IgA In Saliva Research Articles

Anti-modified protein antibodies can be detected in saliva, but not in intestinal secretions of seropositive rheumatoid arthritis patients - evidence of site-specific mucosal autoantibody secretion in RA.

Anti-citrullinated protein antibodies (ACPA) have been detected in sputum and saliva, indicating that anti-modified protein antibodies (AMPA) can be produced at mucosal sites in rheumatoid arthritis (RA) patients. However, the body's largest mucosal compartment, the gut, has not yet been examined. We therefore investigated the presence of several AMPA (antibodies against citrullinated (ACPA), carbamylated (anti-CarP) and acetylated (AAPA) proteins) at different mucosal sites, including the intestinal tract. Paired fecal/ileal wash, saliva and serum samples of RA patients and healthy volunteers were collected in two independent cohorts. Data involving feces was replicated in a third cohort. In these secretions AMPA were analyzed using in-house ELISA with unmodified peptides as control. In fecal samples total IgA and anti-E. coli IgA were measured. ACPA, anti-CarP and AAPA IgA were measurable in saliva of seropositive RA patients (prevalence 9-40%). No AMPA could be detected in feces. IgA was present since total IgA and anti-E. coli IgA was detectable in feces of ACPA-positive RA patients and healthy donors. Results were confirmed in another cohort using colonoscopically collected ileal wash samples. Our study shows the presence of ACPA, anti-CarP and AAPA IgA in saliva of ACPA-seropositive RA patients. However, no AMPA could be detected in feces/ileal wash samples of these patients, although our assays were able to measure other antigen-specific antibodies. These data suggest that mucosal autoantibody secretion may occur in the oral mucosa of RA patients, while no evidence could be found for this process in the lower gastro-intestinal tract.

Quantity and Quality of Naturally Acquired Antibody Immunity to the Pneumococcal Proteome Throughout Life.

Young children and older adults are susceptible for invasive pneumococcal disease (IPD) caused by Streptococcus pneumoniae. Pneumococcal protein-specific antibodies play a protective role against IPD; however, not much is known about the pace of acquisition, maturation, and maintenance of these antibodies throughout life. Immunoglobulin G (IgG) and IgA levels, avidity, and/or specificity to the pneumococcal proteome in serum and saliva from healthy young children, adults, and older adults, with known carriage status, were measured by enzyme-linked immunosorbent assay (ELISA) and 2-dimensional western blotting against ΔcpsTIGR4. Eleven-month-old children, the youngest age group tested, had the lowest pneumococcal proteome-specific IgG and IgA levels and avidity in serum and saliva, followed by 24-month-old children and were further elevated in adult groups. Among adult groups, the parents had the highest serum and saliva IgG and IgA antibody levels. In children, antibody levels and avidity correlated with daycare attendance and presence of siblings, posing as proxy for exposure and immunization. Immunodominance patterns slightly varied throughout life. Humoral immunity against the pneumococcal proteome is acquired through multiple episodes of pneumococcal exposure. Low-level and low-avidity antiproteome antibody profiles in young children may contribute to their IPD susceptibility, while in overall antiproteome antibody-proficient older adults other factors likely play a role.

Serum and Salivary IgG and IgA Response After COVID-19 Messenger RNA Vaccination

There is still considerable controversy in the literature regarding the capacity of intramuscular messenger RNA (mRNA) vaccination to induce a mucosal immune response. To compare serum and salivary IgG and IgA levels among mRNA-vaccinated individuals with or without previous SARS-CoV-2 infection. In this cohort study, SARS-CoV-2-naive participants and those with previous infection were consecutively included in the CoviCompare P and CoviCompare M mRNA vaccination trials and followed up to day 180 after vaccination with either the BNT162b2 (Pfizer-BioNTech) vaccine or the mRNA-1273 (Moderna) vaccine at the beginning of the COVID-19 vaccination campaign (from February 19 to June 8, 2021) in France. Data were analyzed from October 25, 2022, to July 13, 2023. An ultrasensitive digital enzyme-linked immunosorbent assay was used for the comparison of SARS-CoV-2 spike-specific serum and salivary IgG and IgA levels. Spike-specific secretory IgA level was also quantified at selected times. A total of 427 individuals were included in 3 groups: participants with SARS-CoV-2 prior to vaccination who received 1 single dose of BNT162b2 (Pfizer-BioNTech) (n = 120) and SARS-CoV-2-naive individuals who received 2 doses of mRNA-1273 (Moderna) (n = 172) or 2 doses of BNT162b2 (Pfizer-BioNTech) (n = 135). The median age was 68 (IQR, 39-75) years, and 228 (53.4%) were men. SARS-CoV-2 spike-specific IgG saliva levels increased after 1 or 2 vaccine injections in individuals with previous infection and SARS-CoV-2-naive individuals. After vaccination, SARS-CoV-2-specific saliva IgA levels, normalized with respect to total IgA levels, were significantly higher in participants with previous infection, as compared with the most responsive mRNA-1273 (Moderna) recipients (median normalized levels, 155 × 10-5 vs 37 × 10-5 at day 29; 107 × 10-5 vs 54 × 10-5 at day 57; and 104 × 10-5 vs 70 × 10-5 at day 180 [P < .001]). In contrast, compared with day 1, spike-specific IgA levels in the BNT162b2-vaccinated SARS-CoV-2-naive group increased only at day 57 (36 × 10-5 vs 49 × 10-5 [P = .01]). Bona fide multimeric secretory IgA levels were significantly higher in individuals with previous infection compared with SARS-CoV-2-naive individuals after 2 antigenic stimulations (median optical density, 0.36 [IQR, 0.16-0.63] vs 0.16 [IQR, 0.10-0.22]; P < .001). The findings of this cohort study suggest that mRNA vaccination was associated with mucosal immunity in individuals without prior SARS-CoV-2 infection, but at much lower levels than in previously infected individuals. Further studies are needed to determine the association between specific saliva IgA levels and prevention of infection or transmission.

