Development of a humanised mouse model of ovalbumin induced allergy and inflammation as a platform to test functional food

Food allergies are most common during infancy Food allergy is a growing health problem with very limited treatment options For infants, cow’s milk protein allergy (CMPA) is the predominant food allergy, occurring in 2 - 7 5 % of population Infant formula which is often cow’s milk based, is the only approved alternative for breastfeeding However the proteins (casein and whey) in cow’s milk result in allergic symptoms in the infants These allergic symptoms can be avoided by providing the infants with hydrolysates of cow’s milk protein instead of the intact protein Whey protein hydrolysates, small bioactive peptide components of milk have been tested in mouse models for their anti-inflammatory/immunomodulatory properties however to date no studies have repeated efficacy of hydrolysates in a humanised mouse model setting In this study a large number of hydrolysates were screened in vitro to identify hydrolysates with anti-inflammatory properties In vitro we demonstrated that 147 hydrolysate significantly reduced DC maturation in a P-PARy dependent manner To mvestigate the anti-inflammatory potential of hydrolysate 147 in a human relevant model of food allergy we developed a novel humanised mouse model of OVA driven food allergy We directly compared two different strains of nonobese diabetic severe combined immunodeficient mice lacking the cytokine receptor common gamma chain (yc7') (NSG-SCF versus NSG- SGM3) expressing human cytokine genes for their suitability as a humanised mouse model Using NSG-SCF mice a human immune system was developed through engraftment of CD34+ hematopoietic stem cells Using an OVA sensitisation and challenge protocol the model of food allergy was developed in these humanised mice The capacity of hydro lysate 147 to mediate anti-inflammatory effects was examined through repeated administration of 147 hydrolysate after sensitisation and before challenge with OVA Both 147 hydrolysate and the wpc80 parent control equally reduced anaphylaxis. In comparison to wpc80 parent control, 147 reduced specific antibody responses, including OVA-specific IgGl, reduced the production of the pro- inflammatory cytokines IL-6, IL-17 and induced higher levels of the anti-inflammatory cytokine IL-10 in spleens and small intestines of OVA treated mice 147 in comparison to wpc80 was more potent at promoting a Thl/Th2 balance in the spleen and most significantly suppressed the accumulation of granulocyte populations in bone marrow and liver while simultaneously reducing the T , B and myeloid cell populations m the bone marrow of OVA mice For the first time, we have shown that a novel milk protein hydrolysate provided efficacy in comparison to its whey parent control in terms of reducing OVA driven allergy in a humanised mouse model This study demonstrated that 147 may prevent the development of other food allergies (egg-ova) in addition to CMPA (but not examined here) In addition this study, presents a framework, from which suitable conditions for testing functional foods, can be adopted in the development of novel agents for the prevention of allergic inflammation

Linocin and OmpW Are Involved In Attachment Of The Cystic Fibrosis-Associated Pathogen Burkholderia Cepacia Complex To Lung Epithelial Cells and Protect Mice Against Infection

Burkholderia cepacia complex (Bcc) causes chronic opportunistic lung infections in people with cystic fibrosis (CF) resulting in a gradual lung function decline and, ultimately, patient death. Bcc is a complex of eighteen species and is rarely eradicated once a patient is colonised, therefore vaccination may represent a better therapeutic option. We developed a new proteomics approach to identify bacterial proteins that are involved in attachment of Bcc to lung epithelial cells. Fourteen proteins were reproducibly identified by 2-DE from four Bcc strains, representative of two Bcc species: B. cenocepacia, the most virulent and B. multivorans, the most frequently acquired. Seven proteins were identified in both species, but only two were common to all four strains, Linocin and OmpW. Both proteins were selected based previously published data on these proteins in other species. The E. coli strains expressing recombinant Linocin and OmpW showed enhanced attachment (4.2- and 3.9-fold) to lung cells, compared to control, confirming that both proteins are involved in host cell attachment. Immunoproteomic analysis using serum from Bcc colonised CF patients confirmed that both proteins elicit potent humoral responses in vivo. Mice immunised with either recombinant Linocin or OmpW were protected from B. cenocepacia and B. multivorans challenge. Both antigens induced potent antigen-specific antibody responses and stimulated strong cytokine responses. In conclusion, our approach identified adhesins that induced excellent protection against two Bcc species and are promising vaccine candidates for a multi-subunit vaccine. Furthermore, it highlights the potential of our proteomics approach to identify potent antigens against other difficult pathogens

Evaluating scenarios to reduce phosphorus transport in surface waters from slurry applications in temperate grasslands

