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.     

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.     

The state of immunity in the liquidators of consequences of the Chernobyl accident with cardio-vascular diseases

The purpose of the work was research and functional reserves immunity in participants in the Chernobyl with cardiovascular disease. A Clinical Lab 49 men aged 44 to 52 years with diseases of the cardiovascular system, participated in the Chernobyl nuclear power plant in 1986 to 1988. As control used data 33 patients with similar pathology, and 16 healthy men. Researched the total number of white blood cells, lymphocytes and transfusions, the absolute number and relative CD4+ and CD8+, CD 16, CD20+, CD95+ lymphocytes peripheral blood, number mononuclear, synthesizing IL-1beta, TNF-alpha, INF-alpha, IL-4, the content of lipids and proteins, the activity of alkaline phosphatase and mieloperoksidazy in neutrophils. To assess the functional reserve immune system blood samples studied people subjected to radiation doses 0.25, 50, 1.0 Gy of in vitro and studied the reaction cytochemical indicators neutrophils before and after the radiative forcing. The liquidators not detected significant changes in the absolute number of leucocytes, but compared with control groups noted significant reduction in the absolute number of CD8+ and CD20+ lymphocytes, increased the number of cells, expression of FAS-antigen, change the number of mononuclear spontaneously synthesizing and produce cytokines, decreased maintenance of cation proteins in neutrophils. Radiation samples peripheral blood liquidators caused the same reaction cytochemical indicators of neutrophils and control groups, the compensatory and adaptive nature of the changes in the immune system, developed in response to complex factors radiation accident.     

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 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.     

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 CD24 antigen discriminates between pre-B and B cells in human bone marrow.

When bone marrow (BM) lymphoid cells from 12 adult healthy donors were labeled by CD24 antibodies and analyzed by flow cytometry, two positive populations of cells were demonstrated in each sample (by a separated bimodal specific immunofluorescence). One population had intermediate CD24-Ag density (termed CD24+ cells) whereas the other had high CD24-Ag density (termed CD24(2+) cells). CD24+ cells represented 5.8 +/- 2.7% of the total lymphoid BM cells and CD24(2+) cells 5.6 +/- 2.5%. Using dual fluorescence analysis on eight samples, all CD24+ cells expressed the CD21 and CD37 mature B cell Ag and also surface IgM (sIgM), but this population lacked CD10 Ag. These cells also expressed CD19 Ag, and at a higher density than CD24(2+) cells. They were also positive for HLA-DR Ag. Conversely, CD24(2+) cells were shown to be early cells of the B cell lineage. While all the CD24(2+) cells were HLA-DR+ and CD19+, 64 +/- 16% of them expressed CD20 Ag (at a lower density than CD24+ cells), 65 +/- 21% CD10 Ag, and 22 +/- 8% were positive for cytoplasmic mu-chains (c mu). None of these cells expressed the CD21 and CD37 mature B cell Ag or sIgM. Additional experiments on four different healthy donors demonstrated that 30 +/- 9% of the CD24(2+) cells expressed the CD34 Ag and that the CD24+ cells did not express it. Thus, the CD24 Ag permits discrimination between two populations of the B cell lineage present in adult BM: 1) A CD24(2+) cell population including "pre" pre-B cells (HLA-DR+, CD19+, CD10+/-, CD20-, CD21-, CD34+, CD37-, c mu-), "intermediate" pre-B cells (HLA-DR+, CD19+, CD10+, CD20+, CD21-, CD34-, CD37-, c mu-), and "true" pre-B cells (HLA-DR+, CD19+, CD10+, CD20+, CD21-, CD34-, CD37-, c mu+). 2) A CD24+ cell population including B cells of the standard phenotype (HLA-DR+, CD19+, CD10-, CD20+, CD21+, CD34-, CD37+, c mu-, sIgM+).     

