Differential Immune Modulation With Carbon-Ion Versus Photon Therapy.

PURPOSE: Radiation therapy (RT) modulates the immune characteristics of the tumor microenvironment (TME). It is not known whether these effects are dependent on the type of RT used. METHODS AND MATERIALS: We evaluated the immunomodulatory effects of carbon-ion therapy (CiRT) compared with biologically equivalent doses of photon therapy (PhRT) on solid tumors. Orthotopic 4T1 mammary tumors in immunocompetent hosts were treated with CiRT or biologically equivalent doses of PhRT. Seventy-two hours after RT, tumors were harvested and the immune characteristics of the TME were quantified by flow cytometry and multiplex cytokine analyses. RESULTS: PhRT decreased the abundance of CD4+ and CD8+ T cells in the TME at all doses tested, with compensatory increases in proliferation. By contrast, CiRT did not significantly alter CD8+ T-cell infiltration. High-dose CiRT increased secretion of proinflammatory cytokines by tumor-infiltrating CD8+ T cells, including granzyme B, IL-2, and TNF-α, with no change in IFN-γ. Conversely, high-dose PhRT increased CD8+ T-cell secretion of IFN-γ only. At most of the doses studied, PhRT increased proliferation of immunosuppressive regulatory T cells; this was only seen with high-dose CiRT. Cytokine analyses of bulk dissociated tumors showed that CiRT significantly increased levels of IFN-γ, IL-2, and IL-1ß, whereas PhRT increased IL-6 levels alone. CONCLUSIONS: At low doses, lymphocytes differ in their sensitivity to CiRT compared with PhRT. Unlike PhRT, low-dose CiRT is generally lymphocyte-sparing. At higher doses, CiRT is a more potent inducer of proinflammatory cytokines and merits further study as a modulator of the immunologic characteristics of the TME.

Effect of 50-Hz Magnetic Fields on Serum IL-1ß and IL-23 and Expression of BLIMP-1, XBP-1, and IRF-4.

Investigations demonstrated that magnetic fields (MFs) change cytokine production and expression of some immune system genes. This alteration can affect the immune system function and may lead to some diseases. Therefore, this study investigated two important inflammatory cytokines, i.e., IL-1ß and IL-23 at two phases of pre- and post-immunization of the immune system. In addition, the expressions of three important genes in the humoral immunity, i.e., B lymphocyte-induced maturation protein-1 (BLIMP-1), X-box-binding protein-1 (XBP-1), and interferon regulatory factor-4 (IRF-4) were evaluated at post-immunization phase. Eighty adult male rats were divided into four experimental groups and a control. The experimental groups were exposed to 50 -Hz MFs with magnetic flux densities of 1, 100, 500, and 2000 µT, 2 h/day for 2 months. The animals were injected by human serum albumin (100 µg/rat) on days 31, 44, and 58 of exposure. The cytokine levels in serum were measured with enzyme-linked immunosorbent assay (ELISA), and the expression of genes was evaluated with reverse transcription quantitative polymerase chain reaction (RT-qPCR). Serum IL-1ß was decreased at pre-immunization phase after exposure to 1 and 100 µT of 50-Hz MFs. In contrast, serum IL-23 was increased at post-immunization phase in 100 µT group. No change was observed in serum IL-1ß and IL-23 in each group at pre-immunization phase compared with post-immunization. Furthermore, exposure to 100 µT downregulated expression of BLIMP-1, XBP-1, and IRF-4. In conclusion, exposure to 50-Hz MFs may decrease inflammation at short time and increase it at longer time exposures. In addition, 50-Hz MF exposure may decrease the humoral immune responses. It seems that 50-Hz MFs cause more alteration in immune system function at lower densities (100 µT).

Effect of high frequency electromagnetic wave stimulation on muscle injury in a rat model.

INTRODUCTION: The aim of this study was to investigate biological changes in tissues with muscle contusion after the application of high frequency (HF) electromagnetic wave. METHODS: An acrylic pipe was placed on the right hind limb and a metallic ball was dropped inside the pipe, which resulted in a muscle contusion. After acquiring the optimal condition for muscle contusion, 20 Sprague-Dawley rats were allocated to the HF treatment (N = 10) and sham groups (N = 10), which then underwent muscle contusion injury at their right thigh. The thickness and circumference of the right thigh and the left thigh (negative control groups) were measured (day 0). HF electromagnetic wave stimulation for three days was performed on the contusion area in the HF group after one day. Thickness was measured at the thickest area of both hind limbs and the circumference was measured every day for three days. The sham group received no treatment, and the circumference and thickness were measured using the same method. After three days, Hematoxylin and eosin and immunohistochemical (IHC) staining for IL-1ß were performed and TUNEL assay was conducted for apoptosis in the skin and muscle layers. RESULTS: The thigh muscle thickness at day 1 was significantly different between groups (P = 0.018) and this difference was observed between both sham and control groups (corrected P = 0.007), and between sham and HF groups (corrected P = 0.043). Thigh circumference was significantly different at day 3 (P = 0.047) and this difference was found between sham and control groups (corrected P = 0.018), and between sham and HF groups (corrected P = 0.032). In the HF group, the inflammatory response was reduced to almost the same level as the control group. Evaluation of IL-1ß level, the inflammatory cytokine, through IHC showed marked localization of IL-1ß in muscle fibers of the sham group. However, significantly less IL-1ß was observed in the muscle of the HF treatment group. There was neither injury nor apoptosis after HF stimulation. CONCLUSIONS: Application of the HF showed therapeutic effect on muscle contusion by reducing muscle swelling. This effect might be caused by the anti-inflammatory action of the HF, which evoked energy into the injured muscle.

