Immunoglobulin A In Saliva Research Articles

Upper Respiratory Tract Infection and Saliva Immunoglobulin-A During an Arduous Military Training Programme

Upper Respiratory Tract Infection and Saliva Immunoglobulin-A During an Arduous Military Training Programme

Upper Respiratory Tract Infection and Saliva Immunoglobulin-A During an Arduous Military Training Programme

Upper Respiratory Tract Infection and Saliva Immunoglobulin-A During an Arduous Military Training Programme

Oral manifestations in selective IgA deficiency

Selective immunoglobulin A (IgA) deficiency is the most common of the primary immunodeficiencies with a frequency of 1/300-1/3000, depending on the screened population. As secretory IgA (SIgA) has a protective role in mucosal surfaces from invasion of microorganisms, it is thought that IgA-deficient subjects are susceptible to periodontal diseases and oral manifestations. Previous studies show contradictory results, concerning the involvement of the individuals' periodontium with IgA deficiency. The aim of this study was to investigate and compare the oral manifestations in IgA-deficient subjects with controls. Eleven selective IgA-deficient subjects aged 3-18 years with serum IgA levels <10 mg dl(-1) and 11 age-sex-matched healthy children as the controls entered the study. Oral mucosal investigation, dental caries, plaque accumulation and periodontal status were assessed. Serum immunoglobulin levels were measured by single radial immunodiffusion (SRID) method. Saliva immunoglobulins and secretory component levels were measured by enzyme linked immunosorbent assay (ELISA) methods. IgA-deficient patients had serum and saliva IgA levels less than 10 mg dl(-1) and 10 microg ml(-1), respectively, but other serum immunoglobulin levels were normal and saliva immunoglobulin M (IgM) levels were increased, compared with controls. There were no significant differences in oral manifestations between IgA-deficient subjects and controls, which may be a result of compensatory increase of saliva IgM or other non-immunological defence factors in saliva. Thus, it is not necessary to evaluate IgA and SIgA in all the patients with oral and dental lesions and it is thought that it is better to investigate other factors.

Effect of age on immunoglobulin A subclass distribution in human parotid saliva.

Two subclasses of immunoglobulin A (IgA) antibodies are produced in humans, IgA1 and IgA2, IgA2 being more resistant to digestion by bacterial proteases than IgA1. The amount of IgA in saliva has been shown to vary with age; however, little is known about the correlation between IgA subclass distribution in saliva and age. The purpose of this study was to determine whether differences exist in the levels and ratio of IgA subclasses in parotid saliva of children and adults. Parotid saliva was obtained from healthy children (age range 6-12 years, n = 14) and adults (age range 22-51 years, n = 20) using Schaefer cups. Samples were analyzed for levels of total IgA, IgA1, and IgA2 by ELISA. IgA and IgA1 levels were significantly higher in adults than in children. However, no differences were seen in the ratio of IgA1 and IgA2 in the two groups of subjects. These findings indicate that levels of IgA increase with age, whereas the IgA subclass ratio is established early in life.

Human salivary immunoglobulin and antigen-specific antibody activity after tonsillectomy.

The importance of the lymphoid tissue collectively known as Waldeyer's ring, which includes the palatine, lingual and nasopharyngeal tonsils, in the induction and contribution of specific antibody responses in human saliva is not clear. The purpose of this study was to determine whether salivary immunoglobulin A (IgA) levels differ in quantity and quality between subjects who have had a tonsillectomy and age, sex and race-matched controls. Parotid saliva, whole saliva, and blood serum samples were collected from 25 volunteer children who had undergone tonsillectomy (T-) within 6-14 months of sampling and from 25 age, sex and race-matched controls. The levels of total IgA (and subclasses) in saliva, and of antigen-specific salivary IgA and serum IgA and IgG antibodies to 4-9 relevant antigens were analyzed by ELISA. No significant difference was observed in the mean total IgA and IgA subclass levels in parotid and whole saliva, although the mean levels for children with a T- were slightly lower. Children with a T- had significantly higher parotid salivary IgA and IgA1 specific/total activity than controls. The total and specific whole saliva IgA and the specific serum IgA or IgG activities were not significantly different from controls. These results indicate an association between the removal of tonsils and increased levels of specific IgA activity in parotid saliva within the first year after a T-.

