Autoantibody-Based Diagnostic Biomarkers: Technological Approaches to Discovery and Validation

Autoantibodies produced against self-antigens, or ‘autoantigens’, result from a loss of self-tolerance triggered by genetic and/or environmental factors which induce the immune system to attack the host’s own cells, resulting in a condition referred to as autoimmunity. In classic autoimmune diseases, it is well established that the pathology relates directly to the autoantibodies. However, it is increasingly recognised that autoantibodies are also found in many other disease areas, including cancers, cardiovascular and neurodegenerative diseases, as well infectious diseases such as malaria, albeit in such diseases it is unclear whether the autoantibodies play a direct role in the pathology or whether they are merely symptomatic of disease. Irrespective of whether they are causative or symptomatic of specific diseases though, there is increasing interest globally in exploring the clinical potential of circulating autoantibodies as diagnostic biomarkers. This chapter provides an overview of the diagnostic utility of autoantibody biomarkers in a range of disease areas and discusses their potential utility in disease staging, treatment monitoring and in prediction of immune-related adverse events. It also provides an overview of traditional and contemporary technological approaches to autoantibody biomarker discovery and validation, focusing on protein microarrays that are ideally suited to this important area of research

Killing two birds with one stone: a chemically plausible scheme for linked nucleic acid replication and coded peptide synthesis

AbstractTo understand how life began, we must explain the origins of nucleic acid replication and genetically coded peptide synthesis. Neither of these is easy to explain individually; here, we propose a chemically plausible scheme for the evolution of a process that simultaneously produced both polymers. Later, two separate machineries could have evolved from the linked process

Quantitative Profiling of Colorectal Cancer-Associated Bacteria Reveals Associations between Fusobacterium spp., Enterotoxigenic Bacteroides fragilis (ETBF) and Clinicopathological Features of Colorectal Cancer

Various studies have presented clinical or in vitro evidence linking bacteria to colorectal cancer, but these bacteria have not previously been concurrently quantified by qPCR in a single cohort. We quantify these bacteria ( Fusobacterium spp ., Streptococcus gallolyticus , Enterococcus faecalis , Enterotoxigenic Bacteroides fragilis (ETBF), Enteropathogenic Escherichia coli (EPEC), and afaC- or pks-positive E . coli ) in paired tumour and normal tissue samples from 55 colorectal cancer patients. We further investigate the relationship between a) the presence and b) the level of colonisation of each bacterial species with site and stage of disease, age, gender, ethnicity and MSI-status. With the exception of S . gallolyticus , we detected all bacteria profiled here in both tumour and normal samples at varying frequencies. ETBF (FDR = 0.001 and 0.002 for normal and tumour samples) and afaC -positive E . coli (FDR = 0.03, normal samples) were significantly enriched in the colon compared to the rectum. ETBF (FDR = 0.04 and 0.002 for normal and tumour samples, respectively) and Fusobacterium spp. (FDR = 0.03 tumour samples) levels were significantly higher in late stage (III/IV) colorectal cancers. Fusobacterium was by far the most common bacteria detected, occurring in 82% and 81% of paired tumour and normal samples. Fusobacterium was also the only bacterium that was significantly higher in tumour compared to normal samples (p = 6e-5). We also identified significant associations between high-level colonisation by Fusobacterium and MSI-H (FDR = 0.05), age (FDR = 0.03) or pks -positive E . coli (FDR = 0.01). Furthermore, we exclusively identified atypical EPEC in our cohort, which has not been previously reported in association with colorectal cancer. By quantifying colorectal cancer-associated bacteria across a single cohort, we uncovered inter- and intra-individual patterns of colonization not previously recognized, as well as important associations with clinicopathological features, especially in the case of Fusobacterium and ETBF

Socio‐economic impact classification of alien taxa (SEICAT)

