recovery due to T-cell migration from the thymus (the maximum is on the 10th day). The secondary thymic atrophy is accompanied by a weakening migration of the T-cells containing TRECs to lymph nodes. A significant TREC increase in the spleen was registered on the 4th day after irradiation. The rest of the recovery period. (up to 60 days) is characterized by the low TREC level. Thus, determination of TREC level allows obtaining additional information about recovery and migratory processes in lymphoid organs during post-radiation regeneration.

[Cytogenetic and immunological characteristics of stimulated human peripheral blood lymphocytes are connected with cell proliferation rates].

In the current study the authors investigated the connection between the proliferation rate of stimulated hu- man peripheral blood lymphocytes with some other characteristics of the population: expression of immunological markers, spontaneous genome damage, radiosensitivity. The population's proliferation index (PI) was taken as a measure of the rate. It was calculated using the composition of a cell population, which was cyto- kinesis-blocked with a cytochalasin B. If the genotoxic action is absent, the PI does not correlate with the spontaneous frequency of cells with micronuclei or with cell radiosensitivity, but is tightly linked with immunological indexes. It has been determined that after stimulation the level of marker-positive cells (CD25, CD69 and Ki67) is closely related to PI and is greater in the populations with lower proliferation rates. Irradiation of a cell culture 48 h after stimulation at a dose of 1 Gy leads to a correlation between PI and radiosensitivity, measured directly after the irradiation and in the same time frame as the PI measured in the non-irradiated population. The irradiated population's PI is not connected with the level of marker expression.

[Individual variability of immunological markers, radiosensitivity and oxidative status in blood lymphocytes of Moscow residents].

Expression of activation (CD69) and proliferation (Ki67) markers, their connection with each other, with the oxidative status (reactive oxygen species--ROS) and with radiosensitivity (determined by micronucleus test) have been studied on stimulated blood lymphocytes from Moscow inhabitants. It was shown that the content of T-lymphocytes with the expressed CD69 and the content of T-lymphocytes with the expressed Ki67 markers correlate (r = 0.571; p = 0.0004). We can suppose that expression of the CD69 marker (24 h after PHA stimulation) is needed for the cell cycle progression, but it is not enough for the high expression of Ki67 markers 48 h after stimulation (DNA synthesis phase). It was discovered that T-lymphocytes with the CD69 marker or T-lymphocytes with the Ki67 marker are connected by the negative correlation with the frequency of irradiated cell with micronucleus (MN) r = -0.487; p = 0.010; r = -0.440; p = 0.008, respectively. So we can suppose that lymphocyte radiosensitivity decreased with the increase of expression activation and proliferation markers. It was shown that radiosensitivity determined by MN test is not connected with the oxidative status determined by the reactive oxygen species content including superoxide anion radicals. It is possible to explain by the fact that the ROS concentration has been determined in non-stimulated lymphocytes, but frequencies of cells with MN - in the stimulated cells 48 h after stimulation. Using separate analysis of individual differences by the studied parameters that were determined in the same people, it was shown that individual differences are high enough in the same cases. For example, the radiosensitivity when cells were irradiated 48 h after stimulation, ROS concentration, cell content with activation and proliferation markers. In conclusion, we can say that we failed to find important correlation between the parameters studied. However, the presence of individual differences in the marker expression, the frequency of MN cells, the oxidative status in the usual inhabitants, typical donors in Moscow, is very important.

[Changes of thymocyte differentiation, proliferation and apoptosis induced by syntetic peptides].

The changes in the processes of differentiation, proliferation and apoptosis were studied in the culture of human cortical thymocytes after cell exposure to T-32 and T-38 bioregulator peptides. T-32 and T-38 peptides were shown to enhance the differentiation of immature cortical thymocytes (CD4+CD8+) into T-regulatory cells by increasing their proliferate activity and decreasing the level of apoptosis. Moreover, these peptides were found to stimulate the proliferative and antiapoptotic activity of mature T-regulatory (CD4+CD25+) cells.

[T-lymphocyte precursor count in the thymus and the development of lymphomas in AKR/J-strain mice]. / Soderzhanie predshestvennikov T-limfotsitov v timuse i razvitie limfom u myshei linii AKR/J.

The content of T-cell precursors is increased in the thymus of AKR/J mice. Thymus also contains cells carrying markers of pre-T (SC-1-antigen) and T-(Thy-1-antigen) cells simultaneously. The level of SC-1+-cells grows just before the lymphoma appearance in the AKR/J thymus. Lymphoma cells express both SC-1 and Thy-1-antigens, i. e. they are a transitional form between pre-T- and T-cells. It is suggested that accumulation of SC-1+-Thy-1+ cells in the mouse thymus is an essential condition for the development of virus-induced lymphomas.

