Mouse IgG3 [HRP]

This study was conducted to assess the presence of West Nile virus (WNV) in Kosovo by serological testing of apparently healthy local horses and free-range chicken, and it attempted to detect viral nucleic acid in birds and mosquitoes. Between January 2018 and June 2019, 260 equine serum samples were collected, additionally 580 adult mosquitoes (53 pools) were grouped in for genera, includingCulexspp. (226 individuals; 26 pools),Aedesspp. (136 individuals; 16 pools),Anophelesspp. (184 individuals; 7 pools), andCulisetaspp. (34 individuals; 4 pools). Fifty domestic birds and 51 wild birds were collected from different regions of Kosovo. Equine and domestic bird serum samples were tested by flavivirus IgG enzyme-linked immunosorbent assay (ELISA), while mosquitoes and bird viscera were tested for WNV RNA by RT-qPCR. All ELISA-positive results were confirmed by plaque reduction neutralization test (PRNT) and eight by virus neutralization test. WNV antibodies were present in 27 out of 260 equine sera (10.38%) and one out of 50 samples in domestic birds by ELISA and PRNT. Eight of 27 positive equine serum samples with high titer neutralized WNV, but not Usutu virus. No WNV RNA was detected in birds or mosquitoes. The occurrence of WNV antibodies in local equines from all regions of Kosovo indicates that the virus is circulating within the country. Public health authorities should therefore plan a risk assessment and disease control program.

Background: Neutrophils accumulate in atherosclerotic plaques. Neutrophil extracellular traps (NET) were recently identified in experimental atherosclerosis and in complex human lesions. However, not much is known about the NET marker citrullinated histone-3 (H3Cit) expression and functionality in human carotid plaques. Moreover, the association between the proatherosclerotic autoantibody anti-apolipoprotein A-1 (anti-ApoA-1 IgG) and NET has never been investigated. Methods: Atherosclerotic plaques have been obtained from 36 patients with severe carotid stenosis that underwent carotid endarterectomy for severe carotid stenosis. Samples were sectioned into upstream and downstream regions from the same artery segment. Plaque composition and expression of NET markers neutrophil elastase (NE) and H3Cit were quantified by immunohistochemistry. H3Cit expression and function was evaluated by immunofluorescence and confocal analysis in a subset of patients. Results: Pathological features of vulnerable phenotypes were exacerbated in plaques developed at downstream regions, including higher accumulation of neutrophils and enhanced expression of NE and H3Cit, as compared to plaques from upstream regions. The H3Cit signal was also more intense in downstream regions, with significant extracellular distribution in spaces outside of neutrophils. The percentage of H3Cit colocalization with CD66b (neutrophils) was markedly lower in downstream portions of carotid plaques, confirming the extrusion of NET in this region. In agreement, the maximum distance of the H3Cit signal from neutrophils, extrapolated from vortex distance calculation in all possible directions, was also higher in downstream plaques. The serum anti-ApoA-1index positively correlated with the expression of H3Cit in downstream segments of plaques. Expression of the H3Cit signal outside of neutrophils and H3Cit maximal distance from CD66b-positive cells increased in plaques from serum positive anti-ApoA-1 patients compared with serum negative patients. Conclusion: NET elements are differentially expressed in upstream versus downstream regions of human carotid plaques and may be influenced by circulating levels of anti-ApoA-1 IgG. These findings could warrant the investigation of NET elements as potential markers of vulnerability.