Comparison of SARS-CoV-2 spike-specific IgA and IgG in nasal secretions, saliva and serum.

Several novel vaccine platforms aim at mucosal immunity in the respiratory tract to block SARS-CoV-2 transmission. Standardized methods for mucosal sample collection and quantification of mucosal antibodies are therefore urgently needed for harmonized comparisons and interpretations across mucosal vaccine trials and real-world data. Using commercial electrochemiluminescence antibody panels, we compared SARS-CoV-2 spike-specific IgA and IgG in paired saliva, nasal secretions, and serum from 1048 healthcare workers with and without prior infection. Spike-specific IgA correlated well in nasal secretions and saliva (r>0.65, p<0.0001), but the levels were more than three-fold higher in nasal secretions as compared to in saliva (p<0.01). Correlations between the total population of spike-specific IgA and spike-specific secretory IgA (SIgA) were significantly stronger (p<0.0001) in nasal secretions (r=0.96, p<0.0001) as opposed to in saliva (r=0.77, p<0.0001), and spike-specific IgA correlated stronger (p<0.0001) between serum and saliva (r=0.73, p<0.001) as opposed to between serum and nasal secretions (r=0.54, p<0.001), suggesting transudation of monomeric spike specific IgA from the circulation to saliva. Notably, spike-specific SIgA had a markedly higher SARS-CoV-2 variant cross-binding capacity as compared to the total population of spike specific IgA and IgG in both nasal secretions, saliva and serum, (all p<0.0001), which emphasizes the importance of taking potential serum derived monomeric IgA into consideration when investigating mucosal immune responses. Taken together, although spike-specific IgA can be reliably measured in both nasal secretions and saliva, our findings imply an advantage of higher levels and likely also a larger proportion of SIgA in nasal secretions as compared to in saliva. We further corroborate the superior variant cross-binding capacity of SIgA in mucosal secretions, highlighting the potential protective benefits of a vaccine targeting the upper respiratory tract.

Manipulation of neonatal ruminal populations at birth results in sustained effects on microbial populations and measures of health and production in merino and suffolk lambs

The efficiency of the microbial-driven fermentation process in the rumen is closely related to the efficiency of the ruminant host. This experiment examined the effect of manipulating the neonatal microbiota on postnatal development, immune function and productivity (liveweight gain and wool growth parameters). The trial examined the differences between naturally inoculated lambs (maternal control, n = 21) and those given ruminal fluid, which had been collected from ruminally-cannulated ewes, fed either a roughage diet (roughage, n = 14), or a grain-based diet (high grain, n = 14). At lambing, newborn lambs were tagged and weighed. Inoculation lambs received 10 ml of rumen fluid per inoculation, daily, for a total of 7 inoculations. Live weight, body condition scores, wool growth (greasy and clean wool weights), wool length, mean fibre diameter, and differential blood cell counts, and IgG and IgA in both blood and saliva) were quantified from birth until week 18 (slaughter). Total feed intake was recorded from weaning. Ruminal fluid was collected by stomach tube from 12 lambs from each treatment group and their dams (n = 7–8 per treatment, with the lower number of ewes coming about through ewes that had twins) at time of weaning (week 10). The pH of the ruminal fluid, the density of ciliated protozoa in ruminal fluid, and the composition of the ruminal bacterial populations were analysed. At slaughter (week 18), ileal samples were taken from the same focal lambs as were used for microbial analysis. The artificial inoculations resulted in significant differences in ruminal bacterial genera to those of naturally inoculated lambs at time of weaning (some 3 months post-lambing). Significant differences were also apparent in ileal bacterial communities at slaughter, between lambs which had received the inoculum from roughage-fed ewes and naturally inoculated lambs. Both the efficiency (growth, condition, feed conversion efficiency, carcass weight) and indices of health (mortalities, red blood cell count, blood haemoglobin concentration, haematocrit, and eosinophil counts) of the inoculated animals, were affected negatively by the inoculations ( < 0.05). There was a significant interaction between inoculum source and breed in terms of effects on all parameters measured. The effects of the high grain inoculation were significantly affected by breed, with high grain merinos found to have reduced mean fibre diameter (P < 0.05) and increased wool growth relative to the maternal control (P = 0.035). This effect was not noted in the suffolks. In summary, we have demonstrated that long-term effects on ruminal and intestinal microbiota can be generated by intervention in the normal process of ruminal inoculation in neonatal lambs. Moreover, there appeared to be genetic differences in response to artificial inoculation. Whilst the effects of artificial inoculation in this trial were negative in terms of indices of health and production, further studies of maternal diet and its effects on the natural establishment of the neonatal microbiota are warranted given the persistence of the effects and the interaction with breed.

Two-trace two-dimensional correlation spectra (2T2D-COS) analysis using FTIR spectra to monitor the immune response by COVID-19.

