Secretory IgA Research Articles

A Randomized Controlled Trial of Dietary Rice Bran Intake on Microbiota Diversity, Enteric Dysfunction, and Fecal Secretory IgA in Malian and Nicaraguan Infants

ABSTRACTBackgroundMalnutrition and diarrhea are leading causes of death in children aged <5 y. Rice bran is a nutrient-dense prebiotic available globally.ObjectivesThe objective of this secondary analysis was to evaluate the effects of daily rice bran supplementation on environmental enteric dysfunction (EED) markers, total fecal secretory IgA (sIgA), and microbiota in infants at high risk of malnutrition.MethodsSix-month-old Malian and Nicaraguan infants were randomly assigned to control or daily rice bran supplementation cohorts (1 to 5 g/d). Feces were collected monthly for 6 mo to evaluate fecal sIgA, markers of EED, and microbiota diversity. Statistical methods included linear mixed models, generalized mixed models, Spearman correlation, and Wilcoxon rank-sum tests.ResultsSix-month-old Malian infants had significantly elevated sIgA (4.0× higher, P < 0.001), fecal myeloperoxidase (31.6× higher, P < 0.001), fecal α1-antitrypsin (1.8× higher, P = 0.006), and lower fecal neopterin (0.13× higher, P < 0.001) than the age-matched Nicaraguan infants. In the Nicaraguan rice bran cohort from 6 to 12 mo of age, there was a significant decrease in sIgA concentrations (0.4×, P < 0.05) and a correlation between sIgA and the EED marker α1-antitrypsin (0.523, P < 0.0001) at 12 mo of age. In Malian infants, daily rice bran ingestion resulted in decreased EED scores (0.71×, P = 0.02) and a stable sIgA concentration over time. The rice bran group of Malian infants also had correlation between sIgA and the EED marker neopterin (0.544, P < 0.001) at 12 mo of age and a significant (P < 0.05) increase in microbiota α-diversity at a younger age (9 mo with rice bran compared with 10 mo in control group), which supports earlier microbiota maturation.ConclusionsThese results support rice bran as a functional food ingredient targeting gut mucosa in children at high-risk of malnutrition.

DYNAMICS OF SIgA CONTENT IN BREAST MILK WHEN RISK FACTORS AND LACTATIONAL CHARACTERISTICS ARE CONSIDERED

The leading role associated with an anti-infective action of breast milk belongs to secretory IgA (SIgA). Therefore, the determination of the level of SIgA in colostrum and milk of mothers with different lactation levels and mothers at risk at different levels and duration of lactation has a practical interest.&#x0D; Aim. The research aims at studying the dynamics of SIgA content in breast milk, taking into account risk factors and features of lactation.&#x0D; Material and Methods. The content of SIgA in colostrum and breast milk of 372 mothers with full lactation and 208 with hypogalactia, of which 72 with early and 146 with late hypogalactia at different times of lactation (1-8 days and 1-3 months) was determined. The SIgA content was also studied in mothers; 65 with preeclampsia, including 12 with early hypogalactia, 24 with late hypogalactia, 23 with full lactation, and 44 with anemia of pregnancy, including 12 with early hypogalactia, 19 with late hypogalactia and 13 with full lactation. The determination of SIgA content in colostrum and milk was performed using the method of simple radial immunodiffusion in a gel by G. Mancini et al.&#x0D; Results and Discussion. The SIgA concentration in colostrum and breast milk decreased during the secretion process. In early hypogalactia, the content of SIgA did not differ significantly from the level of SIgA in colostrum and breast milk with full lactation in the early neonatal period and at 1-3 months of secretion. In the late hypogalactia, SIgA content did not differ significantly from its level in the control group. Due to the lower amount of milk received by the mother's child with hypogalactia, the supply of children with SIgA is insufficient, and the deficit is higher the sooner the hypogalactia develops. Similar changes in the dynamics of SIgA content have been found in mothers with preeclampsia and at different levels of lactation. The exception was the content of SIgA in colostrum and milk of mothers with anemia of pregnancy and early hypogalactia, which in the first five days of lactation was lower than in mothers with full lactation.&#x0D; Conclusion. The SIgA concentration in colostrum and breast milk decreases during the secretion process. The level of lactation does not affect the dynamics of SIgA content, except for the SIgA content in colostrum and breast milk of mothers with anemia of pregnancy and hypogalactia in the early stages of secretion. Despite the absence of a significant difference in SIgA levels in mothers with different levels of lactation due to less colostrum and milk received by children with early and late hypogalactia, the supply of SIgA is insufficient, and the cumulative deficit is higher the faster the hypogalactia develops.

