Quantification of IgA and IgG and specificities of antibodies to viral proteins in parotid saliva at different stages of HIV-1 infection

Paired sera and parotid saliva from 75 HIV-1-infected patients, divided in three equal groups with CD4⁺ cell counts > 500, 200–500 and < 200/mm³, respectively, were analysed for IgG, IgA and secretory IgA (sIgA) concentrations and for IgG and IgA antibody directed to HIV-1. Twenty-nine age-matched HIV⁻ subjects were used as controls. In serum the concentrations of immunoglobulins were significantly increased in HIV-infected subjects compared with controls, and a progressive increase of IgA and sIgA was noticed while the CD4⁺ cell count decreased. In contrast, concentrations of IgA and sIgA were not different in parotid saliva between the four subject groups. By an ELISA test directed towards HIV-1 proteins, 73 of the 75 serum specimens from the HIV-infected subjects (97%) and 43 of the corresponding saliva (57%) were found positive for specific IgA antibodies to HIV-1, with an even distribution among the three groups of patients. By Western blotting multiple specificities of IgA to HIV-1 proteins were not frequently found in patients. By contrast, in spite of an IgG concentration in saliva about 100 times lower than that of IgA, reactivities were significantly higher for IgG than for IgA antibodies, especially to env and to pol HIV-1 products. Altogether, these data suggest that the regulation of IgA production in HIV-infected subjects is independent in serum and in parotid saliva. This imbalance of IgA/IgG antibodies to HIV-1 at the mucosal level appears to be a specific feature of HIV-1 infection, and may raise important issues in terms of local protection after immunization.     

A novel method to diagnose the infection of enterovirus A71 in children by detecting IgA from saliva

Enterovirus A71 (EV-A71) is one of the main pathogens causing hand, foot, and mouth disease, and often causes diseases of the central nervous system. Early diagnosis is important to prevent EV-A71 outbreaks. The detection of serum immunoglobulin M (IgM) is widely used for the early diagnosis of EV-A71 in clinics, especially in rural areas. However, this technique requires the extraction of blood from children who have thin blood vessels and who might fear the use of needles. Therefore, difficulties in the detection process are often encountered. This study developed a noninvasive method to detect EV-A71-specific immunoglobulin A (IgA) in saliva for the diagnosis of EV-A71 infection. The sensitivity and specificity of IgA detection did not differ significantly compared with IgM detection. IgA antibodies were present in saliva for a relatively shorter period than IgM antibodies were present in serum. The sensitivity of IgA detection was higher than that of IgM detection for secondary EV-A71 infections. These results suggest that the detection of EV-A71-specific IgA in the saliva allows the effective early diagnosis of EV-A71 and may be suitable for detecting EV-A71 infections in children.     

Epigenetic alterations in Sjögren’s syndrome patient saliva

Epigenetic mechanisms have been implicated in the pathogenesis of Sjögren’s syndrome (SS). Extensive alterations in DNA methylation have been described in minor salivary gland (MSG) epithelial cells and lymphocytes derived from SS patients compared to sicca controls. In an effort to identify novel potential epigenetic markers that could prove useful in diagnosis and disease monitoring, we explored whether DNA methylation differences can also be detected in saliva from SS patients compared to sicca controls. We performed DNA methylation analysis by methylation-sensitive restriction digestion followed by quantitative real-time polymerase chain reaction of selected genomic loci in saliva samples of 16 SS patients and 10 sicca controls with negative MSG biopsy. We identified reduced DNA methylation of the imprinting control region (ICR) of the H19 locus in SS patient saliva compared to sicca controls. Levels of saliva H19 ICR methylation were negatively correlated with C4 serum complement levels. Consistent with the reduced methylation of the ICR, H19 RNA levels were increased in SS patient peripheral blood mononuclear cells (PBMCs), while no significant change was observed in MSG H19 RNA levels compared to sicca controls. Our findings support that H19 ICR methylation could be a useful molecular epigenetic marker in monitoring patients with SS, highlighting saliva as a valuable biological sample in SS research and clinical practice. The role of H19 in SS pathogenesis remains to be addressed.     

不同血清型乙型肝炎患者血清和唾液中HBV-DNA含量相关性研究

目的探讨不同血清型乙型肝炎患者血清和唾液中HBV-DNA的差异以及相关性.方法选取60例乙型肝炎患者,分别采用实时荧光定量PCR法同时检测血清与唾液HBV-DNA,用ELISA检测血清HBV标志物,动力学方法检测血清ALT值,并分析其相互关系.结果60例乙型肝炎患者血清与唾液HBV-DNA阳性率分别为93.3％和63.3％(x2=15.91,P＜0.01);HBeAg阳性患者33例,HBeAg阴性患者26例,其血清HBV-DNA阳性率分别为97.15％和88.5％(x2=0.64,P＞0.05);ALT值的四分位间距分别为69.0(45.0～220.5)和38.5 (24.0～103.5)(t=1.36,P=0.180);血清HBV-DNA对数值的四分位间距分别为7.14(6.01～7.62)和5.09(2.30～6.83)(t=3.70,P＜0.05);唾液HBV-DNA对数值的四分位间距分别为4.63(1.19～5.29)和0.00(0.00～4.6)(t=3.25,P＜0.05).对60例乙型肝炎患者血清与唾液HBV-DNA病毒含量的对数值进行分析,发现两者呈显著的正相关(r=0.83,P＜0.05),回归方程为y=0.88x+2.82.结论HBeAg阳性的患者与HBeAg阴性患者相比较,虽然血清中病毒阳性率以及ALT值无差异,但病毒含量有差异.乙型肝炎患者血清与唾液中的病毒含量呈正相关,血清中病毒含量高于唾液,提示血清的传染性比唾液强.     

