IgG In Saliva Research Articles

Changes in Immunoglobulins G and A in the Saliva and Serum of Horses with Equine Gastric Ulcer Syndrome (EGUS) and Their Relationship with Other Immune and Redox Status Biomarkers

Equine Gastric Ulcer Syndrome (EGUS) is a widespread disease with a very high prevalence and importance in horses. It includes two conditions: Equine squamous gastric disease (ESGD), linked to acid disturbances, and Equine Glandular Gastric Disease (EGGD), associated with inflammation and immune system issues. In saliva, increased levels of immunological markers have been observed in this disease, indicating immune system involvement. In addition, changes in biomarkers of oxidative stress have been described. In horses, IgG and IgA are proteins that appear in saliva and serum and are considered to be the major proteins in specific immunity; however, their levels in horses with EGUS have not been previously studied. This report aims to evaluate IgG and IgA concentrations in the saliva and serum of horses with EGUS, measured by automated assays, and compare them with other biomarkers of the immune and oxidative stress system. This work validated the IgG and IgA assays in saliva and showed that IgA significantly increased in the saliva of horses with EGUS. When the correlation between IgG and IgA and other biomarkers of immune system issues and oxidative stress such as ADA, S100 A8-A9 (calprotectin), S100 A12 (calgranulin), uric acid, FRAS, and AOPP was studied, IgA in saliva showed a significant moderate correlation with adenosine deaminase, indicating its possible involvement in the immune reaction occurring in EGUS.

Cellular and humoral response to SARS-CoV-2 vaccine BNT162b2 in adults with Chronic Kidney Disease G4/5.

The mRNA vaccines have proven to be very effective in preventing severe disease and death from SARS-CoV-2 in the general population. However, in patients with chronic kidney disease (CKD) in dialysis or with kidney transplants (KT) the vaccine responses vary, with severe breakthrough infections as a consequence. In this intervention study we investigated the magnitude and quality of the responses to mRNA vaccination administered prior to kidney replacement therapy (KRT). Twenty patients with CKD G4/5 and nine healthy controls were followed for 12 months after receiving two doses of BNT162b2 four weeks apart and a booster dose after 3–6 months. Induction of anti-Spike and anti-RBD IgG in plasma followed the same kinetics in CKD patients and controls, with a trend towards higher titers in controls. In accordance, there was no differences in the establishment of Spike-specific memory B-cells between groups. In contrast, the CKD patients showed lower levels of anti-Spike IgG in saliva and Spike-specific CD8+ T-cells in blood, possibly influencing the capacity of viral clearance which can contribute to an elevated risk of severe breakthrough infections. In conclusion, we found that CKD patients, despite having a reduced mucosal and cytotoxic immunity to BNT162b2, demonstrated a serological response in plasma similar to healthy controls. This suggests that immunization prior to RRT is efficient and motivated. (EudraCT-nr 2021-000988-68).

Minding the margins: Evaluating the impact of COVID-19 among Latinx and Black communities with optimal qualitative serological assessment tools.

COVID-19 disproportionately affected minorities, while research barriers to engage underserved communities persist. Serological studies reveal infection and vaccination histories within these communities, however lack of consensus on downstream evaluation methods impede meta-analyses and dampen the broader public health impact. To reveal the impact of COVID-19 and vaccine uptake among diverse communities and to develop rigorous serological downstream evaluation methods, we engaged racial and ethnic minorities in Massachusetts in a cross-sectional study (April-July 2022), screened blood and saliva for SARS-CoV-2 and human endemic coronavirus (hCoV) antibodies by bead-based multiplex assay and point-of-care (POC) test and developed across-plate normalization and classification boundary methods for optimal qualitative serological assessments. Among 290 participants, 91.4% reported receiving at least one dose of a COVID-19 vaccine, while 41.7% reported past SARS-CoV-2 infections, which was confirmed by POC- and multiplex-based saliva and blood IgG seroprevalences. We found significant differences in antigen-specific IgA and IgG antibody outcomes and indication of cross-reactivity with hCoV OC43. Finally, 26.5% of participants reported lingering COVID-19 symptoms, mostly middle-aged Latinas. Hence, prolonged COVID-19 symptoms were common among our underserved population and require public health attention, despite high COVID-19 vaccine uptake. Saliva served as a less-invasive sample-type for IgG-based serosurveys and hCoV cross-reactivity needed to be evaluated for reliable SARS-CoV-2 serosurvey results. The use of the developed rigorous downstream qualitative serological assessment methods will help standardize serosurvey outcomes and meta-analyses for future serosurveys beyond SARS-CoV-2.

