Individual immunological parameters in clean-up team members and patients with sequelae of acute radiation sickness 5 years after the effects of the Chernobyl accident]

A study was made of deviations, beyond 1 sigma and 1.5 sigma of a mean value (M) of a donor group, in individual immunological parameters (for instance, the number of CD5+, CD2+, CD4+, CD8+, CD25+ and B-cells; alpha 1-thymosin concentration; and autoantibody titers to antigens of epithelial reticulum cell cytoplasm) in patients suffered acute radiation sickness (ARS) and liquidators of Chernobyl NPP accident. The radiation damage to the immune system was reliably detected in the affected subjects examined: they exhibited a decrease in the alpha 1-thymosin level below M = -1.5 sigma and in absolute B cellularity below M = -1 sigma; and increase in the number of CD25+ cells and in the level of serum autoantibodies to antigens of thymus epithelial reticulum cell cytoplasm. When several parameters selected were examined simultaneously the frequency of recording the deviations in merely one of them markedly increased.     

The number of nucleoli in peripheral blood lymphocytes of the Chernobyl accident liquidators]

The number of nucleoli in lymphocyte nuclei was compared in the peripheral blood of the Chernobyl liquidators and non-irradiated persons (control). The former was significantly distinguished from the latter (p < 0.01) by the parameter "the number of nucleoli in lymphocytes", mean numbers of nucleoli per nucleus in these being 1.18 and 1.12, respectively. The increased number of nucleoli in lymphocytes of the Chernobyl liquidators may be associated with cytogenetic radiation effects.     

The connection between molecular-cellular parameters and immune status of liquidators after Chernobyl accident

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.     

Several indices of the peripheral immune system in clean up team workers and patients who survived acute radiation disease 5 years after effects of the radiation accident factors]

A group of liquidators and patients survived acute radiation sickness (ARS) was studied five years after the effect of radiation factors resulted from the Chernobyl A.P.S. disaster. Studied were the number of T-lymphocytes (SD2+, SD3+, SD5+), subpopulations SD16+ and SD25+, the content of SD16+ and SD25+ lymphocytes and B-cells, as well as the levels of serum IgM, A, G; determined was the concentration of serum alpha 1-thymosin. A decrease was noted in the number of T-cells of the examined patients, with respect to all markers studied: the number of SD4+ cells was found decreased in patients with ARS sequelae and normal in liquidators. The decrease in serum alpha 1-thymosin was a function of dose. Dynamics of changes in the T-cells of the examinees may reflect the new trends in the radiation response of T-lymphocytes which were not directly related to initial damages to the latter. The injury to the stroma of the thymus and especially to its epithelial cells may be the basis for later postirradiation damages to the immune system. The decrease in the concentration of alpha 1-thymosin in the blood serum of the examinee is the manifestation of the thymus hypofunction. In liquidators and patients with the ARS sequelae the number of SD25+ cells increased markedly indicating the existence of the source of endogenous cell activation. The content of B-lymphocytes in all studied groups of patients was reduced which may serve as a criterion of chronic stress.     

The epidemiological analysis of monitoring of the immune status in liquidators of consequences of the Chernobyl accident for early identification of risk groups and diagnostics of oncological diseases. Report 1

Ionizing radiation is one of major factors of risk of oncological diseases. A question about the frequency of malignant neoplasms (MN) and their early identification in the liquidators of consequences of the Chernobyl accident remains opened. In the present work, the results of long-term immunological monitoring of the liquidators of consequences of the failure at the Chernobyl Nuclear Power Plant (ChN PP) living in the Northwest region of Russia are analyzed; we also heve made an attempt to reveal the predictors of oncological diseases in this group of individuals. The frequency of the newly revealed MN cases in a cohort of the persons who took part in liquidation of consequences of the ChNPP failure and were followed-up in 1999-2009, has made up 89 cases per 1005 persons (8.856%), which somewhat exceeds general population indicators. Regarding the frequency of separate MN localizations, lung cancer, cancer of stomach and cancer of prostate gland predominated, which corresponds to the world's tendency of MN prevalence. It has been established that as early as 1-3 years before diagnosis of MN is confirmed in liquidators, a number of changes in the immune status comes to light: drop in percentage of CD3+ and CD4(+)-T-lymphocytes, B-lymphocytes to a lesser extent, decrease in the CD4+/CD8+ index, increase of the relative and absolute content of CD16(+)-lymphocytes, increase of absolute content of CD8(+)-T-lymphocytes, prevalence of CD3+16/56+(NK-T) cell over CD3-16/56+(NK) cells, rise of the activity of phagocytes. Patients with the presence of one or several of the above-mentioned signs should be attributed to the MN risk group for determination of tumor markers, thorough examination and dynamic observation.     