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 2. Dependence of the rate and changes in the immune status on radiation accident risk factors

It was demonstrated that malignant neoplasms (MNs) most frequently develop in the liquidators involved in the cleanup activities after the Chernobyl nuclear accident (ChNA) in 1986 (43.75%) and the liquidators involved in long-term activities, one-quarter of whom also participated in the cleanup in 1986. The specific features of the immune status (IS) depending on the period and year of the cleanup activities in the ChNA area were determined. In the cohort with a confirmed MN diagnosis, the observed deviations in the people who participated in these activities only in 1986 and on a permanent basis, including 1986, coincided in their pattern but differed in the magnitude of changes in immunological characteristics. Long-term participation during the extreme period and, correspondingly, higher exposure caused an increase in the contents of CD8⁺ T cells, CD16⁺ lymphocytes, and activated T lymphocytes, as well as decreases in the total immunoregulatory index, percentage of CD3-16/56⁺ (NK), and total IgE and a more pronounced deficiency in B lymphocytes. Differences between the groups of liquidators who participated in the cleanup activities in 1986 and 1987 were detected. The liquidators-1987 display the most pronounced deviations in the IS characteristics. Permanent presence in the ChNA area in 1986 and 1987 caused a similar effect but the magnitude of changes in the content of CD4⁺ T lymphocytes (↓), content of CD8⁺ T lymphocytes (↑), and immunoregulatory index (↓) was considerably higher in liquidators-1987. The permanent cleanup activities in Chernobyl in 1987 caused a deficiency in CD4⁺ T lymphocytes; more pronounced increase in the content of CD8⁺ T lymphocytes; decrease in CD4⁺/CD8⁺ index; change in the ratio of NK-T to NK cells; elevation in the counts of CD95⁺, HLA-DR⁺, and activated T lymphocytes; and decrease in the level of total IgE. Any increase in the expression of cell activation markers was undetectable in the liquidators-1986 involved in the cleanup activities on a long-term basis. The specific features of the IS changes depending on the dose of external γ-irradiation were determined. It was shown that the MN rate in liquidators increases with age relative to the number of examined people in each age cohort. The differences in the IS age-related dynamics in liquidators with and without MNs were detectable in stable contents of CD3⁺, CD4⁺, and CD8⁺ T lymphocytes; immunoregulatory index; and contents of CD95⁺ cells and serum IgA at ages of 40 to 70 years old with a subsequent decrease in these parameters and elevation in the content of CD8⁺ T lymphocytes with age in the absence of MNs; continuous increase in CD3-16/56⁺ (NK) cells in the presence of MNs; and decreases in these values after 70 years in the absence of MNs. In both groups of liquidators older than 70 years (with and without MNs), a deficiency in the T-cell component (CD3⁺ and CD4⁺ T lymphocytes and CD4⁺/CD8⁺ ratio) and an increase in the counts of CD8⁺ T lymphocytes are characteristic of the IS. A growing deficiency in CD4⁺ T lymphocytes during the monitoring on the background of increasing content of CD8⁺ T lymphocytes, which additionally contributes to the weakening of immune regulation due to progressing abnormalities in the ratio of regulatory T-lymphocyte subpopulations, can be regarded as one of the characteristics that suggest an adverse life expectancy prognosis for people with MNs.     

Effect of Cytomegalovirus Co-Infection on Normalization of Selected T-Cell Subsets in Children with Perinatally Acquired HIV Infection Treated with Combination Antiretroviral Therapy

Background

We examined the effect of cytomegalovirus (CMV) co-infection and viremia on reconstitution of selected CD4+ and CD8+ T-cell subsets in perinatally HIV-infected (PHIV+) children ≥ 1-year old who participated in a partially randomized, open-label, 96-week combination antiretroviral therapy (cART)-algorithm study.

Methods

Participants were categorized as CMV-naïve, CMV-positive (CMV+) viremic, and CMV+ aviremic, based on blood, urine, or throat culture, CMV IgG and DNA polymerase chain reaction measured at baseline. At weeks 0, 12, 20 and 40, T-cell subsets including naïve (CD62L+CD45RA+; CD95-CD28+), activated (CD38+HLA-DR+) and terminally differentiated (CD62L-CD45RA+; CD95+CD28-) CD4+ and CD8+ T-cells were measured by flow cytometry.

Results

Of the 107 participants included in the analysis, 14% were CMV+ viremic; 49% CMV+ aviremic; 37% CMV-naïve. In longitudinal adjusted models, compared with CMV+ status, baseline CMV-naïve status was significantly associated with faster recovery of CD8+CD62L+CD45RA+% and CD8+CD95-CD28+% and faster decrease of CD8+CD95+CD28-%, independent of HIV VL response to treatment, cART regimen and baseline CD4%. Surprisingly, CMV status did not have a significant impact on longitudinal trends in CD8+CD38+HLA-DR+%. CMV status did not have a significant impact on any CD4+ T-cell subsets.

