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1.
Sci Rep ; 14(1): 21655, 2024 09 17.
Artigo em Inglês | MEDLINE | ID: mdl-39289450

RESUMO

Using a modified proximity extension assay, total and immunoglobulin (Ig) class-specific anti-SARS-CoV-2 antibodies were sensitively and conveniently detected directly from ø1.2 mm discs cut from dried blood and saliva spots (DBS and DSS) without the need for elution. For total Ig detection, antigen probes were prepared by conjugating recombinant spike protein subunit 1 (S1-RBD) to a pair of oligonucleotides. To detect isotype-specific antibody reactivity, one antigen probe was replaced with oligonucleotide-conjugated antibodies specific for antibody isotypes. Binding of pairs of oligonucleotide-conjugated probes to antibodies in patient samples brings oligonucleotides in proximity. An added DNA polymerase uses a transient hybridization between the oligonucleotides to prime synthesis of a DNA strand, which serves as a DNA amplicon that is quantified by real-time PCR. The S1-RBD-specific IgG, IgM, and IgA antibodies in DBS samples collected over the course of a first and second vaccination exhibited kinetics consistent with previous reports. Both DBS and DSS collected from 42 individuals in the autumn of 2023 showed significant level of total S1-RBD antibodies with a correlation of R = 0.70. However, levels in DSS were generally 10 to 100-fold lower than in DBS. Anti-S1-RBD IgG and IgA in DSS demonstrated a correlation of R = 0.6.


Assuntos
Anticorpos Antivirais , COVID-19 , Imunoglobulina A , Imunoglobulina G , Imunoglobulina M , SARS-CoV-2 , Saliva , Humanos , Saliva/imunologia , SARS-CoV-2/imunologia , Imunoglobulina M/imunologia , Imunoglobulina M/sangue , Imunoglobulina G/imunologia , Imunoglobulina G/sangue , Imunoglobulina A/imunologia , Imunoglobulina A/sangue , Anticorpos Antivirais/imunologia , Anticorpos Antivirais/sangue , COVID-19/imunologia , COVID-19/diagnóstico , COVID-19/virologia , Glicoproteína da Espícula de Coronavírus/imunologia , Teste em Amostras de Sangue Seco/métodos
2.
Sci Rep ; 14(1): 21788, 2024 09 18.
Artigo em Inglês | MEDLINE | ID: mdl-39294156

RESUMO

Oral fluids provide ready detection of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) and host responses. This study sought to evaluate relationships between oral virus, oral and systemic anti-SARS-CoV-2-specific antibodies, and symptoms. Oral fluids (saliva/throat wash (saliva/TW)) and serum were collected from asymptomatic and symptomatic, nasopharyngeal (NP) SARS-CoV-2 RT-qPCR+ human participants (n = 45). SARS-CoV-2 RT-qPCR and N-antigen detection by immunoblot and lateral flow assay (LFA) were performed. RT-qPCR for subgenomic RNA (sgRNA) was sequence confirmed. SARS-CoV-2-anti-S protein RBD LFA and ELISA assessed IgM and IgG responses. Structural analysis identified host salivary molecules analogous to SARS-CoV-2-N-antigen. At time of enrollment (baseline, BL), LFA-detected N-antigen in 86% of TW and was immunoblot-confirmed. However, only 3/17 were saliva/TW qPCR+ . Sixty percent of saliva and 83% of TW demonstrated persistent N-antigen at 4 weeks. N-antigen LFA signal in three anti-spike sero-negative participants suggested potential cross-detection of 4 structurally analogous salivary RNA binding proteins (alignment 19-29aa, RMSD 1-1.5 Angstroms). At enrollment, symptomatic participants demonstrated replication-associated sgRNA junctions, were IgG+ (94%/100% in saliva/TW), and IgM+ (63%/54%). At 4 weeks, SARS-CoV-2 IgG (100%/83%) and IgM (80%/67%) persisted. Oral and serum IgG correlated 100% with NP+ PCR status. Cough and fatigue severity (p = 0.010 and 0.018 respectively), and presence of weakness, nausea, and composite upper respiratory symptoms (p = 0.037, 0.005, and 0.017, respectively) were negatively associated with saliva IgM but not TW or serum IgM. Throat wash IgM levels were higher in women compared to men, although the association did not reach statistical significance (median: 290 (female) versus 0.697, p = 0.056). Important to transmission and disease course, oral viral replication and persistence showed clear relationships with select symptoms and early oral IgM responses during early infection. N-antigen cross-reactivity may reflect mimicry of structurally analogous host proteins.