1 Many alien taxa are known to cause socio‐economic impacts by affecting the different constituents of human well‐being (security; material and non‐material assets; health; social, spiritual and cultural relations; freedom of choice and action). Attempts to quantify socio‐economic impacts in monetary terms are unlikely to provide a useful basis for evaluating and comparing impacts of alien taxa because they are notoriously difficult to measure and important aspects of human well‐being are ignored. 2 Here, we propose a novel standardised method for classifying alien taxa in terms of the magnitude of their impacts on human well‐being, based on the capability approach from welfare economics. The core characteristic of this approach is that it uses changes in peoples' activities as a common metric for evaluating impacts on well‐being. 2 Impacts are assigned to one of five levels, from Minimal Concern to Massive, according to semi‐quantitative scenarios that describe the severity of the impacts. Taxa are then classified according to the highest level of deleterious impact that they have been recorded to cause on any constituent of human well‐being. The scheme also includes categories for taxa that are not evaluated, have no alien population, or are data deficient, and a method for assigning uncertainty to all the classifications. To demonstrate the utility of the system, we classified impacts of amphibians globally. These showed a variety of impacts on human well‐being, with the cane toad (Rhinella marina) scoring Major impacts. For most species, however, no studies reporting impacts on human well‐being were found, i.e. these species were data deficient. 2 The classification provides a consistent procedure for translating the broad range of measures and types of impact into ranked levels of socio‐economic impact, assigns alien taxa on the basis of the best available evidence of their documented deleterious impacts, and is applicable across taxa and at a range of spatial scales. The system was designed to align closely with the Environmental Impact Classification for Alien Taxa (EICAT) and the Red List, both of which have been adopted by the International Union of Nature Conservation (IUCN), and could therefore be readily integrated into international practices and policies

A vision for global monitoring of biological invasions

Managing biological invasions relies on good global coverage of species distributions. Accurate information on alien species distributions, obtained from international policy and cross-border co-operation, is required to evaluate trans-boundary and trading partnership risks. However, a standardized approach for systematically monitoring alien species and tracking biological invasions is still lacking. This Perspective presents a vision for global observation and monitoring of biological invasions. We show how the architecture for tracking biological invasions is provided by a minimum information set of Essential Variables, global collaboration on data sharing and infrastructure, and strategic contributions by countries. We show how this novel, synthetic approach to an observation system for alien species provides a tangible and attainable solution to delivering the information needed to slow the rate of new incursions and reduce the impacts of invaders. We identify three Essential Variables for Invasion Monitoring; alien species occurrence, species alien status and alien species impact. We outline how delivery of this minimum information set by joint, complementary contributions from countries and global community initiatives is possible. Country contributions are made feasible using a modular approach where all countries are able to participate and strategically build their contributions to a global information set over time. The vision we outline will deliver wide-ranging benefits to countries and international efforts to slow the rate of biological invasions and minimize their environmental impacts. These benefits will accrue over time as global coverage and information on alien species increases

Spatiotemporal Variability in Allee Effects of Invading Gypsy Moth Populations

The Allee threshold, the critical population density separating growth from decline in populations experiencing strong Allee effects, can vary over space and time but few empirical studies have examined this variation. A lack of geographically extensive, long-term studies on low density population dynamics makes studying variability in Allee effects difficult. We used North American gypsy moth population data from 1996-2016 to quantify Allee thresholds in 11 regions of the invasion front. Allee thresholds spanned a continuum from being undetectable due to strong population growth at all densities, to being unmeasurable because populations declined across all densities. The lag-1 temporal autocorrelation in Allee thresholds tended to be negative and spatial synchrony in Allee thresholds extended no further than adjacent regions. This work furthers understanding of spatiotemporal variation in Allee effects using extensive empirical data at the range edge of an invasive insect

What factors increase the vulnerability of native birds to the impacts of alien birds?

