An armed anti-immunoglobulin light chain nanobody protects mice against influenza A and B infections.

The immune system eliminates pathogen intruders such as viruses and bacteria. To recruit immune effectors to virus-infected cells, we conjugated a small molecule, the influenza neuraminidase inhibitor zanamivir, to a nanobody that recognizes the kappa light chains of mouse immunoglobulins. This adduct was designed to achieve half-life extension of zanamivir through complex formation with the much-larger immunoglobulins in the circulation. The zanamivir moiety targets the adduct to virus-infected cells, whereas the anti-kappa component simultaneously delivers polyclonal immunoglobulins of indeterminate specificity and all isotypes. Activation of antibody-dependent cell-mediated cytotoxicity and complement-dependent cytotoxicity promoted elimination of influenza neuraminidase-positive cells. A single dose of the conjugate protected mice against influenza A or B viruses and was effective even when given several days after infection with a lethal dose of virus. In the absence of circulating immunoglobulins, we observed no in vivo protection from the adduct. The type of conjugates described here may thus find application for both anti-influenza prophylaxis and therapy.

Targeted delivery of an anti-inflammatory corticosteroid to Ly6C/G-positive cells abates severity of influenza A symptoms.

The distribution of Ly6C/G-positive cells in response to an infection of the mouse respiratory tract with influenza A virus was followed noninvasively over time by immuno-positron emission tomography. We converted nanobodies that recognize Ly6C and Ly6G, markers of neutrophils and other myeloid cells, as well as an influenza hemagglutinin-specific nanobody, into 89Zr-labeled PEGylated positron emission tomography (PET) imaging agents. The PET images showed strong accumulation of these imaging agents in the lungs of infected mice. Immunohistochemistry of influenza virus-infected mice and control mice, injected with a biotinylated and PEGylated version of the Ly6C/G-specific nanobody, showed the presence of abundant Ly6C/G-positive myeloid cells and positivity for Ly6C/G on bronchial epithelium in influenza virus-infected mice. This is consistent with focal inflammation in the lungs, a finding that correlated well with the immuno-PET results. No such signals were detected in control mice. Having shown by PET the accumulation of the Ly6C/G-specific nanobody in infected lungs, we synthesized conjugates of Ly6C/G-specific nanobodies with dexamethasone to enable targeted delivery of this immunosuppressive corticosteroid to sites of inflammation. Such conjugates reduced the weight loss that accompanies infection, while the equivalent amount of free dexamethasone was without effect. Nanobody-drug conjugates thus enable delivery of drugs to particular cell types at the appropriate anatomic site(s). By avoiding systemic exposure to free dexamethasone, this strategy minimizes its undesirable side effects because of the much lower effective dose of the nanobody-dexamethasone conjugate. The ability to selectively target inflammatory cells may find application in the treatment of other infections or other immune-mediated diseases.

Vascular Risk Score and Associations With Past, Current, or Future Migraine in Women: Cohort Study.

BACKGROUND AND OBJECTIVES-: Migraine has consistently been associated with an increased risk of cardiovascular disease (CVD) events. It remains, however, unclear to what extent cardiovascular risk profiles might be linked with migraine activity status, and how these profiles relate to the development of migraine. METHODS-: We used data from a cohort study of female health professionals (Women's Health Study, n=27,539, age ≥45 years at baseline) without a history of CVD or other major diseases and who provided a blood sample at baseline. Framingham risk scores (FRS) estimating the ten-year risk of coronary heart disease calculated at baseline were used to create vascular risk categories. The presence or development of self-reported migraine was assessed by questionnaires. Women were classified as having 'no migraine', 'history of migraine' (experienced migraine in the past but did not experience any migraine attacks in the year before enrollment), 'migraine at baseline' (active), or 'incident migraine' (first report of migraine during follow-up but not at baseline). We used multinomial logistic regression models to calculate odds ratios (ORs) for the association between FRS categories and migraine status. RESULTS-: Of the 27,539 participants, a total of 21,927 women did not report migraine, 1,500 women reported a history of migraine, 3,579 had migraine at baseline, and 533 reported migraine for the first time during follow-up. The odds of the probability of having a history of migraine at baseline (versus never migraine) was 76% higher among those with FRS ≥10% compared with FRS ≤1% after adjustment (OR=1.76, 95%CI: 1.39-2.23). In contrast, having FRS ≥10% was inversely associated with migraine at baseline (OR=0.64, 95%CI: 0.52-0.80), and with newly reported migraine during follow-up (OR=0.42, 95%CI: 0.22-0.81) when compared with women with FRS category ≤1% and those not reporting migraine. A similar inverse association pattern was observed for FRS categories 5-9% and 2-4%. DISCUSSION-: High FRS categories were only observed among women with a history of migraine but not with active migraine at baseline or incident migraine after baseline. Our results suggest that the life course of migraine should be considered when studying associations with the vascular system. Our data further suggest that a relatively healthy vascular system, as assessed by the FRS, is associated with active migraine status or developing migraine in the future.