Conclusions

In this cohort of PHIV+ children, the normalization of naïve and terminally differentiated CD8+ T-cell subsets in response to cART was detrimentally affected by the presence of CMV co-infection. These findings may have implications for adjunctive treatment strategies targeting CMV co-infection in PHIV+ children, especially those that are now adults or reaching young adulthood and may have accelerated immunologic aging, increased opportunistic infections and aging diseases of the immune system.     

Clinical immunological disorders in children from various observation cohorts exposed to radiation factor during various stages of oncogenesis

The immune status disorders and features depending on the radiation impact type in various cohorts of radiation observations long after the Chernobyl (CNPP) disaster and the possible role of these disorders in development of chronic somatic pathology in children are shown. Lymphocyte depletion, T-cell immunity component disorders in the form of cell contraction with CD3, CD4, CD8 markers and the B-cell immunity component disorders in the form of reducing the quantity of CD10, CD23 marker cells were observed in children subject to combined chronic irradiation by 131I, 137Cs, 90Sr radionuclides. The descendants of irradiated parents (the 1st generation; children of the Chernobyl accident consequences liquidators, children of the citizens of radiation contaminated territories with various 137Cs levels) had immunity disorders of different type. A change in the total amount of NK-cells (CD16(+)-lymphocytes) is the general sign for all radiation risk groups; however, people subject to direct radiation impact demonstrated reduction of the antitumor protection potency, whereas descendants of irradiated ones demonstrated its activation with typically increasing number of CD16(+)-lymphocytes. In all radiation risk groups, a tendency to reduction of a number of cells involved in the leukocytal activation with the "pluripotential activation" marker (CD38 marker cells), proliferating cells (CD71 marker cells) and the increase of relative amount of cells with apoptosis marker (CD95(+)-lymphocytes). Immune disorder markers under the radiation impact in various cohorts of children's observation are suggested: antigens: CD4, CD8, CD10, CD23, CD16, CD38, CB71, CD95.     

Dendritic cells in the human decidua

Dendritic cells (DCs) in the pregnant human uterine mucosa have been poorly characterized, although they are likely to regulate immune responses to both placental trophoblast cells and uterine infections. In this study an HLA-DR+, CD11c+ lin- (CD3-, CD19-, CD56-, CD14-) population has been identified by three-color flow cytometry. The cell isolates were prepared either by collagenase digestion or mechanically from first-trimester decidual tissue. The decidual DCs comprised approximately 1.7% of CD45+ cells in the isolates and had the phenotype of immature myeloid DCs. No CD1a+ Langerhans cells or CD123+ plasmacytoid DCs were detected. The decidual DCs were DC-SIGN-, DEC-205+, CD40+. Two subsets could be distinguished on the basis of relative expression of HLA-DR, which also differed in expression of DC-activation markers. The DCs were identified in situ by immunohistology by DEC-205 staining. Cells with dendritic processes were found scattered through both the decidua basalis (in which trophoblast cells are infiltrating) and the decidua parietalis. They were also visible in endothelial-lined spaces. This is the first study to identify and describe the phenotype and distribution of human decidual DCs.     

Modulation of human lymphocyte marker expression by gamma irradiation and mitomycin C.

gamma irradiation (GR) or mitomycin C (MC) treatment of stimulator cells is frequently used to achieve unidirectionality of response in the mixed lymphocyte reaction. As GR differs from MC in the pathways used to block lymphocyte replication, this study analyzes the effects of these modalities upon the expression of various differentiation and Class II major histocompatibility antigens on lymphocytes cultured for 24, 48, and 72 hr. There was a decrease in the mean density of HLA-DR expression on CD3+ and on CD8+ cells at 24, 48, and 72 hr after exposure to GR (42 and 35, 60 and 69, and 26 and 49%, respectively) or to MC (26 and 11, 26 and 18, and 46 and 30%, respectively). There was a parallel decrease in the levels of the corresponding cell subsets when compared with control cultured cells not exposed to GR or MC. In contrast, the density of HLA-DR markers on CD3-negative cells was increased at 24, 48, and 72 hr of culture following exposure to GR (73, 82, and 102%, respectively) or to MC (9, 45, and 80%, respectively). There was a more profound decrease in CD3+, CD8+, and CD19+ cell subset levels and in the density of the corresponding markers in GR-treated cells than in those of cells exposed to MC when the results were compared with those of untreated cultured control cells. Although GR appears to exert a more profound effect than MC, the results indicate that both modalities have the capacity to reduce the density of polymorphic determinants on Class II (HLA-D region)-encoded molecules on T (CD3+ and CD8+) and B (CD19+) cells which are known to trigger potent MLR responses. Both modalities may therefore affect profoundly the relative strength of MLC responses and the derived measurements of the degree of HLA Class II compatibility between stimulator and responder cells.     