Detection of SARS-CoV-2-specific mucosal antibodies in saliva following concomitant COVID-19 and influenza vaccination in the ComFluCOV trial

The ComFluCOV trial randomized 679 participants to receive an age-appropriate influenza vaccine, or placebo, alongside their second COVID-19 vaccine. Concomitant administration was shown to be safe, and to preserve systemic immune responses to both vaccines. Here we report on a secondary outcome of the trial investigating SARS-CoV-2-specific mucosal antibody responses. Anti-spike IgG and IgA levels in saliva were measured with in-house ELISAs. Concomitant administration of an influenza vaccine did not affect salivary anti-spike IgG positivity rates to Pfizer/BioNTech BNT162b2 (99.1 cf. 95.6%), or AstraZeneca ChAdOx1 (67.8% cf. 64.9%), at 3-weeks post-vaccination relative to placebo. Furthermore, saliva IgG positively correlated with serum titres highlighting the potential utility of saliva for assessing differences in immunogenicity in future vaccine studies. Mucosal IgA was not detected in response to either COVID-19 vaccine, reinforcing the need for novel vaccines capable of inducing sterilising immunity or otherwise reducing transmission. The trial is registered as ISRCTN 14391248.

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.

Detection of Specific Immunoglobulins in the Saliva of Patients With Mild COVID-19.

Saliva has many advantages over blood as a biofluid, so using it for measuring and monitoring antibody responses in COVID-19 would be highly valuable. To assess the value of saliva-based IgG and IgM/IgA antibody testing in COVID-19, this cross-sectional pilot study evaluated the accuracy of salivary and serum IgG and IgM/IgA for detecting mild COVID-19 and their correlation. Fifty-one patients with mild COVID-19 (14-28 days post-symptom onset) were included in the study. Enzyme-linked immunosorbent assays (ELISA) were used to measure IgG and IgM/IgA responses to SARS-CoV-2 spike protein in both serum and saliva samples using a slightly modified protocol for saliva samples. Saliva-based IgG testing had 30% sensitivity and 100% specificity, with a positive predictive value (PPV) of 100% and a negative predictive value (NPV) of 50%. Saliva-based IgM/IgA testing had 13.2% sensitivity and 100% specificity, with a PPV of 100% and an NPV of 28.3%. Blood and saliva IgG values were positively correlated. Saliva currently has limited diagnostic value for COVID-19 testing, at least for mild disease. Nevertheless, the significant positive correlation between blood and saliva IgG titers indicates that saliva might be a complementary biofluid for assessing systemic antibody responses to the virus, especially if the assay is further optimized across the full disease spectrum.

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.

Mucosal response of inactivated and recombinant COVID-19 vaccines in Congolese individuals.

