RÉSUMÉ
It is widely accepted that the endocannabinoid system (ECS) is a modulator of neuroinflammation associated with neurodegenerative disorders, including Alzheimer's disease (AD). Thus, expression of the cannabinoid receptor 2 (CB2) is induced in plaque-associated microglia and astrocytes in brain tissues from AD patients and in genetic mouse models expressing pathogenic variants of the amyloid precursor protein (APP). However, the exact mechanism of CB2 signaling in this mouse model remains elusive, because the genetic deletion of CB2 and the pharmacological activation of CB2 both reduced neuroinflammation. Here, we demonstrate that CB2 deletion also improved cognitive and learning deficits in APP/PS1*CB2-/- mice. This was accompanied by reduced neuronal loss and decreased plaque levels and coincided with increased expression of Aß degrading enzymes. Interestingly, plaque-associated microglia in APP/PS1*CB2-/- mice showed a less activated morphology, while plaques were smaller and more condensed than in APP/PS1 mice. Taken together, these results indicate a beneficial effect of CB2-deficiency in APP transgenic mice. CB2 appears to be part of a protective system that may be detrimental when engaged continuously.
Sujet(s)
Maladie d'Alzheimer/anatomopathologie , Maladie d'Alzheimer/physiopathologie , Régulation de l'expression des gènes/génétique , Plaque amyloïde/étiologie , Récepteur cannabinoïde de type CB2/déficit , Facteurs âges , Maladie d'Alzheimer/génétique , Peptides bêta-amyloïdes/métabolisme , Précurseur de la protéine bêta-amyloïde/génétique , Animaux , Modèles animaux de maladie humaine , Endocannabinoïdes/métabolisme , Humains , Apprentissage du labyrinthe/physiologie , Souris , Souris de lignée C57BL , Souris transgéniques , Microglie/métabolisme , Microglie/anatomopathologie , Neurones/métabolisme , Neurones/anatomopathologie , Plaque amyloïde/anatomopathologie , Préséniline-1/génétique , Récepteur cannabinoïde de type CB2/génétique , Transduction du signal/physiologieRÉSUMÉ
Cerebral malaria is a severe and often fatal complication of Plasmodium falciparum infection. It is characterized by parasite sequestration, a breakdown of the blood-brain barrier, and a strong inflammation in the brain. We investigated the role of the cannabinoid receptor 2 (CB2), an important modulator of neuroinflammatory responses, in experimental cerebral malaria (ECM). Strikingly, mice with a deletion of the CB2-encoding gene (Cnr2(-/-)) inoculated with Plasmodium berghei ANKA erythrocytes exhibited enhanced survival and a diminished blood-brain barrier disruption. Therapeutic application of a specific CB2 antagonist also conferred increased ECM resistance in wild type mice. Hematopoietic derived immune cells were responsible for the enhanced protection in bone marrow (BM) chimeric Cnr2(-/-) mice. Mixed BM chimeras further revealed that CB2-expressing cells contributed to ECM development. A heterogeneous CD11b(+) cell population, containing macrophages and neutrophils, expanded in the Cnr2(-/-) spleen after infection and expressed macrophage mannose receptors, arginase-1 activity, and IL-10. Also in the Cnr2(-/-) brain, CD11b(+) cells that expressed selected anti-inflammatory markers accumulated, and expression of inflammatory mediators IFN-γ and TNF-α was reduced. Finally, the M2 macrophage chemokine CCL17 was identified as an essential factor for enhanced survival in the absence of CB2, because CCL17 × Cnr2 double-deficient mice were fully susceptible to ECM. Thus, targeting CB2 may be promising for the development of alternative treatment regimes of ECM.