Molecular and cellular consequences of the Chernobyl accident

In this paper the results of the Chernobyl accident investigation 5-10 and 24 years after are summarized. The genomic instability, adaptive response formation, genome damage and oxidative status have been investigated. The studies were performed on cells in culture, mice, children and adults living in contaminated areas and liquidators. On cells in culture after exposition in the accident zone and culturing thereafter in laboratory conditions the cell proliferative activity decrease; the late cell death, the frequency of cells with micronuclei and giant cells increasing have been observed. In the progeny of exposed cells the enhancement of radiosensitivity has been noticed. So we can suppose that in cultured cells exposition in the zone of the accident the genomic instability is induced which results in many disturbances. At the organism level in mice exposed in the Chernobyl zone the radiosensitivity increase and the decrease of endotheliocytes density in brain tissue has been observed. On the stimulated by PHA blood lymphocytes of children the increase of the frequency of cells with micronuclei more than 2 time have been noticed. In all groups investigated, the decrease of individuals with significant adaptive response was observed. In children and adults inhabitants the increase of radiosensitivity after low dose of irradiation has been noticed. 24-year after the accident it was discovered that in liquidators lymphocytes the frequency of cells with micronuclei, with chromosome type aberrations, with DNA double strand breaks have been increased; the reactive oxygen species (ROS) were decreased in comparison with the control population. We can suppose that genomic instability induced in residents of contaminated regions and liquidators long after the accident results in the genetic apparatus damage, radiosensitivity enhancement, hypoxia that represent risk factors and increase the probability of tumour and non-tumour diseases. The development of these pathological processes may happen in much more remote periods.     

Co-stimulatory and adhesion molecules of dendritic cells in rheumatoid arthritis

Dendritic cells (DCs) in the rheumatoid arthritis (RA) joint mediate the immunopathological process and act as a potent antigen presenting cell. We compared the expression of co-stimulatory and adhesion molecules on DCs in RA patients versus controls with traumatic joint lesions and evalulated the correlation between the immunophenotypical presentation of DCs and the clinical status of the disease. Samples of peripheral venous blood, synovial fluid (SF) and synovial tissue (ST) were obtained from 10 patients with RA at the time of hip or knee replacement and from 9 control patients with knee arthroscopy for traumatic lesions. Clinical status was appreciated using the DAS28 score. Blood, SF and dissociated ST cell populations were separated by centrifugation and analyzed by flow cytometry. Cells phenotypes were identified using three-color flow cytometry analysis for the following receptors HLA-DR, CD80, CD83, CD86, CD11c, CD18, CD54, CD58, CD3, CD4, CD8, CD19, CD20, CD14, CD16, CD56. HLA-DR molecules, co-stimulatory receptors CD80, CD86, CD83 and adhesion molecules CD18, CD11c, CD54, CD58, were analyzed by two-color immunofluorescence microscopy on ST serial sections. In patients with active RA (DAS28>5.1) we found a highly differentiated subpopulation of DCs in the ST and SF that expressed an activated phenotype (HLA-DR, CD86+, CD80+, CD83+, CD11c+, CD54+, CD58+). No differences were found between circulating DCs from RA patients and control patients. Our data suggest an interrelationship between clinical outcome and the immunophenotypical presentation of DCs. Clinical active RA (DAS28>5.1) is associated with high incidence of activated DCs population in the ST and SF as demonstrated by expression of adhesion and co-stimulatory molecules.     