Adjuvant activity of monophosphoryl lipid A for nasal and oral immunization with soluble or liposome-associated antigen.

The effectiveness of monophosphoryl lipid A (MPL) as a mucosal adjuvant was investigated following oral or intranasal (i.n.) administration of an aqueous adjuvant formulation of MPL (MPL-AF) added to soluble antigen or liposomal antigen or incorporated into liposomal antigen membranes. Groups of BALB/c female mice were immunized with 50 to 100 microg of free or liposomal Streptococcus mutans crude glucosyltransferase (C-GTF) with or without MPL-AF added to the vaccine or incorporated into the liposomal membrane. Plasma, saliva, vaginal wash, and fecal extract samples were collected biweekly following immunization and assessed for antigen-specific antibody activity by enzyme-linked immunosorbent assay (ELISA). Mice immunized by the i.n. route had higher levels of salivary, plasma, and vaginal immunoglobulin A (IgA) anti-C-GTF responses and higher levels of plasma IgG anti-C-GTF than the orally immunized groups. A second administration of the vaccine 14 weeks after the initial immunization resulted in an anamnestic response to C-GTF resulting in 10- and 100-fold increases in saliva and plasma IgA and plasma IgG, respectively (in the i.n. immunized groups). Mice receiving a second i.n. immunization with liposomal antigen and MPL-AF had higher salivary IgA anti-C-GTF responses than mice immunized with antigen plus MPL-AF or liposomal antigen (P < 0.05). Plasma IgG anti-C-GTF activity was highest in mice immunized by the i.n. route with antigen formulations containing MPL-AF (P < 0.05). These results demonstrate the effectiveness of MPL-AF as an adjuvant for potentiating mucosal and systemic immune responses to liposomal C-GTF following i.n. immunization.

Correlations between antibody immune responses at different mucosal effector sites are controlled by antigen type and dosage.

Monitoring specific secretory immunoglobulin A (IgA) responses in the intestines after mucosal immunization or infection is impeded by the fact that sampling of small intestinal secretions requires invasive methods not feasible for routine diagnostics. Since IgA plasma cells generated after intragastric immunization are known to populate remote mucosal sites as well, secretory IgA responses at other mucosal surfaces may correlate to those in the intestines and could serve as proxy measures for IgA secretion in the gut. To evaluate the practicability of this approach, mice were immunized intragastrically with 0.2, 2, and 20 mg of ovalbumin plus 10 microg of cholera toxin, and the antigen-specific local secretory IgA responses in duodenal, ileal, jejunal, rectal, and vaginal secretions, saliva, urine, and feces, as well as serum IgG and IgA responses were analyzed by enzyme-linked immunosorbent assay. Correlation analysis revealed significant relationships between serum IgG and IgA, urinary IgA, salivary IgA, and secretory IgA in duodenal, jejunal, ileal, and rectal secretions for the 0.2-mg but not for the 20-mg ovalbumin dose. Fecal samples were poor predictors for intestinal antiovalbumin IgA responses, and no correlations could be established for cholera toxin, neither between local anti-cholera toxin levels nor to the antiovalbumin responses. Thus, specific IgA in serum, saliva, or urine can serve as a predictor of the release of specific IgA at intestinal surfaces after intragastric immunization, but the lack of correlations for high ovalbumin doses and for cholera toxin indicates a strong dependency on antigen type and dosage for these relationships.

Protective immunity to rotavirus shedding in the absence of interleukin-6: Th1 cells and immunoglobulin A develop normally.