Assuntos
Anticorpos Antivirais , COVID-19 , SARS-CoV-2 , Saliva , Humanos , COVID-19/imunologia , COVID-19/virologia , COVID-19/diagnóstico , SARS-CoV-2/imunologia , Saliva/virologia , Saliva/imunologia , Feminino , Masculino , Anticorpos Antivirais/sangue , Anticorpos Antivirais/imunologia , Adulto , Pessoa de Meia-Idade , Imunoglobulina G/sangue , Imunoglobulina G/imunologia , Imunoglobulina M/sangue , Imunoglobulina M/imunologia , Fosfoproteínas/imunologia , RNA Viral , Nasofaringe/virologia , Proteínas do Nucleocapsídeo de Coronavírus/imunologia , Idoso
3.
PLoS One ; 19(9): e0307936, 2024.
Artigo em Inglês | MEDLINE | ID: mdl-39226256

RESUMO

mRNA-based COVID-19 vaccines have played a critical role in reducing severe outcomes of COVID-19. Humoral immune responses against SARS-CoV-2 after vaccination have been extensively studied in blood; however, limited information is available on the presence and duration of SARS-CoV-2 specific antibodies in saliva and other mucosal fluids. Saliva offers a non-invasive sampling method that may also provide a better understanding of mucosal immunity at sites where the virus enters the body. Our objective was to evaluate the salivary immune response after vaccination with the COVID-19 Moderna mRNA-1273 vaccine. Two hundred three staff members of the U.S. Centers for Disease Control and Prevention were enrolled prior to receiving their first dose of the mRNA-1273 vaccine. Participants were asked to self-collect 6 saliva specimens at days 0 (prior to first dose), 14, 28 (prior to second dose), 42, and 56 using a SalivaBio saliva collection device. Saliva specimens were tested for anti-spike protein SARS-CoV-2 specific IgA and IgG enzyme immunoassays. Overall, SARS-CoV-2-specific salivary IgA titers peaked 2 weeks after each vaccine dose, followed by a sharp decrease during the following weeks. In contrast to IgA titers, IgG antibody titers increased substantially 2 weeks after the first vaccine dose, peaked 2 weeks after the second dose and persisted at an elevated level until at least 8 weeks after the first vaccine dose. Additionally, no significant differences in IgA/IgG titers were observed based on age, sex, or race/ethnicity. All participants mounted salivary IgA and IgG immune responses against SARS-CoV-2 after receiving the mRNA-1273 COVID-19 vaccine. Because of the limited follow-up time for this study, more data are needed to assess the antibody levels beyond 2 months after the first dose. Our results confirm the potential utility of saliva in assessing immune responses elicited by immunization and possibly by infection.


Assuntos
Anticorpos Antivirais , Vacinas contra COVID-19 , COVID-19 , Imunoglobulina A , Imunoglobulina G , SARS-CoV-2 , Saliva , Vacinação , Humanos , Saliva/imunologia , Feminino , Masculino , Adulto , SARS-CoV-2/imunologia , COVID-19/imunologia , COVID-19/prevenção & controle , Anticorpos Antivirais/imunologia , Anticorpos Antivirais/sangue , Vacinas contra COVID-19/imunologia , Vacinas contra COVID-19/administração & dosagem , Pessoa de Meia-Idade , Imunoglobulina A/imunologia , Imunoglobulina A/análise , Imunoglobulina G/imunologia , Imunoglobulina G/sangue , Vacina de mRNA-1273 contra 2019-nCoV , Adulto Jovem , Imunidade nas Mucosas/imunologia , Glicoproteína da Espícula de Coronavírus/imunologia
4.
Folia Med (Plovdiv) ; 66(4): 515-520, 2024 Aug 31.
Artigo em Inglês | MEDLINE | ID: mdl-39257271

RESUMO

INTRODUCTION: Malnutrition in children is epidemic in developing countries. Several health issues and consequences are believed to develop due to this phenomenon. Children's oral health is also affected by malnutrition. The main aspects of oral health status are caries experience, the existence of cariogenic bacteria, and salivary immunoglobulin A.


Assuntos
Cárie Dentária , Saúde Bucal , Saliva , Humanos , Cárie Dentária/epidemiologia , Criança , Saliva/imunologia , Masculino , Feminino , Desnutrição/epidemiologia , Imunoglobulina A Secretora/metabolismo , Imunoglobulina A/metabolismo
5.
J Infect ; 89(4): 106237, 2024 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-39121969

RESUMO

BACKGROUND: Unlike adults, children experienced stronger and longer vector replication in plasma and shedding in saliva following rVSVΔG-ZEBOV-GP vaccination. The resulting risks of immunosuppression or immune hyperactivation leading to increased Adverse Events (AEs) and altered antibody responses are concerns that have been addressed in the present manuscript. METHODS: Children aged 1-12 years living in Gabon received either rVSVΔG-ZEBOV-GP (ERVEBO®) vaccine or the varicella-zoster virus (VZV) vaccine (VZV). The concentration of rVSVΔG vector in blood and saliva, the occurrence of AEs up to day 28; the anti-rVSVΔG-ZEBOV-GP and anti-VZV IgG antibody titres, neutralising and avidity functions of anti-rVSVΔG-ZEBOV-GP by day 365; were assessed in serum. (PACTR202005733552021) FINDINGS: In the rVSVΔG-ZEBOV-GP group, 70% and 7% of children had >0 copies/ml of rVSVΔG respectively in plasma by day 3 and in saliva by day 14 after vaccination, with no detection on day 28. Significantly higher but transient AEs occurred in the rVSVΔG-ZEBOV-GP group. Both vaccines induced seroconversion on day 28 and sustainable IgG antibody titres by day 365. Avidity and neutralisation functions of the anti-rVSVΔG-ZEBOV-GP antibodies peaked at day 28 and were maintained by day 365. INTERPRETATION: The replication and shedding do not affect the favourable risk-benefit balance of the rVSVΔG-ZEBOV-GP in children.