Specific changes in auditory cognitive evoked potentials in those who participated in the liquidation of the chernobyl accident: I. Analysis of the early N1 component

The amplitude and temporal parameters of the N1 component of the auditory cognitive evoked potential (EP) were analyzed in ten people who participated in the elimination of the consequences of the Chernobyl accident (the liquidators) and ten healthy subjects aged 47 ± 6.0 and 50.5 ± 4.0 years, respectively. In the liquidators of the Chernobyl accident, the amplitudes of the N1 component of the auditory EP were decreased in all cortical areas. This decrease was the greatest in the central and frontal areas. The changes in response to stimuli of different significances in the liquidators were inverted as compared to the healthy subjects of the same age. The differences between the control group and the liquidators were the most pronounced in the temporal characteristics of the N1 component. The latent periods (LPs) for all stimuli presented under all experimental conditions were significantly shorter in the group of liquidators, and this effect was even stronger for the significant stimulus in the case of stimulus counting. In the liquidators, the maximum changes in the LPs of the N1 component were observed in the frontal area of the left hemisphere, and they were associated with an inversion of the LP asymmetry. Changes in this parameter were smaller in the central areas and were similar in the parietal areas in both groups studied. Changes in the amplitude and temporal characteristics of the N1 component of the auditory cognitive EP observed in the liquidators of the Chernobyl accident indicate the impairments of their involuntary attention and its capacity due to weakening of inhibition processes as compared to healthy subjects. This effect is similar to that found in elderly subjects. Our data support the hypothesis on the accelerated aging of the brain in the liquidators of the Chernobyl accident due to the effect of low doses of irradiation. The data also suggest that irradiation results in pathological aging.     

Analysis of the cellular mechanism of antitumor responses and autoimmunity in patients treated with CTLA-4 blockade

We have demonstrated previously that the administration of CTLA-4 blockade has mediated objective cancer regression and autoimmunity in patients with metastatic melanoma. To explore the mechanism of these in vivo effects, we have studied the changes in lymphocyte phenotype and function in patients receiving anti-CTLA-4 Ab (MDX-010). Patients with stage IV melanoma or renal cell cancer were treated every 3 wk with an anti-CTLA-4 Ab with or without peptide immunization. Pheresis samples were analyzed using flow cytometry to determine lymphocyte cell surface markers. Gene expression analyses and proliferation assays were conducted on purified T cell subsets. Anti-CTLA-4 Ab did not inhibit the suppressive activity of CD4+CD25+ cells in vitro or in vivo. In addition, there was no decrease in the expression of CD4+CD25+ cells in whole PBMC, nor a decrease in Foxp3 gene expression in the CD4+ or CD4+CD25+ purified cell populations posttreatment. The percentage of CD4+, CD8+, CD4+CD25+, and CD4+CD25- T cells in PBMC expressing the activation marker HLA-DR increased following anti-CTLA-4 Ab administration. Therefore, our results suggest that the antitumor effects of CTLA-4 blockade are due to increased T cell activation rather than inhibition or depletion of T regulatory cells.     

In vitro suppression of lymphocyte activation in patients with seasonal allergic rhinitis and pollen-related asthma by cetirizine or azelastine in combination with ginkgolide B or astaxanthin

Novel strategies are evaluated for management of allergic rhinitis and asthma in patients co-afflicted with both disorders. It is hypothesized that the platelet activating factor receptor antagonist ginkgolide B (GB) and the carotenoid antioxidant astaxanthin (ASX) interact with antihistamines cetirizine dihydrochloride (CTZ) and azelastine (AZE) to potentiate their ability to downregulate potentially pathological immune activation. Peripheral blood mononuclear cells from asthmatics and healthy subjects, cultured 24 hours with 50 μg/ml phytohemaglutinin (PHA) or PHA plus each drug are analyzed by flow cytometry for expression of CD25+ or HLA-DR+ by CD3+ (T cells). Results are reported as stimulation indices for CD3+CD25+ (SICD3+CD25+) and CD3+HLA-DR+ (SICD3+HLADR+) cells in cultures treated with PHA alone, versus cultures treated with both PHA and drugs. Optimal suppression of activated cells was observed in cultures stimulated with ASX 10-6 M + CTZ 10-6 M (SICD3+CD25+, p = 0.016; SICD3+HLADR, p = 0.012); ASX 10-6 M + AZE 10-6 M (SICD3+CD25+, p = 0.012; SICD3+HLADR, p = 0.015); GB 10-6 M + CTZ 10-6 M (SICD3+CD25+, p = 0.024, SICD3+HLADR+, p = 0.019). Results demonstrate improved activity of antihistamines by 2 phytochemicals, suggesting dosing strategies for animal trials of ASX- or GB-augmented formulations for seasonal allergic rhinitis and asthma.     

