Hodgkin's lymphoma (HL) patients were investigated before and during chemical and radiation therapy. The properties of peripheral blood lymphocytes of the HL patients before treatment have been compared with healthy donors and the patients during the treatment. The genetic damage--frequency of cells with micronuclei (MN), the level of DNA single- and double-strand breaks (SSB and DSB), DNA-protein cross-links (DPC) have been studied. Biochemical and physiological parameters have been compared as well: the concentration of reactive oxygen species (ROS), the ability to the adaptive response induction. The radiosensitivity of lymphocytes in vitro exposed to the 1 Gy irradiation has also been determined (by MN test). It was shown that in Hodgkin's lymphoma patients' lymphocytes (in comparison with healthy donors) the frequency of cells with MN does not change, the level of SSBs and DSBs increases, the amount of DPC does not change, and ROS concentration (on average) significantly increases because of the part of the population that have high ROS content. The ROS concentration decreases to control level, the frequency of cells with MN increases, the level of DSBs does not change but the level of DPCs (which prevents the determination of DSB) increases in the patients during treatment. It was also discovered that lymphocyte radiosensitivity correlates with the MN cells frequency before treatment and the ROS concentration. These results make it possible to suppose that the high MN frequency and high ROS concentration in Hodgkin's lymphoma patient lymphocytes (before treatment) can serve as prognostic factors for the effectiveness of radio and chemical therapy.

The molecular-cellular parameters complex has been studied on the blood lymphocytes of malignant Hodgkin's lymphoma (HL) patients: the frequency of cells with micronuclei (MN) and chromosome aberrations; the level of DNA single and double strand breaks - OR and DR DNA (DNA comet assay), oxidative status--the content of reactive oxygen species (ROS) by using nonfluorescent dye that is oxygenated in the cells to fluorescent reagent and detection of fluorescence intensity after there. It was shown that the patients with LH had the increased level of DR and OR DNA, the increased frequency of cells with chromosome aberrations and the number of aberrations per cell was increased too. The concentration of ROS is increased too for the most individuals with intoxication. In the process of the chemical and radiation therapy the increase of OR DNA level, the frequency of the cell with MN has been registered. The ROS concentration correlates with the level of DNA-strand breaks. So the blood lymphocytes of HL patients before treatment differ from the lymphocytes of healthy donors. The damage of genome and the change of oxidative status have been observed that can be additive markers for the HL diagnosis, their sensitivity to the treatment and the characteristic of lymphocytes changes by this disease.

The genome damage (the frequencies of cells with micronuclei (MN), chromosome aberrations, the level of DNA double-strand breaks (DSB DNA), the concentration of reactive oxygen species (ROS) and 28 immunological parameters have been studied on the blood lymphocytes of Chernobyl accident liquidators. The purpose of this article was the investigation of cytogenetic, molecular changes of blood lymphocytes of irradiated individuals 24 years after accident, examination it there are correlation between genome damage and immunological parameters. It was shown that in lymphocytes of liquidators the frequencies of cells with MN and with all type of chromosome aberrations didn't differ from the lymphocytes of nonirradiated individuals, but the frequency of chromosome aberration type was increased, the level of DSB DNA was increased too. The concentration of ROS is decreased. The percent of cytotoxic CD8(+)-T-lymphocytes, natural killer cells (CD16(+)-lymphocytes), CD3+ CD16+ CD56+ (NK-T-cells), that posses antivirus and antitumor activity--HLA-DR+, regulatory T-lymphocytes (CD4+ CD25+high) in liquidators significantly increases. The level of serum immunoglobulin (Ig A) significantly increases too. The index of immune regulation, meaning of phagocyte neutrophil (FAN) and macrophage activity decreases. In liquidators there are significant correlation between the frequencies of cells with MN and the content of regulatory T-lymphocytes (p < 0.05), between the concentrations of ROS and activated T-lymphocytes. More connection is on the tendency level (p < 0.10): the frequency of chromosome aberrations, the DSB DNA level with natural killer cells and regulatory T-lymphocytes; the frequency of cells with MN and DSB DNA and FAM. We can suppose that genomic instability induced by the liquidators of Chernobyl accident consequences 24 years ago manifests now as increased genome damage and oxidative status decrease that can result in imbalance of cells and humoral immune status, disturbancies of health.

Using a DNA-comet assay was shown that irradiation of human blood lymphocytes at G1 cell cycle with a low conditioning dose (5 cGy) induces an adaptive response (AR) manifested in reduction of the double-strand DNA (DSB) amount induced by challenging dose at 10 Gy. 24 h after conditioning irradiation (48 h after PHA addition) in cells irradiated at both conditioning and challenging doses a relative DBS amount was approximately 24% less in comparison to versus a control irradiated at challenging dose only. 48 h after adapting irradiation this index increased to approximately 35%, while 72 h after was decreased to approximately 29%. AR observed by us during 72 h after its induction did not accompanied by statistically significant changes in DBS repair enhancing. It is possible to assume that basic role in AR forming in lymphocytes under experimental conditions used by us playing the processes preventing radiation-induced DBS formation (antioxidant defense system activation, chromatin conformation changes ets).