There is a growing trend in using saliva for SARS-CoV-2 detection with reasonable accuracy. We have studied the responses of IgA, IgG, and IgM in human saliva by directly comparing disease with control analyzing two-trace two-dimensional correlation spectra (2T2D-COS) employing Fourier transform infrared (FTIR) spectra. It explores the molecular-level variation between control and COVID-19 saliva samples. The advantage of 2T2D spectra is that it helps in discriminating remarkably subtle features between two simple pairs of spectra. It gives spectral information from highly overlapped bands associated with different systems. The clinical findings from 2T2D show the decrease of IgG and IgM salivary antibodies in the 50, 60, 65, and 75-years COVID-19 samples. Among the various COVID-19 populations studied the female 30-years group reveals defense mechanisms exhibited by IgM and IgA. Lipids and fatty acids decrease, resulting in lipid oxidation due to the SARS-CoV-2 in the samples studied. Study shows salivary thiocyanate plays defense against SARS-CoV-2 in the male population in 25 and 35 age groups. The receiver operation characteristics statistical method shows a sensitivity of 98% and a specificity of 94% for the samples studied. The measure of accuracy computed as F score and G score has a high value, supporting our study's validation. Thus, 2T2D-COS analysis can potentially monitor the progression of immunoglobulin's response function to COVID-19 with reasonable accuracy, which could help diagnose clinical trials. KEY MESSAGES: The molecular profile of salivary antibodies is well resolved and identified from 2T2D-COS FTIR spectra. The IgG antibody plays a significant role in the defense mechanism against SARS-CoV-2 in 25-40years. 2T2D-COS reveals the absence of salivary thiocyanate in the 40-75years COVID-19 population. The receiver operation characteristic (ROC) analysis validates our study with high sensitivity and specificity.

Psycho-emotional portrait of primary school-age children with recurrent respiratory infections

Respiratory infections occupy a leading place in the structure of the population’s morbidity, especially among children. At the same time, recurrent respiratory infections (RRIs) impose the main burden on society and families. Most of the factors contributing to the manifestation of the RRI phenomenon have already been well-studied. However, the influence of psychological and emotional aspects on a child’s health has remained largely overlooked. Aim. to study the psychological characteristics of children aged 5–7 years in comparison to episodically ill peers and their correlation with local immunity indicators. Materials and Methods. The study involved 80 children, with 60 in the main group (children with recurrent respiratory infections) and 20 in the control group (episodically ill children). For psychological testing (psychometrics), the following methods were used: Blob Tree test (P. Wilson) to determine motivation and adaptation of children in a group setting. The projective „Family Sociogram‟ test (E.G. Aidemiller) to determine the place of the subject in the system of interpersonal relationships and to assess the nature of communication within the family. A modified version of A. Etkind’s Color Relations Test was used to determine individual typological personality characteristics and attitudes towards significant people and the surrounding environment on both a conscious and partially unconscious level. Clinical laboratory examination included measuring levels of secretory IgA and lysozyme in unstimulated saliva. Results. When analyzing the data obtained, a statistically significant difference was found between the results of children in the main and control groups. Conclusions. During color selection, children with RRIs more often associated themselves with the color red (40.7 %), their mothers with purple (40.7 %), and kindergarten or school with yellow (29.7 %) compared to episodically ill peers (p &lt; 0.05). Children with RRIs in the „Family Sociogram‟ test more often than episodically ill peers demonstrated reduced self-esteem (73 % vs. 30 %, p = 0.038). In the Blob Tree test, children with RRIs differ in their first choice (current position in the group) from episodically ill peers (p &lt; 0.0001). The level of sIgA and lysozyme in saliva in the group of children with RRIs correlates with the sensory choice of color for the mother (p &lt; 0.0001).

Antibodies against Neisseria meningitidis serogroups A, C, W and Y in serum and saliva of Norwegian adolescents

IntroductionThe incidence of invasive meningococcal disease (IMD) among Norwegian 16–19-year-olds was 1–7/100,000 in the decade before the COVID-19 pandemic, with serogroup Y (MenY) dominance. In contrast to many other European countries, meningococcal vaccines are not part of the national immunisation program (NIP) in Norway. This cross-sectional study aimed to measure the degree of natural immunity against Neisseria meningitidis among adolescents in Norway to evaluate the need for introducing tetravalent meningococcal conjugate vaccine (MCV4) in the NIP. Materials and methodsSerum and saliva samples were collected from students in upper and lower secondary schools in Norway in 2018. Samples were analysed for meningococcal capsular polysaccharide (PS)-specific antibodies using a bead-based multiplex immunoassay. PS-specific antibody levels were linked to data on meningococcal carriage, vaccination status and risk factors for carriage (assessed with questionnaire) and analysed by linear regression of log transformed concentrations. A subset of samples from unvaccinated individuals was analysed for serum bactericidal antibodies (SBA). ResultsA total of 1344 participants, median age 16 years (range 12–24), were included in the study. Overall, 60.9% of the participants were female and 1137 (84.6%) were not vaccinated with MCV4. PS-specific antibody concentrations in serum and saliva were low among unvaccinated individuals for all serogroups and only 6.7–20.0% of the subpopulation with high PS-specific antibodies assessed with SBA had protective levels. Unvaccinated MenY carriers had higher levels of MenY anti-PS IgG in serum and IgA in saliva than those not carrying MenY. Use of Swedish snus was associated with lower anti-PS IgG levels in serum and waterpipe use with lower anti-PS IgG levels in saliva. ConclusionUnvaccinated adolescents in Norway have a low degree of natural immunity against the serogroups of N. meningitidis predominating among cases of IMD in this age group. Therefore, introduction of MCV4 for adolescents in the NIP is recommended.