O-149 Personalized probiotics and food supplementation according to microbiome miRNA test improves the conception rate in multiple IVF failure patients

Abstract Study question Can the microbiota play a role in the development of infertility by affecting the epigenetic, immunologic, and/or biochemical functions of the host? Summary answer The customization of diet and a nutraceutical and probiotic supplementation according microbiome miRNA signature could increase the pregnancy and live birth rates. What is known already The normal gut microbiota has an impact on the normal function of the male and female reproductive tract by regulating the immunological changes associated with conception. Several miRNAs -a class of small noncoding RNA molecules that control gene expression at the post-transcriptional level- have been described to be associated with dysbiosis and an immune imbalance. We previously demonstrated that women with unexplained infertility could have a gut and vaginal microbiome imbalance with increased intestinal permeability linked to an overexpression of miR-21 and miR-155. Study design, size, duration Observational study including 287 multiple IVF failure infertile patients. They had an average age of 40y and they performed 4 IVF-ET within 10 years on average. Swab samples were collected from the vagina and rectum. Participants/materials, setting, methods Microbial composition by NGS and miRNA expression by real time PCR of vaginal and rectal samples were measured. Immunometabolic markers from blood (TPO, vitamin D and B12, insulin, LDL) were analyzed. Patients followed a customized food supplementation, according to the tested biomarkers, for 90 days. Main results and the role of chance We found that 80% of women showed increased miRNA levels associated with gut and vaginal microbiome imbalances. Deficient values of Vitamin D and/or B were found in 73.2% of tested patients, whereas 56.1% showed high LDL and or insulin values and 19.9% showed positive anti thyroid TPO antibodies. Based upon the elevated levels of miRNAs and immunometabolic markers (insulin, vitamin D, autoantibodies, LDL cholesterol and secretory IgA), patients completed a 60-90 day therapy prior to their next IVF-ET attempt with no clinical side effects. BMI measurements before and after the therapy were not significantly different. After normalization of the blood biomarkers, each patient underwent a next IVF -ET attempt. The total pregnancy rate resulted in 75% (215/287) of positive chemical pregnancy (hCG) and 73% (157/215) of clinically confirmed pregnancy (ultrasound). The live birth rate per intended egg retrieval (all embryos transfers) was 60% (129/215); 27 pregnancies resulted in first trimester losses and one fetal death after a severe preeclampsia. Limitations, reasons for caution A future study should be performed testing miRNA expression before and after the food supplementation to achieve a direct correlation between the microRNA test and the supplementation in a placebo control study in order to evaluate the efficacy of the personalized supplementation on improving pregnancy and live birth rates. Wider implications of the findings Immune system and microbiome modulation could be adress by customizing different strains of probiotics and nutraceuticals selected for their immunomodulatory capacity. An anti-inflammatory dose-specific effect to maintain a delicate immune balance and favor endometrial mucous vascularization and the subsequent placentation, should be personalized to achieve a healthy pregnancy. Trial registration number CEI 1590

Enteric viruses replicate in salivary glands and infect through saliva.

Enteric viruses like norovirus, rotavirus and astrovirus have long been accepted as spreading in the population through fecal–oral transmission: viruses are shed into feces from one host and enter the oral cavity of another, bypassing salivary glands (SGs) and reaching the intestines to replicate, be shed in feces and repeat the transmission cycle1. Yet there are viruses (for example, rabies) that infect the SGs2,3, making the oral cavity one site of replication and saliva one conduit of transmission. Here we report that enteric viruses productively and persistently infect SGs, reaching titres comparable to those in the intestines. We demonstrate that enteric viruses get released into the saliva, identifying a second route of viral transmission. This is particularly significant for infected infants, whose saliva directly transmits enteric viruses to their mothers’ mammary glands through backflow during suckling. This sidesteps the conventional gut–mammary axis route4 and leads to a rapid surge in maternal milk secretory IgA antibodies5,6. Lastly, we show that SG-derived spheroids7 and cell lines8 can replicate and propagate enteric viruses, generating a scalable and manageable system of production. Collectively, our research uncovers a new transmission route for enteric viruses with implications for therapeutics, diagnostics and importantly sanitation measures to prevent spread through saliva.

High ambient humidity aggravates ammonia-induced respiratory mucosal inflammation by eliciting Th1/Th2 imbalance and NF-κB pathway activation in laying hens

Ammonia (NH3) is an irritant and harmful gas. Its accumulation in the poultry house poses detrimental effects on the respiratory mucosal system of birds. In this process, the relative humidity of the poultry house also plays an important role in potentiating the adverse effects of NH3 on the respiratory status of birds, causing severe physiological consequences. In this study, the combined effects of NH3 and humidity on the respiratory mucosal barrier of laying hens was studied. The gene expression of tight junction proteins, mucin, inflammatory cytokines secreted by Th1/Th2 cells, and proteins related to the Nuclear factor-κB (NF-κB) signaling pathway were detected by qRT-PCR. In addition, the contents of mucin and secretory immunoglobulin A (SIgA) in bronchoalveolar lavage fluid (BALF) were determined. The results showed that treatment with NH3 alone or NH3 and humidity led to morphological changes in the respiratory tract, decreased the gene expressions of tight junction protein, and increased the expression of mucin. Also, the expression of interleukin-4 (IL-4) and IL-10 were increased, whereas, the expression of interferon-γ (IFN-γ) and IL-2 was decreased in laying hens treated with NH3 and humidity. Furthermore, the activation of inhibitor kappa B kinase β (I-KK-β) and the degradation of inhibitor of NF-κB α (I-κB-α) contributed to the activation of the NF-κB pathway, such that the downstream genes, cycooxygenase 2 (COX2) and inducible nitric oxide synthase (iNOS) were significantly increased. In conclusion, NH3 damaged the mucosal barrier and induced an imbalance in the mucosal immunity, leading to respiratory tract inflammation. Thus, the relative humidity of the environment aggravates the adverse effects of NH3 in poultry.