The efficacy of immunization against an airborne pathogen depends in part on its ability to induce antibodies at the major entry site of the virus, the mucosa. Recent studies have revealed that mucosal immunity is poorly activated after vaccination with messenger RNA vaccines, thus failing in blocking virus acquisition upon its site of initial exposure. Little information is available about the induction of mucosal immunity by inactivated and recombinant coronavirus disease 2019 (COVID-19) vaccines. This study aims to investigate this topic. Saliva and plasma samples from 440 healthy Congolese were collected including (1) fully vaccinated 2 month postvaccination with either an inactivated or a recombinant COVID-19 vaccine and (2) nonvaccinated control group. Total anti-severe acute respiratory syndrome coronavirus 2 receptor-binding domain IgG and IgA antibodies were assessed using in-house enzyme-linked immunosorbent assays for both specimens. Altogether, the positivity of IgG was significantly higher in plasma than in saliva samples both in vaccinated and nonvaccinated control groups. Inversely, IgA positivity was slightly higher in saliva than in plasma of vaccinated group. The overall IgG and IgA levels were respectively over 103 and 14 times lower in saliva than in plasma samples. We found a strong positive correlation between IgG in saliva and plasma also between IgA in both specimens (r = .70 for IgG and r = .52 for IgA). Interestingly, contrary to IgG, the level of salivary IgA was not different between seropositive control group and seropositive vaccinated group. No significant difference was observed between recombinant and inactivated COVID-19 vaccines in total IgG and IgA antibody concentration release 2 months postvaccination both in plasma and saliva. Inactivated and recombinant COVID-19 vaccines in use in the Republic of Congo poorly activated mucosal IgA-mediated antibody response 2 months postvaccination.

Mucosal antibody responses to SARS-CoV-2 booster vaccination and breakthrough infection.

Antibodies on mucosal surfaces of the upper respiratory tract have been shown to be important for protection from infection with SARS-CoV-2. Here we investigate the induction of serum IgG, saliva IgG, and saliva sIgA after COVID-19 mRNA booster vaccination or breakthrough infections.

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.

Detection of anti-SARS-CoV-2 salivary antibodies in vaccinated adults.

Since the introduction of efficient anti-SARS-CoV-2 vaccines, the detection of antibodies becomes useful for immunological monitoring and COVID-19 control. Therefore, this longitudinal study aimed to evaluate the detection of SARS-CoV-2 antibodies in the serum and saliva of COVID-19-vaccinated adults. The study included 13 not vaccinated and 35 vaccinated participants with two doses of CoronaVac (Sinovac/Butantan) vaccine who subsequently received BNT162b2 (Pfizer-BioNTech) vaccine as a booster dose. Vaccinated participants donated saliva and serum in three different time points. Enzyme-linked immunosorbent assay was used for antibody detection. In our results, the serum neutralizing antibodies (NAb) were detected in 34/35 samples after second dose and in 35/35 samples one and five months after the booster dose. In saliva, NAb were detected in 30/35 samples after second dose and in 35/35 of samples one and five months after the booster dose. IgA was detected in 19/34 saliva samples after second dose, in 18/35 one month after the booster and in 30/35 five months after. IgG in saliva was detected in 1/34 samples after second dose, 33/35 samples one month after the booster dose and in 20/35 five months after. A strong correlation was found between IgG and neutralizing activity in saliva, and salivary IgA would be a sign of recent exposure to the virus. In conclusion, saliva can be suitable for monitoring antibodies anti-SARS-CoV-2 after vaccination. Heterologous vaccination contributed to increase anti-SARS-CoV-2 antibodies in the Brazilian health context. Complementary studies with large groups are mandatory to conclude the interest in following mucosal immunity.

Diverging humoral and cellular immune responses due to Omicron-a national study from the Faroe Islands.