CATION COMPOSITION OF ORAL SECRETION IN PRIMARY SCHOOL-AGED CHILDREN

SUMMARY. Recurrent respiratory infections (RRIs) currently have the greatest medical and social burden among infectious diseases worldwide, with the highest prevalence and incidence rates of acute respiratory infections in children. Recent studies indicate the role of inadequate immune response and the development of recurrent acute respiratory infections (ARI) in children due to deficiencies in essential trace elements in the body, which are crucial for the proper functioning of physical barriers and the immune system. The hypothesis of our study assumed that, considering the recurrence of at least 6 episodes of ARI per year, children with RRIs may have changes in salivary ion levels, which could serve as a basis for local immune suppression of the upper respiratory mucosa. The aim – to investigate the levels of metals in the saliva of children of primary school age with recurrent respiratory infections in comparison with episodically ill peers and establish the relationship of these changes with the state of local immunity. Material and Methods. In the actual study, 40 children participated, with 30 of them (the main group) being children with recurrent respiratory infections and 10 other children who had episodic illnesses were assigned to the control group. The levels of metal cations in the saliva were examined using inductively coupled plasma atomic emission spectrometry, including essential metals such as copper, manganese, zinc, calcium, cobalt, potassium, magnesium, sodium, phosphorus, selenium, iron, chromium, and sulfur. Additionally, levels of conditionally essential metals like lithium, nickel, and boron were measured. The levels of secretory IgA and lysozyme in saliva were determined using the enzyme-linked immunosorbent assay method. Results. The main group was not homogeneous; therefore, we divided it into two clusters. Respondents in one cluster had significantly lower relative levels of essential ion content in saliva. Specifically, calcium was lower in 64 % (p=0.003), copper in 76.5 % (p=0.001), manganese in 41.1 % (p=0.008), magnesium in 64 % (p=0.02), and zinc in 58.8 % (p=0.027). This fact can be explained by their increased losses due to frequent inflammatory processes in the upper respiratory tract, resulting in higher daily replenishment needs compared to their peers. Correlation analysis revealed a connection between illness incidence and the levels of copper (p=0.008), sodium (p=0.00005), and phosphorus (p=0.028), indicating potential associations between saliva metal profiles and the frequency of ARI in children with RRIs. The presence of dental caries correlated with lithium (p=0.02), copper (p=0.032), and nickel levels (p=0.037). Integral indicators of local immunity, such as lysozyme and secretory IgA, exhibited correlations with metal levels in saliva (Li, Cu, Ni, Na, Co, P, Zn), confirming the hypothesis of their role in regulating immune reactions in the oral cavity. Conclusions. Cluster analysis of the metal profiles of the group of children with RRI shows its heterogeneity, 43 % of representatives of this cohort had significantly lower levels of calcium (p=0.003), copper (p=0.001), manganese (p=0.008), magnesium (p=0.02) and zinc (p=0.027) than the control group. Which may indicate the presence of deficient conditions in some children with PRI as a result of the increased need for their recovery. The presence of caries, the level of SIgA, lysozyme (clinical indicators of local immunity of the oral cavity) have certain relationships with the levels of essential and conditionally essential levels of salivary metals (Li, Cu, Ni, Na, Co, P, Zn), which indicates the benefit of the connection between the metal profile of saliva and the state of local immunity.

The emergence of SARS-CoV-2 lineages and associated saliva antibody responses among asymptomatic individuals in a large university community.

SARS-CoV-2 (CoV2) infected, asymptomatic individuals are an important contributor to COVID transmission. CoV2-specific immunoglobulin (Ig)-as generated by the immune system following infection or vaccination-has helped limit CoV2 transmission from asymptomatic individuals to susceptible populations (e.g. elderly). Here, we describe the relationships between COVID incidence and CoV2 lineage, viral load, saliva Ig levels (CoV2-specific IgM, IgA and IgG), and ACE2 binding inhibition capacity in asymptomatic individuals between January 2021 and May 2022. These data were generated as part of a large university COVID monitoring program in Ohio, United States of America, and demonstrate that COVID incidence among asymptomatic individuals occurred in waves which mirrored those in surrounding regions, with saliva CoV2 viral loads becoming progressively higher in our community until vaccine mandates were established. Among the unvaccinated, infection with each CoV2 lineage (pre-Omicron) resulted in saliva Spike-specific IgM, IgA, and IgG responses, the latter increasing significantly post-infection and being more pronounced than N-specific IgG responses. Vaccination resulted in significantly higher Spike-specific IgG levels compared to unvaccinated infected individuals, and uninfected vaccinees' saliva was more capable of inhibiting Spike function. Vaccinees with breakthrough Delta infections had Spike-specific IgG levels comparable to those of uninfected vaccinees; however, their ability to inhibit Spike binding was diminished. These data are consistent with COVID vaccines having achieved hoped-for effects in our community, including the generation of mucosal antibodies that inhibit Spike and lower community viral loads, and suggest breakthrough Delta infections were not due to an absence of vaccine-elicited Ig, but instead limited Spike binding activity in the face of high community viral loads.

Local and Systemic Immunity During Five Vaccinations Against SARS-CoV-2 in Zanubrutinib-Treated Patients With Chronic Lymphocytic Leukemia.