Assuntos
Anticorpos Antivirais , Vacinas contra Ebola , Humanos , Gabão , Pré-Escolar , Anticorpos Antivirais/sangue , Masculino , Feminino , Criança , Lactente , Vacinas contra Ebola/imunologia , Vacinas contra Ebola/efeitos adversos , Vacinas contra Ebola/administração & dosagem , Saliva/imunologia , Saliva/virologia , Ebolavirus/imunologia , Ebolavirus/genética , Imunoglobulina G/sangue , Doença pelo Vírus Ebola/prevenção & controle , Replicação Viral , Imunogenicidade da Vacina , Anticorpos Neutralizantes/sangue , Vacinação , Eliminação de Partículas Virais
6.
J Dent Res ; 103(10): 988-998, 2024 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-39101654

RESUMO

Most patients diagnosed with oral squamous cell carcinoma (OSCC) present with locally advanced stages, which are typically associated with poor outcomes. Although immunotherapy offers potential improvements in patient survival, its efficacy is hampered by low response rates. The microbiome is widely involved in tumor immunity and may play a role in immunotherapy. This study aimed to investigate the potential association between the oral (salivary) microbiome and immunotherapy response in patients with OSCC. Salivary metagenome sequencing was performed on 47 patients with OSCC undergoing neoadjuvant immunotherapy (NAIT) in a clinical trial (NCT04649476). Patients were divided into responders and nonresponders based on their pathological responses. The results showed that the species richness of the salivary microbiome was lower in the nonresponders before NAIT than in the responders. Differential analysis revealed that nonresponders exhibited a lower relative abundance of 34 bacterial species and a higher relative abundance of 4 bacterial species. Notably, low levels of Eubacterium infirmum, Actinobaculum, and Selenomas (EAS) in the saliva may be associated with the nonresponse of patients with OSCC to NAIT. A nomogram based on EAS was developed and validated to determine the efficacy of NAIT. The area under the curve for the training cohort was 0.81 (95% confidence interval, 0.66 to 0.81). Quantitative polymerase chain reaction confirmed that low levels of salivary EAS effectively identified nonresponders to NAIT. Furthermore, the low abundance of salivary EAS was closely correlated with a low density of intratumoral CD4+, CD14+, CD68+, and FOXP3+ cells. Metabolic functional annotation revealed numerous biosynthetic processes associated with EAS that were more active in responders. In summary, this study provides valuable data resources for the salivary microbiome and reveals that nonresponders have different salivary microbiome profiles than responders do before NAIT. Low salivary EAS levels can serve as potential biomarkers for distinguishing nonresponders from responders.


Assuntos
Carcinoma de Células Escamosas , Imunoterapia , Microbiota , Neoplasias Bucais , Terapia Neoadjuvante , Saliva , Adulto , Idoso , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Carcinoma de Células Escamosas/terapia , Carcinoma de Células Escamosas/imunologia , Carcinoma de Células Escamosas/microbiologia , Imunoterapia/métodos , Neoplasias Bucais/terapia , Neoplasias Bucais/microbiologia , Neoplasias Bucais/imunologia , Saliva/microbiologia , Saliva/imunologia , Resultado do Tratamento
7.
Clin Exp Dent Res ; 10(4): e945, 2024 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-39104139

RESUMO

OBJECTIVES: This study aimed to assess the intricate relationship between salivary IgA antibody levels to PAc (361-386) (PPA), mutans streptococci colonization, and root caries development in older adults. MATERIALS AND METHODS: This study included 307 participants aged 76 years residing in Niigata city, Japan. Clinical oral examinations were performed at baseline in 2004 and 1 year later, during which the total number of untreated and treated root caries was assessed using the root decayed, filled tooth (DFT) index. The stimulated saliva samples were collected using the spitting method during the baseline survey. Salivary IgA antibody levels to amino acid residues 361-386 of Streptococcus mutans PAc were quantified using an enzyme-linked immunosorbent assay. Statistical analyses, including the χ2 test, Mann-Whitney U test, and logistic regressions, were performed to examine the association of increased root DFT with the independent variables. RESULTS: Among the 307 participants (53.1% men), the mean root DFT at baseline was 3.77 ± 3.66, and 36.5% of the study sample exhibited increased root DFT after 1 year with a mean increment of 0.36 ± 0.48. Participants with increase in root DFT after 1 year had significantly higher rates of low PPA levels (≤ 25th percentile) than those without increased root DFT (p = 0.020). Low PPA levels (≤ 25th percentile) were significantly more likely to have an increased risk of root caries development compared with PPA levels > 25th percentile (adjusted OR: 1.88, 95% CI: 1.09-3.25). CONCLUSION: Low PPA levels and root caries incidence correlated significantly, suggesting that low levels of salivary IgA antibody to PAc (361-386) may serve as a risk factor for increased root caries in older adults.