Immunity following infection and vaccination with the SARS-CoV-2 Omicron variant is poorly understood. The aim was to investigate immunity assessed with antibody response, neutralizing antibodies (NAbs), and IFN-γ release under different scenarios: in vaccinated and unvaccinated individuals with and without SARS-CoV-2 infection with the Omicron variant. This nationwide single-center study was conducted between January and March 2022, where all convalescent individuals were infected with the Omicron variant and included six study groups: unvaccinated-naïve, unvaccinated convalescent, vaccinated-naïve (second dose), vaccinated-naïve (third dose), vaccinated convalescent (second dose), and vaccinated convalescent (third dose). Antibody responses were assessed by determining receptor binding domain-specific antibodies and NAbs levels in serum, and IgG in saliva. T-cell responses in whole blood were measured as IFN-γ levels released after stimulation with spike peptides. We found that the humoral response against the spike protein was higher among vaccinated-naïve than unvaccinated convalescent. Unvaccinated with and without infection had comparable low humoral responses, while those vaccinated with a second or third dose, independent of infection status, had increasingly higher levels. Only 22% of the unvaccinated convalescent individuals mounted consistent detectable humoral responses following Omicron infection. However, 98% had spike peptide T-cell responses assessed by IFN-γ release. In conclusion, primary Omicron infection mounts a low humoral immune response, significantly enhanced by prior vaccination. Omicron infection induced a robust T-cell response in both unvaccinated and vaccinated, demonstrating that the evasive immune potential of primary Omicron infection affects humoral immunity more significantly than T-cell immunity. IMPORTANCE The immunity following infection and vaccination with the SARS-CoV-2 Omicron variant is poorly understood. We investigated immunity assessed with antibody and T-cell responses under different scenarios in vaccinated and unvaccinated individuals with and without Omicron infection. We found that the humoral response was higher among vaccinated-naïve than unvaccinated convalescent. Unvaccinated with and without infection had comparable low humoral responses, whereas vaccinated with a second or third dose, independent of infection status, had increasingly higher levels. Only a minor fraction of unvaccinated individuals had detectable humoral responses following Omicron infection, while almost all had positive T-cell responses. In conclusion, primary Omicron infection mounts a low humoral immune response, enhanced by prior vaccination. Omicron infection induced a robust T-cell response in both unvaccinated and vaccinated, demonstrating that immune evasion of primary Omicron infection affects humoral immunity more than T-cell immunity.

B-072 Analysis of Anti-SARS-Cov-2 S1-RBD and N IgG in Saliva and Serum

Abstract Background Mucosal antibody testing may be a significant and more practical tool for measuring humoral immunity since it is non-invasive. The aim of this study was to evaluate the SARS-CoV-2-specific antibody prevalence in saliva compared to serum in 2022. Methods From February to December 2022, the paired serum and saliva specimens were collected twice over a period. The saliva specimens were collected using a Salivette Cortisol (Sarstedt AG &amp; Co. KG, Germany) device according to manufacturers’ instructions. Antibody tests were performed using nucleocapsid protein-based Elecsys anti-SARS-CoV-2 (anti-N IgG) and spike protein targeted SARS-CoV-2 IgG II Quant (anti-S1-RBD IgG). Results 162 (97.6%) of 166 had anti-S1-RBD IgG in their serum. At the 1st visit and 2nd visit, 79 (47.5%) and 97 (58.4%) patients, respectively, had anti-N IgG in their serum. At 1st visit and 2nd visit, respectively, 14 (8.4%) and 13 (7.8%) patients had anti-S1-RBD IgG in their saliva. Four (2.4%) and five (3.0%) patients had anti-N IgG in their saliva during the 1st visit and 2nd visit, respectively. The correlation coefficient of the results of anti-S1-RBD IgG and anti-N IgG quantitative results between at 1st visit and 2nd visit were low (r = 0.1721 to 0.6448). All patients in the anti-S1-RBD IgG seronegative group (N = 4) had anti-N IgG in their serum, but neither anti-S1-RBD IgG nor anti-N IgG were detected in their saliva. All anti-N IgG seronegative patients (N = 68) had anti-S1-RBD IgG in serum, while 3 patients had anti-S1-RBD IgG and 7 patients had anti-N IgG in saliva. Conclusion Mucosal antibodies were detected in certain seronegative patients. The assessment of SARS-CoV-2-specific saliva antibodies should be regarded as a supplemental non-invasive diagnostic to serum to evaluate the prevalence and transmission of COVID-19.

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.

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.

Detection of SARS-CoV-2 IgG antibodies and inflammatory cytokines in saliva-a pilot study.