Role of Interleukin-6 in the Antigen-Specific Mucosal Immunoglobulin A Responses Induced by CpG Oligodeoxynucleotide-Loaded Cationic Liposomes.

An advantage of mucosal vaccines over conventional parenteral vaccines is that they can induce protective immune responses not only at mucosal surfaces but also in systemic compartments. Despite this advantage, few live attenuated or inactivated mucosal vaccines have been developed and applied clinically. We recently showed that the intranasal immunization of ovalbumin (OVA) with class B synthetic oligodeoxynucleotides (ODNs) containing immunostimulatory CpG motif (CpG ODN)-loaded cationic liposomes synergistically exerted both antigen-specific mucosal immunoglobulin A (IgA) and systemic immunoglobulin G (IgG) responses in mice. However, the mechanism underlying the mucosal adjuvant activity of CpG ODN-loaded liposomes remains unknown. In the present study, we showed that the intranasal administration of CpG ODN-loaded cationic liposomes elicited interleukin (IL)-6 release in nasal tissues. Additionally, pre-treatment with an anti-IL-6 receptor (IL-6R) antibody attenuated antigen-specific nasal IgA production but not serum IgG responses. Furthermore, the intranasal administration of OVA and CpG ODN-loaded cationic liposomes increased the number of IgA+/CD138+ plasma cells and IgA+/B220+ B cells in the nasal passages. This increase was markedly suppressed by pre-treatment with anti-IL-6R blocking antibody. In conclusion, IL-6 released by CpG ODN-loaded cationic liposomes at the site of administration may play a role in the induction of antigen-specific IgA responses by promoting differentiation into IgA+ plasma cells for IgA secretion from B cells.

Exercise Performance and Immune Competence [EPIC]: Background of Natural Immunity, Immune Diversity and Immuno-iatrogenesis

Impeccable Metabolic Maneuvers Upbring Natural Immunity To You [IMMUNITY]. Homeostatic competence of human body is an output of meticulous and interdependent/integrated/impeccable metabolic functions to resist or overcome diseases and facilitate a lengthy lifespan along the healthy ageing trajectory. Principles of homeostasis are seldom employed in the clinical applications. Perhaps this has led to almost an irreparable derailment of ethics and core competence of medical profession. Ivan Dominic Illich emphasized his criticisms on iatrogenesis and argued that modern medicine had hubristically taken on a mission to eradicate pain, sickness, even death but expropriated the autonomous power of humans to cope with their ailments. It is also quite obvious that the medical profession has failed to learn about ‘healthy individuals’ elaborately. Healthy individuals possess quintessential epigenetic efficiency and homeostatic competence built through several years of disciplined lifestyle, and especially their fitness characteristics are incomparably superior to individuals who never or rarely display propensity to improve their health standards. The disastrous perception of Immunology that ‘Best Homeostasis’ and ‘Worst Allostasis’ are same, hence all the individuals regardless of their health status are equally a potential source of disease transmission during an infectious disease outbreak, has caused severe disruption in various aspects of human life across the globe. Even the healthiest/fittest athletes and outstanding exercisers could asymptomatically transmit an infection to unhealthy people [who led an unhealthy lifestyle] whose disease could be anywhere in the allostasis spectrum ranging from early-stage co-morbidities to severe functional deterioration? Under the aegis of homeostasis and non-specific antibodies [Secretory IgA], a long-lasting Natural Immunity [NI] should be possible excluding the need of ‘specific antibodies’. Expanded Trained Immunity [ETI] proposed by Cassone has been found not only in innate immune system but also in nonimmune cells [both cells of hematopoietic and nonhematopoietic origin] associated with immunological memories from previous microbial encounters. Interesting discoveries about NI and ETI could emerge if they are linked with homeostasis and allostasis to develop objective immunologic tests to differentiate the healthy and unhealthy individuals, and enhance the clarity of immunology. Exercises are strongly linked to immunity; thus, measurements of exercise performance standards could accurately reveal the status of NI of the individuals if exercise performance data are systematically interpreted and correlated with immunologic function. Exercise performance data can be gathered using easily accessible non-invasive inexpensive tools. From the perspectives of homeostasis, we may not have distinct departments in the body to curb/defeat immunological or other non-immunological diseases, but any disease could be a threat to recovery to pre-existing conditions or even survival, if the homeostatic functions are impaired by iatrogenesis. KEYWORDS: Homeostasis, Innate Immunity, Adaptive Immunity, Exercise, Epigenetics, Trained Immunity, Secretory IgA, Allostasis, Immunosenescence, Iatrogenesis.

Secretory Cells Are the Primary Source of pIgR in Small Airways.