Patients with chronic lymphocytic leukemia (CLL) are vulnerable to coronavirus disease 2019 (COVID-19) and are at risk of inferior response to severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) vaccination, especially if treated with the first-generation Bruton's tyrosine kinase inhibitor (BTKi) ibrutinib. We aimed to evaluate the impact of the third-generation BTKi, zanubrutinib, on systemic and mucosal response to SARS-CoV-2 vaccination. Nine patients with CLL with ongoing zanubrutinib therapy were included and donated blood and saliva during SARS-CoV-2 vaccination, before vaccine doses 3 and 5 and 2 - 3 weeks after doses 3, 4, and 5. Ibrutinib-treated control patients (n = 7) and healthy aged-matched controls (n = 7) gave blood 2 - 3 weeks after vaccine dose 5. We quantified reactivity and neutralization capacity of SARS-CoV-2-specific IgG and IgA antibodies (Abs) in both serum and saliva, and reactivity of T cells activated with viral peptides. Both zanubrutinib- and ibrutinib-treated patients had significantly, up to 1,000-fold, lower total spike-specific Ab levels after dose 5 compared to healthy controls (P < 0.01). Spike-IgG levels in serum from zanubrutinib-treated patients correlated well to neutralization capacity (r = 0.68; P < 0.0001) and were thus functional. Mucosal immunity (specific IgA in serum and saliva) was practically absent in zanubrutinib-treated patients even after five vaccine doses, whereas healthy controls had significantly higher levels (tested in serum after vaccine dose 5) (P < 0.05). In contrast, T-cell reactivity against SARS-CoV-2 peptides was equally high in zanubrutinib- and ibrutinib-treated patients as in healthy control donors. In our small cohort of zanubrutinib-treated CLL patients, we conclude that up to five doses of SARS-CoV-2 vaccination induced no detectable IgA mucosal immunity, which likely will impair the primary barrier defence against the infection. Systemic IgG responses were also impaired, whereas T-cell responses were normal. Further and larger studies are needed to evaluate the impact of these findings on disease protection.

Effects of 24-week prebiotic intervention on self-reported upper respiratory symptoms, gastrointestinal symptoms, and markers of immunity in elite rugby union players

ABSTRACT Objectives Elite rugby union players face numerous physiological and psychological stressors which can increase upper respiratory and gastrointestinal illness risk, and in turn can compromise training and competitive performance. This study aimed to investigate the effect of daily prebiotic supplementation on upper respiratory symptoms, gastrointestinal symptoms, and markers of immune function in elite rugby union players. Methods Thirty-three elite rugby union players were randomly assigned to consume a prebiotic (2.8 g/day galactooligosaccharide) or placebo (2.8 g/day maltodextrin), daily for 168 days under double-blind conditions. Participants completed daily and weekly questionnaires for self-reported upper respiratory and gastrointestinal symptoms respectively. Blood and saliva samples were collected at 0, 84, and 168 days for assessment of plasma TNF-α and CRP, and saliva IgA respectively. Results The prebiotic group experienced a 2-day reduction in upper respiratory symptom duration (P = 0.045). Gastrointestinal symptom severity and incidence were lower in the prebiotic group compared to the placebo group (P < 0.001, P = 0.041) respectively. Salivary immunoglobulin A secretion rate was 42% greater in the prebiotic group compared to the placebo group at day 168 (P = 0.004), no differences in CRP and TNF-α were found (P > 0.05). Conclusion A 168-day dietary prebiotic intervention reduced the duration of upper respiratory symptoms and reduced the incidence and severity of gastrointestinal symptoms in elite rugby union players. These findings suggest that seasonal prebiotic interventions may be beneficial for reducing illness in elite rugby union players, improving their availability to train and compete. Key points Elite athletes are susceptible to upper respiratory symptoms and gastrointestinal symptoms which may impact upon training availability and competition performance. For the first time, this study shows that a dietary prebiotic intervention can reduce the duration of upper respiratory symptoms by 2 days in elite rugby union players. Dietary prebiotic supplementation can improve the incidence and severity of gastrointestinal symptoms experienced by elite rugby union players. Prebiotic supplementation was able to increase salivary IgA secretion after 168 days. These findings can inform practice suggesting that seasonal prebiotic use has the potential to modulate immune function and reduce illness in elite rugby union, which may improve a player’s availability to train and compete. The mechanisms by which prebiotics reduce URS and GIS require further research exploration.

Detection of anti-SARS-CoV-2 antibodies in tears: Ocular surface immunity to COVID-19

PurposeTo evaluate the presence of SARS-COV-2 specific IgA and IgG antibodies in tears of unvaccinated and anti-COVID-19 vaccinated subjects with previous history of SARS-COV-2 infection. To compare results in tears with those in saliva and serum and correlate with clinical data and vaccination regimens. MethodsCross-sectional study including subjects with a previous history of SARS-CoV-2 infection, both unvaccinated and vaccinated against COVID-19. Three samples were collected: tears, saliva and serum. IgA and IgG antibodies against S-1 protein of SARS-CoV-2 were analyzed with a semi-quantitative ELISA. Results30 subjects, mean age 36.4 ± 10, males 13/30 (43.3%) with history of mild SARS-CoV-2 infection were included. 13/30 (43.3%) subjects had received a 2-dose regimen and 13/30 (43.3%) a 3-dose regimen of anti-COVID-19 vaccine, 4/30 (13.3%) subjects were unvaccinated. All the participants with full anti-COVID-19 vaccination (2-or 3-doses) presented detectable anti-S1 specific IgA in all three biofluids, tears, saliva and serum. Among unvaccinated subjects, specific IgA was detected in 3/4 subjects in tears and saliva, whereas IgG was not detected. Considering IgA and IgG antibodies titers, no differences were observed between the 2- and 3-dose vaccination regimen. ConclusionsSARS-CoV-2-specific IgA and IgG antibodies were detected in tears after mild COVID-19, highlighting the role of the ocular surface as a first line of defense against infection. Most naturally infected unvaccinated individuals exhibit long-term specific IgA in tears and saliva. Hybrid immunization (natural infection plus vaccination) appears to enhance mucosal and systemic IgG responses. However, no differences were observed between the 2- and 3-dose vaccination schedule.