Assuntos
Cárie Radicular , Saliva , Streptococcus mutans , Humanos , Cárie Radicular/imunologia , Cárie Radicular/epidemiologia , Idoso , Feminino , Masculino , Saliva/imunologia , Saliva/microbiologia , Saliva/química , Streptococcus mutans/imunologia , Streptococcus mutans/isolamento & purificação , Fatores de Risco , Japão/epidemiologia , Imunoglobulina A Secretora/análise , Imunoglobulina A Secretora/imunologia , Ensaio de Imunoadsorção Enzimática , Imunoglobulina A/análise , Imunoglobulina A/imunologia , Imunoglobulina A/metabolismo , Anticorpos Antibacterianos/análise , Anticorpos Antibacterianos/imunologia , Índice CPO
8.
Front Immunol ; 15: 1454018, 2024.
Artigo em Inglês | MEDLINE | ID: mdl-39136018

RESUMO

Cells exposed to stressors of various origin activate protective mechanisms that include the expression of heat shock proteins (Hsps)/molecular chaperones belonging to several families. Well-characterized inducible Hsp70 is present in all human cell-types and biological fluids, including blood, urine, and saliva. The presence of anti-Hsp70 autoantibodies in the serum of healthy individuals has already been confirmed, and their elevated titers positively correlated with the severity of several pathological conditions, including coeliac disease and dermatitis herpetiformis - a cutaneous manifestation of coeliac disease. Here, using an indirect enzyme-linked immunosorbent assay, we demonstrate, for the first time, that anti-Hsp70 autoantibodies are present in the saliva and urine of healthy individuals. Although the occurrence of anti-Hsp70 autoantibodies in the biological fluids of healthy individuals is intriguing, their physiological role is currently unknown. It is believed that antibodies reacting with self-molecules present in the serum of healthy individuals are part of natural autoantibody pool with multiple regulatory functions. On the other hand, some autoantibodies (e.g., typical of autoimmune bullous skin diseases or systemic lupus erythematosus) may be present before the onset of the disease and serve as specific predictive biomarkers. Therefore, we would like to initiate a discussion or future research direction on the use of anti-Hsp70 autoantibodies as a potential "biomarker" in the diagnosis or prediction of autoimmune diseases. Our findings can be considered in biomedical research to develop noninvasive, inexpensive and easy-to-use tests. Nevertheless, large-scale comparative studies should be initiated, involving the collection and analysis of biological samples such as saliva or urine from patients suffering from autoimmune diseases or other inflammatory or neoplastic diseases, to determine whether the levels of anti-Hsp70 autoantibodies are indeed elevated and whether they correlate with the clinical picture of any disease or established biomarkers.


Assuntos
Autoanticorpos , Proteínas de Choque Térmico HSP70 , Saliva , Humanos , Saliva/imunologia , Saliva/metabolismo , Proteínas de Choque Térmico HSP70/imunologia , Autoanticorpos/imunologia , Autoanticorpos/sangue , Feminino , Adulto , Masculino , Biomarcadores/urina , Pessoa de Meia-Idade , Ensaio de Imunoadsorção Enzimática , Voluntários Saudáveis
9.
Brain Behav Immun ; 122: 555-564, 2024 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-39168271

RESUMO

Situational factors can increase people's vulnerability to intergroup bias, including prejudicial attitudes, negative stereotyping, and discrimination. We proposed that increases in inflammatory activity that coincide with acute illness may represent a hitherto unstudied situational factor that increases intergroup bias. The current study experimentally manipulated increases in inflammatory activity by administering the seasonal influenza vaccine or a saline placebo. We quantified inflammatory activity by assessing change in salivary pro-inflammatory cytokines and assessed intergroup bias using a resume evaluation task and self-reported ethnocentrism. Primary analyses focused on a subsample of 117 participants who provided high quality data; robustness analyses included various permutations of lower quality participants. Findings revealed that changes in the cytokine interleukin-1ß (IL-1ß) in response to the vaccine were associated with greater intergroup bias. Among participants who received the vaccine, IL-1ß change was negatively associated with evaluation of a Latina (but not a White woman) applicant's competency and recommended starting salary. Moreover, IL-1ß change was positively associated with ethnocentrism. Overall, results provide support for the hypothesis that acute illness, via the mechanistic role of inflammatory cytokines, affects social cognition in ways that can increase intergroup bias.