Publication history: Accepted - 7 April 2022; Published online - 30 May 2022.This study evaluates a range of scenarios to reduce soluble reactive phosphorus (SRP) losses using the surface runoff phosphorus transport model (Surphos) to simulate the application of liquid manure (slurry) to grassland catchments. Surphos was applied using data from two contrasting sites in the Republic of Ireland and Northern Ireland. It explored scenarios that investigated changes to the timing of slurry applications, based both on policy (i.e. a “closed” period where regulations prohibit any slurry spreading) and on climate-based restrictions, where soil moisture and antecedent rainfall were important factors. The observed data showed a considerable spatial variability in runoff at both sites, which resulted in a corresponding variable range of SRP losses predicted by the model. However, at both sites the model results showed that maintaining a closed period led to a greater reduction in SRP losses than opening this period up to slurry applications under climate-based restrictions.This work was supported in part by the Department of Agriculture, Environment and Rural Affairs (DAERA) under an Evidence and Innovation Grant [Award No. 17-4-08]

Development of a humanised mouse model of ovalbumin induced allergy and inflammation as a platform to test functional food

Food allergies are most common during infancy Food allergy is a growing health problem with very limited treatment options For infants, cow’s milk protein allergy (CMPA) is the predominant food allergy, occurring in 2 - 7 5 % of population Infant formula which is often cow’s milk based, is the only approved alternative for breastfeeding However the proteins (casein and whey) in cow’s milk result in allergic symptoms in the infants These allergic symptoms can be avoided by providing the infants with hydrolysates of cow’s milk protein instead of the intact protein Whey protein hydrolysates, small bioactive peptide components of milk have been tested in mouse models for their anti-inflammatory/immunomodulatory properties however to date no studies have repeated efficacy of hydrolysates in a humanised mouse model setting In this study a large number of hydrolysates were screened in vitro to identify hydrolysates with anti-inflammatory properties In vitro we demonstrated that 147 hydrolysate significantly reduced DC maturation in a P-PARy dependent manner To mvestigate the anti-inflammatory potential of hydrolysate 147 in a human relevant model of food allergy we developed a novel humanised mouse model of OVA driven food allergy We directly compared two different strains of nonobese diabetic severe combined immunodeficient mice lacking the cytokine receptor common gamma chain (yc7') (NSG-SCF versus NSG- SGM3) expressing human cytokine genes for their suitability as a humanised mouse model Using NSG-SCF mice a human immune system was developed through engraftment of CD34+ hematopoietic stem cells Using an OVA sensitisation and challenge protocol the model of food allergy was developed in these humanised mice The capacity of hydro lysate 147 to mediate anti-inflammatory effects was examined through repeated administration of 147 hydrolysate after sensitisation and before challenge with OVA Both 147 hydrolysate and the wpc80 parent control equally reduced anaphylaxis. In comparison to wpc80 parent control, 147 reduced specific antibody responses, including OVA-specific IgGl, reduced the production of the pro- inflammatory cytokines IL-6, IL-17 and induced higher levels of the anti-inflammatory cytokine IL-10 in spleens and small intestines of OVA treated mice 147 in comparison to wpc80 was more potent at promoting a Thl/Th2 balance in the spleen and most significantly suppressed the accumulation of granulocyte populations in bone marrow and liver while simultaneously reducing the T , B and myeloid cell populations m the bone marrow of OVA mice For the first time, we have shown that a novel milk protein hydrolysate provided efficacy in comparison to its whey parent control in terms of reducing OVA driven allergy in a humanised mouse model This study demonstrated that 147 may prevent the development of other food allergies (egg-ova) in addition to CMPA (but not examined here) In addition this study, presents a framework, from which suitable conditions for testing functional foods, can be adopted in the development of novel agents for the prevention of allergic inflammation

Development of a humanised mouse model of ovalbumin induced allergy and inflammation as a platform to test functional food