Loss of secretory IgA (SIgA) is common in chronic obstructive pulmonary disease (COPD) small airways and likely contributes to disease progression. We hypothesized that loss of SIgA results from reduced expression of pIgR (polymeric immunoglobulin receptor), a chaperone protein needed for SIgA transcytosis, in the COPD small airway epithelium. pIgR-expressing cells were defined and quantified at single-cell resolution in human airways using RNA in situ hybridization, immunostaining, and single-cell RNA sequencing. Complementary studies in mice used immunostaining, primary murine tracheal epithelial cell culture, and transgenic mice with secretory or ciliated cell-specific knockout of pIgR. SIgA degradation by human neutrophil elastase or secreted bacterial proteases from nontypeable Haemophilus influenzae was evaluated invitro. We found that secretory cells are the predominant cell type responsible for pIgR expression in human and murine airways. Loss of SIgA in small airways was not associated with a reduction in secretory cells but rather a reduction in pIgR protein expression despite intact PIGR mRNA expression. Neutrophil elastase and nontypeable H. influenzae-secreted proteases are both capable of degrading SIgA invitro and may also contribute to a deficient SIgA immunobarrier in COPD. Loss of the SIgA immunobarrier in small airways of patients with severe COPD is complex and likely results from both pIgR-dependent defects in IgA transcytosis and SIgA degradation.

Comparative Profiles of SARS-CoV-2 Spike-Specific Human Milk Antibodies Elicited by mRNA- and Adenovirus-Based COVID-19 Vaccines.

Background: Numerous COVID-19 vaccines are authorized globally. To date, ∼71% of doses comprise the Pfizer/BioNTech vaccine, and ∼17% the Moderna/NIH vaccine, both of which are messenger RNA (mRNA) based. The chimpanzee Ad-based Oxford/AstraZeneca (AZ) vaccine comprises ∼9%, while the Johnson & Johnson/Janssen (J&J) human adenovirus (Ad26) vaccine ranks fourth at ∼2%. No COVID-19 vaccine is yet available for children 0-4. One method to protect this population may be passive immunization through antibodies (Abs) provided in the milk of a lactating vaccinated person. Our early work and other reports have demonstrated that unlike the post-SARS-CoV-2 infection milk Ab profile, which is rich in specific secretory (s)IgA, the vaccine response is highly IgG dominant. Results: In this report, we present a comparative assessment of the milk Ab response elicited by Pfizer, Moderna, J&J, and AZ vaccines. This analysis revealed 86-100% of mRNA vaccine recipient milk exhibited Spike-specific IgG endpoint titers, which were 12- to 28-fold higher than those measured for Ad vaccine recipient milk. Ad-based vaccines elicited Spike-specific milk IgG in only 33-38% of recipients. Specific IgA was measured in 52-71% of mRNA vaccine recipient milk and 17-23% of Ad vaccine recipient milk. J&J recipient milk exhibited significantly lower IgA than Moderna recipients, and AZ recipients exhibited significantly lower IgA titers than Moderna and Pfizer. Less than 50% of milk of any group exhibited specific secretory Ab, with Moderna recipient IgA titers measuring significantly higher than AZ. Moderna appeared to most frequently elicit greater than twofold increases in specific secretory Ab titer relative to prevaccine sample. Conclusion: These data indicate that current Ad-based COVID-19 vaccines poorly elicit Spike-specific Ab in milk compared to mRNA-based vaccines, and that mRNA vaccines are preferred for immunizing the lactating population. This study highlights the need to design vaccines better aimed at eliciting an optimal milk Ab response.

Protective Effect of Levilactobacillus brevis Against Yersinia enterocolitica Infection in Mouse Model via Regulating MAPK and NF-κB Pathway

Although the use of the probiotic bacterium Lactobacillus for the treatment and prevention of diseases caused by various pathogenic bacteria has received increasing attention in recent years, its mechanism remains incompletely understood. Levilactobacillus brevis 23017 is a select probiotic strain that can regulate the immunity of host animals and resist pathogen infections. In this study, we analyzed the effect of L. brevis 23017 on Yersinia enterocolitica intestinal infection in a BALB/c mouse model and discussed its underlying mechanism. We found that in the mouse model, L. brevis 23017 prevented the damage of villi in the small intestine and decelerated weight loss after Y. enterocolitica infection. Moreover, we focused on the mechanism of the protective effect of L. brevis 23017 from the perspective of the damage and repair of the intestinal mucosal barrier. We observed that L. brevis 23017 maintained a normal mucosal barrier by altering the expression of tight junction proteins. Notably, our results indicated that L. brevis 23017 effectively promoted the secretion of the intestine-specific secretory immunoglobulin A (SIgA) by B cells via regulating cytokines and oxidative damage levels. This mechanism may be the reason for its protective role in Y. enterocolitica infection. In addition, our results demonstrated that the mechanism of L. brevis 23017 was related to antibacterial colonization and inflammation regulation and closely related to antioxidative stress and SIgA promotion. The protective effect of L. brevis 23017 on mice was related to the signaling pathway protein p38 MAPK and the phosphorylation levels of NF-κB. Our study provided novel insight into the mechanism of Lactobacillus against pathogenic bacterial infections. Such insight is of great importance for the prevention, diagnosis, and treatment of related diseases.

