How fast does wasp venom immunotherapy affect a regulatory T cell subpopulation (CD4+ CD25+ Foxp3+) and the synthesis of interleukins 10, 21 and transforming growth factor ß1?

INTRODUCTION: The literature describes the influence of venom immunotherapy (VIT) on the subpopulation of T regulatory cells (CD4+ CD25+ Foxp3+) and the synthesis of IL-10, TGF-ß1 as well as many other cytokines at various times after immunotherapy. AIM: To assess changes in the percentage of cells of CD4+ and CD25+ in peripheral blood and serum concentrations of IL-10, IL-21 and TGF-ß1 in the early stages of VIT. MATERIAL AND METHODS: The study included 18 patients who were allergic to wasp venom and who in the past underwent systemic anaphylactic reaction after stinging, meeting the criteria to qualify for VIT. The immunoenzymatic method (ELISA) was used to assess concentrations of cytokines IL-10, IL-21 and TGF-ß1 and the surface antigens CD4 and CD25 on the cells. The concentrations were determined by flow cytometry method at baseline (before VIT) and after 2.5 and 24 h from the VIT starting point. RESULTS: The mean values of the activity of T lymphocytes CD4+ CD25+ FoxP3+ and concentrations of the cytokines IL-10, IL-21 and TGF-ß1 are shown in table. CONCLUSIONS: A 24-hour activation assessment of serum concentrations of cytokines IL-10, IL-21 and TGF-ß1 during the first day of the Hymenoptera venom immunotherapy by ultra-rush protocol does not show the significant dynamics of change of the examined parameters.

Effect of internal contamination with tritiated water on the neoplastic colonies in the lungs, innate anti-tumour reactions, cytokine profile, and haematopoietic system in radioresistant and radiosensitive mice.

Tritium is a potentially significant source of internal radiation exposure which, at high levels, can be carcinogenic. We evaluated whether single intraperitoneal injection of BALB/c and C57BL/6 mice with tritiated water (HTO) leading to exposure to low (0.01 or 0.1 Gy) and intermediate (1.0 Gy) cumulative whole-body doses of ß radiation is immunosuppressive, as judged by enhancement of artificial tumour metastases, functioning of NK lymphocytes and macrophages, circulating cytokine's levels, and numbers of bone marrow, spleen, and peripheral blood cells. We demonstrate that internal contamination of radiosensitive BALB/c and radioresistant C57BL/6 mice with HTO at all the absorbed doses tested did not affect the development of neoplastic colonies in the lungs caused by intravenous injection of syngeneic cancer cells. However, internal exposure of BALB/c and C57BL/6 mice to 0.1 and 0.01 Gy of ß radiation, respectively, up-regulated cytotoxic activity of and IFN-γ synthesis in NK lymphocytes and boosted macrophage secretion of nitric oxide. Internal contamination with HTO did not affect the serum levels of pro- (IL-1ß, IL-2, IL-6, TNF-α,) and anti-inflammatory (IL-1Ra, IL-4, IL-10) cytokines. In addition, exposure of mice of both strains to low and intermediate doses from the tritium-emitted ß-particles did not result in any significant changes in the numbers of bone marrow, spleen, and peripheral blood cells. Overall, our data indicate that internal tritium contamination of both radiosensitive and radioresistant mice leading to low and intermediate absorbed ß-radiation doses is not immunosuppressive but may enhance some but not all components of anticancer immunity.

Cancer immunotherapy: how low-level ionizing radiation can play a key role.

The cancer immunoediting hypothesis assumes that the immune system guards the host against the incipient cancer, but also "edits" the immunogenicity of surviving neoplastic cells and supports remodeling of tumor microenvironment towards an immunosuppressive and pro-neoplastic state. Local irradiation of tumors during standard radiotherapy, by killing neoplastic cells and generating inflammation, stimulates anti-cancer immunity and/or partially reverses cancer-promoting immunosuppression. These effects are induced by moderate (0.1-2.0 Gy) or high (>2 Gy) doses of ionizing radiation which can also harm normal tissues, impede immune functions, and increase the risk of secondary neoplasms. In contrast, such complications do not occur with exposures to low doses (≤0.1 Gy for acute irradiation or ≤0.1 mGy/min dose rate for chronic exposures) of low-LET ionizing radiation. Furthermore, considerable evidence indicates that such low-level radiation (LLR) exposures retard the development of neoplasms in humans and experimental animals. Here, we review immunosuppressive mechanisms induced by growing tumors as well as immunomodulatory effects of LLR evidently or likely associated with cancer-inhibiting outcomes of such exposures. We also offer suggestions how LLR may restore and/or stimulate effective anti-tumor immunity during the more advanced stages of carcinogenesis. We postulate that, based on epidemiological and experimental data amassed over the last few decades, whole- or half-body irradiations with LLR should be systematically examined for its potential to be a viable immunotherapeutic treatment option for patients with systemic cancer.

