Consistent with humoral reactions to the rotavirus vaccine (38), we did not get evidence for an association between antigen-specific immune-mediator concentrations and plasma sCD14, a biomarker of monocyte activation frequently used while an indirect indication of microbial translocation. were quantified in tradition supernatants by ELISA to determine antigen-specific immune function. The effect of stunting status and early-life exposures (anthropometry, swelling at 18 months, maternal health during pregnancy, household WASH) on immune function was tested in logit and censored log-normal (tobit) regression models. Results Children who were stunted (n = 44) experienced higher proportions (86.4% vs. 65.2%; 88.6% and affects 144 million children under 5 years old, mainly in LMIC (8). Stunted children possess high symptomatic and asymptomatic pathogen carriage (9) and are at greater risk of infectious mortality than their non-stunted peers (10). However, linear growth deficits are more likely an indicator than a cause of infectious susceptibility among undernourished children (11). There is some evidence that systemic inflammatory mediators are consistently higher (e.g., 6 weeks to 6 months (12), birth to 12 months (13)) in stunted versus non-stunted children, positively associated with circulating pathogen-associated molecular patterns (PAMPs) and anti-pathogen antibodies (14) but negatively associated with linear growth factors (12, 13, 15). Environmental enteropathy, a disorder which develops in the context of asymptomatic enteropathogen carriage, recurrent illness, intestinal damage and inflammation, is definitely thought to underlie the relationship between stunting and systemic swelling (9, 16), primarily translocation of PAMP from microbes in the gut into blood circulation (9, 14). For example, Rabbit polyclonal to GNRH concentrations of lipopolysaccharide (LPS, a component of gram-negative bacteria also termed endotoxin) in plasma samples from a cohort of babies in rural Gambia were found to be twice the top research limit for babies in high-income settings and negatively associated with their linear and ponderal growth (14). However, no studies to date have directly assessed whether immune cells from stunted children respond in a different way to PAMP relative to those of non-stunted children, how these practical variations originate or whether they relate to subsequent illness risk and severity. The goal of this study was to assess blood immune cell function among 18-month-old children in rural Zimbabwe, which has an estimated 23.5% prevalence of stunting among children under 5 years old (8), high rates of adverse birth results (17) and endemic pathogen exposure (18, 19). We focused on PAMPs derived from two gram-negative bacteria generally isolated from stool samples collected from children in LMIC (purified LPS from and whole heat-killed (urinary microscopy), and HIV status (the Zimbabwe National HIV screening algorithm (22)) were measured, blood and stool samples collected and maternal and household characteristics assessed by questionnaire at baseline. Birthweight (Tanita TBA-354 scales; Amsterdam, Netherlands) and delivery characteristics were recorded at birth. Child anthropometry and symptoms of illness (7-day time caregiver recall questionnaire) and blood and stool sampling were carried out whatsoever postnatal visits. TBA-354 In the 18-month check out, child haemoglobin was measured by HemoCue and median size determined from three measurements. Plasma and Stool ELISA Biomarkers of systemic swelling (capsular polysaccharide-reactive protein (CRP), sCD14) and environmental enteropathy (intestinal fatty acid-binding protein (IFABP)) were quantified in plasma by enzyme-linked immunosorbent assay (ELISA) (23). Biomarkers of intestinal swelling and environmental enteropathy (myeloperoxidase (MPO), neopterin, -1-antitrypsin (AAT)) were measured by stool ELISA (23). Whole Blood Ethnicities Blood was collected from each child by venipuncture at their 18-month check TBA-354 out. Six hundred millilitres of blood was diluted in RPMI 1640 medium supplemented with 1% v/v penicillinCstreptomycin (GIBCO, Amarillo, USA) and cultured in 24-well plates for 24 h under three parallel conditions (one tradition well/condition; culture conditions for the study were pre-prepared and stored at -80C until use to minimise inter-plate variance): without stimulus (Press); 0.25 EU/mL ultrapure lipopolysaccharide from (LPS); and 1 108 cells/mL heat-killed serovar Typhimurium (HKST; Invivogen, Toulouse, France). Ethnicities were managed at 37C, 6% CO2, using the CO2Gen Compact system (Oxoid, Basingstoke, UK). Cell-free tradition supernatants were harvested and stored at -80C. Supernatants were transferred to Zvitambo Institute laboratory, Harare, where TNF, IL-6, IL-8, IL-12p70, hepcidin, sCD163, MPO (Bio-Techne DuoSet, Abingdon, UK) and IFN (PBL VeriKine, Piscataway, USA) concentrations were quantified in duplicate TBA-354 ELISA relative to eight-point standard curves. Mediators were chosen to reflect important domains of innate immune cell function: pro-inflammatory cytokines (TNF and IL-6) that result in the liver acute-phase response; IFN known for its antiviral and antibacterial action (24); IL-8, a pro-inflammatory cytokine and neutrophil chemoattractant; sCD163, a bacterial sensor shed by triggered monocytes/macrophages (25); IL-12p70 which promotes antibacterial/antiviral T-cell function; hepcidin which inhibits cellular iron efflux to regulate iron homeostasis and/or defend against extracellular pathogens (26); and MPO, an enzyme which catalyses bactericidal.
Consistent with humoral reactions to the rotavirus vaccine (38), we did not get evidence for an association between antigen-specific immune-mediator concentrations and plasma sCD14, a biomarker of monocyte activation frequently used while an indirect indication of microbial translocation