Measuring salivary cortisol in studies of child development: watch out--what goes in may not come out of saliva collection devices

Technical advances that enable the noninvasive measurement of biomarkers in saliva have spawned a generation of investigations that integrate biological variables into behavioral and developmental research. This study examines whether the collection of saliva, using common absorbent devices compromises the measurement of cortisol when saliva specimens have low sample volume. Within subjects (n = 20), saliva samples were prepared to experimentally represent a gradient of lower to higher sample volumes. One aliquot was immediately frozen (no treatment control) and the remaining aliquots were absorbed ("collected") using one of three collection techniques employed in studies of child development (e.g., braided cotton dental rope, Salivette cotton pledget, or hydrocellulose microsponge). The sample volume recovered from each device relative to the initial volume available to be absorbed, and cortisol level recovered from each device relative to the untreated-control condition were measured. Results reveal that for certain collection devices (1) the percent volume recovered is related to the initial volume available to be absorbed, (2) a substantial percentage of cortisol in saliva specimens can remain in absorbent material, and (3) the percent of cortisol recovered can be associated with the initial sample volume available to be absorbed. When research participants, such as young children, produce low volume saliva specimens, some absorbent devices may have the potential to introduce error variance in the measurement of salivary cortisol.     

Alternative Recruitment Strategies Influence Saliva Sample Return Rates in Community‐Based Genetic Association Studies

Collection of saliva for DNA extraction has created new opportunities to recruit participants from the community for genetic association studies. However, sample return rates are variable. No prior study has specifically addressed how study design impacts sample return. Using data from three large‐scale genetic association studies we compared recruitment strategy and sample return rates. We found highly significant differences in sample return rates between the studies. In studies that recruited retrospectively, overall returns were much lower from families with a self‐limiting condition who provided samples at a research centre or home visit, than adult elderly individuals with a chronic disease who provided samples by post (59% vs. 84%). Prospective recruitment was associated with high agreement to participate (72%), but subsequent low return of actual saliva samples (42%). A telephone call had marginal effect on recruitment in a retrospective family study, but significantly improved returns in a prospective family study. We found no effect upon DNA yield comparing observed versus unobserved sample collection, or between male and female adult participants. Overall, study design significantly impacts upon response rates for genetic association studies recruiting from the community. Our findings will help researchers in constructing and costing a recruitment protocol.     

The Effect of Stress and Meditation on Salivary Protein and Bacteria: A Review and Pilot Study

Abstract

Dentally-induced stress and relaxation-induced anxiety reduction have been correlated with salivary changes in dental patients in two recent studies. In two subsequent studies, test anxiety-induced stress and relaxation-induced anxiety reduction were correlated with salivary changes in dental students. In another study using the resazurin dye indicator, increased salivary bacterial levels were correlated with an increased dental caries incidence. As a result of these findings, it was decided to reinvestigate the effects of stress and relaxation on salivary changes and in addition to examine the effects of those conditions on salivary bacteria. The hypotheses under consideration were: (1) Salivary changes from stress to relaxation will be from opaque to translucent and from high to low protein levels; and (2) salivary bacteria will increase under the condition of stress and decrease under the condition of relaxation. The subjects were twelve dental students. Stress and relaxation were evaluated before and after meditation by verbal reports and examination of saliva for opacity, translucency, protein and bacteria (resazurin dye method). There were significant anxiety-reduction changes by the end of the meditation sessions (p < 0.001) as measured by increased salivary translucency, decreased salivary protein and reduced subjective evaluation of stress. Using the resazurin dye method, bacterial levels showed a significant decrease by the end of the meditation sessions (p < 0.001). The results support hypothesis 1 and reaffirm previous findings in regard to the effectiveness of: (1) salivary changes as measures of stress and relaxation; and (2) meditation to induce deep relaxation. The finding of high bacteria levels under stress and lower bacterial levels under relaxation supports hypothesis 2 and indicates that stress may contribute to dental caries and relaxation may have an anti-caries effect.     

Salivary prekallikrein output during the ranger training‐induced stress

Saliva can be considered a mirror of the body, reflecting its general conditions. The present study was conducted to identify the changes in specific neurotransmitter and prekallikrein levels in saliva and the correlations between changes in neuroendocrine factors in plasma and saliva after intensive physical training of rangers of Japan Ground Self Defence Force (JGSDF). The subjects were 14 young uniformed males (aged 20–26 years old) who underwent ranger training under the control of the 4th Division of the JGSDF. Before training began and after 30 days of hard training, the levels of prekallikrein, β‐endorphin, serotonin, noradrenaline and dopamine in saliva and plasma were routinely measured. The level of prekallikrein in saliva was significantly increased after training, and the level of β‐endorphin in saliva showed an upward trend after the training. The level of prekallikrein in plasma showed a significant decrease after the training, while the level of β‐endorphin was unchanged. Average concentrations of values in serotonin, noradrenaline and dopamine in plasma were significantly increased after the training. These findings suggest that the comparison of levels of prekallikrein and β‐endorphin in saliva and plasma may give useful insights for the estimation of stress‐induced analgesia. Copyright © 2004 John Wiley & Sons, Ltd.     

Persistence of free HBV DNA in body secretions and liver despite loss of serum HBV DNA after interferon-induced seroconversion

Following interferon therapy, a chronic hepatitis B (HBV) carrier lost all serum markers of active viral replication and became anti-HBe positive but remained positive for free and replicative HBV-DNA in semen, saliva, urine, and liver four months later. At 12 months, when he also developed anti-HBs, urine and saliva analysed for free HBV-DNA were positive. Despite histological remission and loss of HBV-DNA from serum, the potential for transmission of HBV and reactivation of disease remain.