Assuntos
Citocinas , Hispânico ou Latino , Vacinas contra Influenza , Interleucina-1beta , Humanos , Feminino , Vacinas contra Influenza/imunologia , Masculino , Adulto , Hispânico ou Latino/psicologia , Citocinas/metabolismo , Interleucina-1beta/metabolismo , Adulto Jovem , Saliva/imunologia , Saliva/química , Candidatura a Emprego , Preconceito
10.
Virol Sin ; 39(4): 675-684, 2024 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-38997087

RESUMO

Norovirus (NoV) infection is a major cause of gastroenteritis worldwide. The virus poses great challenges in developing vaccines with broad immune protection due to its genetic and antigenic diversity. To date, there are no approved NoV vaccines for clinical use. Here, we aimed to develop a broad-acting quadrivalent NoV vaccine based on a chimpanzee adenovirus vector, AdC68, carrying the major capsid protein (VP1) of noroviral GI and GII genotypes. Compared to intramuscular (i.m.), intranasal (i.n.), or other prime-boost immunization regimens (i.m. â€‹+ â€‹i.m., i.m. â€‹+ â€‹i.n., i.n. â€‹+ â€‹i.m.), AdC68-GI.1-GII.3 (E1)-GII.4-GII.17 (E3), administered via i.n. â€‹+ â€‹i.n. induced higher titers of serum IgG antibodies and higher IgA antibodies in bronchoalveolar lavage fluid (BALF) and saliva against the four homologous VP1s in mice. It also significantly stimulated the production of blocking antibodies against the four genotypes. In response to re-stimulation with virus-like particles (VLP)-GI.1, VLP-GII.3, VLP-GII.4, and VLP-GII.17, the quadrivalent vaccine administered according to the i.n. â€‹+ â€‹i.n. regimen effectively triggered specific cell-mediated immune responses, primarily characterized by IFN-γ secretion. Furthermore, the preparation of this novel quadrivalent NoV vaccine requires only a single recombinant adenovirus to provide broad preventive immunity against the major GI/GII epidemic strains, making it a promising vaccine candidate for further development.


Assuntos
Adenoviridae , Anticorpos Antivirais , Infecções por Caliciviridae , Vetores Genéticos , Camundongos Endogâmicos BALB C , Norovirus , Pan troglodytes , Vacinas Virais , Animais , Norovirus/imunologia , Norovirus/genética , Anticorpos Antivirais/sangue , Anticorpos Antivirais/imunologia , Camundongos , Infecções por Caliciviridae/prevenção & controle , Infecções por Caliciviridae/imunologia , Vacinas Virais/imunologia , Vacinas Virais/genética , Vacinas Virais/administração & dosagem , Vetores Genéticos/genética , Vetores Genéticos/imunologia , Adenoviridae/genética , Adenoviridae/imunologia , Feminino , Proteínas do Capsídeo/genética , Proteínas do Capsídeo/imunologia , Imunoglobulina G/sangue , Gastroenterite/prevenção & controle , Gastroenterite/virologia , Gastroenterite/imunologia , Anticorpos Neutralizantes/sangue , Anticorpos Neutralizantes/imunologia , Imunoglobulina A/sangue , Genótipo , Saliva/imunologia , Saliva/virologia , Líquido da Lavagem Broncoalveolar/imunologia , Líquido da Lavagem Broncoalveolar/virologia
11.
Transl Psychiatry ; 14(1): 305, 2024 Jul 24.
Artigo em Inglês | MEDLINE | ID: mdl-39048549

RESUMO

We recently indicated that four-week probiotic supplementation significantly reduced depression along with microbial and neural changes in people with depression. Here we further elucidated the biological modes of action underlying the beneficial clinical effects of probiotics by focusing on immune-inflammatory processes. The analysis included a total of N = 43 participants with depression, from which N = 19 received the probiotic supplement and N = 24 received a placebo over four weeks, in addition to treatment as usual. Blood and saliva were collected at baseline, at post-intervention (week 4) and follow-up (week 8) to assess immune-inflammatory markers (IL-1ß, IL-6, CRP, MIF), gut-related hormones (ghrelin, leptin), and a stress marker (cortisol). Furthermore, transcriptomic analyses were conducted to identify differentially expressed genes. Finally, we analyzed the associations between probiotic-induced clinical and immune-inflammatory changes. We observed a significant group x time interaction for the gut hormone ghrelin, indicative of an increase in the probiotics group. Additionally, the increase in ghrelin was correlated with the decrease in depressive symptoms in the probiotics group. Transcriptomic analyses identified 51 up- and 57 down-regulated genes, which were involved in functional pathways related to enhanced immune activity. We identified a probiotic-dependent upregulation of the genes ELANE, DEFA4 and OLFM4 associated to immune activation and ghrelin concentration. These results underscore the potential of probiotic supplementation to produce biological meaningful changes in immune activation in patients with depression. Further large-scale mechanistic trials are warranted to validate and extend our understanding of immune-inflammatory measures as potential biomarkers for stratification and treatment response in depression. Trial Registration: www.clinicaltrials.gov , identifier: NCT02957591.