Sujet(s)
Barrière hémato-encéphalique/immunologie , Chimiokine CCL17/immunologie , Paludisme cérébral/immunologie , Plasmodium berghei/immunologie , Récepteur cannabinoïde de type CB2/immunologie , Animaux , Arginase/génétique , Arginase/immunologie , Barrière hémato-encéphalique/parasitologie , Barrière hémato-encéphalique/anatomopathologie , Chimiokine CCL17/génétique , Modèles animaux de maladie humaine , Prédisposition aux maladies , Femelle , Interleukine-10/génétique , Interleukine-10/immunologie , Lectines de type C/génétique , Lectines de type C/immunologie , Macrophages/immunologie , Macrophages/anatomopathologie , Paludisme cérébral/génétique , Paludisme cérébral/anatomopathologie , Mâle , Récepteur du mannose , Lectines liant le mannose/génétique , Lectines liant le mannose/immunologie , Souris , Souris knockout , Granulocytes neutrophiles/immunologie , Granulocytes neutrophiles/anatomopathologie , Récepteur cannabinoïde de type CB2/génétique , Récepteurs de surface cellulaire/génétique , Récepteurs de surface cellulaire/immunologieRÉSUMÉ
The endocannabinoid system (ECS) is a retrograde messenger system, consisting of lipid signaling molecules that bind to at least two G-protein-coupled receptors, Cannabinoid receptor 1 and 2 (CB1 and 2). As CB2 is primarily expressed on immune cells such as B cells, T cells, macrophages, dendritic cells, and microglia, it is of great interest how CB2 contributes to immune cell development and function in health and disease. Here, understanding the mechanisms of CB2 involvement in immune-cell function as well as the trafficking and regulation of CB2 expressing cells are crucial issues. Up to now, CB2 antibodies produce unclear results, especially those targeting the murine protein. Therefore, we have generated BAC transgenic GFP reporter mice (CB2-GFPTg) to trace CB2 expression in vitro and in situ. Those mice express GFP under the CB2 promoter and display GFP expression paralleling CB2 expression on the transcript level in spleen, thymus and brain tissue. Furthermore, by using fluorescence techniques we show that the major sources for GFP-CB2 expression are B cells in spleen and blood and microglia in the brain. This novel CB2-GFP transgenic reporter mouse line represents a powerful resource to study CB2 expression in different cell types. Furthermore, it could be used for analyzing CB2-mediated mobilization and trafficking of immune cells as well as studying the fate of recruited immune cells in models of acute and chronic inflammation.
Sujet(s)
Chromosomes artificiels de bactérie/génétique , Protéines à fluorescence verte/métabolisme , Souris transgéniques , Récepteur cannabinoïde de type CB2/génétique , Animaux , Lymphocytes B/métabolisme , Encéphale/métabolisme , Protéines à fluorescence verte/sang , Protéines à fluorescence verte/génétique , Souris , Régions promotrices (génétique) , Récepteur cannabinoïde de type CB2/sang , Récepteur cannabinoïde de type CB2/métabolisme , Rate/métabolisme , Thymus (glande)/métabolismeRÉSUMÉ
Several studies have indicated that the cannabinoid receptor 2 (CB2) plays an important role in neuroinflammation associated with Alzheimer's disease (AD) progression. The present study examined the role of CB2 in microglia activation in vitro as well as characterizing the neuroinflammatory process in a transgenic mouse model of AD (APP/PS1 mice). We demonstrate that microglia harvested from CB2(-/-) mice were less responsive to pro-inflammatory stimuli than CB2(+/+) microglia, based on the cell surface expression of ICAM and CD40 and the release of chemokines and cytokines CCL2, IL-6, and TNFα. Transgenic APP/PS1 mice lacking CB2 showed reduced percentages of microglia and infiltrating macrophages. Furthermore, they showed lowered expression levels of pro-inflammatory chemokines and cytokines in the brain, as well as diminished concentrations of soluble Aß 40/42. The reduction in neuroinflammation did not affect spatial learning and memory in APP/PS1*CB2(-/-) mice. These data suggest a role for the CB2 in Alzheimer's disease-associated neuroinflammation, independent of influencing Aß-mediated pathology and cognitive impairment.