Expression of functional kappa-opioid receptors on murine dendritic cells.

Endogenous and exogenous opioids are known to exert direct effects on the immune system and the expression of functional opioid receptors has been reported for several immune cell types. Since dendritic cells are important inducers and regulators of immune responses, we investigated whether murine dendritic cells express functional kappa-opioid receptors. FACScan analysis and radioligand binding studies revealed the expression of kappa-opioid receptors by murine dendritic cells, which by RT-PCR were also shown to express kappa-opioid mRNA. In a primary allogenic mixed-lymphocyte reaction the kappa-agonists dynorphin A and U50488H suppressed the capacity of dendritic cells to induce T-cell proliferation in a concentration-dependent manner. Preincubation with the kappa-specific antagonist nor-binaltrophimine abolished the observed effect, indicating specificity. In contrast, antigen uptake by dendritic cells as well as phenotypic maturation of dendritic cells were not influenced by the kappa-agonists dynorphin A and U50488H. In summary our data demonstrate that dendritic cells express functional kappa-opioid receptors and that specific agonists exert a direct effect on these cells. Therefore, dendritic cells might be involved in the interaction of the neuroendocrine hormones and the immune system.

Toxicological safety evaluation of gadobutrol.

OBJECTIVES: Gadobutrol (Gadovist/Gadavist, Bayer Pharma AG, Berlin, Germany) is a nonionic, macrocyclic, gadolinium-based contrast agent for magnetic resonance imaging of the central nervous system as well as liver and kidneys and for contrast enhancement in magnetic resonance angiography. For risk assessment of the single diagnostic use in humans, the toxicity of this compound was evaluated with a series of preclinical studies. MATERIALS AND METHODS: Preclinical studies into acute, repeated-dose, reproductive, and developmental toxicity as well as genotoxicity, local tolerance, contact-sensitizing potential, and antigenicity were performed. RESULTS: In rodents, lethality was observed after a single intravenous administration of 20 mmol/kg, representing doses at least 2 orders of magnitude higher than the standard single diagnostic dose in humans (0.1 mmol/kg). The no observed adverse effect levels after repeated (daily) administrations over the course of 4 weeks exceeded the single diagnostic dose in humans by a factor of 12 in rats and 10 in dogs (calculated on the basis of body weight), and no unexpected organ toxicity was observed. The most salient finding of repeated dosing in both rats and dogs was vacuolization of renal tubular epithelium without concomitant effect on kidney function, which represents a well-known finding for this class of compounds. Gadobutrol was not teratogenic in rats, rabbits, and monkeys even when given repeatedly during organogenesis at maximum dose levels tested, being 25 to 100 times (based on body weight) above the diagnostic dose in humans. No indications of potential genotoxic, contact allergenic, or immunotoxic effects were observed. In local tolerance testing, gadobutrol was well tolerated after intravenous administration. CONCLUSIONS: Gadobutrol was well tolerated with high safety margins between the single diagnostic dose of 0.1 mmol/kg in humans and the doses showing effects in animal studies.

Chemoimmunotherapy for melanoma with dacarbazine and 2,4-dinitrochlorobenzene elicits a specific T cell-dependent immune response.

An empirically established chemoimmunotherapy for metastatic melanoma combines the systemic administration of the chemotherapeutic agent dacarbazine (DTIC) with the epifocal application of the contact sensitizer 2,4-dinitrochlorobenzene (DNCB) on cutaneous metastases. Although this therapy yields high response rates resulting in prolonged survival, the mechanisms involved remain unknown. Here, we investigated whether treatment of tumor-bearing mice with DTIC and DNCB resulted in a specific immune response against the tumor. Subcutaneous (s.c.) tumors and lung metastases were induced in C57BL/6 mice by injecting syngeneic B16-melanoma cells s.c. or into the lateral tail vein, respectively. Mice were treated with intraperitoneal injections of DTIC followed by epifocal application of DNCB. This therapeutic approach significantly reduced the growth of s.c. tumors as well as lung metastases. Our data showed that the effector mechanisms involved are dependent on T cells. No therapeutic effect was observed in immunodeficient RAG-1(-/-) mice, or when the contact sensitizer DNCB was replaced by skin irritants (croton oil or tributyltin). Splenic lymphocytes obtained from treated mice displayed a three-fold higher specific cytolytic activity against B16 cells than in tumor-bearing controls. Both CD8(+) and CD4(+) T cells were able to lyse B16 cells. No changes were observed in natural killer (NK) cell activity. Likewise, tumor-infiltrating lymphocytes (TIL) of treated mice showed higher cytolytic activity than that of controls. Analysis of cytokine expression in s.c. tumors revealed increased mRNA levels of interferon-gamma (IFN-gamma), tumor necrosis factor-alpha (TNF-alpha), and interleukin-6 (IL-6) in treated tumors. Together, these findings demonstrate the ability of DTIC/DNCB treatment to induce an effective T cell-dependent host immune response against a syngeneic tumor.