Anti-inflammatory signaling in microglia exacerbates Alzheimer's disease-related pathology.

In this issue of Neuron, Chakrabarty et al. (2015) and Guillot-Sestier et al. (2015) reveal that the anti-inflammatory cytokine IL-10 inhibits Aß clearance by microglia, worsening cognitive decline in mouse models of Alzheimer's disease (AD). These studies provide further support that pro-inflammatory signaling is an innate immune defense mechanism in AD.

IL-10 alters immunoproteostasis in APP mice, increasing plaque burden and worsening cognitive behavior.

Anti-inflammatory strategies are proposed to have beneficial effects in Alzheimer's disease. To explore how anti-inflammatory cytokine signaling affects Aß pathology, we investigated the effects of adeno-associated virus (AAV2/1)-mediated expression of Interleukin (IL)-10 in the brains of APP transgenic mouse models. IL-10 expression resulted in increased Aß accumulation and impaired memory in APP mice. A focused transcriptome analysis revealed changes consistent with enhanced IL-10 signaling and increased ApoE expression in IL-10-expressing APP mice. ApoE protein was selectively increased in the plaque-associated insoluble cellular fraction, likely because of direct interaction with aggregated Aß in the IL-10-expressing APP mice. Ex vivo studies also show that IL-10 and ApoE can individually impair glial Aß phagocytosis. Our observations that IL-10 has an unexpected negative effect on Aß proteostasis and cognition in APP mouse models demonstrate the complex interplay between innate immunity and proteostasis in neurodegenerative diseases, an interaction we call immunoproteostasis.

Immunoproteomics for serological diagnosis of hypersensitivity pneumonitis caused by environmental microorganisms.

Diagnosis of immunoallergenic pathologies due to microorganisms such as hypersensitivity pneumonitis includes detection of circulating specific antibodies. Detection of precipitins has classically been performed using immunoprecipitation techniques with crude antigenic extracts from microorganisms implicated as etiologic agents. However, these techniques lack standardization because of the different composition of fungal antigenic extracts from one batch to another. Therefore, there is high interest in developing standardized serological diagnostic methods using recombinant antigens. Immunoproteomics have proved to be useful for identifying the immunogenic proteins in several microorganisms linked to hypersensitivity pneumonitis. With this approach, the causative microorganisms are first isolated from the environment of patients. Then the proteins are separated by two-dimensional electrophoresis and revealed by Western blotting with sera of different patients suffering from the disease compared to sera of asymptomatic exposed controls. Immunoreactive proteins are identified by mass spectrometry. Identified immunoreactive proteins found to be specific markers for the disease could be subsequently produced as recombinant antigens using various expression systems to develop ELISA tests. Using recombinant antigens, standardized ELISA techniques can be developed, with sensitivity and specificity reaching 80% and 90%, respectively, and more if using a combination of several antigens. Immunoproteomics can be applied to any environmental microorganisms, with the aim of proposing panels of recombinant antigens able to improve the sensitivity and standardization of serologic diagnosis of hypersensitivity pneumonitis, but also other mold-induced allergic diseases such as allergic broncho pulmonary aspergillosis or asthma.

Gene expression in splenic CD4 and CD8 cells from BALB/c mice immunized with Porphyromonas gingivalis.

BACKGROUND: T cells are fundamental in the pathogenesis of periodontal disease. Suppression of cell-mediated responses is associated with disease progression together with the concomitant increase in plaque pathogens including Porphyromonas gingivalis. The aim of the present study was to examine gene expression in T cells in response to P. gingivalis in mice. METHODS: BALB/c mice were given weekly intraperitoneal injections of P. gingivalis outer-membrane antigens with Freund's incomplete adjuvant for 3 weeks, whereas control mice received phosphate buffered saline (PBS) and adjuvant only. Splenic CD4 and CD8 subpopulations were isolated by magnetic cell separation and their responses investigated using microarray analysis. RESULTS: Most genes coded for enzymes concerned with metabolic pathways. Only five and 28 genes, respectively, were upregulated in CD4 and CD8 cells extracted from P. gingivalis-immunized mice, including immunoglobulin (Ig) heavy-chain genes for IgG1 and IgG2a in CD4 cells. In contrast, 1,141 and 1,175 genes, respectively, were downregulated. A total of 60 and 65 genes, respectively, coded for immune response proteins or those relevant to periodontal disease pathogenesis. The overlap of genes in the two subsets was 21%. One of the major effects, apart from T-cell function suppression, was the shift away from Th1 responses, although there was also a downregulation of two genes and upregulation of one Th2-response gene. Genes downregulated included those encoding cytokines, proteins involved in Ig binding, antigen presentation, innate immunity, extracellular matrix, and cell adhesion molecules that could result in dysregulation in the progressive periodontal lesion. CONCLUSIONS: Early findings in humans demonstrated that periodontopathic bacteria induce immunosuppressive effects on T cells. The present study has shown that P. gingivalis had a predominant downregulatory effect on gene expression in CD4 and CD8 T cells in mice.

Identification of basophils by a mAb directed against pro-major basic protein 1.

BACKGROUND: Basophils possess characteristics of both mast cells and eosinophils, and all 3 cell types often are found together, particularly during allergic reactions. A mAb (J175-7D4) generated against the recombinant pro-form of human eosinophil granule major basic protein 1 (rproMBP1) appeared to stain only basophils in tissue specimens. OBJECTIVE: We investigated J175-7D4 to characterize its specificity for basophils. METHODS: Fluid-phase immunoprecipitation, Western blotting, and immunocytochemistry and immunohistochemistry were used to establish the specificity of J175-7D4. RESULTS: First, J175-7D4 binds to various glycosylated and proteolytically processed forms of rproMBP1, but not to major basic protein. Second, cells transfected with the rproMBP1 gene and human placental tissue (known to express the pro-form of major basic protein 1 [proMBP1]) stain specifically with J175-7D4. In contrast, although mature eosinophils contain substantial major basic protein, they lack proMBP1 and do not stain. Neutrophils, lymphocytes, monocytes, and skin mast cells also are not stained. However, blood basophils are stained by J175-7D4, anti-IgE, Wright-Giemsa (metachromatically), and a previously characterized basophil-specific mAb, 2D7. Finally, formaldehyde-fixed, paraffin-embedded basophils are identically detected by J175-7D4 and 2D7, and J175-7D4 also recognizes putative basophils in formaldehyde-fixed, paraffin-embedded tissue specimens from inflammatory dermatoses, such as atopic dermatitis and delayed pressure urticaria. CONCLUSION: The J175-7D4 mAb recognizes proMBP1 as a novel marker for human basophils. J175-7D4 should prove useful for characterizing basophil involvement in human health and disease.