[Radiation and immunity. Contemporary view of old problems]. / Radiatsiia i immunitet. Sovremennyi vzgliad na starye problemy.

Some key problems of radiation immunology which were formulated during last decades have been considered. The possibility or their decision appeared at recent time proceeding from the new view on the ionizing irradiation as a source of biologically significant signals. From that point of view some experimental data concerning radiation effects on the maturation and selection of lymphocytes, contact interaction of immune system cells and cytokine network were reviewed. It was concluded that the disturbances of the spatial and temporary organization and integrative functions of immune system were the results of the non-adequate radiation-induced signalling.

[Radiation and immunity. Interference of ionizing radiation with key immune processes]. / Radiatsiia i immunitet. Vmeshatel'stvo ioniziruiushchikh izluchenii v kliuchevye immunnye protsessy.

Recent evidences of the interference of ionizing radiation into the intimate immune processes are presented in the review. gamma-irradiation induces some events in V-gene rearrangement in T cells. Low-dose irradiation can result in the thymocyte differentiation and activation. Immediately after irradiation stromal thymic cell activity are stimulated and in the later stages it is depressed. Irradiation induces an expression of some functionally important molecules on a surface of the cells of immune system and thus influences the processes of cell interaction, costimulation, adhesion and transvascular migration. It is shown that many events of the signal transduction pathways resulting in radiation-induced and activation-induced apoptosis of lymphocyte are general. This data evidence that the possibility of the choice between the death and activation exists for the cells which are undergone to action of ionizing radiation.

[The effect of the local irradiation of areas of the thymus, hypothalamus/hypophysis and gonads in mice on the cellularity of the autologous and transplanted thymus]. / Vliianie lokal'nogo oblucheniia oblastei timusa, gipotalamusa/gipofiza i gonad myshei na kletochnost' autologichnogo i transplantirovannogo timusa.

Mice were grafted with syngeneic neonatal thymus 1-3 days or 7.5-8 months after local irradiation of thymus, hypothalamus/hypophysis and gonads. Thymus weight and thymocyte number were measured in autologous and grafted thymus 35-40 days after the transplantation. Thymus transplantation induces significant decrease in weight and cellularity of both autologous and grafted thymus. Irradiation of autologous and grafted thymus in doses of 1 and 10 Gy abrogates this effect. Irradiation of hypothalamus/hypophysis and gonads induces decrease of weight and cellularity of autologous and grafted thymus. Irradiation of gonads does not abrogate inhibitory effect of thymus transplantation. Local irradiation of all three regions decreases titre of serum thymic activity and influence concentrations of some hypophysis and adrenal hormones.

[Effect of ionizing radiation on the thymus stroma of embryos and newborn mice]. / Deistvie ioniziruiushchei radiatsii na stromu timusa émbrionov i novorozhdennykh myshei.

The lobes of thymus of newborn mice irradiated on 17 day of embryonic life or on 1 day after the birth were transplanted under the renal capsule of adult mice. The recipients were thymectomized, lethally irradiated and reconstituted by syngeneic bone marrow cells treated by monoclonal anti-Thy-1-antibody and complement 1 month before the transplantation of thymus. Colonization of the thymus transplants, number and functional activity of T cells in peripheral lymphoid organs of recipients were measured 30 day after the transplantation of thymus. The colonization of the thymus transplants was decreased after irradiation of the thymus donors in doses of 6 and 8 Gy. The number of L3T4+ cells (T helpers) in the lymph nodes of the recipients and the level of the humoral immune response on thymus-dependent antigen were decreased after the exposure of the thymus donors to dose of 1 Gy or higher and the level of graft-versus-host reactivity of lymph node cells--after the dose of 2 Gy and higher. The degree of the suppression of the humoral immune response was higher when the donors of thymus were irradiated in embryonal period of development than after the birth. Thus the damage of the components of thymic microenvironment responsible for the T cell development can be arisen by relatively low doses of ionizing irradiation.

[Functionally active T-cells and T-lymphocyte precursors in the thymus of newborn mice irradiated in the intrauterine period of development]. / Funktsional'no aktivnye T-kletki i predshestvenniki T-limfotsitov v timuse novorozhdennykh myshei, obluchennykh vo vnutriutrobnom periode razvitiia.