We investigated whether interleukin-6 (IL-6) was required for the development of immunoglobulin A (IgA)- and T-helper 1 (Th1)-associated protective immune responses to rotavirus by using adult IL-6-deficient mice [BALB/c and (C57BL/6 x O1a)F(2) backgrounds]. Naive IL-6(-) mice had normal frequencies of IgA plasma cells in the gastrointestinal tract. Consistent with this, total levels of IgA in fecal extracts, saliva, and sera were unaltered. In specific response to oral infection with rhesus rotavirus, IL-6(-) and IL-6(+) mice exhibited efficient Th1-type gamma interferon responses in Peyer's patches with high levels of serum IgG2a and intestinal IgA. Although there was an increase in Th2-type IL-4 in CD4(+) T cells from IL-6(-) mice following restimulation with rotavirus antigen in the presence of irradiated antigen-presenting cells, unfractionated Peyer's patch cells failed to produce a significant increase in IL-4. Moreover, virus-specific IgG1 in serum was not significantly increased in IL-6(-) mice in comparison with IL-6(+) mice. Following oral inoculation with murine rotavirus, IL-6(-) and IL-6(+) mice mediated clearance of rotavirus and mounted a strong IgA response. When IL-6(-) and IL-6(+) mice [(C57BL/6 x O1a)F(2) background] were orally inoculated with rhesus rotavirus and later challenged with murine rotavirus, all of the mice maintained high levels of IgA in feces and were protected against reinfection. Thus, IL-6 failed to provide unique functions in the development of IgA-secreting B cells and in the establishment of Th1-associated protective immunity against rotavirus infection in adult mice.

Targeted salivary gland immunization with plasmid DNA elicits specific salivary immunoglobulin A and G antibodies and serum immunoglobulin G antibodies in mice.

For the development of vaccines against oral and pharyngeal pathogens invading the mucosal epithelia, both secretory and serum immunoglobulin A (IgA) and IgG antibodies and cytotoxic T lymphocytes (CTL) have been induced. We used a novel approach, targeted salivary gland (TSG) immunization, using plasmid pcDNA3/fimA, coding for Porphyromonas gingivalis fimbriae. Expression of subunit protein, fimbrillin, was observed in eukaryotic cells growing in vitro following transfection with pcDNA3/fimA. In this study, we obtained good humoral and cell-mediated immune responses in BALB/c mice by TSG administration using the above-mentioned DNA vaccine. The production of fimbria-specific IgA and IgG antibodies in saliva and serum IgG antibody was significantly stimulated by TSG immunization. Injection of DNA vaccine into salivary gland elicited high-level production of antigen-specific IgG antibody, similar to that induced following intramuscular immunization. The major IgG subclass that recognized fimbriae was IgG2a in serum from pcDNA3/fimA-immunized mice. Reverse transcription-PCR analysis of mononuclear cells from salivary glands showed that levels of Th2 cytokine-specific mRNA were higher in the immunized group than in the nonimmunized group. In addition, TSG DNA immunization resulted in the generation of antigen-specific CTL in spleen. These results indicate that TSG immunization with plasmid DNA may represent a genetic immunization strategy against infection by oral and pharyngeal pathogens that may invade local, mucosal surfaces.

Simultaneous induction of specific immunoglobulin A--producing cells in major and minor salivary glands after tonsillar application of antigen in rabbits.

The immunoglobulin A (IgA)-producing cells in the stroma of major salivary glands are induced by antigenic stimulation of the mucosal immune system. Whether such cells also are induced in minor salivary glands by this stimulation remains to be determined. After application of sheep red blood cells to the palatine tonsils every 3 days for 6 weeks, anti-sheep red blood cell IgA was detected in saliva both by agglutination tests and by enzyme-linked immunosorbent assay. Using enzyme-linked immunospot assay, an increase in the number of anti-sheep red blood cell IgA-producing cells was found in minor as well as in major salivary glands of the sixth week of application; such cells constituted 4.9% to 5.9% of the total number of IgA-producing cells in these tissues. Tonsillar application of whole cells of formalin-killed Streptococcus sobrinus induced anti-S. sobrinus IgA in saliva. The number of anti-S. sobrinus IgA-producing cells in the above glands simultaneously increased over 6 weeks, and reached 5.2-5.6% of the total number of IgA-producing cells.

Early appearance of antibodies to simian immunodeficiency virus in saliva and serum of infected macaques.