Characterization of immunity status in exposed residents of the Techa riverside villages 50 years after the onset of radiation exposure

The goal of the study was to assess the state of immunity in exposed residents of the Techa riverside villages 50 years, or more, after the onset of radiation exposure. 127 chronically exposed persons and 55 unexposed persons were studied. The mean dose to red bone marrow (RBM) was 0.69 Sv in exposed subjects, the mean dose to soft tissue was 0.07 Sv, the mean dose rate amounted to 0.10 Sv/yr to RBM and 0.02 Sv/yr to soft tissues in 1950. The state of the basic links of the immunity system (cellular, humoral, mononuclear phagocyte system, cytokine spectrum, etc.) was assessed using conventional methods. Exposed persons manifested a significant reduction in the absolute counts of CD3+, CD4+, CD 11b+, CD16+ lymphocytes in the peripheral blood, as well as an increase in the relative counts of CD8+. The group comprised of the Techa riverside residents demonstrated an increased immunoregulatory index (exposed individuals: 1.47; controls: 1.71, p = 0.001). An increased production of Immunoglobulin A and increased proportions of CD25+ lymphocytes were revealed in exposed individuals. Changes in the phagocytic activity of neutrophils and monocytes were insignificant, and were primarily associated with changes in the proportions of pagocytes in the peripheral blood stream. The state of the immunity in chronically exposed individuals at late time after the begin of exposure is characterized by a number of specific features reflected primarily on the cellular immunity. No relationship between immunity changes and accumulated exposure dose and dose rate were noted over the period of maximum radiation exposures (1950).     

Dynamics of immune status indices of liquidators in the remote period after the accident at the Chernobyl NPP

In the report there are present the results of monitoring of the state of immune status in the remote period after the accident at the Chernobyl Nuclear Power Plant among three groups of liquidators, which were from Moscow and the Moscow district, the Northwestern region of Leningrad district and from Krasnoyarsk territory. During the monitoring of the liquidators from different regions there was found phenotype of immune status with regional characteristics. In the Moscow district the clinical displays of immune insufficiency prevail and in the North western region the modifications of immune status prevail among liquidators. In all the groups we could see quantitative and immunoregulatory disbalance with the increase of parameters of cellular activity such as CD95+, HLA-DR+, T-activated lymphocytes, fluctuation cytotoxic cells (CD8+, CD16+), persevering disimmunoglobulinemia with differently directed modification serum of immunoglobulins at decrease B-lymphocytes. By the end of the twenty-year period after the accident we can see a similar phenotype of immune status in all the groups with increasing of the marker of late activation (HLA-DR+) and apoptotic activity (CD95+) and it is potentially dangerous regarding cancerogenesis. According to the basis of long-term monitoring there has been discovered the immune characteristic of proliferative syndrome for the first time. The monitoring of 2004 has revealed a huge similarity immune status at non-malignant growth at significant decrease CD25+ in the Northwestern region and some differences in common IgE.     

Elevated frequencies of hypoxanthine phosphoribosyltransferase lymphocyte mutants are detected in Russian liquidators 6 to 10 years after exposure to radiation from the Chernobyl nuclear power plant accident