On the blood lymphocytes of prostate cancer (PCa) patients before and during radiotherapy: DNA damage by DNA commet assay (DNA double strand breaks - DSB); the frequency of cells with micronuclei (MN) with cytokinetic cytochalasin block; the adaptive response induction by the additional irradiation of PHA stimulated lymphocytes in the doses of 0.05 and 1.0 Gy 24 h and 48 h after stimulation were studied. Changes of these parameters with the decreasing of prostate specific antigen (PSA) have been compared. PSA decreasing is an adequate of the radiotherapy efficiency. It was shown that in oncological patients the DSB level and the frequency of cells with MN have been increased. During radiotherapy (in 3 months) the DNA DSB level and the frequency of cells with MN is enhancing. The degree and direction change of these parameters coincide. It was discovered the significant correlations between the enhancing of DNA DSB level and the cell frequency with MN during therapy and degree of the PSA level decreasing. Then it was shown that when the cell frequency with MN before treatment is higher the radiotherapy efficiency is worse. These results can have great significance for the evaluation of the prognosis of the treatment efficiency. The investigation of lymphocytes for the adaptive response ability has shown that in the patients with the pronounced adaptive response before radiotherapy the decrease of PSA level during treatment was not significant (in mean 3.5-3.6 ng/ml); when the adaptive response is absent or the phenomenon of enhanced radiosensitivity was observed the PSA level (in the most cases) was decreased very essential (in mean 0.07 ng/ml). We can suppose that prognosis of the treatment efficiency of the prostate cancer patients with the pronounced adaptive response in blood lymphocytes will be worse.

У лимфоцитов крови больных раком предстательной железы (РПЖ) до и во время лучевого лечения изучали: поврежденность ДНК методом ДНК-комет (двунитевые разрывы ДНК – ДР ДНК); частоту клеток с микроядрами (МЯ) при использовании цитокинетического блока цитохалазином В; индукцию адаптивного ответа при дополнительном облучении ФГА стимулированных лимфоцитов в дозах 0.05 и 1.0 Гр через 24 и 48 ч после стимуляции. Изменения этих показателей сравнивали со степенью снижения уровня простатспецифического антигена (ПСА), что является адекватным критерием эффективности лечения. Обнаружено, что в лимфоцитах онкологических больных повышен выход ДР разрывов ДНК и частота клеток с МЯ. При проведении лучевой терапии (в течение 3 мес) уровень ДР ДНК и частота клеток с МЯ возрастают. Степень и направленность изменения этих показателей совпадают. Были обнаружены достоверные корреляционные зависимости между степенью увеличения выхода ДР ДНК и частоты клеток с МЯ во время лечения и степенью снижения уровня ПСА. Показано, что чем выше исходная частота лимфоцитов с МЯ (до лечения), тем хуже эффективность лечения. Эти результаты могут иметь большое значение для определения прогноза эффективности лечения. Исследование способности лимфоцитов к индукции адаптивного ответа показало, что у больных с выраженным адаптивным ответом до лечения отмечается менее значительное снижение уровня ПСА во время лечения (в среднем до 3.5–3.6 нг/мл); при отсутствии адаптивного ответа или индукции повышенной радиочувствительности уровень ПСА в большинстве случаев снижается весьма существенно (в среднем до 0.07 нг/мл). Можно полагать, что прогноз эффективности лечения больных РПЖ с адаптивным ответом будет хуже.

Методом ДНК-комет было показано, что облучение в малой адаптирующей дозе (5 сГр) лимфоцитов крови человека в стадии G1 клеточного цикла индуцирует адаптивный ответ (АО), проявляющийся в снижении индукции двунитевых разрывов ДНК (ДР ДНК) проявляющим облучением в дозе 10 Гр. Через 24 ч после адаптирующего облучения (48 ч после добавления ФГА) в облученных в адаптирующей и проявляющих дозах клетках относительное количество ДР ДНК меньше на 24% по сравнению с контролем, облученным только в проявляющей дозе. Через 48 ч после адаптирующего облучения этот показатель увеличивается до 35%, а через 72 ч снижается до 29%. Наблюдаемый нами в течение 72 ч после его инициации AO не сопровождается статистически достоверным изменением эффективности репарации ДР ДНК. Можно полагать, что основную роль в формировании АО в лимфоцитах, при использованных условиях эксперимента, играют процессы, предотвращающие образование радиоиндуцированных ДР ДНК (активизация систем антиоксидантной защиты, изменения конформации хроматина и т.д.).