The pandemic caused by SARS-CoV-2 virus continues to have a profound effect worldwide. However, COVID-19 induced oral facial manifestations have not been fully described. We conducted a prospective study to demonstrate feasibility of anti-SARS-CoV-2 IgG and inflammatory cytokine detection in saliva. Our primary objective was to determine whether COVID-19 PCR positive patients with xerostomia or loss of taste had altered serum or saliva cytokine levels compared to COVID-19 PCR positive patients without those oral symptoms. Our secondary objective was to determine the correlation between serum and saliva COVID-19 antibody levels. For cytokine analysis, saliva and serum were obtained from 17 participants with PCR-confirmed COVID-19 infection at three sequential time points, yielding 48 saliva samples and 19 paired saliva-serum samples from 14 of the 17 patients. For COVID-19 antibody analyses, an additional 27 paired saliva-serum samples from 22 patients were purchased. The saliva antibody assay had 88.64% sensitivity [95% Confidence Interval (CI) 75.44%, 96.21%] to detect SARS-CoV-2 IgG antibodies compared to serum antibody. Among the inflammatory cytokines assessed - IL-6, TNF-α, IFN-γ, IL-10, IL-12p70, IL-1β, IL-8, IL-13, IL-2, IL-5, IL-7 and IL-17A, xerostomia correlated with lower levels of saliva IL-2 and TNF-α, and elevated levels of serum IL-12p70 and IL-10 (p<0.05). Loss of taste was observed in patients with elevated serum IL-8 (p<0.05). Further studies are needed to construct a robust saliva-based COVID-19 assay to assess antibody and inflammatory cytokine response, which has potential utility as a non-invasive monitoring modality during COVID-19 convalescence.

Detection of immunoglobulins specific to SARS-CoV-2 virus S-protein in mixed saliva

BACKGROUND: Since the beginning of the COVID-19 pandemic, scientists have been concerned about finding an effective alternative to blood-based diagnostics of antibodies to SARS-CoV-2. According to the results of numerous studies by domestic and foreign authors, saliva is a valuable diagnostic material. Mixed saliva implies the possibility of painless and non-invasive sampling and opens up great opportunities for diagnosing COVID-19 in vulnerable populations, such as children.&#x0D; AIM: Development of a method for detecting immunoglobulins specific to the SARS-CoV-2 virus in saliva after infection with COVID-19 and vaccination.&#x0D; MATERIALS AND METHODS: The study involved 43 people aged 20 to 67 years, divided into 3 groups: recovered from COVID-19; those who believe they have not had COVID-19; vaccinated with various drugs against COVID-19. Samples of unstimulated mixed saliva and capillary blood from a finger were obtained from each subject, which were subsequently examined by methods of qualitative and quantitative enzyme immunoassay.&#x0D; RESULTS: A high level of correlation between the content of local IgG and IgA in saliva was demonstrated, while the relationship between the content of IgA in blood and saliva was shown to be of medium strength. A higher number of seropositive patients was demonstrated in the group vaccinated with vaccines against SARS-CoV-2, compared with those who recovered from COVID-19, which is consistent with the anamnesis data.&#x0D; CONCLUSIONS: This work demonstrates and confirms that saliva in the oral fluid is a valuable diagnostic material for the study of local antibodies. The data obtained will allow the development of unique test systems for studying humoral immunity to the SARS-CoV-2 virus, which can become an alternative replacement for the usual determination of virus-specific immunoglobulins in the blood.

The relationship between the intensity of Gasterophilus intestinalis larvae infection and the serum and salivary humoral immune response in horses

Infection with Gasterophilus intestinalis (botfly) larvae often occurs in horses. The aim of the study was to isolate the larvae of G. intestinalis and evaluate the serum and salivary humoral immune response using self-developed ELISA in G. intestinalis infected horses. Blood serum or saliva samples were taken from 125 infected horses and 54 uninfected slaughtered horses. The antigens from G. intestinalis larvae were used for development of ELISA in order to evaluate the intensity of G. intestinalis IgG, IgM, and IgA antibody reactivity in the serum or saliva of naturally infected horses and horses without larvae in the gastrointestinal tract (control group). Serum antibodies against second and third larvae’s stadium antigens reacted significantly more intensively in infected than in healthy horses in IgG (p ≤ 0.001; p ≤ 0.05, respectively) and IgA (p ≤ 0.05;p ≤ 0.001, respectively) classes. Salivary IgG and IgA specific’s antibody reactivity was significantly higher in horses with moderate (p ≤ 0.01) and severe infection (p ≤ 0.001) compared to the healthy horses. The determination of the G. intestinalis IgG and IgA antibody activity in saliva and serum may be used for detecting horses moderately and severely infested with larvae.