Correlation between salivary immunoglobulin A and interleukin-1beta in smokers with dental caries.

Introduction. Dental caries is one of the most common infectious diseases of the oral cavity, and is an inflammatory disease caused by several factors. Interleukin-1β (IL-1β) is a major mediator of acute inflammation and essential for the development of specific immune responses. The objective of this study was to assess the levels of secretory IgA (s-IgA) and IL-1β in the saliva of smokers with dental caries, and to discover the correlation between these parameters and dental caries. Methods. Saliva samples were collected from 30 smokers, age range 21-70 years, with dental caries, in addition to 18 healthy non-smoker volunteers, age range 21-65 years. s-IgA and IL-1β levels in the saliva samples were estimated by enzyme-linked immunosorbent assay (ELISA). Results. The mean saliva IgA levels between smokers with dental caries group and healthy subjects were not significantly different (p=0.077), while the saliva levels of IL-1β were higher in the smokers in dental caries group, with a significant difference of p<0.05. No significant associations were found between s-IgA and IL-1β levels, and other parameters such as age, sex, body mass index (BMI) and C-reactive protein (CRP) (p>0.05). There were highly positive associations and significant differences between IL-1β and CRP levels in the two groups under the study (p=0.006). Conclusions. Our study revealed a significant increase in IL-1β levels in saliva of smokers with dental caries, and a positive association between IL-1β levels and caries disease. There is no significant relationship between elevated IL-1β levels and s-IgA in smokers with dental caries.

NEI supplementation improves alum-based immunity in CFTR KO mice

Abstract Cystic fibrosis (CF) is a genetic disorder that causes severe damage to the lungs and digestive system. CF affects the cells that produce mucus, sweat, and digestive enzymes and is caused by mutations in the Cystic Fibrosis Transmembrane Conductance Regulator (CFTR). Expression and function of CFTR have been extensively studied on epithelial cells and several recent publications have shown that CFTR also regulates the functions of myeloid cells. We addressed whether CFTR is expressed by B cells and the role of CFTR in B cell functions. We found that CFTR is expressed on murine and human B cells. Compared to control wild-type (WT) mice, CFTR KO exhibited reduced frequency of B cells in lymphoid tissues and lower levels of serum immunoglobulin isotypes. Upon systemic immunization with an alum-based vaccine, CFTR KO mice developed lower antibody responses than control mice. Interestingly, vaccine supplementation with an inhibitor of the serine protease elastase (NEI) enhanced the titers of vaccine-specific serum antibody responses in CFTR KO mice and CFTR heterozygote (CFTR Het) mice. We have previously reported that NEI supplementation could promote mucosal immunity in mice immunized with the alum-based injected vaccine. In this regard, CFTR KO and CFTR Het mice immunized with NEI-supplemented alum-based injected developed higher IgA and IgG responses in saliva than control WT mice. These results provide new insights on the regulatory role of CFTR for B cell differentiation and function. They also suggest that vaccine supplementation with NEI could improve protection of individuals leaving with CF against respiratory pathogens. NIH R01 DK101323

Detección de anticuerpos anti-SARS-CoV-2 en lágrimas: inmunidad de la superficie ocular frente a COVID-19

PurposeTo evaluate the presence of SARS-CoV-2 specific IgA and IgG antibodies in tears of unvaccinated and anti-COVID-19 vaccinated subjects with previous history of SARS-CoV-2 infection. To compare results in tears with those in saliva and serum and correlate with clinical data and vaccination regimens. MethodsCross-sectional study including subjects with a previous history of SARS-CoV-2 infection, both unvaccinated and vaccinated against COVID-19. Three samples were collected: tears, saliva and serum. IgA and IgG antibodies against S-1 protein of SARS-CoV-2 were analyzed with a semi-quantitative ELISA. ResultsThirty subjects, mean age 36.4±10, males 13/30 (43.3%) with history of mild SARS-CoV-2 infection were included. 13/30 (43.3%) subjects had received a 2-dose regimen and 13/30 (43.3%) a 3-dose regimen of anti-COVID-19 vaccine, 4/30 (13.3%) subjects were unvaccinated. All the participants with full anti-COVID-19 vaccination (2-or 3-doses) presented detectable anti-S1 specific IgA in all 3biofluids, tears, saliva and serum. Among unvaccinated subjects, specific IgA was detected in 3/4 subjects in tears and saliva, whereas IgG was not detected. Considering IgA and IgG antibodies titers, no differences were observed between the 2- and 3-dose vaccination regimen. ConclusionsSARS-CoV-2-specific IgA and IgG antibodies were detected in tears after mild COVID-19, highlighting the role of the ocular surface as a first line of defense against infection. Most naturally infected unvaccinated individuals exhibit long-term specific IgA in tears and saliva. Hybrid immunization (natural infection plus vaccination) appears to enhance mucosal and systemic IgG responses. However, no differences were observed between the 2- and 3-dose vaccination schedule.