This study was conducted to determine whether the frequency of hypoxanthine phosphoribosyltransferase (HPRT) deficient lymphocyte mutants would detect an effect of radiation exposure in a population of Russians who were exposed to low levels of radiation while working in 1986 and 1987 as liquidators cleaning up after the Chernobyl nuclear power reactor accident. The HPRT lymphocyte cloning assay was performed on peripheral blood lymphocytes collected between 1992 and 1996 from 142 liquidators and 66 Russian controls, and between 1989 and 1993 from 231 American controls. Russian and American controls were not significantly different for either cloning efficiency or mutant frequency (MF); inclusion of both sets of controls in the analysis increased the ability to detect a Chernobyl exposure effect in the liquidators. After adjusting for age and smoking, the results revealed no significant difference in cloning efficiency of Chernobyl liquidators relative to Russian controls but a significant, 24% increase in liquidator HPRT mutant frequency over Russian controls (90% confidence interval was 7% to 45% increase). The analytical method also accounted for differences in precision of the individual estimates of log CE and log MF and accommodated for outliers. The increase in HPRT mutant frequency of liquidators is an attribute of the exposed population as a whole rather than of individuals. These results demonstrate that, under appropriate circumstances, the HPRT specific locus mutation assay of peripheral blood lymphocytes can be used to detect a semi-acute, low dose radiation exposure of a population, even 6 to 10 years after the exposure.     

The significance of cytogenetic investigation for the estimation of Chernobyl accident consequences

The results of the cytogenetic investigation of people, which were exposed to radiation in the result of the Chernobyl NPP accident, were presented. Also the possibilities of the application of cytogenetic findings for dose estimations and for the prediction of the radiation influence consequences were examined. During the period of time since 1986 till 2004 the cytogenetic investigations of 1724 liquidators participating in the liquidation works after the Chernobyl accident were carried out. The radiation dose estimated by the frequency of dicentrics in 1986 was about 0.16 Gy. The doses for liquidators were determined by the frequency of translocations (FISH method) during the period from 1992 till 1995. For liquidators who worked in Chernobyl only in 1986 the average dose of radiation was about 0.19 Gy and for liquidators who worked repeatedly during the period from 1986 till 1995 - 0.39 Gy. There was shown that during the whole period of investigation (1986-2004) the frequency of dicentrics in peripheral blood lymphocytes was significantly higher than the control level. The cytogenetic investigation of Bryansk region inhabitants which was carried out in 1992-1994 discovered heightened value in 5 times than the control one. Findings are of great importance for the prediction of ill effects of radiation and for the development of sensitive criterions for early exposure disturbances in state of health.     

The epidemiological analysis of monitoring of the immune status in liquidators of consequences of the Chernobyl accident for early identification of risk groups and diagnostics of oncological diseases. Report 2. Dependence of frequency and changes in the immune status on risk factors of radiation accident

Malignant neoplasms (MN) have been found to develop most frequently in the liquidators of entry into the ChNPP zones in 1986 (43.75%), as well as among the liquidators who worked for long, one quarter of whom participated in liquidation of the consequences of failure (LCF) in 1986. Specific features of the immune status depending on the timing of participation in LCF and the year of entry into the ChN PP zone have been established. Changes in the immune system in the persons with a confirmed diagnosis of MN who took both a non-permanent and permanent part in liquidating the consequences of the ChNPP failure in 1986 had the same character of deviations and differed in the magnitudes of deviations of immunological parameters. Continuous participation in the period of extreme conditions and a greater exposure to the radiation factor led to the increased content of CD8(+)-T-cells, CD16(+)-lymphocytes and activated T-lymphocytes, as well as to the reduced index of immune regulation, decreased content ofCD3-16/56+(NK)-cells (%) and the total IgE and to a greater deficiency of B-lymphocytes. Distinctions in the groups of liquidators who participated in LCF in 1986 and 1987 have been revealed. The greatest deviations in the IS indicators were found in liquidators-87. A similar effect came to light in case of a continuance in the ChNPP zones in 1986 and 1987; however, the degree of deviation of the content of CD4(+)-T-lymphocytes (41), CD8(+)-T-lymphocytes (1) and the immune regulation index (41) were remarkably higher in liquidators-87. A continuous stay in the ChNPP zones in 1987 led to the deficiency of CD4(+)-T-lymphocytes, increased values of CD8(+)-T-lymphocytes, a decreased index of CD4+/CD8+, as well as to the change in the ratio between NK-T and NK cells, increased numbers of CD95+, HLA-DR+ and activated T-lymphocytes, and a lower level of the total IgE. Long-term participation in LCF didn't cause any enhanced expression of cellular activation markers in liquidators-86. Specific features of changes in IS depending on a dose of external gamma-irradiation have been established. Increase in the frequency of MN among liquidators, in relation to the number of examinees in each age group, with age has been revealed. Distinctions in the age dynamics of IS in liquidators in the presence and in the absence of MN manifested themselves in a stable level of values of CD3+, CD4+, CD8(+)-T-lymphocytes, immune regulation index, CD95+, serum IgA at the age between 40 and 70 years old with a subsequent reduction in indicators and increase in the content of CD8(+)-T-lymphocytes with age in the absence of MN; continuous increase of CD3-16/56(+)-NK-cells in the presence of MN and decrease in the values after 70 in the absence of MN. Also revealed in IS of the both age groups of liquidators over 70 with and without MN was the deficiency of the T-cell component (CD3+, CD4(+)-T-lymphocytes, CD4+/CD8+ index) and the increase in absolute values of CD8(+)-T-lymphocytes. The growing deficiency of CD4(+)-T-lymphocytes during monitoring against the background of ever rising values of CD8(+)-T-lymphocytes leading to the weakening of the immune regulation due to progressing disorders of the T-lymphocyte regulatory subpopulation distribution can serve an indicator for the adverse prognosis of the life expectancy in the presence of MN.     