Genetic effects of bystander factors from the blood sera of people irradiated as the result of the Chernobyl accident

The purpose of this work was the analysis of the effects of bystander factors from blood sera of people affected by the Chernobyl accident on human keratinocyte cell culture (HPV-G cells). A new method was developed for evaluation of the bystander factor presence in vivo in blood of the people irradiated by the Chernobyl accident. Affected population groups included liquidators of the Chernobyl accident and people living and working in areas of the Gomel region contaminated by radionuclides. The analysis has shown that bystander factors persist in Chernobyl liquidator blood samples for more than 20 years since irradiation. The data suggest that blood sera contain bystander factors, which are able to induce micronuclei and decrease the metabolic activity of HPV-G cells.     

Genetic effects in the liquidators of consequences of Chernobyl Nuclear Power Plant accident

The purpose of the present research was the estimation of probable genetic consequences at the liquidators of the consequences of Chernobyl accident in 1986-1987. The research is made on two groups of the liquidators. The first group included the liquidators taking place on the account in the branch register and working now at the enterprises of a nuclear industry. The second group included 902 liquidators of consequences of Chernobyl accident in 1986 constantly living in the Ryazan area and which are taking place on permanent observation the account in regional hospital. For an estimation of probable genetic effects analyzed the data on frequency and outcomes pregnancy of the wives of the liquidators, on condition and on diseases of newborn, on switching intrauterine development defects (IDD). The analysis carried out depending on dozes of an irradiation: up to 5 cGy; 5-10 cGy and 10-25 cGy. Received materials testify, that at the liquidators, at a doze of an external irradiation 10-25 cGy, the determined effects--period long sterility, kept at a part them till 3 years come to light. The set of the received data, such as depending from the dose increase of frequency of spontaneous abortions and of inherent defects of development of newborn, the increase of frequency diseases of newborn and share newborn with low weight, allows to make a conclusion about an induction of genetic effects in sexual cells of the liquidators of consequences of Chernobyl accident at dozes of an external irradiation more than 10 cGy. Taking into account high biological efficiency of alpha-radiation (K = 20), and of beta-radiation (K = 2-4), the equivalent effective doze male gonads (testes) in 3-5 times is higher, than estimated only from external gamma-radiation.     

Changes in the expression of membrane markers and in the number of human blood monocytes after single and repeated courses of visible and infrared light at therapeutic doses

An attempt has been made to prove that the immunomodulating effect of therapeutic doses of polychromatic visible + infrared polarized (VIP) light at its application to a small body surface area is connected with a transcutaneous photomodification of a small amount of blood in superficial skin microvessels. For this purpose, in parallel experiments, using monoclonal antibodies, the membrane phenotype of circulating blood mononuclears was studied after irradiation of volunteers, of samples of their blood in vitvo, and of a mixture of the irradiated and non-irradiated autologous blood in a 1:10 volume ratio, thereby modeling events in vivo, when a small amount of the transcutaneously photomodified blood in the vascular bed contacts its main circulating volume. In this variant of experiment, a great similarity has been established of changes in expression of mononuclear membrane markers (CD3, CD4, CD8, CD20, CD16, HLA-DR and to a lesser degree of CD25); the ability has been proven of the photomodified blood to "translate" the light-induced changes to a much higher volume of non-irradiated blood, which might represent a mechanism of the systemic immunomodulating effect of phototherapy. Under conditions in vivo and in vitro, the most "reactive" were HLA-DR+, CD20+, CD16+, CD4+, and 0-cells. An increase of the total number of lymphocytes and monocytes has been shown by the end of the 10-day-long phototherapeutic course. The regulatory character of the single and course sessions of the VIP light on the blood immunocompetent cells is substantiated: depending on the initial state of the immune system, the VIP light can produce both stimulating and inhibitory effect on lymphoid cell subpopulations, which opens large possibilities of using this method for correction of immunological disturbances in diseases of different etiopathogenesis.