Innate and adaptive immunity in amyotrophic lateral sclerosis: evidence of complement activation.

Increasing evidence suggests a role for the immune system in amyotrophic lateral sclerosis (ALS). To determine the extent of the immune activation in ALS we analyzed the expression and cellular distribution of components of innate and adaptive immunity in spinal cord (SC) and motor cortex (MCx) from patients with rapid and slow sporadic ALS and controls. High levels of mRNA and protein of classical complement pathway, C1q and C4, as well as the downstream complement components C3 and C5b-9 were found in all ALS samples. Furthermore, we found higher numbers of activated microglia, reactive astrocytes, dendritic cells (DCs) and CD8(+) T-cells in ALS than in control tissue. Rapid ALS cases had more dendritic cells than slow ALS cases, whereas slow ALS cases had more activated microglia than rapid cases. Our findings demonstrate a persistent and prominent activation of both innate and adaptive immunity in ALS. We propose a complement-driven immune response which may contribute to the progression of the inflammation and ultimately lead to even more motor neuron injury.

Microscopic detection of viable Staphylococcus epidermidis in peri-implant tissue in experimental biomaterial-associated infection, identified by bromodeoxyuridine incorporation.

Infection of biomedical devices is characterized by biofilm formation and colonization of surrounding tissue by the causative pathogens. To investigate whether bacteria detected microscopically in tissue surrounding infected devices were viable, we used bromodeoxyuridine (BrdU), a nucleotide analogue that is incorporated into bacterial DNA and can be detected with antibodies. Infected human tissue was obtained postmortem from patients with intravascular devices, and mouse biopsy specimens were obtained from mice with experimental biomaterial infection. In vitro experiments showed that Staphylococcus epidermidis incorporated BrdU, as judged from staining of the bacteria with anti-BrdU antibodies. After incubation of bacteria with BrdU and subsequent staining of microscopic sections with anti-BrdU antibodies, bacteria could be clearly visualized in the tissue surrounding intravascular devices of deceased patients. With this staining technique, relapse of infection could be visualized in mice challenged with a low dose of S. epidermidis and treated with dexamethasone between 14 and 21 days after challenge to suppress immunity. This confirms and extends our previous findings that pericatheter tissue is a reservoir for bacteria in biomaterial-associated infection. The pathogenesis of the infection and temporo-spatial distribution of viable, dividing bacteria can now be studied at the microscopic level by immunolabeling with BrdU and BrdU antibodies.

[An experience of psychological support at home for aged and dependent people and their families. Assesment of the PsyAdom programme]. / Une expérience de soutien psychologique à domicile pour des personnes âgées et leurs aidants familiaux. Evaluation du programme PsyAdom.

The PsyAdom project has been initiated by the Clinical Psychologist Working in Gerontology Association. It consists in setting up an at home psychological follow-up for aged dependent subjects and their caregivers. Families are often isolated, and being kept up in their homes involves a risk of exhaustion. Psychological help is usually offered outside home, and cannot be available or very costly for some people. Eighteen couples were included in the study. Anxiety of the patients and caregivers was assessed by the Goldberg's anxiety scale, depression by the MADRS, experienced stress by the Mini-Zarit scale and attitude towards psychological help by the NPI. A qualitative assessment of the intervention was performed by recording semi-directed interviews of the involved psychologists to assess the patients and caregivers acceptance of the intervention, their compliance with participating, and reasons for stopping the intervention. No significant difference was found in the caregivers' burden and anxiety following the intervention. However, the results showed a lower level of depression in the caregivers, and a better attitude of the patients towards the caregivers. Analysis of the initial application for help and acceptance of the follow-up shows that possibility and relevance of psychological intervention at home depend on accurate indications. Initial application should be very unequivocal and the therapy protocol clearly explicited from the start to allow a good understanding of the goals of the therapy by the patients and caregivers.

The functional and morphological characteristics of sciatic nerve degeneration and regeneration after crush injury in rats.