An epidemiological analysis of monitoring of the immune status of chernobyl nuclear accident liquidators for early detection of risk groups and diagnosis of cancer diseases. Communication 1

Ionizing radiation is one of the major risk factors for cancer diseases. The question on the rate of malignant neoplasms (MNs) and their early detection in Chernobyl nuclear accident (ChNA) liquidators is still open. In this work, the results of a long-term immunological monitoring of the ChNA liquidators who live in the northwestern region of Russia have been analyzed with an attempt to detect the predictors of cancer in this population cohort. The rate of newly detected MN cases in the cohort of followup ChNA liquidators monitored in 1990–2009 was 8.856% (89 cases per 1005 persons), which is somewhat higher as compared with the average rate for the total population. As for the rate of individual MN types, lung, gastric, and prostate cancers were prevalent, which matches the worldwide trend in MN abundance. It was shown that 1-3 years before the MN diagnosis, liquidators displayed detectable changes in their immune status, including a decrease in the percentages of CD3⁺ and CD4⁺ T lymphocytes, and, to a lesser degree, B lymphocytes; a decrease in the CD4⁺/CD8⁺ ratio; increase in the relative and absolute contents of CD16⁺ lymphocytes; increase in the absolute CD8⁺ T lymphocyte counts; prevalence of CD3⁺16/56⁺ (NK-T) cells over CD3-16/56⁺ (NK) cells; and increase in the activity of phagocytes. The patients who displayed one or several of the listed characteristics should be ascribed to the MN risk group to be assessed for cancer markers, be more comprehensively examined, and be the subjects of dynamic observation.     

The molecular and cellular consequences of the chernobyl accident

In this paper the results of research at 5–10 and 24 years after the Chernobyl accident are summarized. These results include the investigation of genomic instability, formation of the adaptive response, genome damage, and oxidative status. The studies were performed on cells in culture, mice, children and adults who lived in the contaminated areas, and liquidators of the consequences of the Chernobyl accident. Inhibition of cell proliferative activity, late cell death, and the increase in micronucleus and giant cell frequency were observed after the exposure of cells in culture in the accident zone followed by their culturing in laboratory conditions. In the progeny of the exposed cells, the effect of enhanced radiosensitivity was detected. Thus, it can be assumed that exposure of parental cells in culture in the area of the accident induced genomic instability that resulted in the development of various abnormalities in progeny cells. At the organism level, the Chernobyl zone exposure of mice caused an increase in radiosensitivity; as well, a decrease in the endotheliocyte density in the cerebral cortex and other brain tissues was observed. In the blood lymphocytes of children stimulated by PHA, a more than two times increase in micronucleus cell frequency was detected. A reduced number of individuals with significant adaptive response was found in both the juvenile and adult groups. In all investigated populations, an increased number of individuals with enhanced radiosensitivity were observed in response to low-dose radiation exposure. At 24 years after the accident liquidators were subjected to examinations, which revealed an increased frequency of cells with micronuclei and chromosome-type aberrations in blood lymphocytes, an elevated level of DNA double strand breaks, and a reduced level of reactive oxygen species compared to those of the control group. This means that the genomic instability that was accumulated by the residents of the contaminated regions and liquidators as a result of the accident leads to damage of the genetic apparatus, an increase in radiosensitivity, and hypoxia as late consequences that all are risk factors and increase the probability of the development of tumor and non-tumor diseases. The development of the above-mentioned pathological processes may occur in the distant future.