Effect of photobiomodulation on endothelial cell exposed to Bothrops jararaca venom.

Bleeding is a common feature in envenoming caused by Bothrops snake venom due to extensive damage to capillaries and venules, producing alterations in capillary endothelial cell morphology. It has been demonstrated, in vivo, that photobiomodulation (PBM) decreases hemorrhage after venom inoculation; however, the mechanism is unknown. Thus, the objective was to investigate the effects of PBM on a murine endothelial cell line (tEnd) exposed to Bothrops jararaca venom (BjV). Cells were exposed to BjV and irradiated once with either 660- or 780-nm wavelength laser light at energy densities of 4 and 5 J/cm(2), respectively, and irradiation time of 10 s. Cell integrity was analyzed by crystal violet and cell viability/mitochondrial metabolism by MTT assay. The release of lactic dehydrogenase (LDH) was quantified as a measure of cell damage. In addition, cytokine IL1-ß levels were measured in the supernatant. PBM at 660 and 780 nm wavelength was able to increase cellular viability and decrease the release of LDH and the loss of cellular integrity. In addition, the concentration of pro-inflammatory cytokine IL1-ß was reduced after PBM by both wavelengths. The data reported herein indicates that irradiation with red or near-infrared laser resulted in protection on endothelial cells after exposure to Bothrops venom and could be, at least in part, a reasonable explanation by the beneficial effects of PBM inhibiting the local effects induced by Bothrops venoms, in vivo.

Systemic response to ultraviolet radiation involves induction of leukocytic IL-1ß and inflammation in zebrafish.

Ultraviolet radiation is a pervasive stimulus with wide-ranging effects on all living forms. The effects of UV vary from physiological to pathological, depending on levels of exposure, but the immune response at the organismal level is not well understood. We use the zebrafish embryo and larva to study immune responses to UV stress in vivo. UV exposure causes inflammation characterized by systemic induction of proinflammatory cytokines. Leukocytes are an important component of this systemic response and upregulate IL-1ß expression proportional to the dose of UV exposure. Increased levels of this proinflammatory cytokine counteract the lethal effect of high doses of UV.

Cytokine changes in response to radio-/chemotherapeutic treatment in head and neck cancer.

BACKGROUND: Radiation and systemic chemotherapy are standard treatment strategies for advanced or metastatic head and neck cancer. However, little is known about the implications and changes in the tumor microenvironment, including the T-helper (TH)1/TH2 balance in response to these treatment regimens. The aim of the current study was to unravel the effects of chemotherapeutic drugs and radiation on cytokine changes. MATERIALS AND METHODS: In this study, the effect of radiation and chemotherapeutic treatment (5-fluorouracil and cisplatin) on eight cell lines was determined. Before and after exposure, cytokine levels in culture supernatants of cell lines were evaluated using the Bio-Plex Assay (Bio-Rad) and the Human TH1/TH2 Cytometric Bead Array (Becton Dickinson). Results were correlated with parallel measurements for cellular proliferation assessed by cytotoxicity assay. RESULTS: Seven out of eight cell lines of primary tumors or metastases demonstrated an enhanced level of the cytokines interleukin (IL)-1ß, IL-6, IL-8, granulocyte colony-stimulating factor (G-CSF), granulocyte-macrophage-colony stimulating factor (GM-CSF) and tumor necrosis factor-α (TNF-α), after sub-lethal radiation doses. Under treatment with low concentrations of 5-fluorouracil and cisplatin, all examined cell lines showed an increasing secretion of the cytokines IL-6 and G-CSF. In contrast, sub-lethal doses of both cytostatic drugs revealed a dose-dependent decrease in secretion IL-1ß. Regarding GM-CSF and TNF-α, we demonstrated an increase in secretion by the primary tumors under low doses of 5-fluorouracil and cisplatin, whereas the metastases showed a sharp drop of GM-CSF and TNF-α secretion. Chemotherapeutic treatment led to no changes of the IL-8 cytokine profile. CONCLUSION: The results suggest complex cytokine changes of the tumor microenvironment and more aberrant expression profiles under treatment with radiation and the chemotherapeutic drugs 5-fluorouracil and cisplatin.