Mice were irradiated in dose of 2 Gy in 14 or 17 days of gestation. Irradiation retarded the increase of cell number in developing thymuses but in the day of birth the number of thymocytes was normalized. In normal development SC-1+ cells (T cell precursors) disappeared from the thymus immediately before the birth. After the irradiation they persisted in the newborn thymus. Mitogenic responses of newborn thymocytes on the action of thymic peptides and T cell mitogens were decreased after the fetal irradiation (adult irradiation enhances mitogenic response of thymocytes). The helper activity of thymocytes in humoral immunity was decreased after the fetal irradiation in more degree and helper activity in splenic colony-formation--in less degree than after the adult irradiation. The consequences of the irradiation in 14 and 17 days of gestation were similar.

[The state of classical and alternative complement activation pathways in persons who participated in the cleanup of the aftereffects of the accident at the Chernobyl Atomic Power Plant]. / Sostoianie klassicheskogo i al'ternativnogo putei aktivatsii T-limfotsitov u likvidatorov posledstvii avarii na ChAES.

The levels of T-cell activation through classical and alternative pathways were studied in persons participated in cleaning-up operations at Chernobyl N. P. P. Proliferation response of T-cells on the action of monoclonal antibody anti-CD3 (classical activation) as well as response on phytohaemagglutinin was partially decreased just as response on autologic erythrocytes in combination with phorbol 12-myristate-13 acetate (CD2-dependent alternative activation) was completely suppressed in all affected persons. Addition of interleukin 2 did not restore T-cell responses on both anti-CD3 and alternative stimulation. The different degree of decrease of T-cell responses on anti-CD3 and CD2-dependent alternative stimulation is not a result of respective alterations of cell surface CD3 and CD2 expression.

[A comparative evaluation of the content of T-lymphocyte subpopulations, alpha 1-thymosin and autoantibodies to epithelial thymic cells in the personnel in the 30-kilometer control zone of the accident at the Chernobyl Atomic Electric Power Station]. / Sravnitel'naia otsenka soderzhaniia subpopuliatsii T-limfotsitov, al'fa 1-timozina i autoantitel k épitelial'nym kletkam timusa u personala 30-kilometrovoi zony kontrolia avarii na Chernobyl'skoi AES.

A significant decrease in mean number of CD5+, CD8(+)-lymphocytes in persons, who worked in 30-km zone of Chernobyl nuclear power station was revealed. A significant increase in percent of CD5+, CD4(+)-cell percents was observed in workers, who worked for 1,2-2,5 years in zone, but absolute number, were decreased comparing a control and data received in people, who have just arrived to work in 30-km zone. The positive correlation exists between the percent of lymphocytes and years of service in 30-km zone. The lower level of alpha 1-thymosine was revealed in serum of the persons, who worked in zone for 4.5-5 years than data received in people, who worked for 0.5 year. Increase level of serum autoantibodies reacting with thymic epithelial cell was detected in men, who worked in zone for 3-3.5 years. In persons, who worked more 5 years and have just arrived in zone identical data were received.

[Dynamics in changes of a1-thymosin level under irradiation of mice thymus in vivo and in vitro: relation to the antibody-antigen production of epithelial thymus reticulum]. / Dinamika izmenenii urovnia a1-timozina pri obluchenii timusa myshei in vivo i in vitro: sviaz s produktsiei autoantitel k antigenam épitelialnogo retikuluma timusa.

In 3- and 9-month experiments with mice, a study was made of the effect of radiation on serum alpha 1-thymosine concentration after whole-body irradiation and local exposure of the thymus at doses of 1-20 Gy. The effect of 137Cs-gamma-rays on the in vitro cultured thymus stroma cells, with respect to alpha 1-thymosine secretion, and the influence of local irradiation of the thymus of production of autoantibodies that react with epithelial thymus cells were studied. Both whole-body irradiation and local exposure of the thymus were shown to cause changes in the alpha 1-thymosine content of the blood plasma. The direction and dynamics of the changes observed are different with whole-body and local exposure. Irradiation of cultured thymus cells of mice causes alterations in alpha 1-thymosine secretion, that is, stimulation at a dose of 1 Gy and inhibition at higher doses. With respect to dose- and time-response, these changes are closer to those observed in alpha 1-thymosine concentration in mouse serum after whole-body irradiation than after local exposure of the thymus in vivo. At remote times after local irradiation of the thymus with doses of 1-10 Gy, autoantibodies are found in mouse serum that react with epithelial cells of the thymus stroma. Autoantibodies are absent at doses of 15 and 20 Gy.

