Evaluation of the action of LPSF/GQ-16 on cytokines and PPAR-γ gene expression after in vitro irradiation of peripheral blood mononuclear cells.

PURPOSE: Patients submitted to radiotherapy (RT) may present in their healthy tissues surrounding the treated tumor, some typical acute inflammatory reactions induced by ionizing radiation (IR). The manifestation of inflammatory processes is a result of exacerbation of the immune system, as a response to radiation exposure, and this can be a limiting factor for RT protocols. To counteract this, some thiazolidinediones, such as LPSF/GQ-16, may be useful for modulating the patient's radioinduced inflammatory response in normal tissues. In this context, the present work aims to evaluate the activity of LPSF/GQ-16 on the levels of cytokines and the expression of the gene PPARγ in mononuclear cells irradiated in vitro, to analyze the immunomodulatory activity of the molecule and its action on radiomitigation. MATERIALS AND METHODS: For this, blood samples from eight donors were collected and irradiated with 2 Gy, then the PBMC (peripheral blood mononuclear cells) were cultured and treated with LPSF/GQ-16. The levels of cytokines TNF-α, IFN-γ, IL-2 and IL-4 were quantified by CBA, while the genes of TNF-α, IFN-γ and PPARγ were analyzed by RT-PCR. RESULTS: LPSF/GQ-16 significantly reduced the expression of proinflammatory cytokines (IFN-γ and TNF-α) in irradiated and nonirradiated groups. There was no significant reduction of anti-inflammatory cytokines (IL-2 and IL-4) by LPSF/GQ-16. The mRNA expression of PPAR-γ, IFN-γ and TNF-α in the presence of LPSF/GQ-16 was higher in the nonirradiated sample. CONCLUSION: LPSF/GQ-16 showed effective activity after irradiation, with an important immunomodulatory activity in irradiated PBMCs.

Micronucleus assay for predicting side effects of radiotherapy for cervical cancer.

Radiotherapy (RT) is an important treatment for cervical cancer. The quality of life of patients undergoing RT may be compromised during and following treatment by nausea, diarrhea, vomiting, burns, erythema and fistula. Cytokinesis-block micronucleus (CBMN) assays may be useful for predicting adverse effects of RT for cancer. The CBMN test is easy to perform and is reproducible for screening subjects exposed to ionizing radiation. We investigated the use of the frequency of micronuclei (MN) from peripheral blood samples, irradiated in vitro, as a possible biomarker to predict the side effects of RT in patients with cervical cancer. We used 10 patients with cervical cancer receiving RT and chemotherapy. We found a strong relation between the frequency of MN and the appearance of acute side effects of RT for cervical cancer. We suggest that the methodology presented here may be useful for predicting side effects of RT for patients affected by cervical cancer and who have undergone chemotherapy.

Evaluation of Th17-related cytokines and IFNγ production from blood mononuclear cells of moderate and severe asthmatic children reveals methylprednisolone does not decrease IL-22 levels.

OBJECTIVE: The aim of this study was to correlate IL-6, IL-17A, IFNγ, and IL-22 production with asthma disease severity and to evaluate if methylprednisolone downregulated cytokine production in peripheral blood mononuclear cells (PBMCs). METHODS: Forty-two children with chronic persistent asthma and 34 non-asthmatic children were selected. Cytokines were quantified by ELISA from serum or PBMCs supernatants, after the PMA and Ionomycin stimulation, with or without methylprednisolone at 100 µM. RESULTS: Our data showed undetectable levels of serum cytokines in most patients and controls. In the PBMCs, we have observed a higher production of IL-17A than IL-22 among asthmatics and controls, although it is not statistically significant. IL-6, IFNγ, and IL-17A levels were significantly reduced after methylprednisolone treatment (p = 0.02, 0.03, and 0.03, respectively) in Severe Persistent Asthma (SPA) and in Moderate Persistent Asthma (MPA), (p = 0.007, 0.01, and 0.007, respectively). However, IL-22 levels were unaffected (SPA, p = 0.12 and MPA, p = 0.93). CONCLUSION: Methylprednisolone downregulated IL-6, IL17A, and IFNγ, but not IL-22, in stimulated PBMCs from asthmatic children indicating that methylprednisolone has no effect on IL-22 production by PBMCs.