Differences in systemic and mucosal SARS-CoV-2 antibody prevalence in a prospective cohort of Dutch children.

BackgroundAs SARS-CoV-2 will likely continue to circulate, low-impact methods become more relevant to monitor antibody-mediated immunity. Saliva sampling could provide a non-invasive method with reduced impact on children. Studies reporting on the differences between systemic and mucosal humoral immunity to SARS-CoV-2 are inconsistent in adults and scarce in children. These differences may be further unraveled by exploring associations to demographic and clinical variables.MethodsTo evaluate the use of saliva antibody assays, we performed a cross-sectional cohort study by collecting serum and saliva of 223 children attending medical services in the Netherlands (irrespective of SARS-CoV-2 exposure, symptoms or vaccination) from May to October 2021. With a Luminex and a Wantai assay, we measured prevalence of SARS-CoV-2 spike (S), receptor binding domain (RBD) and nucleocapsid-specific IgG and IgA in serum and saliva and explored associations with demographic variables.FindingsThe S-specific IgG prevalence was higher in serum 39% (95% CI 32 – 45%) than in saliva 30% (95% CI 24 – 36%) (P ≤ 0.003). Twenty-seven percent (55/205) of children were S-specific IgG positive in serum and saliva, 12% (25/205) were only positive in serum and 3% (6/205) only in saliva. Vaccinated children showed a higher concordance between serum and saliva than infected children. Odds for saliva S-specific IgG positivity were higher in girls compared to boys (aOR 2.63, P = 0.012). Moreover, immunocompromised children showed lower odds for S- and RBD-specific IgG in both serum and saliva compared to healthy children (aOR 0.23 – 0.25, P ≤ 0.050).ConclusionsWe showed that saliva-based antibody assays can be useful for identifying SARS-CoV-2 humoral immunity in a non-invasive manner, and that IgG prevalence may be affected by sex and immunocompromisation. Differences between infection and vaccination, between sexes and between immunocompromised and healthy children should be further investigated and considered when choosing systemic or mucosal antibody measurement.

Poly(hydrophobic Amino Acids) and Liposomes for Delivery of Vaccine against Group A Streptococcus.

Adjuvants and delivery systems are essential components of vaccines to increase immunogenicity against target antigens, particularly for peptide epitopes (poor immunogens). Emulsions, nanoparticles, and liposomes are commonly used as a delivery system for peptide-based vaccines. A Poly(hydrophobic amino acids) delivery system was previously conjugated to Group A Streptococcus (GAS)-derived peptide epitopes, allowing the conjugates to self-assemble into nanoparticles with self adjuvanting ability. Their hydrophobic amino acid tail also serves as an anchoring moiety for the peptide epitope, enabling it to be integrated into the liposome bilayer, to further boost the immunological responses. Polyleucine-based conjugates were anchored to cationic liposomes using the film hydration method and administered to mice subcutaneously. The polyleucine-peptide conjugate, its liposomal formulation, and simple liposomal encapsulation of GAS peptide epitope induced mucosal (saliva IgG) and systemic (serum IgG, IgG1 and IgG2c) immunity in mice. Polyleucine acted as a potent liposome anchoring portion, which stimulated the production of highly opsonic antibodies. The absence of polyleucine in the liposomal formulation (encapsulated GAS peptide) induced high levels of antibody titers, but with poor opsonic ability against GAS bacteria. However, the liposomal formulation of the conjugated vaccine was no more effective than conjugates alone self-assembled into nanoparticles.