Assuntos
Probióticos , Humanos , Probióticos/uso terapêutico , Probióticos/administração & dosagem , Masculino , Feminino , Adulto , Pessoa de Meia-Idade , Grelina/sangue , Hidrocortisona/sangue , Inflamação/imunologia , Método Duplo-Cego , Saliva/química , Saliva/imunologia , Biomarcadores/sangue , Leptina/sangue , Depressão/imunologia , Depressão/terapia , Suplementos Nutricionais
12.
Methods Mol Biol ; 2821: 205-216, 2024.
Artigo em Inglês | MEDLINE | ID: mdl-38997491

RESUMO

Enzyme-linked immunosorbent assay (ELISA) detects qualitatively and quantitatively the presence of antibodies or antigens in a sample. Due to its simplicity, high sensitivity, and user-friendliness, the test is widely used in laboratory research, clinical diagnoses, and food testing. This chapter describes the indirect semiquantitative ELISA protocol used to monitor antibody levels in animals and analyze the titer levels of specific antibodies against a target antigen in serum and saliva.


Assuntos
Anticorpos , Ensaio de Imunoadsorção Enzimática , Saliva , Ensaio de Imunoadsorção Enzimática/métodos , Saliva/imunologia , Animais , Anticorpos/imunologia , Anticorpos/sangue , Antígenos/imunologia , Humanos
13.
Sci Rep ; 14(1): 17308, 2024 07 27.
Artigo em Inglês | MEDLINE | ID: mdl-39068230

RESUMO

The SARS-CoV-2 outbreak has provoked more than 6 million deaths worldwide. The scarcity of effective treatments and its virulence converted the vaccines into an essential tool to face it. The most used vaccines were the mRNA, adenovirus vector, and inactivated whole-virus. However, nowadays, infants aged < 6 months are not eligible for any vaccines against COVID-19, and their immunization relies on passive immunity. In this research, we investigated the humoral and cellular immune response generated on newborns of SARS-CoV-2 vaccinated mothers with mRNA or viral vector (VV) vaccine employing Fourier transformed infrared (FTIR) spectroscopy in saliva samples. For this purpose, saliva samples of newborns and their mothers were collected; the population was divided into two groups, VV and mRNA, which were subdivided into three subgroups: before pregnancy (BP), at the first (FTP) and second (STP) trimesters of pregnancy. The samples were analyzed using FTIR spectroscopy, and the bands associated with the humoral and cellular immune responses, such as IgG, IgA, and IFN-γ were analyzed. The integrated areas were calculated and compared to elucidate the quantity of those immunoglobins and the cytokine. Likewise, the correlation of the humoral and cellular immune response between the newborns and their mothers and the correlation between cellular and humoral immune response was also evaluated. The VV vaccine produced a significant humoral and cellular immune response in newborns and their mothers when they received it at the STP compared with the mRNA vaccine, evidencing statistical significance. However, no correlation was observed between newborns and their mothers when the vaccine was applied in this trimester of pregnancy. When administered BP, the mRNA vaccine generated more humoral immunity in newborns and their mothers. Nevertheless, compared with the VV vaccine, it only showed statistical significance in the mothers, highlighting that IgG showed a moderate positive correlation between the newborns and their mothers.


Assuntos
Vacinas contra COVID-19 , COVID-19 , SARS-CoV-2 , Vacinação , Humanos , Feminino , Espectroscopia de Infravermelho com Transformada de Fourier/métodos , Recém-Nascido , COVID-19/prevenção & controle , COVID-19/imunologia , Gravidez , Vacinação/métodos , SARS-CoV-2/imunologia , Vacinas contra COVID-19/imunologia , Adulto , Mães , Anticorpos Antivirais/imunologia , Anticorpos Antivirais/sangue , Anticorpos Antivirais/análise , Imunidade Humoral , Saliva/imunologia , Imunidade Celular , Imunoglobulina G/sangue , Imunoglobulina G/imunologia , Imunoglobulina A/imunologia , Imunoglobulina A/análise , Interferon gama/metabolismo , Vacinas de mRNA/imunologia
14.
PLoS One ; 19(7): e0307568, 2024.
Artigo em Inglês | MEDLINE | ID: mdl-39052608

RESUMO

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.


Assuntos
COVID-19 , Hispânico ou Latino , SARS-CoV-2 , Humanos , COVID-19/epidemiologia , COVID-19/diagnóstico , COVID-19/imunologia , COVID-19/sangue , Feminino , Masculino , Adulto , SARS-CoV-2/imunologia , SARS-CoV-2/isolamento & purificação , Estudos Transversais , Pessoa de Meia-Idade , Anticorpos Antivirais/sangue , Anticorpos Antivirais/imunologia , Vacinas contra COVID-19/imunologia , Massachusetts/epidemiologia , Saliva/virologia , Saliva/imunologia , Negro ou Afro-Americano , Teste Sorológico para COVID-19/métodos , Idoso
15.
Vet Parasitol ; 330: 110224, 2024 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-38861910