     

Phenotypic characteristics and tendency to apoptosis of peripheral blood mononuclear cells from HIV+ long term non progressors

The aim of this study was to analyze (i) phenotype, (ii) in vitro spontaneous and induced apoptosis, (iii) glutathione (GSH) intracellular content and (iv) inhibitors of apoptosis of potential therapeutical use in peripheral blood mononuclear cells (PBMC) from HIV+ long term non progressors (LTNP), in comparison with progressors (HIV+P) and seronegative controls (HIV-). Three groups of subjects were studied: 15 HIV+P (patients losing >150 CD4+/year), 9 LTNP (subjects infected by HIV for at least 7 years without clinical and immunological signs of progression, with a mean of 898 CD4+/microL) and 18 HIV-. All subjects were living in a large community for former drug addicts, and were matched for age and sex. We used flow cytometry for analyzing PBMC phenotype and apoptosis; high performance liquid chromatography for measuring intracellular GSH content. PBMC phenotype of LTNP shared characteristics with those of both HIV- and HIV+P. Indeed, LTNP showed a normal number CD4+ cells (an inclusion criteria), but significantly increased numbers of CD8+ lymphocytes, activated T cells, CD19+, CD5+ B lymphocytes and CD57+ cells, as well as a decrease in CD19+, CD5- B lymphocytes and CD16+ cells. In LTNP, spontaneous apoptosis was similar to that of HIV- and significantly lower than that of HIV+P. Adding interleukin-2 (IL-2) or nicotinamide (NAM) significantly decreased spontaneous apoptosis in LTNP and HIV+P. Pokeweed mitogen-induced apoptosis was also similar in LTNP and HIV-, but significantly lower than that of HIV+P. In HIV+P, but also in LTNP, spontaneous apoptosis was inversely correlated to the absolute number and percentage of CD4+ cells and directly correlated to the number and percentage of activated T cells present in peripheral blood. GSH intracellular content was greatly decreased in PBMC from HIV+P and slightly, but significantly, reduced in LTNP. Adding 2-deoxy-D-ribose, an agent provoking apoptosis through GSH depletion, to quiescent PBMC resulted in similar levels of massive cell death in the three groups. This phenomenon was equally prevented in the three groups by N-acetyl-cysteine but not by IL-2. A complex immunological situation seems to occur in LTNP. Indeed, PBMC from LTNP are characterized by a normal in vitro tendency to undergo apoptosis despite the presence of a strong activation of their immune system, unexpectedly similar to that of HIV+P. Our data suggest that NAM and IL-2 are possible candidates for reducing spontaneous apoptosis in HIV infection.     

Effect of chronic psychoemotional stress on subpopulation spectrum of T-lymphocytes in immunocompetent organs in male mice

Subpopulation spectrum of T lymphocytes (CD3+, CD4+, CD8+, CD25+, and CD3(+)25+) in thymus, spleen and inguinal lymphatic nodes have been studied in male mice after 20 days of psychoemotional stress produced by social defeats in daily agonistic confrontations. A reduction of total number of cells, of absolute numbers of all researched subpopulations of lymphocytes and % CD3+ cells in thymus of submissive mice was shown in comparison with intact animals. Reduction of total number of splenocytes and absolute numbers of CD4+ and CD8+ lymphocytes has been observed in a spleen of submissive mice. Besides, % CD3+, CD25+, CD4+ and CD25+ cells were increased in these animals in comparison with intact mice. The absolute number of cells with CD8 phenotype was increased in inguinal lymphatic nodes. The data obtained suggest that the chronic psychoemotional stress is accompanied by serious changes of the cellular link of immunity. The effect of chronic emotional social stress on mutual interaction of the central and peripheral links of immunity has been discussed.     

IL-6/BSF-2 selectively stimulates the GO----S progression of CD8+ lymphocytes.