중·고 탁구선수의 시합 중 코티졸, DHEA과 IgA 반응과 스트레스 지각에 관한 연구

The purpose of this study was to investigate changes of stress-related hormones, immune response and stress perception level during competition among middle and high school elite table tennis players. The study subjects were 10 female players and 10 male players. Stress-related hormone and immune response measured at rest, before and after game by saliva collected from them. Stress perception level used visual analogue scale (VAS). The results of this study were as follows: Saliva cortisol according to gender differences showed significant interaction effects of between group×time (p=.039). Also, school levels didn’t show significant interaction effects while in the main effects verification was significant differences among group (p=.044) and time (p=.000). Saliva dehydroepiandrosterone (DHEA) according to gender differences showed significant differences in time (p=.015) which is, male players were significantly increases between measurement time but female players were significantly decreases at after game. School levels didn’t show significant interaction effects while in the main effects verification was significant differences among group (p=.007) and time (p=.016). Saliva IgA according to gender differences showed no significant interaction effects, in the main effects verification was significant differences between time (p=.040). In the school levels was no significant changes in the interaction effects and main effects. Stress perception according to gender differences was no significant interaction effects, but in the main effects verification was significant differences among group (p=.040) and time (p=.000). Stress perception according to school levels showed significant the interaction effects of between group×Time (p=.005). In the correlation between cortisol, DHEA and VAS, there showed significant difference. Therefore, the present study will hopefully provide basic data for the development of programs to minimize the stress of table tennis players. And further studies are needed to understand its topic.

Results of the study of mucosal immunity indices in patients with cancer of the oral cavity and oropharynx during radiotherapy or chemoradiotherapy therapy and immunotherapy with α/β-defensins.

The aim of the study was to investigate the concentration of interferon (INF) -a, INF- g, interleukin (IL) -6, and secretory IgA (sIgA) in saliva during various regimens of antitumour treatment and immunotherapy (IT) with a/b-defensins in patients with cancer of the oral cavity and oropharynx in order to find ways to increase the effectiveness and improvement of the tolerability of antitumour treatment on the base of the identification of biomarkers for the evaluation of the antitumour effect and the prediction of complications. We have studied the changes in the immunity indices of 105 patients who were diagnosed with squamous cell carcinoma of the oral cavity or oropharynx for the first time. The patients received radiotherapy (RT) or chemoradiotherapy and IT with a/b-defensins in different doses (40 and 60 mg) at the 1st phase of the special treatment. A determined drop in the concentration of INF-a after cytostatic treatment, and the additional use of IT with a/b-defensins in different doses do not produce the protective effect on the production of INF-a. Regarding INF- g, a more than two-fold decrease in the concentration of INF- g in the saliva of patients in group receiving a double dose of an immunotherapeutic agent along with radiation therapy (RT) was noted, which may indicate an adjuvant effect of a/b-defensins in relation to RT, enhancing its antitumour influence, and thereby ensuring the regression of neoplasia. In case of an increased dose of a/b-defensins use during RT, there was found immunomodulatory effect in relation to IL-6. In the group of patients who received RT and a higher dose of the immune agent, the "scissors phenomenon" was noted - a simultaneous decrease in the concentration of INF- g and an increase in the concentration of sIgA in saliva, which, taking into account the reduced risk of mucositis and better regression of the tumour, shows the meaningful adjuvant and immunomodulating effects of a/b-defensin therapy in the study group. High-dose IT with a/b-defensins against the background of cytostatic therapy in patients with cancer of the oral cavity and oropharynx potentially leads to an adjuvant and immunomodulatory effect with a decrease in the concentration of INF- g and a parallel increase in the concentration of sIgA in saliva, i.e., reconstruction of the immune response from Th1- to Th2-profile - the profile associated with the tumour regression. With the development of the radio-induced mucositis in these patients, a decrease in concentration of sIgA in saliva with a tendency to a progressive decrease of this index with the increase of mucositis severity was noted. The data obtained allow us to consider INF- g and sIgA as biomarkers of the effectiveness of traditional anticancer therapy during the use of a/b-defensins, and sIgA as a biomarker of the risk of developing radio-induced mucositis in patients with cancer of the oral cavity and oropharynx, which should be verified in further clinical studies with better design.

Comparative ELISA Detection of SARS-COV-2 Monoclonal Antibodies in Patients` Serum, Saliva, and Nasal Fluid in Iraq