Simian immunodeficiency virus (SIV) infection in macaques is an important animal model for human immunodeficiency virus infection in humans. This study evaluated the temporal development of antibodies to SIV in the parotid saliva of macaques inoculated with the virus and compared these findings with the development of antibodies to SIV in the animals' sera. Three animals (ages, 14, 18, and 18 years) were inoculated with the macrophagetropic strain SIVmac239. Prior to inoculation and at consecutive weekly intervals during a four-week period following the initial virus inoculations, parotid saliva and serum were collected from each animal. A fourth animal (age, 9 years) served as a negative control, and the fifth and sixth animals (ages, 2 and 22 years) served as positive controls (6 and 18 months postinoculation, respectively) with SIVmac239. Saliva and serum samples were reacted against SIV antigen in Western blots (immunoblots) prepared in the standard fashion to determine the presence of antibodies. The reactions of these antigen-antibody complexes with biotinylated anti-human immunoglobulin A (IgA), IgM, and IgG and biotinylated anti-human secretory component (SC) determined the class of antibody present or the presence of SC in the original parotid saliva or serum samples. In infected animals, the IgM to SIV was detectable in serum and saliva at 13 days, and antiviral IgA and IgG in serum and saliva were detectable at 20 to 27 days postinoculation. The antibody to SC reacted to saliva from only two animals at 20 and 27 days, and long-term positive controls were positive for SC in saliva, indicating that either secretory IgA or secretory IgM was present in these samples. Antibodies to SIVmac239 antigens have therefore been detected in saliva as early as 13 days postinfection. Saliva may be as useful as serum as a diagnostic specimen and/or disease-monitoring method in this important animal model.

Intrafamilial similarity in immunoblot profiles of salivary immunoglobulin A antibody activity to oral streptococci

Analysis of the salivary immunoglobulin A (IgA) antibody activity to antigens separated from oral streptococci was carried out on 49 individuals in 11 families. The aim of this study were to i) study the human salivary IgA activity within families to antigens separated from reference strains of Streptococcus mutans, Streptococcus sobrinus and Streptococcus parasanguis and ii) to search for potential differences in the salivary IgA activity to the streptococcal antigens several years apart. The immunoblot revealed similarity in saliva IgA response within the families. A majority of the bands, approximately 7 (median) for each extract was found in both child and parent. A few bands (approximately 3) were found in parents but not in children. Only one to two bands were found in children's saliva without a corresponding band in any of the parents. The antibody activity of saliva samples obtained several years apart was essentially unchanged, especially considering the total number of bands. Differences in the relative intensity of the bands could be seen, and a few bands appeared only in some immunoblots for a specific subject. For 3 of 4 children participating in the longitudinal study, more bands against mutans streptococci appeared with age.

Immunoglobulin A antibodies to mutans streptococci in human saliva and serum comparing fresh and subcultivated strains and activity in repeated saliva samples

The aims of this study were i) to characterize and compare the sodium dodecyl sulfate-polyacrylamide gel electrophoresis protein patterns of reference Streptococcus mutans and Streptococcus sobrinus strains, subcultivated for years, with fresh isolates of the same serotype; ii) to study possible differences between the human salivary immunoglobulin A (IgA) activity to reference strains and to fresh bacterial isolates of the saliva donors; iii) to examine for potential differences in the salivary IgA activity to the streptococcal antigens during 1 week; and iv) to map, in the same individuals, the serum IgA activity against the selected bacteria. S. mutans reference and fresh isolated strains showed a similar protein pattern with few exceptions. The immunoblot also revealed similarity in saliva IgA response, with only one subject's saliva displaying clearly one band's difference. For S. sobrinus a larger discrepancy was seen. The antibody activity during the one week interval was essentially unchanged. When incubated with serum, a different immunoblot profile was seen compared with saliva, although most bands revealed by saliva were also displayed by serum.

Induction of specific immunoglobulin A in the small intestine, colon-rectum, and vagina measured by a new method for collection of secretions from local mucosal surfaces.