RESUMO

The identification and characterization of tick proteins allow us to discover new physiological targets useful for the development of tick control methods. Bm05br (Brazil Rhipicephalus microplus protein 05) is a protein with unknown function identified in the saliva of R. microplus. Rs05br (Brazil Rhipicephalus sanguineus protein 05), a protein with 99 % similarity to Bm05br, was identified in Rhipicephalus linnaei egg, larval, and nymphal stages, as well as in adult saliva. To improve the knowledge about both proteins, immunological characterization was performed, including antigenicity analysis, vaccination trials, and artificial feeding. The sequence and antigenicity analysis of Bm05br and Rs05br proteins showed that R. linnaei could serve as a tick model for cross-protection studies. The recombinant Bm05br protein was immunogenic. Anti-Bm05br antibodies recognized the homologous protein Rs05br in different stages, organs, and in the saliva of R. linnaei. Although rBm05br did not induce a protective response against infestation in R. linnaei in this study, further experiments could be developed taking into account new formulations and animal models for vaccination. These results also serve as a reference for future research on the function of these proteins in R. microplus and R. linnaei physiology, as well as other species of the genus Rhipicephalus.


Assuntos
Proteínas de Artrópodes , Rhipicephalus , Infestações por Carrapato , Animais , Rhipicephalus/imunologia , Rhipicephalus/química , Proteínas de Artrópodes/imunologia , Proteínas de Artrópodes/genética , Proteínas de Artrópodes/química , Infestações por Carrapato/veterinária , Infestações por Carrapato/parasitologia , Infestações por Carrapato/imunologia , Infestações por Carrapato/prevenção & controle , Feminino , Proteínas Recombinantes/imunologia , Coelhos , Larva/imunologia , Saliva/imunologia , Saliva/química , Sequência de Aminoácidos
16.
Infect Immun ; 92(7): e0007724, 2024 Jul 11.
Artigo em Inglês | MEDLINE | ID: mdl-38869295

RESUMO

The interplay between host nutritional immune mechanisms and bacterial nutrient uptake systems has a major impact on the disease outcome. The host immune factor calprotectin (CP) limits the availability of essential transition metals, such as manganese (Mn) and zinc (Zn), to control the growth of invading pathogens. We previously demonstrated that the competition between CP and the human pathogen group A streptococcus (GAS) for Zn impacts GAS pathogenesis. However, the contribution of Mn sequestration by CP in GAS infection control and the role of GAS Mn acquisition systems in overcoming host-imposed Mn limitation remain unknown. Using a combination of in vitro and in vivo studies, we show that GAS-encoded mtsABC is a Mn uptake system that aids bacterial evasion of CP-imposed Mn scarcity and promotes GAS virulence. Mn deficiency caused by either the inactivation of mtsC or CP also impaired the protective function of GAS-encoded Mn-dependent superoxide dismutase. Our ex vivo studies using human saliva show that saliva is a Mn-scant body fluid, and Mn acquisition by MtsABC is critical for GAS survival in human saliva. Finally, animal infection studies using wild-type (WT) and CP-/- mice showed that MtsABC is critical for GAS virulence in WT mice but dispensable in mice lacking CP, indicating the direct interplay between MtsABC and CP in vivo. Together, our studies elucidate the role of the Mn import system in GAS evasion of host-imposed metal sequestration and underscore the translational potential of MtsABC as a therapeutic or prophylactic target.


Assuntos
Complexo Antígeno L1 Leucocitário , Manganês , Infecções Estreptocócicas , Streptococcus pyogenes , Manganês/metabolismo , Infecções Estreptocócicas/microbiologia , Infecções Estreptocócicas/imunologia , Infecções Estreptocócicas/metabolismo , Streptococcus pyogenes/metabolismo , Streptococcus pyogenes/patogenicidade , Streptococcus pyogenes/imunologia , Animais , Humanos , Camundongos , Complexo Antígeno L1 Leucocitário/metabolismo , Virulência , Proteínas de Bactérias/metabolismo , Proteínas de Bactérias/genética , Interações Hospedeiro-Patógeno/imunologia , Saliva/microbiologia , Saliva/imunologia , Modelos Animais de Doenças
17.
Plant Cell Physiol ; 65(8): 1213-1223, 2024 Sep 03.
Artigo em Inglês | MEDLINE | ID: mdl-38877965

RESUMO

Plants and insects have co-existed for almost 400 million years and their interactions can be beneficial or harmful, thus reflecting their intricate co-evolutionary dynamics. Many herbivorous arthropods cause tremendous crop loss, impacting the agro-economy worldwide. Plants possess an arsenal of chemical defenses that comprise diverse secondary metabolites that help protect against harmful herbivorous arthropods. In response, the strategies that herbivores use to cope with plant defenses can be behavioral, or molecular and/or biochemical of which salivary secretions are a key determinant. Insect salivary secretions/oral secretions (OSs) play a crucial role in plant immunity as they contain several biologically active elicitors and effector proteins that modulate plants' defense responses. Using this oral secretion cocktail, insects overcome plant natural defenses to allow successful feeding. However, a lack of knowledge of the nature of the signals present in oral secretion cocktails has resulted in reduced mechanistic knowledge of their cellular perception. In this review, we discuss the latest knowledge on herbivore oral secretion derived elicitors and effectors and various mechanisms involved in plant defense modulation. Identification of novel herbivore-released molecules and their plant targets should pave the way for understanding the intricate strategies employed by both herbivorous arthropods and plants in their interactions.