Marie and Irène Curie, two brilliant women who pioneered the development of nuclear chemistry, radiotherapy, and radiobiology.

OBJECTIVE: To describe the contributions of Marie Sklodowska-Curie and her daughter Irene Curie to radiation science and how these studies in physics, chemistry and medicine led to the need to study radiobiological effects. CONCLUSIONS: The seminal discoveries of Maria Sklodowska-Curie in radiation physics and chemistry provided the basis for later investigations of the effects of ionizing radiation on cells and tissues and the role of radioactivity in mutagenesis and carcinogenesis. Both Maria and Irene contributed to the development of radiotherapy and the use of x-rays and radioisotopes in medical diagnosis. Their legacy is not confined to their own generation but can be traced through grand- and great-grand-students of these pioneering women.

Time to rejuvenate ultra-low dose whole-body radiotherapy of cancer.

The results of clinical trials performed from the 1930s until the end of the 20th century in which total-body ultra-low level ionizing radiation (TB-LLR) was used demonstrate that this form of treatment can be equal or superior to other systemic anti-neoplastic modalities in terms of the rates of remissions, toxicity, and side effects. In this review, we provide the rationale for TB-LLR and analyze the results of reliable clinical trials in patients with predominantly lymphoproliferative disorders but also advanced solid cancers. The doses used in these trials did not exceed 0.1-0.2 Gy per fraction and cumulative totals ranged from 1 to 4 Gy. Based on the reviewed results we conclude that it is appropriate to revive interest in and resume clinical investigations of TB-LLR in order to refine and improve the effectiveness of such treatment, whether employed alone or in combination with other anticancer strategies.

A derivative of vitamin B3 applied several days after exposure reduces lethality of severely irradiated mice.

Most, if not all, of the hitherto tested substances exert more or less pronounced pro-survival effects when applied before or immediately after the exposure to high doses of ionizing radiation. In the present study we demonstrate for the first time that 1-methyl nicotinamide (MNA), a derivative of vitamin B3, significantly (1.6 to 1.9 times) prolonged survival of BALB/c mice irradiated at LD30/30 (6.5 Gy), LD50/30 (7.0 Gy) or LD80/30 (7.5 Gy) of γ-rays when the MNA administration started as late as 7 days post irradiation. A slightly less efficient and only after the highest dose (7.5 Gy) of γ-rays was another vitamin B3 derivative, 1-methyl-3-acetylpyridine (1,3-MAP) (1.4-fold prolonged survival). These pro-survival effects did not seem to be mediated by stimulation of haematopoiesis, but might be related to anti-inflammatory and/or anti-thrombotic properties of the vitamin B3 derivatives. Our results show that MNA may represent a prototype of a radioremedial agent capable of mitigating the severity and/or progression of radiation-induced injuries when applied several hours or days after exposure to high doses of ionizing radiation.

Bringing Radiation Exposures and Associated Health Risks into Perspective-Development of an App.

The NATO HFM 291 research task group (RTG) on "Ionizing Radiation Bioeffects and Countermeasures" represents a group of scientists from military and civilian academic and scientific institutions primarily working in the field of radiobiology. Among other tasks, the RTG intends to extend their work on risk estimation and communication to bridge the gap in appropriate judgment of health risks given a certain radiation exposure. The group has no explicit psychological background but an expertise in radiobiology and risk assessment. The group believes that, as one of the essential first steps in risk communication, it is required to put radiation risk into perspective. Radiation risk requires a weight in comparison to already-known risks. What we envision is to Compare Radiation exposure-associated health Risks (CRRis App) with daily life health risks caused by other common exposures such as cigarette smoking, driving a car, etc. Within this paper, we provide (1) an overview of health risks after radiation exposure, (2) an explanation of the task and concept of an envisioned CRRis App, (3) an overview of existing software tools related to this issue, (4) a summary of inputs and discussions with experts in the field of radiation protection and risk communication during the ConRad conference, and finally, (5) identification of the next steps in the development of the App.

Enhanced cytotoxic activity of macrophages and suppressed tumor metastases in mice irradiated with low doses of X- rays.