Precise management of COVID-19 infection and global pandemic requires a precise assay of SARS-COV-2 antibodies. Spike protein and RBD specific serum monoclonal antibodies has been known to be elicited with the currently approved mRNA vaccine for COVID-19. Since combating SARS-CoV-2 infection begins as early at the viral entry port, mediated by mucosal immunity; whether the currently approved vaccines have the capacity to elicit mucosal immunity is still to be studied. Method; We conducted our research to detect SARS-Cov-2 IgA-RBD and IgG-NCP, in a set of a randomized three patients’ groups (total 55 subjects, randomized into 20 (Non previously infected control, 20 fully vaccinated people from AlSalam teaching hospital vaccination ward upon receiving the vaccine doses, and 15 patients newly recovered from SARS-CoV-2 infection from AlShifaa quarantine hospital after remission from COVID-19 infection and preparing to discharge) in three fluid samples serum, saliva and nasal fluid, for a total of three samples per participant, to make a total of 165 samples to be tested, using a commercial immunoassay kit (Anti-SARS-CoV-2 ELISA RBD IgA, and Anti-SARS- CoV-2 NCP ELISA IgG) the study was designed to assess the immunoglobulins levels in each sample from different health status and anatomical locations to give the statistical difference and the correct conclusion regarding this study. Statistical analysis was performed using SPSS version 24, as well as Microsoft excel version (16.0.15028), test equations were performed via Mann-Whitney U Test, as well as The Kruskal-Wallis H test, to evaluate the non-gaussian distribution among the study group, for each of the outcomes, p &lt; 0.05 was undertaken as statistically significant. Results; Our work showed that two doses of mRNA BNT162b2 vaccine Comirnaty (Pfizer/BioNTech, New York, NY, USA), elicited a robust anti SARS-CoV-2 nasal and salivary IgA and IgG monoclonal antibody protection (as compared to control 0.625 COI, the vaccinated subjects resulted in a higher median salivary concentration of IgG NCP values as 1.69 COI, with a p-value of 0.0001. same goes for median salivary concentration of IgA RBD with value of 2.875 ± 1.276, as compared to control median values of 0.667 ± 0.208), (as compared to control 0.462 ± 0.2369, the vaccinated subjects resulted in a higher median nasal concentration of IgG NCP values as 1.944 ± 0.694, with a p-value of 0.0001. Same goes for median nasal concentration of IgA RBD with value of 1.941 ± 0.53, as compared to control median values of 0.681 ± 0.226). Two injections 25 days a part shown to trigger a stronger titer of protective immunity as compared to single early shot, still it`s less robust compared to the titers measured for the recovered subjects from COVID-19 infection. Taking into consideration that there was a lack of trustable ELISA based assays with specially designed kit for this purpose, focusing on nasal and salivary secretions, the current study that our pre investigational and investigational data are consistent. The results of this study indicates that a protective antigen specific nasal and salivary monoclonal antibody, can be triggered following proper vaccination regimen with mRNA COVID-19 vaccine, so this paves the way for using mucosal protective antibodies detection kits, as a suitable alternative option as a less invasive method compared to serum investigations, for detecting the protection against SARS-Cov-2 infection, induced by vaccine. The vaccinated subjects are significantly better tested via IgA in Saliva as compared to IgG, because of the higher median values for the IgA vs IgG, 2.875 ± 1.276 and 2.083 ± 1.610 respectively with a p-value of 0.008. in a similar state the IgA is the best test for the convalescent subjects in term of nasal fluid samples investigations, as the median value are comparatively higher than median value of IgG, 2.024 ± 0.520 and 1.786 ± 1.115 respectively.

Short-Lived Antibody-Mediated Saliva Immunity against SARS-CoV-2 after Vaccination.

Knowledge about the effect of vaccination against severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) on immunity reflected in the saliva is sparse. We examined the antibody response in saliva compared to that in serum 2 and 6 months after the first vaccination with the BNT162b2 vaccine. Four hundred fifty-nine health care professionals were included in a prospective observational study measuring antibody levels in saliva and corresponding serum samples at 2 and 6 months after BNT162b2 vaccination. Vaccinated, previously SARS-CoV-2-infected individuals (hybrid immunity) had higher IgG levels in saliva at 2 months than vaccinated, infection-naive individuals (P < 0.001). After 6 months, saliva IgG levels declined in both groups (P < 0.001), with no difference between groups (P = 0.37). Furthermore, serum IgG levels declined from 2 to 6 months in both groups (P < 0.001). IgG antibodies in saliva and serum correlated in individuals with hybrid immunity at 2 and 6 months (ρ = 0.58, P = 0.001, and ρ = 0.53, P = 0.052, respectively). In vaccinated, infection-naive individuals, a correlation was observed at 2 months (ρ = 0.42, P < 0.001) but not after 6 months (ρ = 0.14, P = 0.055). IgA and IgM antibodies were hardly detectable in saliva at any time point, regardless of previous infection. In serum, IgA was detected at 2 months in previously infected individuals. BNT162b2 vaccination induced a detectable IgG anti-SARS-CoV-2 RBD response in saliva at both 2 and 6 months after vaccination, being more prominent in previously infected than infection-naive individuals. However, a significant decrease in salivary IgG was observed after 6 months, suggesting a rapid decline in antibody-mediated saliva immunity against SARS-CoV-2, after both infection and systemic vaccination. IMPORTANCE Knowledge about the persistence of salivary immunity after SARS-CoV-2 vaccination is limited, and information on this topic could prove important for vaccine strategy and development. We hypothesized that salivary immunity would wane rapidly after vaccination. We measured anti-SARS-CoV-2 IgG, IgA, and IgM concentrations in saliva and serum in both previously infected and infection-naive individuals, 2 and 6 months after first vaccination with BNT162b2, in 459 hospital employees from Copenhagen University Hospital. We observed that IgG was the primary salivary antibody 2 months after vaccination in both previously infected and infection-naive individuals, but dropped significantly after 6 months. Neither IgA nor IgM was detectable in saliva at either time point. Findings indicate that salivary immunity against SARS-CoV-2 rapidly declines following vaccination in both previously infected and infection-naive individuals. We believe this study shines a light on the workings of salivary immunity after SARS-CoV-2 infection, which could prove relevant for vaccine development.