cGAS-STING signalling regulates microglial chemotaxis in genome instability.

Defective DNA damage signalling and repair is a hallmark of age-related and genetic neurodegenerative disease. One mechanism implicated in disease progression is DNA damage-driven neuroinflammation, which is largely mediated by tissue-resident immune cells, microglia. Here, we utilise human microglia-like cell models of persistent DNA damage and ATM kinase deficiency to investigate how genome instability shapes microglial function. We demonstrate that upon DNA damage the cytosolic DNA sensing cGAS-STING axis drives chronic inflammation and a robust chemokine response, exemplified by production of CCL5 and CXCL10. Transcriptomic analyses revealed that cell migratory pathways were highly enriched upon IFN-ß treatment of human iPSC-derived microglia, indicating that the chemokine response to DNA damage mirrors type I interferon signalling. Furthermore, we find that STING deletion leads to a defect in microglial chemotaxis under basal conditions and upon ATM kinase loss. Overall, this work provides mechanistic insights into cGAS-STING-dependent neuroinflammatory mechanisms and consequences of genome instability in the central nervous system.

Newborn Screening for Presymptomatic Diagnosis of Complement and Phagocyte Deficiencies.

The clinical outcomes of primary immunodeficiencies (PIDs) are greatly improved by accurate diagnosis early in life. However, it is not common to consider PIDs before the manifestation of severe clinical symptoms. Including PIDs in the nation-wide newborn screening programs will potentially improve survival and provide better disease management and preventive care in PID patients. This calls for the detection of disease biomarkers in blood and the use of dried blood spot samples, which is a part of routine newborn screening programs worldwide. Here, we developed a newborn screening method based on multiplex protein profiling for parallel diagnosis of 22 innate immunodeficiencies affecting the complement system and respiratory burst function in phagocytosis. The proposed method uses a small fraction of eluted blood from dried blood spots and is applicable for population-scale performance. The diagnosis method is validated through a retrospective screening of immunodeficient patient samples. This diagnostic approach can pave the way for an earlier, more comprehensive and accurate diagnosis of complement and phagocytic disorders, which ultimately lead to a healthy and active life for the PID patients.

Parallel barcoding of antibodies for DNA-assisted proteomics.

DNA-assisted proteomics technologies enable ultra-sensitive measurements in multiplex format using DNA-barcoded affinity reagents. Although numerous antibodies are available, nowadays targeting nearly the complete human proteome, the majority is not accessible at the quantity, concentration, or purity recommended for most bio-conjugation protocols. Here, we introduce a magnetic bead-assisted DNA-barcoding approach, applicable for several antibodies in parallel, as well as reducing required reagents quantities up to a thousand-fold. The success of DNA-barcoding and retained functionality of antibodies were demonstrated in sandwich immunoassays and standard quantitative Immuno-PCR assays. Specific DNA-barcoding of antibodies for multiplex applications was presented on suspension bead arrays with read-out on a massively parallel sequencing platform in a procedure denoted Immuno-Sequencing. Conclusively, human plasma samples were analyzed to indicate the functionality of barcoded antibodies in intended proteomics applications.

Magnetic bead assisted labeling of antibodies at nanogram scale.

There are currently several initiatives that aim to produce binding reagents for proteome-wide analysis. To enable protein detection, visualization, and target quantification, covalent coupling of reporter molecules to antibodies is essential. However, current labeling protocols recommend considerable amount of antibodies, require antibody purity and are not designed for automation. Given that small amounts of antibodies are often sufficient for downstream analysis, we developed a labeling protocol that combines purification and modification of antibodies at submicrogram quantities. With the support of magnetic microspheres, automated labeling of antibodies in parallel using biotin or fluorescent dyes was achieved.

Hot and Cold natures and some parameters of neuroendocrine and immune systems in traditional Iranian medicine: a preliminary study.

AIM: The purpose of this study was to assess differences in persons of a Hot or Cold nature (according to traditional Iranian medicine), in terms of changes in their neuroendocrine and immune systems. MATERIALS AND METHODS: Thirty-seven (37) male volunteers (20-40 years old) were divided into two groups, by whether they had a Hot or Cold nature. In addition, the Warmth/Coldness ratio of all the volunteers was assessed. Plasma concentrations of epinephrine, norepinephrine and cortisol, and also the concentrations of interferon (IFN)-gamma and interleukin (IL)-4 produced by peripheral blood mononuclear cells stimulated by mitogen were measured. RESULTS: The results showed that norepinephrine/epinephrine and norepinephrine/cortisol ratios were significantly higher, and that there was a borderline significantly increased IL-4/IFN-gamma ratio in the Hot nature group compared with those in the Cold nature group. In addition, there was a significant linear positive correlation between the norepinephrine/epinephrine and Warmth/Coldness ratios and a significant nonlinear association between the IL-4/IFN-gamma and Warmth/Coldness ratios. CONCLUSIONS: It can be deduced that the persons of a Hot nature had more sympathetic nervous system activity, less adrenal sympathetic, adrenal corticosteroid, and parasympathetic nervous system activities and more deviation of the immune system toward T-helper (Th)2 responses than the persons of a Cold nature. Moreover, the activity of the sympathetic nervous system was increased and adrenal sympathetic was decreased with an increasing Warmth/Coldness ratio. Furthermore, when the person's nature veered toward extreme Warmth or extreme Coldness, the deviation of the immune system toward Th2-like responses was greater, but this increased deviation was much more marked when veering toward extreme Warmth than toward extreme Coldness.