Assuntos
Herbivoria , Insetos , Imunidade Vegetal , Animais , Insetos/fisiologia , Plantas/imunologia , Plantas/metabolismo , Saliva/imunologia , Saliva/metabolismo , Defesa das Plantas contra Herbivoria
18.
Braz Oral Res ; 38: e043, 2024.
Artigo em Inglês | MEDLINE | ID: mdl-38747830

RESUMO

This cross-sectional study evaluated the association between salivary immunoglobulins, plaque index, and gingival index in Brazilian children with and without type 1 diabetes mellitus (DM1). The Strengthening the Reporting of Observational Studies in Epidemiology (STROBE) checklist for the reporting of observational studies was followed. The DM1 group had 38 children, and an equal number of volunteers matched by sex and age were recruited as controls. Clinical examination was performed for plaque index and gingival index determination. Non-stimulated whole saliva was collected. Concentrations of IgA, IgG, and IgM were determined by ELISA test. Data were tested by the Kolmogorov-Smirnov, Mann-Whitney, and Spearman tests and a multiple linear regression model (p<0.05) was performed. Gingival index was higher in the Control (DM1: 0.16±0.17; Control: 0.24±0.23, p=0.040). In DM1, there was a correlation between IgA and age (rho=0.371, p=0.024), IgM and IgG (rho=0.459, p=0.007), and IgM and gingival index (rho=0.394, p=0.014). In DM1, multiple linear regression showed that age (p=0.041; ß=0.363), gingival index (p=0.041; ß=0.398), and plaque index (p=0.008; ß=-0.506) were good predictors of IgA levels in saliva. Thus, IgA was the only researched immunoglobulin that was directly associated with plaque and gingival indices in Brazilian children with DM1, but not in control subjects.


Assuntos
Índice de Placa Dentária , Diabetes Mellitus Tipo 1 , Imunoglobulina A , Índice Periodontal , Saliva , Humanos , Diabetes Mellitus Tipo 1/imunologia , Masculino , Feminino , Saliva/química , Saliva/imunologia , Estudos Transversais , Criança , Brasil/epidemiologia , Estudos de Casos e Controles , Imunoglobulina A/análise , Imunoglobulina G/análise , Estatísticas não Paramétricas , Imunoglobulina M/análise , Valores de Referência , Ensaio de Imunoadsorção Enzimática , Adolescente , Modelos Lineares , Fatores Etários , Imunoglobulinas/análise
20.
J Allergy Clin Immunol ; 154(3): 690-697.e4, 2024 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-38821318

RESUMO

BACKGROUND: Reaction threshold and severity in food allergy are difficult to predict, and noninvasive predictors are lacking. OBJECTIVE: We sought to determine the relationships between pre-challenge levels of peanut (PN)-specific antibodies in saliva and reaction threshold, severity, and organ-specific symptoms during PN allergic reactions. METHODS: We measured PN-specific antibody levels in saliva collected from 127 children with suspected PN allergy before double-blind, placebo-controlled PN challenges in which reaction threshold, severity, and symptoms were rigorously characterized. Low threshold (LT) PN allergy was defined as reaction to <300 mg of PN protein cumulatively consumed. A consensus severity grading system was used to grade severity. We analyzed associations between antibody levels and reaction threshold, severity, and organ-specific symptoms. RESULTS: Among the 127 children, those with high pre-challenge saliva PN IgE had higher odds of LT PN allergy (odds ratio [OR] 3.9, 95% CI 1.6-9.5), while those with high saliva PN IgA:PN IgE ratio or PN IgG4:PN IgE ratio had lower odds of LT PN allergy (OR 0.3, 95% CI 0.1-0.8; OR 0.4, 95% CI 0.2-0.9). Children with high pre-challenge saliva PN IgG4 had lower odds of severe PN reactions (OR 0.4, 95% CI 0.2-0.9). Children with high saliva PN IgE had higher odds of respiratory symptoms (OR 8.0, 95% CI 2.2-26.8). Saliva PN IgE modestly correlated with serum PN IgE levels (Pearson r = 0.31, P = .0004). High and low saliva PN IgE levels further distinguished reaction threshold and severity in participants stratified by serum PN IgE, suggesting endotypes. CONCLUSIONS: Saliva PN antibodies could aid in noninvasive risk stratification of PN allergy threshold, severity, and organ-specific symptoms.


Assuntos
Imunoglobulina E , Hipersensibilidade a Amendoim , Saliva , Índice de Gravidade de Doença , Humanos , Hipersensibilidade a Amendoim/imunologia , Saliva/imunologia , Feminino , Masculino , Imunoglobulina E/imunologia , Imunoglobulina E/sangue , Criança , Pré-Escolar , Alérgenos/imunologia , Arachis/imunologia , Imunoglobulina A/imunologia , Método Duplo-Cego , Imunoglobulina G/imunologia , Imunoglobulina G/sangue , Adolescente
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