Roles of Secretory Immunoglobulin A in Host-Microbiota Interactions in the Gut Ecosystem.

In the gastrointestinal tract (GIT), the immune system interacts with a variety of microorganisms, including pathogens as well as beneficial symbionts that perform important physiological functions for the host and are crucial to sustain intestinal homeostasis. In normal conditions, secretory immunoglobulin A (SIgA) is the principal antibody produced by B cells in the GIT mucosa. Polyreactivity provides certain SIgA molecules with the ability of binding different antigens in the bacterial surface, such as O-antigens and teichoic acids, while cross-species reactivity allows them to recognize and interact with different types of bacteria. These functions may be crucial in allowing SIgA to modulate the complex gut microbiota in an efficient manner. Several studies suggest that SIgA can help with the retention and proliferation of helpful members of the gut microbiota. Gut microbiota alterations in people with IgA deficiency include the lack of some species that are known to be normally coated by SIgA. Here, we discuss the different ways in which SIgA behaves in relation to pathogens and beneficial bacteria of the gut microbiota and how the immune system might protect and facilitate the establishment and maintenance of certain gut symbionts.

Cholecystokinin Outcome on Markers of Intestinal IgA Antibody Response.

Cholecystokinin 8 (CCK8) is an entero-octapeptide that participates in crosstalk with components of intestinal immunity via the CCK receptor (CCKR), but its role in modulation of the IgA response is not fully known under physiological conditions. Male eight-week-old BALB/c mice each were intraperitoneally injected once during 7 days with CCK8, devazapide (CCKR1 antagonist), L365,260 (CCKR2 antagonist) or vehicle (sham group). In intestinal lavages, total and secretory IgA (SIgA) were determined by ELISA; in lamina propria, IgA+ B lymphocytes and IgA+ plasma cells were analyzed by flow cytometry; mRNA levels of polymeric immunoglobulin receptor (pIgR) in epithelial cells and α chain, interleukins (ILs) in lamina propria cells were assessed by qRTPCR. Regarding the sham conditions, IgA+ plasma-cell percentage and IL-2, IL-5, IL-10 and transforming growth factor-β (TGF-β) mRNA levels were either increased by CCK8 or decreased by both CCKR antagonists. For IgA/SIgA responses, IL-4/IL-6 mRNA levels were decreased by all drugs and pIgR mRNA was increased by CCK8 and reduced by L365,260. IgA+ B cell percentage and α chain mRNA levels were elicited by CCK8 and L365,260. Data suggested a presumable differential role of CCK/CCKR on the IgA-response; outcome of L365,260 on the elicitation of IgA+ B cells and α chain mRNA needs further examination.

Divergent age-related humoral correlates of protection against respiratory syncytial virus infection in older and young adults: a pilot, controlled, human infection challenge model

Respiratory viral infections are typically more severe in older adults. Older adults are more vulnerable to infection and do not respond effectively to vaccines due to a combination of immunosenescence, so-called inflamm-ageing, and accumulation of comorbidities. Although age-related changes in immune responses have been described, the causes of this enhanced respiratory disease in older adults remain poorly understood. We therefore performed volunteer challenge with respiratory syncytial virus (RSV) in groups of younger and older adult volunteers. The aim of this study was to establish the safety and tolerability of this model and define age-related clinical, virological, and immunological outcomes. In this human infection challenge pilot study, adults aged 18-55 years and 60-75 years were assessed for enrolment using protocol-defined inclusion and exclusion criteria. Symptoms were documented by self-completed diaries and viral load determined by quantitative PCR of nasal lavage. Peripheral blood B cell frequencies were measured by enzyme-linked immunospot and antibodies against pre-fusion and post-fusion, NP, and G proteins in the blood and upper respiratory tract were measured. The study was registered with ClinicalTrials.gov, NCT03728413. 381 adults aged 60-75 years (older cohort) and 19 adults aged 18-55 years (young cohort) were assessed for enrolment using protocol-defined inclusion and exclusion criteria between Nov 12, 2018, and Feb 26, 2020. 12 healthy volunteers aged 60-75 years and 21 aged 18-55 years were inoculated intranasally with RSV Memphis-37. Nine (67%) of the 12 older volunteers became infected, developing mild-to-moderate upper respiratory tract symptoms that resolved without serious adverse events or sequelae. Viral load peaked on day 6 post-inoculation and symptoms peaked between days 6 and 8. Increases in circulating IgG-positive and IgA-positive antigen-specific plasmablasts, serum neutralising antibodies, and pre-F specific IgG were similar younger and older adults. However, in contrast to young participants, secretory IgA titres in older volunteers failed to increase during infection and, unlike serum IgG, did not correlate with protection. Better understanding of age-related differences in clinical outcomes and immune correlates of protection can overcome reduction in vaccine efficacy with advancing age. We identify correlates of protection in older adults, revealing previously unrecognised factors which might have implications for targeted vaccine discovery and drug development in this vulnerable group. Medical Research Council and GlaxoSmithKline EMINENT Consortium.

The microbiota in respiratory tract infections: from association to intervention.