Tissue-engineered mucosa is a suitable model to quantify the acute biological effects of ionizing radiation.

The aim of this study was to evaluate the suitability of tissue-engineered mucosa (TEM) as a model for studying the acute effects of ionizing radiation (IR) on the oral mucosa. TEM and native non-keratinizing oral mucosa (NNOM) were exposed to a single dose of 16.5Gy and harvested at 1, 6, 24, 48, and 72h post-irradiation. DNA damage induced by IR was determined using p53 binding protein 1 (53BP1), and DNA repair was determined using Rad51. Various components of the epithelial layer, basement membrane, and underlying connective tissue were analyzed using immunohistochemistry. The expression of cytokines interleukin-1ß (IL-1ß) and transforming growth factor beta 1 (TGF-ß1) was analyzed using an enzyme-linked immunosorbent assay. The expression of DNA damage protein 53BP1 and repair protein Rad51 were increased post-irradiation. The expression of keratin 19, vimentin, collage type IV, desmoglein 3, and integrins α6 and ß4 was altered post-irradiation. Proliferation significantly decreased at 24, 48, and 72h post-irradiation in both NNOM and TEM. IR increased the secretion of IL-1ß, whereas TGF-ß1 secretion was not altered. All observed IR-induced alterations in TEM were also observed in NNOM. Based on the similar response of TEM and NNOM to IR we consider our TEM construct a suitable model to quantify the acute biological effects of IR.

Saponins from the roots of Platycodon grandiflorum suppress ultraviolet A-induced matrix metalloproteinase-1 expression via MAPKs and NF-κB/AP-1-dependent signaling in HaCaT cells.

Saponins from the roots of Platycodon grandiflorum (CKS) have been shown to exhibit many pharmacological activities, including anti-cancer and anti-inflammatory activities and antioxidant effects. However, anti-skin photoaging effects of CKS have not yet been reported. In this study, we investigated the protective effects of CKS against UVA damage on immortalized human keratinocytes (HaCaT). We then explored the inhibitory effects of CKS on UVA-induced MMP-1 and investigated the molecular mechanism underlying those effects. CKS increased the cell viability and inhibited reactive oxygen species (ROS) production in HaCaT cells exposed to UVA irradiation. Pre-treatment of HaCaT cells with CKS inhibited UVA-induced production of MMP-1 and MMP-9. In addition, CKS decreased UVA-induced expression of the inflammatory cytokines IL-1ß and IL-6. Western blot analysis further revealed that CKS markedly suppressed the enhancement of collagen degradation in UVA-exposed HaCaT cells. CKS also suppressed UVA-induced activation of NF-κB or c-Jun and c-Fos, and the phosphorylation of MAPKs, which are upstream modulators of NF-κB and AP-1.

Anti-ulcerogenic effect of aqueous propolis extract and the influence of radiation exposure.

PURPOSE: To study the effect of aqueous propolis extract (AEP) against indomethacin (Indo)-induced gastric ulcers in irradiated and non-irradiated rats. MATERIALS AND METHODS: Animals were irradiated at different radiation dose levels before the induction of ulcers. AEP was injected orally 1 hour before induction of gastric ulcers and the effects compared with those of lansoprazole (Lanso), which was used as a reference anti-ulcerogenic drug. RESULTS: Pretreatment of rats, either irradiated or non-irradiated, with AEP effectively protected against Indo-induced gastric ulceration. This was associated with a reduction in acid output and peptic activity and an increase in the secretion of mucin. The mucosal prostaglandin E(2) (PGE(2)) level was also increased. The levels of tumor necrosis factor-alpha (TNF-α) and interleukin-1beta (IL-1ß) were suppressed to the same extent after treatment. Both propolis and Lanso were effective in reducing the number of gastric lesions as well as the plasma level of malondialdehyde (MDA). CONCLUSIONS: These findings indicate that the gastroprotective effect of AEP could be of value in the management of excessive gastric damage induced by radiation exposure.

Positive affect and inflammation during radiation treatment for breast and prostate cancer.

There is growing evidence that positive affect may influence health and immune function, although few studies have examined links between positive affect and immune processes in clinical populations. The purpose of this study was to examine whether positive affect is associated with changes in proinflammatory cytokines in cancer patients undergoing radiation treatment. Subjects were 50 individuals with early-stage breast and prostate cancer who completed psychosocial questionnaires and provided blood samples at seven time points before, during, and after radiation treatment. Positive affect was assessed before treatment onset using the CES-D (Center for Epidemiological Studies Depression Scale). Blood samples were assayed for serum levels of proinflammatory cytokines IL-1beta and IL-6. Patients with higher levels of positive affect before treatment exhibited higher mean levels of IL-1beta and IL-6 during radiation treatment (all ps<.05). Results suggest that positive affect enhances the acute inflammatory response to radiation treatment, perhaps facilitating tissue repair processes.