[An immunomorphological study of the effect of ionizing radiation on the nonlymphoid structures of the mouse thymus using monoclonal antibodies]. / Immunomorfologicheskoe izuchenie vliianiia ioniziruiushchei radiatsii na nelimfoidnye struktury timusa myshei s pomoshch'iu monoklonal'nykh antitel.

Expression of cytoskeleton components revealed with monoclonal antibodies was increased in epithelial cells of mouse thymus one day after total gamma-irradiation with doses of 0.5-4.0 Gy, while after the doses of 10 and 15 Gy it was decreased. The antigen expression was reversed to initial level 14 days after the dose of 4 Gy; thymocyte depletion was progressed and continuous massifs of epithelial cells was formed. The oval cysts and immunoglobulin deposits were appeared in thymus two months after irradiation with the dose of 4 Gy.

[Changes in the immune system of the victims of the action of the factors in the accident at the Chernobyl Atomic Electric Power Station. Their manifestations, nature and the possible sequelae]. / Izmeneniia v immunnoi sisteme postradavshikh ot deistviia faktorov avarii na ChAES. Proiavleniia, priroda, vozmozhnye posledstviia.

The hypothesis is formulated, which explains genesis of long-lasting disturbances in the immune system of the persons affected by factors of Chernobyl disaster. Immunological alterations which are displayed at the late time after action of radiation in doses 0.5 Gy or lower are not a result of direct damage of the cells of immune system by irradiation. Their development is more probably a result of appearance of some systemic conditions and factors in affected organism--such as hormonal disbalance and especially autoantibodies of different specificities, including those reactive with thymic epithelial cells. Autoantibodies of the last type induce the decrease of thymic hormone secretion which results in functional deficiency of T lymphocytes. This chain of events is similar to those occurring in aging and does not directly causes development of the clinical displays of immunodeficiency. Only irradiation in doses of 4-6 Gy or higher inducing the structural damage of thymic microenvironment can rouse the long-lasting T cell immunodeficiency with the clinical manifestations.

[The action of laser radiation on thymic hormone production]. / Deistvie lazernogo izlucheniia na vyrabotku gormonov timusa.

Irradiation of the epithelial cells from human thymus (cell line HTSC) by arsenid-gallium laser (0.89 nm, 1500 Hz) induces an enhancement of thymic hormone production. The concentration of alpha-1-thymosin was substantially increased on the days 1 and 5 in culture after irradiation. The increase of thymulin level in cultural supernatants was less pronounced and displayed after irradiation during 3-4 minutes. Local laser irradiation of thymus region of rats (5 minutes, 10 times) induce the increase of serum concentration of alpha-1-thymosin and the decrease of serum level of thymulin 2 days after irradiation.

[Serum levels of thymus hormones and autoantibodies to the thymus epithelium in inhabitants of regions contaminated with radionuclides]. / Soderzhanie gormonov timusa i autoantitel k épiteliiu timusa v syvorotke krovi zhitelei raionov, zagriaznennykh radionuklidami.

Increased level of serum thymic activity and normal serum concentration of alpha-1-thymosin were revealed in inhabitants of region polluted by radionuclides after Chernobyl accident (settlement Vyshkov, Bryansk region). In the same population the serum level of autoantibodies, reacting with thymic epithelial cell line was elevated. Similar but less prominent alterations were found in population of radiologically pure region (settlement Potchep). Revealed autoantibodies had mainly IgM isotype. The negative correlation exists between levels of autoantibodies and serum thymic activity. Sera of persons from the control group and one half of Vyshkov inhabitants reacted with thymic epithelial cells of both human and mouse origin; in other half of Vyshkov inhabitants species-specific autoantibodies were revealed. We propose that correlated increase of antithymic autoantibody level and decrease of serum thymic activity are the consequence and indicators of unfavorable ecological conditions; however the contribution of radiation factor to their induction can not be estimated on the basis of presented data.

[Quantitative interrelationships between the T- and B-cell populations of mouse lymphoid organs]. / Kolichestvennye vzaimootnosheniia populiatsii T- i B-kletok limfoidnykh organov myshei.

The number of thymocytes and splenic T-lymphocytes correlate negatively. After the treatment of mice with cortisol, this negative correlation changes for a positive one. It is proposed that the negative correlation between cortisone-sensitive thymocytes and splenic T cells is mediated by cortisone-resistant thymocytes. The negative correlations were also found between amounts of T-cells in mesenteric lymph nodes and in Peyer's patches, the positive correlations being seen between numbers of splenic and lymph nodes T-lymphocytes. No correlations between B-cell populations in different organs were revealed. Apparently, correlations between lymphocyte populations reflect some aspect of hierachic structure of the lymphoid system.