We showed in our previous report that a single exposure of mice to 0.1 or 0.2 Gy X-rays led to the significant inhibition of the development of artificial tumor metastases in the lungs and that the effect was related to the enhanced activity of natural killer cells. In the present study, a possible involvement of cytotoxic macrophages in the anti-metastatic effect of the low-level X-ray exposures was investigated. We now demonstrate that irradiation of mice with either of the two low doses of X-rays significantly stimulates the macrophage-mediated cytolysis of the susceptible tumor targets and that the effect coincides with the enhanced production of nitric oxide in the collected effector cells. We also show that suppression of the in vivo function of macrophages by carrageenan eliminates the inhibitory effect of the two low doses of X-rays on the development of pulmonary tumor colonies as well as significantly suppresses the macrophage-mediated cytotoxicity and nitric oxide production. Finally, aminoguanidine added to the culture medium of the assayed macrophages not only shuts down the nitric oxide synthesis in these cells but also significantly suppresses their cytolytic activity. Overall, the obtained results indicate that inhibition of the tumor metastases by a single exposure of mice to 0.1 or 0.2 Gy X-rays results, to a large extent, from the radiation-induced stimulation of the cytocidal activity of macrophages which secrete enhanced amounts of nitric oxide.

Single low doses of X rays inhibit the development of experimental tumor metastases and trigger the activities of NK cells in mice.

There is evidence indicating that low-level exposures to low- LET radiation may inhibit the development of tumors, but the mechanism of this effect is virtually unknown. In the present study, BALB/c mice were irradiated with single doses of 0.1 or 0.2 Gy X rays and injected intravenously 2 h later with syngeneic L1 sarcoma cells. Compared to the values obtained for sham-irradiated control mice, the numbers of pulmonary tumor colonies were significantly reduced in the animals exposed to either 0.1 or 0.2 Gy X rays. Concurrently, a significant stimulation of NK cell-mediated cytotoxic activity was detected in splenocyte suspensions obtained from irradiated mice compared to sham-exposed mice. Intraperitoneal injection of the NK-suppressive anti-asialo GM1 antibody totally abrogated the tumor inhibitory effect of the exposures to 0.1 and 0.2 Gy X rays. These results indicate that single irradiations of mice with either 0.1 or 0.2 Gy X rays suppress the development of experimental tumor metastases primarily due to the stimulation of the cytolytic function of NK cells by radiation.

Epidemiological evidence of childhood leukaemia around nuclear power plants.

A few reports of increased numbers of leukaemia cases (clusters) in children living in the vicinity of nuclear power plants (NPP) and other nuclear installations have triggered a heated debate over the possible causes of the disease. In this review the most important cases of childhood leukaemia clusters around NPPs are described and analyzed with special emphasis on the relationship between the environmental exposure to ionizing radiation and the risk of leukaemia. Since, as indicated, a lifetime residency in the proximity of an NPP does not pose any specific health risk to people and the emitted ionizing radiation is too small to cause cancer, a number of hypotheses have been proposed to explain the childhood leukaemia clusters. The most likely explanation for the clusters is 'population mixing', i.e., the influx of outside workers to rural regions where nuclear installations are being set up and where local people are not immune to pathogens brought along with the incomers.

Commentary: ethical issues of current health-protection policies on low-dose ionizing radiation.

The linear no-threshold (LNT) model of ionizing-radiation-induced cancer is based on the assumption that every radiation dose increment constitutes increased cancer risk for humans. The risk is hypothesized to increase linearly as the total dose increases. While this model is the basis for radiation safety regulations, its scientific validity has been questioned and debated for many decades. The recent memorandum of the International Commission on Radiological Protection admits that the LNT-model predictions at low doses are "speculative, unproven, undetectable and 'phantom'." Moreover, numerous experimental, ecological, and epidemiological studies show that low doses of sparsely-ionizing or sparsely-ionizing plus highly-ionizing radiation may be beneficial to human health (hormesis/adaptive response). The present LNT-model-based regulations impose excessive costs on the society. For example, the median-cost medical program is 5000 times more cost-efficient in saving lives than controlling radiation emissions. There are also lives lost: e.g., following Fukushima accident, more than 1000 disaster-related yet non-radiogenic premature deaths were officially registered among the population evacuated due to radiation concerns. Additional negative impacts of LNT-model-inspired radiophobia include: refusal of some patients to undergo potentially life-saving medical imaging; discouragement of the study of low-dose radiation therapies; motivation for radiological terrorism and promotion of nuclear proliferation.

Effect of low doses of low-let radiation on the innate anti-tumor reactions in radioresistant and radiosensitive mice.