Food allergies are most common during infancy Food allergy is a growing health problem with very limited treatment options For infants, cow’s milk protein allergy (CMPA) is the predominant food allergy, occurring in 2 - 7 5 % of population Infant formula which is often cow’s milk based, is the only approved alternative for breastfeeding However the proteins (casein and whey) in cow’s milk result in allergic symptoms in the infants These allergic symptoms can be avoided by providing the infants with hydrolysates of cow’s milk protein instead of the intact protein Whey protein hydrolysates, small bioactive peptide components of milk have been tested in mouse models for their anti-inflammatory/immunomodulatory properties however to date no studies have repeated efficacy of hydrolysates in a humanised mouse model setting In this study a large number of hydrolysates were screened in vitro to identify hydrolysates with anti-inflammatory properties In vitro we demonstrated that 147 hydrolysate significantly reduced DC maturation in a P-PARy dependent manner To mvestigate the anti-inflammatory potential of hydrolysate 147 in a human relevant model of food allergy we developed a novel humanised mouse model of OVA driven food allergy We directly compared two different strains of nonobese diabetic severe combined immunodeficient mice lacking the cytokine receptor common gamma chain (yc7\u27) (NSG-SCF versus NSG- SGM3) expressing human cytokine genes for their suitability as a humanised mouse model Using NSG-SCF mice a human immune system was developed through engraftment of CD34+ hematopoietic stem cells Using an OVA sensitisation and challenge protocol the model of food allergy was developed in these humanised mice The capacity of hydro lysate 147 to mediate anti-inflammatory effects was examined through repeated administration of 147 hydrolysate after sensitisation and before challenge with OVA Both 147 hydrolysate and the wpc80 parent control equally reduced anaphylaxis. In comparison to wpc80 parent control, 147 reduced specific antibody responses, including OVA-specific IgGl, reduced the production of the pro- inflammatory cytokines IL-6, IL-17 and induced higher levels of the anti-inflammatory cytokine IL-10 in spleens and small intestines of OVA treated mice 147 in comparison to wpc80 was more potent at promoting a Thl/Th2 balance in the spleen and most significantly suppressed the accumulation of granulocyte populations in bone marrow and liver while simultaneously reducing the T , B and myeloid cell populations m the bone marrow of OVA mice For the first time, we have shown that a novel milk protein hydrolysate provided efficacy in comparison to its whey parent control in terms of reducing OVA driven allergy in a humanised mouse model This study demonstrated that 147 may prevent the development of other food allergies (egg-ova) in addition to CMPA (but not examined here) In addition this study, presents a framework, from which suitable conditions for testing functional foods, can be adopted in the development of novel agents for the prevention of allergic inflammation

Effect of angiotensin-converting enzyme inhibitor and angiotensin receptor blocker initiation on organ support-free days in patients hospitalized with COVID-19

IMPORTANCE Overactivation of the renin-angiotensin system (RAS) may contribute to poor clinical outcomes in patients with COVID-19. Objective To determine whether angiotensin-converting enzyme (ACE) inhibitor or angiotensin receptor blocker (ARB) initiation improves outcomes in patients hospitalized for COVID-19. DESIGN, SETTING, AND PARTICIPANTS In an ongoing, adaptive platform randomized clinical trial, 721 critically ill and 58 non–critically ill hospitalized adults were randomized to receive an RAS inhibitor or control between March 16, 2021, and February 25, 2022, at 69 sites in 7 countries (final follow-up on June 1, 2022). INTERVENTIONS Patients were randomized to receive open-label initiation of an ACE inhibitor (n = 257), ARB (n = 248), ARB in combination with DMX-200 (a chemokine receptor-2 inhibitor; n = 10), or no RAS inhibitor (control; n = 264) for up to 10 days. MAIN OUTCOMES AND MEASURES The primary outcome was organ support–free days, a composite of hospital survival and days alive without cardiovascular or respiratory organ support through 21 days. The primary analysis was a bayesian cumulative logistic model. Odds ratios (ORs) greater than 1 represent improved outcomes. RESULTS On February 25, 2022, enrollment was discontinued due to safety concerns. Among 679 critically ill patients with available primary outcome data, the median age was 56 years and 239 participants (35.2%) were women. Median (IQR) organ support–free days among critically ill patients was 10 (–1 to 16) in the ACE inhibitor group (n = 231), 8 (–1 to 17) in the ARB group (n = 217), and 12 (0 to 17) in the control group (n = 231) (median adjusted odds ratios of 0.77 [95% bayesian credible interval, 0.58-1.06] for improvement for ACE inhibitor and 0.76 [95% credible interval, 0.56-1.05] for ARB compared with control). The posterior probabilities that ACE inhibitors and ARBs worsened organ support–free days compared with control were 94.9% and 95.4%, respectively. Hospital survival occurred in 166 of 231 critically ill participants (71.9%) in the ACE inhibitor group, 152 of 217 (70.0%) in the ARB group, and 182 of 231 (78.8%) in the control group (posterior probabilities that ACE inhibitor and ARB worsened hospital survival compared with control were 95.3% and 98.1%, respectively). CONCLUSIONS AND RELEVANCE In this trial, among critically ill adults with COVID-19, initiation of an ACE inhibitor or ARB did not improve, and likely worsened, clinical outcomes. TRIAL REGISTRATION ClinicalTrials.gov Identifier: NCT0273570