An adjuvant strategy enabled by modulation of the physical properties of microbial ligands expands antigen immunogenicity.

Activation of the innate immune system via pattern recognition receptors (PRRs) is key to generate lasting adaptive immunity. PRRs detect unique chemical patterns associated with invading microorganisms, but whether and how the physical properties of PRR ligands influence the development of the immune response remains unknown. Through the study of fungal mannans, we show that the physical form of PRR ligands dictates the immune response. Soluble mannans are immunosilent in the periphery but elicit a potent pro-inflammatory response in the draining lymph node (dLN). By modulating the physical form of mannans, we developed a formulation that targets both the periphery and the dLN. When combined with viral glycoprotein antigens, this mannan formulation broadens epitope recognition, elicits potent antigen-specific neutralizing antibodies, and confers protection against viral infections of the lung. Thus, the physical properties of microbial ligands determine the outcome of the immune response and can be harnessed for vaccine development.

Self-perceived attention deficit and hyperactivity symptom levels and risk of non-migraine and migraine headaches among university students: A cross-sectional study.

OBJECTIVES: The aim was to evaluate the association of self-perceived levels of attention deficit and hyperactivity symptoms with non-migraine and migraine headaches among university students. We also evaluated their association with migraine aura. METHODS: Study subjects were all participants in the internet-based Students Health Research Enterprise. Scores were built to evaluate global attention and hyperactivity symptom levels, self-perceived attention deficit levels and self-perceived hyperactivity symptom levels based on the Adult Attention Deficit and Hyperactivity Disorder Self-Report Scale (ASRS v1.1.). We used standardised questions to classify headache and group participants into "no headache," "non-migraine headache," "migraine without aura" or "migraine with aura". RESULTS: A total of 4816 students were included (mean age 20.3 ± 2.8 years; 75.5% women). Compared with participants without headache, we found significant associations between global ADHD scores and migraine. Students in the highest quintile of global ASRS scores had adjusted odds ratio (aOR) of 1.95 (95% CI 1.56-2.45) when compared to the lowest. This association was mainly driven by an association between self-perceived hyperactivity and migraine with aura. The aOR for migraine with aura was 2.83 (95% CI 2.23-3.61) for students in the highest quintile of hyperactivity. No significant association was found for any attention and hyperactivity symptom level measure and non-migraine headache and between self-perceived levels of attention deficit and migraine. CONCLUSIONS: Among students in higher education in France, self-perceived levels of attention deficit and hyperactivity symptoms were selectively associated with migraine. The association was strongest for the hyperactivity domain and migraine with aura.

Screen time exposure and reporting of headaches in young adults: A cross-sectional study.

Objective The objective of this article is to investigate whether excessive screen time exposure is associated with non-migraine headache and migraine in young adults. Background Increased levels of television time have been associated with increased risk of headache. However, time spent using newer electronic devices with a screen (smartphone, tablet) has not been examined yet. Methods We conducted a cross-sectional study among 4927 participants of the French i-Share cohort. Demographic characteristics, screen time exposure (computers, tablets, smartphones and television) as well as headache/migraine symptoms were recorded in a standardized questionnaire. Multinomial logistic regression models were used to evaluate the association between screen time exposure and headache status. Results Participants had a mean age of 20.8 years and 75.5% were female. The multivariable model showed that students in the highest screen time exposure quintile had an increased risk for migraine. The odds ratio (OR) (95% confidence interval (CI)) was 1.37 (1.14 to 1.66) for migraine when compared with students without headache and with low screen time exposure. This association was somewhat stronger for migraine without aura (OR = 1.50, 95% CI 1.19 to 1.89). We found no significant association between screen time exposure and non-migraine headache. Conclusion High levels of screen time exposure are associated with migraine in young adults. No significant association was found with non-migraine headache.