BACKGROUND: Peripheral nerve damage induces a sequence of degeneration and regeneration events with a specific time course that leads to (partial) functional recovery. Quantitative electrophysiological analysis of degeneration and recovery over time is essential to understand the process. NEW METHOD: The presented ex vivo neurophysiological method evaluates functional recovery of the propagation of the compound action potential after crush injury of the rat sciatic nerve. A 32 channel electrode array was used to monitor compound action potential propagation at time points between 1h and 35 days after semi-quantitative crush injury of the rat sciatic nerve. RESULTS: The compound action potential was characterized by four measures: the latency, the duration, the amplitude and a measure that combined time and location. These four parameters reflected the subsequent steps in early axonal degradation, the transition to rapid degeneration followed by sprouting and the long period of remyelination that accompanied regeneration. COMPARISON WITH EXISTING METHODS: The neurophysiology measures of the compound action potential were compared with the morphology of the nerve at representative time points and analysis of functional recovery of action potential propagation was compared with a behavioral test: the foot flick test. CONCLUSIONS: Our data suggests that the ex vivo electrophysiological method is complementary to the classical behavioral foot flick test in that it allows a detailed time analysis of the degeneration and early regeneration phases at a high spatial and temporal sensitivity. The results were well-matched with observations made with immunohistochemical and morphological methods.

C6 deficiency does not alter intrinsic regeneration speed after peripheral nerve crush injury.

Peripheral nerve injury leads to Wallerian degeneration, followed by regeneration, in which functionality and morphology of the nerve are restored. We previously described that deficiency for complement component C6, which prevents formation of the membrane attack complex, slows down degeneration and results in an earlier recovery of sensory function after sciatic nerve injury compared to WT animals. In this study, we determine whether C6(-/-) rats have an intrinsic trait that affects sciatic nerve regeneration after injury. To study the contribution of complement activation on degeneration and regeneration with only minimal effect of complement activation, a crush injury model with only modest complement deposition was used. We compared the morphological and functional aspects of crushed nerves during degeneration and regeneration in C6(-/-) and WT animals. Morphological changes of myelin and axons showed similar degeneration and regeneration patterns in WT and C6(-/-) injured nerves. Functional degeneration and regeneration, recorded by ex vivo electrophysiology and in vivo foot flick test, showed that the timeline of the restoration of nerve conduction and sensory recovery also followed similar patterns in WT and C6(-/-) animals. Our findings suggest that C6 deficiency by itself does not alter the regrowth capacity of the peripheral nerve after crush injury.

Toll-like receptor signaling in amyotrophic lateral sclerosis spinal cord tissue.

Increasing evidence indicates that inflammatory responses could play a critical role in the pathogenesis of motor neuron injury in amyotrophic lateral sclerosis (ALS). Recent findings have underlined the role of Toll-like receptors (TLRs) and the receptor for advanced glycation endproducts (RAGE) in the regulation of both innate and adaptive immunity in different pathologies associated with neuroinflammation. In the present study we investigated the expression and cellular distribution of TLR2, TLR4, RAGE and their endogenous ligand high mobility group box 1 (HMGB1) in the spinal cord of control (n=6) and sporadic ALS (n=12) patients. The immunohistochemical analysis of TLR2, TLR4 and RAGE showed increased expression in reactive glial cells in both gray (ventral horn) and white matter of ALS spinal cord. TLR2 was predominantly detected in cells of the microglia/macrophage lineage, whereas the TLR4 and RAGE was strongly expressed in astrocytes. Real-time quantitative PCR analysis confirmed the increased expression of both TLR2 and TLR4 and HMGB1 mRNA level in ALS patients. In ALS spinal cord, HMGB1 signal is increased in the cytoplasm of reactive glia, indicating a possible release of this molecule from glial cells. Our findings show increased expression of TLR2, TLR4, RAGE and HMGB1 in reactive glia in human ALS spinal cord, suggesting activation of the TLR/RAGE signaling pathways. The activation of these pathways may contribute to the progression of inflammation, resulting in motor neuron injury. In this context, future studies, using animal models, will be important to achieve a better understanding of these signaling pathways in ALS in view of the development of new therapeutic strategies.