In order study patterns of local antibody responses following mucosal immunization of mice via different routes, a method for collection of secretions directly from mucosal surfaces was developed. Mice were immunized on days 0, 10, 17, and 24 by administration of cholera toxin into the oral cavity, stomach, colon-rectum, or vagina. At sacrifice on day 32, absorbent wicks were placed in the oral cavity and, via an applicator tube, into the vagina and distal colon-rectum and along the entire small intestine after flushing of luminal contents. Protein was quantitatively extracted from wicks, and specific anti-cholera toxin immunoglobulin A (IgA) and IgG were measured by enzyme-linked immunosorbent assay. Concentrations of specific IgA in secretions at various mucosal sites were dramatically influenced by the route of immunization. Oral immunization effectively induced IgA in saliva, and the intragastric route was optimal for induction of IgA in the small intestine. High levels of specific IgA appeared on the colonic-rectal mucosal surface only after rectal delivery of antigen. Oral, gastric, and rectal immunizations also produced distant responses in the vagina. Following vaginal immunization, however, neither local nor distant IgA responses were detected. These results suggest that vaccines intended for protection of colonic-rectal and vaginal mucosal surfaces might best be administered by the rectal route.

Secretory immunoglobulin A in childhood: does the saliva value reflect the bronchial value?

Immunoglobulin A (IgA) can be found in different body secretions and plays a major role in the local immune response. It inhibits bacterial adherence, neutralizes toxins and protects the mucosa from penetrating antigens and allergens. Whereas measurement of IgA in saliva mostly does not show any problems, difficulties in assessing IgA in bronchial fluids often occur mainly due to variable dilutional effects. Aim of the present study was to find out whether saliva IgA predicts bronchial IgA. In 15 children aged 4 months to 14 years (mean 53.5 months; SEM 12.3) with chronic cough (n = 10), mediastinal mass (n = 1), recurrent airways obstructions (n = 2) and inspiratory stridor (n = 2) we performed a diagnostic rigid bronchoscopy and assessed IgA by means of bronchial lavage (BL). We attempted to control for uncertain dilution by the use of albumin as a denominator and to present our data as ratios of IgA to albumin. As various disease states alter the integrity of the alveolar-capillary membrane and influence the concentration of albumin in the epithelial lining fluid we developed an optical score to describe the state of the bronchial mucosa. Measurement of saliva IgA is easy and can be done without dilutional effects. The mean value of IgA in saliva was 65.49 mg/l (SEM 14.75; range 3.5-227), the one of IgA in bronchial lavage fluid 30.75 mg/l (SEM 7.11; range 3.5-100). IgA-albumin ratio ranged from 0.006 to 1.46 (mean 0.36, SEM 0.12). Saliva IgA did neither significantly correlate with bronchial IgA nor with bronchial IgA-albumin ratio.(ABSTRACT TRUNCATED AT 250 WORDS)

Salivary Immunoglobulin A Production in Chronic Bronchitis Patients Given an Orally Administered Bacterial Extract

The concentration of secretory immunoglobulin A (IgA) has been measured by the immunodiffusion method in 28 chronic bronchitis patients (group A) and in 11 comparable patients receiving corticosteroid therapy (0.5 mg/kg/day prednisone; group B). The measurements have been taken before and after oral administration of Broncho-Vaxom which is an extract of bacteria that usually cause infection of the respiratory tract. The mean concentration of secretory IgA in the saliva was increased by over 130% after the 10-day administration of the preparation to group A patients. This increase was statistically significant between the 12th and 40th day after the beginning of the treatment (p < 0.001). It fell to initial levels after 1 month in 15 patients (group A2) who received a single treatment course. In 13 patients (group A1) who received a second treatment course, beginning 1 month after termination of the first course, the high concentration of IgA in saliva persisted for at least 3 months. The saliva IgA was increased by over 35% in 11 chronic bronchitis patients (group B) who received corticosteroid therapy. In this group of patients the rise of IgA levels began later than in group A. The IgA fell to initial level after 1 month in 5 patients (group B2) who received a single treatment course. In 6 patients (group B1) who received a second treatment course the high concentration persisted longer. The saliva level of IgA in groups A and A1 was higher than in groups B and B1 (p < 0.05 and p < 0.001, respectively). No differences of IgA in the saliva between groups A2 and B2 were found during the study.