The respiratory microbiota has a role in respiratory tract infection (RTI) pathogenesis. On the mucosa, the respiratory microbiota interacts with potential pathogenic viruses, bacteria and the host immune system, including secretory IgA (sIgA). This review discusses the role of the respiratory microbiota and its interaction with the (mucosal) immune system in RTI susceptibility, as well as the potential to exploit the microbiota to promote health and prevent RTIs. Recent studies confirm that specific microbiota profiles are associated with RTI susceptibility and during susceptibility and found accompanying RTIs, although clear associations have not yet been found for SARS-CoV-2 infection. sIgA plays a central role in RTI pathogenesis: it stands under control of the local microbiota, while at the same time influencing bacterial gene expression, metabolism and defense mechanisms. Respiratory microbiota interventions are still newly emerging but promising candidates for probiotics to prevent RTIs, such as Corynebacterium and Dolosigranulum species, have been identified. Improved understanding of the respiratory microbiota in RTIs and its interplay with the immune system is of importance for early identification and follow-up of individuals at risk of infection. It also opens doors for future microbiota interventions by altering the microbiota towards a healthier state to prevent and/or adjunctively treat RTIs.

Oil-in-ionic liquid nanoemulsion-based intranasal delivery system for influenza split-virus vaccine.

Effective antigen delivery and immune stimulation in nasal mucosa determine the success of mucosal immunity. Here, an oil-in-ionic liquid (o/IL) nanoemulsion formulated with choline and niacin IL ([Cho][Nic]), squalene, and Tween 80 surfactant is explored as a vaccine delivery system for intranasal mucosal immunization. Compared to the o/w emulsion counterpart without the ILs, the o/IL manoemulsion showed a reduced and more uniform size of approximately 168nm and significantly improved stability. Studies in mice model showed that when was used as an intranasal vaccine delivery system for influenza split-virus antigens, the antigens in the o/IL nanoemulsion induced strong mucosal immune responses with secretory IgA titers 25- and 5.8-fold higher than those of naked and commercial MF59-adjuvanted antigens, respectively. The o/IL nanoemulsion system also induced stronger systemic humoral responses. The excellent mucosal adjuvant effects of the o/IL nanoemulsion mainly benefited from the prolonged retention of antigens in the nasal cavity, enhanced antigen permeation into the submucosa, and the consequently promoted proliferation of CD11b cells and CD4+ T cells in nasal mucosa-associated lymphoid tissue. Moreover, when used as an injection adjuvant, the o/IL nanoemulsion also induced stronger humoral immune responses than MF59. Thus, the [Cho][Nic]-based o/IL nanoemulsion vaccine delivery system can serve as a promising adjuvant platform.

Early Weaning Stress Induces Intestinal Microbiota Disturbance, Mucosal Barrier Dysfunction and Inflammation Response Activation in Pigeon Squabs.

Early weaning stress has been reported to impair intestinal health in mammals. Like mammals, weaning of the pigeon squab, an altricial bird, is associated with social, environmental and dietary stress. However, understanding of weaning stress on intestinal functions is very limited in altricial birds, especially in squabs. This study was aimed to evaluate the effects of early weaning stress on intestinal microbiota diversity, architecture, permeability, the first line defense mechanisms, mucosal barrier functions, and immune cell responses. A total of 192 newly hatched squabs were randomly allocated into two groups, one weaned on day 7 and the other remained with the parent pigeons. Mucosal tissue and digesta in ileum, as well as blood samples, were collected from squabs (n = 8) on days 1, 4, 7, 10, and 14 postweaning. Our results showed that weaning stress induced immediate and long-term deleterious effects on both growth performance and intestinal barrier functions of squabs. Early weaning significantly increased ileal bacterial diversity and alters the relative abundance of several bacteria taxa. Weaning stress can also cause morphological and functional changes in ileum, including an atrophy in villi, an increase in permeability, and a variation in the mRNA expression of genes encoding mucins, immunoglobulins, tight junction proteins, toll-like receptors, and cytokines, as well as the concentration of secretory IgA. We concluded that the impaired intestinal barrier functions accompanied with early weaning stress seems to be the main reason for the poor growth rate after weaning in squabs. In addition, the disturbance of intestinal microbiota of early weaning stress in squabs coincided with dysfunction of intestinal mucosal barrier and activation of inflammation cell responses that were possibly mediated via the activation of toll-like receptors.

Особенности иммунитета и возможности коррекции дезадаптации иммунного ответа у больных острыми респираторными вирусными инфекциями из организованных коллективов