IL-6 preferentially promotes the DNA synthesis of human peripheral blood CD8+, rather than CD4+, lymphocytes in presence of PHA: this effect is observed in serum-free cultures of greater than 99% purified CD8+ lymphocytes. However, IL-6 is able to stimulate DNA synthesis of CD8+ lymphocytes triggered by a mitogenic anti-CD2 mAb, but not by anti-CD3 mAb: these results suggest that IL-6 selectively induces activation of CD8+ lymphocytes through the CD2 rather than the CD3 pathway. Limiting dilution analysis indicates that accessory cells are not required to mediate the action of IL-6 on CD8+ cells. Furthermore, this action is not blocked by addition of mAb neutralizing either IL-2 or IL2R, thus suggesting that IL-6 does not act via IL-2. CD8+ lymphocytes grown in the presence of PHA + IL-6 incorporate (3H)-thymidine to the same extent as those stimulated with PHA + IL-2, but do not increase in number until day 6 of culture. It is hence apparent that the stimulating activity of IL-6 on CD8+ lymphocytes is restricted to the GO----S phase progression, but does not lead to mitosis. IL-6 receptors are expressed on resting CD4+ and CD8+ lymphocytes: their expression is significantly enhanced on both activated CD4+ and CD8+ cells. Scatchard analysis of (125I)-IL-6 binding data showed the presence of high (Kd, 3 x 10(-10) M) and low (Kd, 6 x 10(-8) M) affinity IL6R on both lymphocyte populations. Similarly, mRNA encoding IL6R was detected in both CD4+ and CD8+ lymphocytes. Thus, our studies indicate that IL-6 directly and selectively stimulates the GO----S progression of CD8+ lymphocytes in the presence of mitogen and absence of IL-2: this phenomenon may be of interest for the elucidation of mechanisms activating cytotoxic T lymphocytes.     

Interrelation between the activity of hepatitis, liver fibrosis and immune status in children with chronic hepatitis B and C

The lesion of the liver in viral hepatitis was found to depend on the state of the immune system. Relationship between the content of lymphocyte subpopulations (CD3+, CD4+, CD8+, CD20+) in the blood and immunoglobulins (IgG, IgM, IgA) with parameters of semi-quantitative evaluation of the activity of hepatitis and the stage of liver fibrosis in children with chronic virus hepatitis B, C, B + C was studied. The characteristic feature of all hepatitis was a decrease in the number of T lymphocytes CD4+ below the normal level and an increase in the content of B lymphocytes. The correlation between the morphological activity of hepatitis and the amount of T lymphocytes CD8+ was established only in chronic hepatitis B. In chronic hepatitis B and B + C the absolute amount of blood lymphocytes decreased with the increase of the age of the patients, but in chronic hepatitis B this was accompanied by the decrease of the morphological activity of hepatitis and in hepatitis B + C by its increase. The amount of lymphocytes CD4+ rose with the increase of liver fibrosis in chronic hepatitis B. In children with chronic hepatitis C and B + C the amount of blood lymphocytes was found to be unrelated to the morphological activity of hepatitis.     

Characterization of interleukin 2-expanded human peripheral blood lymphocytes: not only NKH-1+-NK cells but also NKH-1+ and NKH-1- T cells are LAK effectors

In vitro culture of human peripheral blood mononuclear cells (PBMC) with interleukin 2 (IL-2) results in the expansion of lymphocytes including lymphokine-activated killer (LAK) cells. Using flow cytometry, studies were undertaken to determine the phenotype and LAK activity of each subset of lymphocytes expanded in vitro as a result of incubation for 2 weeks with 2500 U/ml of recombinant IL-2. Such expanded PBMC, when examined by two-color staining with various combinations of anti-CD3, 4, 8, 16, and NKH-1 monoclonal antibodies, consisted of the following six subgroups of cells: (1) CD3+4+8-, (2) CD3+4-8+, (3) CD3+4-8-, (4) CD3-16+NKH-1+, (5) CD3-16-NKH-1+, and (6) CD3-16-NKH-1-. Of the six subgroups, all five subgroups that could be tested, i.e., CD3+ T cells (CD3+4+8-, CD3+4-8+, CD3+4-8-), CD16+ natural killer (NK) cells (CD3-16+NKH-1+), and CD3-16-NKH-1- non-T non-NK cells, possessed LAK activity. Both NKH-1- as well as NKH-1+ T and non-T cells possessed LAK activity.     