BALB/c and C57BL/6 mice differ in their Th1/Th2 lymphocyte and M1/M2 macrophage phenotypes, radiosensitivity, and post-irradiation tumor incidence. In this study we evaluated the effects of repeated low-level exposures to X-rays on the development of artificial tumor colonies in the lungs of animals from the two strains and cytotoxic activities of natural killer (NK) cells and macrophages obtained from these mice. After ten daily irradiations of BALB/c or C57BL/6 mice with 0.01, 0.02, and 0.1 Gy X-rays NK cell-enriched splenocytes collected from the animals demonstrated significant and comparable up-regulation of their anti-tumor cytotoxic function. Likewise, peritoneal macrophages collected from the two irradiated strains of mice exhibited the similarly stimulated cytotoxicities against susceptible tumor cells and produced significantly more nitric oxide. These results were accompanied by the significantly reduced numbers of the neoplastic colonies induced in the lungs by intravenous injection of syngeneic tumor cells. The obtained results indicate that ten low-level irradiations with X-rays stimulate the generally similar anti-tumor reactions in BALB/c and C57BL/6 mice.

Anti-neoplastic and immunostimulatory effects of low-dose X-ray fractions in mice.

PURPOSE: The exploration of immune mechanisms of the tumour-inhibitory effect of exposures to low-level fractions of X-rays. MATERIALS AND METHODS: BALB/c mice were exposed to whole-body daily irradiations with 0.01, 0.02, or 0.1 Gy X-rays per day for 5 days/week for two weeks. Then, mice were intravenously injected with L1 tumour cells, killed 14 days later, and neoplastic colonies were counted in the lungs. Natural killer (NK) cell-enriched splenocytes and activated peritoneal macrophages (Mϕ) were collected and cytotoxic activities of these cells against susceptible tumour targets were assayed. Concanamycin A (CMA) and antibody against the ligand for the Fas receptor (FasL) were used to inhibit the NK cell-mediated cytotoxicity. Production of nitric oxide (NO) was quantified using the Griess reagent. Secretion of interferon-γ (IFN-γ), interleukin-1ß (IL-1ß), interleukin-12 (IL-12), and tumour necrosis factor-α (TNF-α) was measured using the enzyme-linked immunosorbent assays. RESULTS: All the exposures to X-rays significantly reduced the number of the induced tumour colonies and enhanced cytotoxic properties of the NK cell-enriched splenocytes and activated Mϕ. CONCLUSION: Suppression of the growth of pulmonary tumour colonies by irradiations of mice with low-dose fractions of X-rays may result from stimulation of anti-tumour reactions mediated by NK cells and/or cytotoxic macrophages.

Immunological mechanism of the low-dose radiation-induced suppression of cancer metastases in a mouse model.

According to the doctrine underlying the current radiation protection regulations each, no matter how small, exposure to ionizing radiation may be carcinogenic. However, numerous epidemiological observations demonstrate that cancer incidence and/or mortality are not elevated among inhabitants of the high- versus low-natural-background radiation areas and homes. Results of our own and other authors' studies described in this paper bear testimony to the possibility that stimulation of the anti-neoplastic immune surveillance mediated by NK lymphocytes and activated macrophages explains, at least partially, the accumulating epidemiological and experimental evidence indicating that low-level exposures to the low-linear energy transfer (LET) radiation inhibit the development of spontaneous and artificial metastases in humans and laboratory animals, respectively. The results presented also suggest the possibility of using low-level X- and gamma-ray exposures to cure cancer and to prevent cancer metastases. For a broader perspective, the results presented may help towards relaxing the current radiation protection regulations, especially as they apply to diagnostic and therapeutic exposures of patients to the indicated forms of radiation.

Production of cytokines by peritoneal macrophages and splenocytes after exposures of mice to low doses of X-rays.

We have shown previously that irradiations of mice with 0.1 or 0.2 Gy of X-rays stimulate anti-tumour cytotoxic activities of peritoneal macrophages and splenocytes enriched for NK lymphocytes and suppress the development of pulmonary tumour colonies. The up-regulated cytotoxicities were related to the production of nitric oxide by macrophages, and perforin and Fas ligand by the splenocytes, but specific blockade of these pathways did not totally suppress the effector cell-mediated cytolysis of the tumour target. Hence, other factors such as cytotoxic/cytostatic cytokines might have been produced by the effector cells. To test this possibility peritoneal macrophages and splenocytes were isolated from BALB/c mice which had been either once or tentimes whole body-irradiated with the total doses of 0.1 and 0.2 Gy of X-rays and assayed for the levels of IL-1beta, IL-2, IL-12, IFN-gamma and TNF-alpha in the incubation medium using the respective ELISA kits. The results demonstrate that both single and multiple exposures to the two low doses of X-rays significantly stimulate secretion of IL-1beta, TNF-alpha and IL-12 by macrophages and IL-2 and IFN-gamma by splenocytes, but the kinetics and magnitude of the induced changes in the production of these cytokines differ between the two irradiation protocols.