IL-17 producing gammadelta T cells are required for a controlled inflammatory response after bleomycin-induced lung injury.

BACKGROUND: gammadelta T cells play a key role in the regulation of inflammatory responses in epithelial tissue, and in adaptive immunity, as gammadelta T cell deficient mice have a severely impaired capacity to clear lung pathogens. gammadelta T cells regulate the initial inflammatory response to microbial invasion and thereby protect against tissue injury. Here we examined the response of gammadelta T cells to lung injury induced by bleomycin, in an effort to study the inflammatory response in the absence of any adaptive immune response to a pathogen. RESULTS: After lung injury by bleomycin, we localized the gammadelta T cells to the lung lesions. gammadelta T cells were the predominant source of IL-17 (as detected by flow cytometry and real-time PCR). Moreover, gammadelta T cell knockout mice showed a significant reduction in cellular infiltration into the airways, reduced expression of IL-6 in the lung, and a significant delay in epithelial repair. CONCLUSION: Mouse gammadelta T cells produce IL-17 in response to lung injury and are required for an organized inflammatory response and epithelial repair. The lack of gammadelta T cells correlates with increased inflammation and fibrosis.

Evaluation of Infrared Thermography for Temperature Measurement in Adult Male NMRI Nude Mice.

Temperature monitoring during critical care provides important data required to guide treatment delivery. Body temperatureis an easily quantified clinical parameter that can yield much information concerning the health of an animal. In researchsettings, temperature has been adopted as a means to judge humane endpoints. Therefore, reliable, noninvasive, and inexpensivemethods for temperature monitoring are becoming a necessity in research laboratories. This study aimed to determinethe accuracy and agreement of using an infrared camera as an alternative method of temperature measurement in mice andto compare the accuracy of this noninvasive method with established subcutaneous, intraperitoneal, and rectal techniques.Measurement of body-surface temperature by using an infrared camera was compared with these 3 established methods inmale NMRI nude mice (n = 10; age, 10 mo); data were obtained 3 times daily over 14 d. Subcutaneous temperatures weremeasured remotely by using a previously implanted subcutaneous temperature transponder, after which temperature wasmeasured by using noncontact infrared thermometry and a rectal probe. Measurements from intraperitoneal data loggers wereobtained retrospectively. The data show that using an infrared camera provides a simple, reliable method for measuring bodytemperature in male NMRI nu/nu mice that minimizes handling and is minimally invasive. Whether infrared thermometry is a useful method for measuring body temperature in furred mice warrants further investigation.

Tumor regression by combination antisense therapy against Plk1 and Bcl-2.

Increased expression of the cell proliferation-associated polo-like kinase 1 (PLK1) and apoptosis-associated BCL-2 genes has been observed in different human malignancies. Inhibition of cell proliferation and reactivation of apoptosis are basic principles in anticancer therapy. The efficiency of this approach is often limited by insuf-ficient targeting and delivery of anticancer drugs into the tumors. Phosphorothioate antisense oligodeoxynucleotides (ODNs) directed against PLK1 and BCL-2 were administered systemically via the tail vein into nude mice bearing A549, MDA-MB-435, and Detroit562 xenografts. To enhance tumor-specific uptake and to reduce systemic toxicity of antisense ODNs membrane electroporation transfer was applied in vivo. Northern and Western blot analyses were used to assess PLK1 and BCL-2 expression. Tumor mass was assessed after resection of tumors. All three cell lines and corresponding xenografts expressed high levels of PLK1 and were sensitive towards antisense PLK1 treatment. Antisense BCL-2 therapy was effective in tumors expressing high levels of BCL-2, but not in A549 cells and corresponding xenografts, which express low levels of BCL-2. Administration of antisense ODNs in a dose of 5 mg/kg, twice weekly during four weeks supported by the membrane electroporation transfer, eradicated 60-100% of the xenografted tumors. Antitumor effect in BCL-2 overexpressing MDA-MB-435 cells was synergistic for BCL-2 and PLK1 combination therapy. This study provides evidence that combined systemic administration of antisense ODNs against proliferation and pro- survival associated targets and in vivo electroporation of tumors represents a promising antitumor therapeutic approach.

Evaluation of a resistance-based model for the quantification of pulmonary arterial hypertension using MR flow measurements.

PURPOSE: To establish an estimate for the mean pulmonary arterial pressure (mPAP) derived from noninvasive data acquired with magnetic resonance (MR) velocity-encoded sequences. MATERIALS AND METHODS: In seven sedated pigs synchronous catheter-based invasive pressure measurements (IPM) and noninvasive MR were acquired in the main pulmonary artery (MPA) at different severities of pulmonary arterial hypertension (PAH) that were caused by infusion of thromboxane A2 (TxA2). The invasively measured mPAP was correlated with the noninvasive MR velocity data and linear combination equations (LCE) were computed. RESULTS: Intravenously applied TxA2 induced a dose dependent level of severity of PAH with an mPAP of up to 54 mmHg without systemic effects. The acceleration time (AT) measured with MR demonstrated the best correlation with the mPAP (r(2) = 0.75). The LCE with the highest correlation (R = 0.945, alpha < 0.01) between IPM and MR revealed a mean difference of 0, a SD of s = 4.66 and a maximal difference of 12.2 mmHg using the Bland-Altman analysis. CONCLUSION: Applying the identified LCE allowed the estimation of the mPAP in an acute and resistance-based model of PAH with high accuracy using noninvasive MR velocity-encoded sequences.

Chemotherapy of glioblastoma in rats using doxorubicin-loaded nanoparticles.

Glioblastomas belong to the most aggressive human cancers with short survival times. Due to the blood-brain barrier, they are mostly inaccessible to traditional chemotherapy. We have recently shown that doxorubicin bound to polysorbate-coated nanoparticles crossed the intact blood-brain barrier, thus reaching therapeutic concentrations in the brain. Here, we investigated the therapeutic potential of this formulation of doxorubicin in vivo using an animal model created by implantation of 101/8 glioblastoma tumor in rat brains. Groups of 5-8 glioblastoma-bearing rats (total n = 151) were subjected to 3 cycles of 1.5-2.5 mg/kg body weight of doxorubicin in different formulations, including doxorubicin bound to polysorbate-coated nanoparticles. The animals were analyzed for survival (% median increase of survival time, Kaplan-Meier). Preliminary histology including immunocytochemistry (glial fibrillary acidic protein, ezrin, proliferation and apoptosis) was also performed. Rats treated with doxorubicin bound to polysorbate-coated nanoparticles had significantly higher survival times compared with all other groups. Over 20% of the animals in this group showed a long-term remission. Preliminary histology confirmed lower tumor sizes and lower values for proliferation and apoptosis in this group. All groups of animals treated with polysorbate-containing formulations also had a slight inflammatory reaction to the tumor. There was no indication of neurotoxicity. Additionally, binding to nanoparticles may reduce the systemic toxicity of doxorubicin. This study showed that therapy with doxorubicin bound to nanoparticles offers a therapeutic potential for the treatment of human glioblastoma.