BackgroundAlthough discovery research has identified the importance of dozens of pro- and anti-inflammatory immune mediators in the pathogenesis, maintenance, exacerbation and resolution of inflammatory diseases, most human cohort studies have incorporated few or no immunological intermediate phenotypes in their analyses. Significant hindrances have been (1) the limited panel of biomarkers known to be readily detected in healthy human populations and (2) the stability, hence utility, of such biomarkers to repeated analysis.MethodsThe frequency and stability of 14 plasma biomarkers linked to in vivo immune regulation of allergic and autoimmune inflammatory disorders was determined in 140 healthy pediatric and adult participants. The impact of initial and multiple subsequent freeze/thaw cycles on pro-inflammatory (CCL2, CXCL10, IL-18, TNFα, IL-6), anti-inflammatory (IL-10, sTNF-RII, IL-1Ra), acute phase proteins (CRP, PTX3) and other biomarkers (sST2, IL-1RAcP) was subsequently quantified.ResultsMultiple biomarkers capable of providing an innate immune signature of inflammation were readily detected directly ex vivo in healthy individuals. These biomarker levels were unaffected when comparing paired data sets from freshly obtained, never frozen plasma or serum and matched aliquots despite extensive freeze/thaw cycles. Neither age nor sex affected stability. Similarly, no quantitative differences were found following repetitive analysis of inflammatory biomarkers in culture samples obtained following in vitro stimulation with TLR and RLR ligands.ConclusionsA broad panel of in vivo and ex vivo cytokine, chemokine and acute phase protein biomarkers that have been linked to human chronic inflammatory disorders are readily detected in vivo and remain stable for analysis despite multiple freeze thaw cycles. These data provide the foundation and confidence for large scale analyses of panels of inflammatory biomarkers to provide better understanding of immunological mechanisms underlying health versus disease.Electronic supplementary materialThe online version of this article (doi:10.1186/s12967-017-1154-3) contains supplementary material, which is available to authorized users.
Food allergies, and peanut allergy in particular, are leading causes of anaphylactic fatalities worldwide. The immune mechanisms that underlie food allergy remain ill-defined and controversial, in part because studies in humans typically focus on analysis of a limited number of prototypical Th1/Th2 cytokines. Here we determine the kinetics and prevalence of a broad panel of peanut-driven cytokine and chemokine responses in humans with current peanut allergy vs those with stable, naturally occurring clinical tolerance to peanut. Our primary focus is identification of novel indicators of immune dysregulation. Antigen-specific cytokine mRNA and protein responses were elicited in primary culture via peanut or irrelevant antigen (Leishmania extract, milk antigens) mediated stimulation of fresh peripheral blood cells from 40 individuals. Peanut extract exposure in vitro induced a broad panel of responses associated with Th2/Th9-like, Th1-like and Th17-like immunity. Peanut-dependent Type 2 cytokine responses were frequently found in both peanut allergic individuals and those who exhibit clinical tolerance to peanut ingestion. Among Th2/Th9-associated cytokines, IL-9 responses discriminated between allergic and clinically tolerant populations better than did commonly used IL-4, IL-5 or IL-13 responses. Comparison with responses evoked by unrelated control antigen-mediated stimulation showed that these differences are antigen-dependent and allergen-specific. Conversely, the intensity of IL-12, IL-17, IL-23 and IFN-γ production was indistinguishable in peanut allergic and peanut tolerant populations. In summary, the ability to generate and maintain cytokine responses to peanut is not inherently distinct between allergic and peanut tolerant humans. Quantitative differences in the intensity of cytokine production better reflects clinical phenotype, with optimally useful indicators being IL-9, IL-5, IL-13 and IL-4. Equivalent, and minimal, Ag-dependent pro-inflammatory cytokine levels in both healthy and peanut allergic volunteers argues against a key role for such cytokines in maintenance of clinical tolerance to food antigens in humans.
OBJECTIVES: The aim of this research is to examine the awareness and use of the Good Food Junction (GFJ), a not-for-profit full service cooperative grocery store in a former food desert in Saskatoon, Canada. METHODS:Through door-to-door sampling, 365 residents in their neighbourhoods surrounding the GFJ grocery store were recruited. Quantitative surveys examined awareness, use and primary use of GFJ, mode of transportation to and from GFJ and primary grocery stores, other food program use and demographic data. Differences between those who had or had not shopped at GFJ were characterized using descriptive statistics and Pearson's chi-square test. Univariate and multivariate logistic regression models were developed to predict shopping at GFJ and the use of GFJ as a primary grocery store. RESULTS:Of those surveyed, 69% had shopped at GFJ. A significant proportion of shoppers were Aboriginal, had an annual household income per person of less than $20,000, and participated in other food-based programs and initiatives. Aboriginal people (OR = 2.0, p = 0.03) and users of neighbourhood-based fruit and vegetable markets (OR = 2.7, p = 0.04) were significantly more likely, but new immigrants to Canada (OR = 0.3, p = 0.05) were significantly less likely to have ever shopped at GFJ. Aboriginal respondents (OR = 2.6, p = 0.04) were significantly more likely to use GFJ as their primary grocery store. CONCLUSION:Our results confirm both that GFJ is able to serve households where food insecurity is likely and, based on the prevalence of users, the importance and need for a full-service supermarket in Saskatoon's inner city.KEY WORDS: Food supply; food deprivation; intervention studies La traduction du résumé se trouve à la fin de l'article.
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