The purpose of the study was to determine the severity of maladaptation disorders of the immune response and the possibility of their correction in patients with acute respiratory viral infections from organized groups. Material and methods. 90 patients with uncomplicated forms of ARVI and a group of conditionally healthy (n = 30) from organized groups were examined. In addition to symptomatic therapy, patients received a combination of rectal and intranasal forms of recombinant IFN-α-2b. Identification of the ARVI pathogen was determined by PCR. ELISA was used to determine serum concentrations of IFN-α and IFN-γ and the ability of blood cells to produce these cytokines ex vivo spontaneously and when stimulated by Newcastle disease virus or phytohemagglutinin. Flow cytometry was used to analysis the subpopulation composition of peripheral blood lymphocytes. The study of the levels of secretory IL-8, IgA, serum IL-8 was carried out by enzyme immunoassay. Results. The etiology of ARVI was deciphered in 62.1% of patients, the main share in the morbidity structure was due to influenza (39.3%) and adenoviruses (35.6%). In the first 3 months of the formation of organized teams in apparently healthy people, a decrease in the level of T-helpers, natural killers and inhibition of the interferon system was revealed, which indicates a violation of adaptive mechanisms and contributes to a decrease in body resistance. An analysis of the cytokine system showed an increase in the concentration of secretory and serum IL-8 in patients with acute respiratory viral infections compared with a group of practically healthy people. In the immune status 1 month after ARVI, there was a significant increase in the number of T-lymphocytes, T-helpers with a tendency to normalize T-lymphocytes carrying IFN-α and -γ receptors, while maintaining a significantly reduced number of B-lymphocytes. The immunomodulatory efficacy of combination therapy with recombinant IFN-α-2b is characterized by an increase in the amount of induced IFN-α during its initial depression and a decrease in initial hyperproduction. Also, at the initial level of induced IFN-α less than 200 pg/ml, there was an increase in the concentration of secretory IL-8 and IgA. Conclusion. In conditionally healthy individuals, in the first 3 months of the formation of organized groups, maladjustment disorders in the immune response were detected in 34% of cases. The interferon-modulating effect of combination therapy with recombinant IFN-α-2b was shown.

Safety and immunogenicity of a live-attenuated influenza virus vector-based intranasal SARS-CoV-2 vaccine in adults: randomised, double-blind, placebo-controlled, phase 1 and 2 trials

BackgroundAll currently available SARS-CoV-2 vaccines are administered by intramuscular injection. We aimed to evaluate the safety and immunogenicity of a live-attenuated influenza virus vector-based SARS-CoV-2 vaccine (dNS1-RBD) administered by intranasal spray in healthy adults.MethodsWe did double-blind, randomised, placebo-controlled phase 1 and 2 trials, followed by a phase 2 extension trial, at a single centre in Jiangsu, China. Healthy adults (≥18 years) who had negative serum or fingertip blood total antibody tests for SARS-CoV-2 (in phases 1 and 2), with no prevalent SARS-CoV-2 infection or history of infection and no SARS-CoV-2 vaccination history (in all three trials reported here), were enrolled. Participants were randomly allocated (4:1 in phase 1, 2:1 in phase 2, and 1:1 in the extension trial) to receive two intranasal doses of the dNS1-RBD vaccine or placebo on days 0 and 14 or, for half of the participants in phase 2, on days 0 and 21. To avoid cross-contamination during administration, vaccine and placebo recipients were vaccinated in separate rooms in the extension trial. The phase 1 primary outcome was safety (adverse events recorded on days 0–44; serious adverse events recorded from day 0 until 12 months after the second dose). In the phase 2 and extension trials, the primary immunogenicity outcomes were SARS-CoV-2-specific T-cell response in peripheral blood (measured by IFN-γ ELISpot), proportion of participants with positive conversion for SARS-CoV-2 receptor-binding domain (RBD)-specific IgG and secretory IgA (s-IgA) antibodies, and concentration of SARS-CoV-2 RBD IgG in serum and SARS-CoV-2 RBD s-IgA in the nasopharynx (measured by ELISA) at 1 month after the second dose in the per-protocol set for immunogenicity. χ2 test and Fisher's exact test were used to analyse categorical data, and t test and Wilcoxon rank sum test to compare the measurement data between groups. These trials were registered with the Chinese Clinical Trial Registry (ChiCTR2000037782, ChiCTR2000039715, and ChiCTR2100048316).FindingsBetween Sept 1, 2020, and July 4, 2021, 63, 724, and 297 participants without a history of SARS-CoV-2 vaccination were enrolled in the phase 1, phase 2, and extension trials, respectively. At least one adverse reaction after vaccination was reported in 133 (19%) of 684 participants in the vaccine groups. Most adverse reactions were mild. No vaccine-related serious adverse event was noted. Specific T-cell immune responses were observed in 211 (46% [95% CI 42–51]) of 455 vaccine recipients in the phase 2 trial, and in 48 (40% [31–49]) of 120 vaccine recipients compared with one (1% [0–5]) of 111 placebo recipients (p<0·0001) in the extension trial. Seroconversion for RBD-specific IgG was observed in 48 (10% [95% CI 8–13]) of 466 vaccine recipients in the phase 2 trial (geometric mean titre [GMT] 3·8 [95% CI 3·4–4·3] in responders), and in 31 (22% [15–29]) of 143 vaccine recipients (GMT 4·4 [3·3–5·8]) and zero (0% [0–2]) of 147 placebo recipients (p<0·0001) in the extension trial. 57 (12% [95% CI 9–16]) of 466 vaccine recipients had positive conversion for RBD-specific s-IgA (GMT 3·8 [95% CI 3·5–4·1] in responders) in the phase 2 trial, as did 18 (13% [8–19]) of 143 vaccine recipients (GMT 5·2 [4·0–6·8]) and zero (0% [0–2]) of 147 placebo recipients (p<0·0001) in the extension trial.InterpretationdNS1-RBD was well tolerated in adults. Weak T-cell immunity in peripheral blood, as well as weak humoral and mucosal immune responses against SARS-CoV-2, were detected in vaccine recipients. Further studies are warranted to verify the safety and efficacy of intranasal vaccines as a potential supplement to current intramuscular SARS-CoV-2 vaccine pools. Steps should be taken in future studies to reduce the potential for cross-contamination caused by the vaccine strain aerosol during administration.FundingNational Key Research and Development Program of China, National Science, Fujian Provincial Science, CAMS Innovation Fund for Medical Sciences, and Beijing Wantai Biological Pharmacy Enterprise.