Biodiversity impacts caused by alien species can be severe, including those caused by alien birds. In order to protect native birds, we aimed to identify factors that influence their vulnerability to the impacts of alien birds. We first reviewed the literature to identify native bird species sustaining such impacts. We then assigned impact severity scores to each native bird species, depending on the severity of the impacts sustained, and performed two types of analyses. First, we used contingency table tests to examine the distribution of impacts across their severity, type and location, and across native bird orders. Second, we used mixed-effects models to test factors hypothesised to influence the vulnerability of native birds to the impacts of alien birds. Ground-nesting shorebirds and seabirds were more prone to impacts through predation, while cavity-nesting woodpeckers and parrots were more prone to impacts through competition. Native bird species were more vulnerable when they occupied islands, warm regions, regions with climatic conditions similar to those in the native range of the invading alien species, and when they were physically smaller than the invading alien species. To a lesser extent, they were also vulnerable when they shared habitat preferences with the invading alien species. By considering the number and type of native bird species affected by alien birds, we demonstrate predation impacts to be more widespread than previously indicated, but also that damaging predation impacts may be underreported. We identify vulnerable orders of native birds, which may require conservation interventions; characteristics of native birds that increase their vulnerability, which may be used to inform risk assessments; and regions where native birds are most vulnerable, which may direct management interventions. The impacts sustained by native birds may be going unnoticed in many regions of the world: there is a clear need to identify and manage them

No saturation in the accumulation of alien species worldwide

Although research on human-mediated exchanges of species has substantially intensified during the last centuries, we know surprisingly little about temporal dynamics of alien species accumulations across regions and taxa. Using a novel database of 45,813 first records of 16,926 established alien species, we show that the annual rate of first records worldwide has increased during the last 200 years, with 37% of all first records reported most recently (1970–2014). Inter-continental and inter-taxonomic variation can be largely attributed to the diaspora of European settlers in the nineteenth century and to the acceleration in trade in the twentieth century. For all taxonomic groups, the increase in numbers of alien species does not show any sign of saturation and most taxa even show increases in the rate of first records over time. This highlights that past efforts to mitigate invasions have not been effective enough to keep up with increasing globalization

in silico verification and parallel reaction monitoring prevalidation of potential prostate cancer biomarkers

Purpose: Targeted proteomics of potential biomarkers is often challenging. Hence, we developed an intermediate workflow to streamline potential urinary biomarkers of prostate cancer (PCa). Materials & methods: Using previously discovered potential PCa biomarkers; we selected proteotypic peptides for targeted validation. Preliminary in silico immunohistochemical and single reaction monitoring (SRM) verification was performed. Successful PTPs were then prevalidated using parallel reaction monitoring (PRM) and reconfirmed in 15 publicly available databases. Results: Stringency-based targetable potential biomarkers were shortlisted following in silico screening. PRM reveals top 12 potential biomarkers including the top ranking seven in silico verification-based biomarkers. Database reconfirmation showed differential expression between PCa and benign/normal prostatic urine samples. Conclusion: The pragmatic penultimate screening step, described herein, would immensely improve targeted proteomics validation of ..

Identification of potential autoantigens in anti-CCP-positive and anti-CCP-negative rheumatoid arthritis using citrulline-specific protein arrays

Abstract The presence or absence of autoantibodies against citrullinated proteins (ACPAs) distinguishes two main groups of rheumatoid arthritis (RA) patients with different etiologies, prognoses, disease severities, and, presumably, disease pathogenesis. The heterogeneous responses of RA patients to various biologics, even among ACPA-positive patients, emphasize the need for further stratification of the patients. We used high-density protein array technology for fingerprinting of ACPA reactivity. Identification of the proteome recognized by ACPAs may be a step to stratify RA patients according to immune reactivity. Pooled plasma samples from 10 anti-CCP-negative and 15 anti-CCP-positive RA patients were assessed for ACPA content using a modified protein microarray containing 1631 different natively folded proteins citrullinated in situ by protein arginine deiminases (PADs) 2 and PAD4. IgG antibodies from anti-CCP-positive RA plasma showed high-intensity binding to 87 proteins citrullinated by PAD2 and 99 proteins citrullinated by PAD4 without binding significantly to the corresponding native proteins. Curiously, the binding of IgG antibodies in anti-CCP-negative plasma was also enhanced by PAD2- and PAD4-mediated citrullination of 29 and 26 proteins, respectively. For only four proteins, significantly more ACPA binding occurred after citrullination with PAD2 compared to citrullination with PAD4, while the opposite was true for one protein. We demonstrate that PAD2 and PAD4 are equally efficient in generating citrullinated autoantigens recognized by ACPAs. Patterns of proteins recognized by ACPAs may serve as a future diagnostic tool for further subtyping of RA patients