Microsatellite mutations show no increases in the children of the Chernobyl liquidators

We performed a study on Belarusian "liquidators", exploring whether increase in the frequencies of germline mutations at microsatellite loci could be found in their progeny. The liquidators, mostly young males, were those involved (during 1986 and 1987) in clean-up operations in the radioactively contaminated area following the Chernobyl nuclear power plant accident in 1986. Many liquidators fathered children during the clean-up period and after the work had been terminated. The numbers of families studied were 64 (liquidators) and 66 (controls). A total of 72 loci (31 autosomal, one X-linked and 40 Y-linked) were used. DNA was isolated from peripheral blood lymphocytes and the microsatellite loci were amplified by the polymerase chain reaction with fluorescence-labelled primers. Mutations were detected as variations in the length of the loci. At the Y-linked loci, the mutation rates (expressed as number of mutations among the total number of loci for the individuals included) are 2.9 x 10(-3) (4/1392) and 2.1 x 10(-3) (3/1458) in the children of exposed and control parents, respectively. This difference is not statistically significant. At the autosomal loci, the corresponding estimates are 5.9 x 10(-3) (11/1862; exposed group) and 8.5 x 10(-3) (18/2108; control). Again, the difference is not significant. The possibility that the Belarusian population might have been unexpectedly exposed to some chemical contaminants in the environment appears unlikely in view of the finding that the spontaneous mutation rates at the same set of loci in several non-Belarusian populations were similar to those in Belarus. The estimated mean radiation dose to the liquidators was small, being about 39 mSv, and this might be one reason why no increases in mutation rates due to radiation could be found.     

Changes in blood leucocyte populations induced by acute maximal and chronic submaximal exercise

Absolute (x 10(3).mm-3) or relative (%) numbers of blood leucocyte types (monocytes, lymphocytes, neutrophils) and lymphocyte subsets (T11+, T4+, T8+, B1+, and NKH1+) reacting with specific monoclonal antibodies were determined at rest, immediately after maximal exercise on a treadmill, in six controls (C), and in six young cyclists before training (BT) and after 5 months of training (AT). Maximal exercise significantly increased the absolute number (mobilization) of virtually all the types of leucocytes and subsets of lymphocytes in C, BT and AT subjects. In these subjects mobilization of natural killer cells (NKH1+) and cytotoxic/suppressor T lymphocytes (T8+) was greater than mobilization of the other leucocyte types and lymphocyte subsets; however, maximal exercise induced no significant changes in the relative numbers of any leucocyte types and lymphocyte subsets, except in the case of T4+ lymphocytes in At cyclists. Chronic submaximal exercise induced increased mobilization of neutrophils and decreased mobilization of lymphocytes during maximal exercise, except in the case of B lymphocytes (B1+) and NKH1+ cells, and decreases in the absolute and relative number of neutrophils at rest. It remains to be seen how these results can explain the modifications of leucocyte activities noted in vitro after isolated or chronic exercise.     

Mechanisms of the natural reactivity of lymphocytes from noninfected individuals to membrane-associated Leishmania infantum antigens

Membrane-associated Leishmania Ags (MLA) or soluble Leishmania Ags were used in vitro to stimulate cord blood or PBMC from healthy donors noninfected by Leishmania parasites. MLA, but not soluble Leishmania Ags, constantly induce strong proliferation of cord blood mononuclear cells and PBMC from noninfected individuals. Responding cells are CD3+, CD4+, TCRalphabeta+, CD45RO+, and CD45RA+ and secrete IFN-gamma and IL-10, but not IL-4. MLA do not activate NK cells nor NKT cells. Membrane Ags also induce purified macrophages from noninfected individuals to secrete IL-10 and TNF-alpha, but have no effect on IL-1alpha or IL-12 secretion. The effects of MLA are proteinase K-sensitive and resistant to lipid extraction. The lymphoproliferative responses are inhibited by anti-HLA-DR Abs and require Ag processing by APCs, excluding that the biological effect of MLA could be attributed to a superantigen. Finally, TCR repertoire analysis shows that the T cell expansion induced by MLA uses TCR with various variable beta segment rearrangements and CDR3 lengths, features much more characteristic to those observed with a polyclonal activator than with a conventional Ag. These results suggest a particular mechanism developed during the host's natural response to Leishmania parasites that allows direct activation of naive CD4 lymphocytes by parasite membrane-associated Ags.