Electroacupuncture at "Zusanli"(ST36) protects intestinal mucosal immune barrier by suppre-ssing apoptosis of intestinal lymphocytes and regulating expression of Bcl-2 and Bax in sepsis rats

To observe the effect of electroacupuncture (EA) at "Zusanli" (ST36) on apoptosis of intestinal T lymphocytes, translocation of intestinal bacteria and expression of intestinal Bcl-2 and Bax proteins and intestinal mucosal immune barrier in sepsis rats, so as to explore its underlying mechanism in relieving sepsis. SD rats were randomly divided into sham operation (n=6), model (n=15), non-meridian and non-acupoint (non-acupoint, n=15) and acupoint EA(n=15) groups by using random number table method. The sepsis model was established by using cecal ligation and perforation(CLP) method. EA (2 Hz, 2 mA) was applied to bilateral ST36 or non-acupoint for 30 min one hour after modeling, once every day for 3 days. The rats' general conditions and fatality rate in 3 days after modeling were recorded. The liver, spleen and mesenteric lymph nodes were taken for bacterial culture to detect the translocation rate of intestinal bacteria. The small intestinal tissue was taken for observing histopathological changes (Chiu's score: 0-5 points) after HE staining, and for determining the expression levels of Bcl-2 and Bax proteins using Western blot. The intestinal mucosa was sampled for detecting the apop-tosis (apoptotic index) of lymphocytes by using terminal deoxynucleoitidyl transferase-mediated deoxyuridine triphosphate biotin nick end labeling (TUNEL) assay, and the counts of CD4+ and CD8+T cells using flow cytometry. The contents of IL-4 in the small intestine and that of secretory IgA (sIgA) in the small intestinal mucus were determined by using ELISA. After modeling, of the 15 rats in each of the 3 groups, 7, 7 and 2 in the model, non-acupoint and EA groups were dead in the first 3 days, with the fatality rate being 46.67% (7/15), 46.67% (7/15) and 13.33% (2/15), respectively (being obviously lower in the EA group than in the former two groups, P<0.05). Compared with the sham operation group, the incidence of intestinal bacterial translocation, apoptotic index, Chiu's score, and Bax expression were significantly increased (P<0.05), and the percentages of CD4+ and CD8+T cells, IL-4 and sIgA contents and Bcl-2 expression considerably decreased (P<0.05) in the model group. In comparison with the model group, modeling-induced increase of incidence of bacterial translocation, apoptotic index and Bax expression, and decrease of percentages of CD4+ and CD8+T cells, IL-4 and sIgA contents and Bcl-2 expression were reversed (P<0.05) in the EA group. EA at ST36 can reduce death rate and intestinal bacteria translocation incidence in sepsis rats, which may be related to its functions in regulating the expression of intestinal Bcl-2 and Bax proteins and inhibiting the apoptosis of intestinal mucosal T lymphocytes, thereby protecting the immune barrier function of intestinal mucosa to reduce the intestinal permeability.

Amelioration of Ovalbumin-Induced Food Allergy in Mice by Targeted Rectal and Colonic Delivery of Cyanidin-3-O-Glucoside.

Targeted rectal and colonic delivery is an effective strategy to exploit the biological functions of polyphenols. This work investigated the anti-food allergy (FA) activity of cyanidin-3-O-glucoside (C3G) delivered by enteric sodium alginate in vivo. The results showed that through targeted rectal and colonic delivery, the C3G showed better results in ameliorating clinical allergic symptoms, diarrhea, and serological indicators including ovalbumin-specific IgE, histamine, and mast cell protease-1. The C3G was more efficient in enhancing the intestinal epithelial barrier by up-regulating the tight junction protein expression and promoting secretory IgA and β-defensin secretion. The improved bioactivity in regulating T helper (Th)1/Th2 immune balance in the intestinal mucosa was also observed. Compared with the intestinal microbiota structure of the model group, targeted rectal and colonic delivery of C3G was able to bring the abundance of Bacteroidota and Firmicutes close to the levels found in normal mice. Furthermore, there was an evident increase in beneficial bacteria in the intestinal flora, such as Lactobacillus and Odoribacter, and a decrease in pathogenic bacteria like Helicobacter and Turicibacter. Therefore, the anti-FA activity of C3G could be increased via targeted rectal and colonic delivery, while the mechanism might be attributed to the regulation of intestinal microecological homeostasis.