Basketball exercise and secretory immunoglobulin A

This study examined saliva levels of immunoglobulin A (IgA) before and after three games and three practice sessions during the basketball season. Saliva was collected from 27 prepubescent boys (10-12 years) in a small Fry league and 23 postpubescent boys (16-18 years) on a high school varsity team. Saliva samples were frozen for later assay using a standard enzyme-linked immunosorbent assay technique. IgA levels were significantly increased after games 1 and 3 in both age groups and after practice 3 in the high school athletes. Over the 2 months of saliva collections the pre-exercise IgA increased significantly with games 2 and 3 higher than game 1, and practice 3 higher than practices 1 and 2, in both age groups. These results indicate that basketball exercise can increase saliva IgA levels and that chronic exercise over the basketball season may increase the resting levels of IgA. These changes may give athletes more protection against respiratory infections both after exercise and in the resting state later in the season.

Glucocorticoid Regulation of the Humoral Immune System. Dexamethasone Stimulation of Secretory Component in Serum, Saliva, and Bile*

Dexamethasone, a potent synthetic glucocorticoid, has been shown to increase polymeric immunoglobulin A (IgA) and antigen-specific IgA antibody levels in serum. This response coincided with a decline in IgA and IgG levels in saliva and vaginal secretions after dexamethasone treatment. To investigate the mechanism(s) of glucocorticoid action responsible for the accumulation of polymeric IgA in serum, we undertook in the present study to determine whether dexamethasone alters the production of secretory component (SC). SC is the receptor responsible for transporting IgA from blood and tissues into bile and external secretions. Dexamethasone administered in vivo increased SC levels in serum in a dose- and time-dependent manner. When bile and saliva samples were analyzed, dexamethasone treatment increased saliva SC levels and the production rate of SC by the liver, but had a pronounced inhibitory effect on both bile and salivary levels of IgA. As judged by HPLC, the majority of SC in blood was associated with polymeric IgA and not with monomeric IgA. Immunoblots showed the presence of a 29/27K doublet band of SC in serum from control- and dexamethasone-stimulated rats. These findings indicate that the elevation of polymeric IgA levels in serum after dexamethasone treatment may be due to decreased clearance of IgA into bile. Further, it demonstrates that liver production rate of SC as well as saliva and serum SC levels increase with glucocorticoid treatment. This findings suggests that glucocorticoids, which are known to stimulate hepatocyte SC synthesis in vitro, increase SC levels in blood, possibly as SC fragments that are capable of binding polymeric IgA and retarding its clearance from blood to bile.

Reduction of saliva immunoglobulin levels by swim training

Saliva immunoglobulin A (IgA) and cortisol levels were measured in 21 male members of a major midwestern swim team. Saliva samples were collected before and after training sessions four times during the fall season; the training intensity was light, moderate, heavy and during the taper period before a major competitive meet. Saliva IgA levels were decreased after each training session, reaching statistical significance with the moderate training intensity. Over the 3-month training period the pre-session and post-session IgA levels both decreased significantly during the heavy and taper training intensities later in the fall season. Cortisol levels were significantly elevated only after the heavy-intensity training session. The Profile of Mood States (POMS) was used to assess the swimmers' overall mood on each test day. No significant correlations were found between the global POMS score and IgA or cortisol. Also, cortisol and IgA were not significantly correlated except after the light training session. Results from this study indicate that acute bouts of exercise can reduce salivary IgA levels and that chronic exercise of high intensity can reduce the resting levels of IgA. These changes may render the athletes more vulnerable to respiratory infections after exercise and even at rest during the later stages of the competitive season.

Degradation of human secretory immunoglobulin a by protease isolated from the anaerobic periodontopathogenic bacterium, Bacteroides gingivalis

This bacterium is implicated in periodontal diseases of human adult type. Secretory immunoglobulin A (IgA) purified from human colostrum (HC-IgA) was incubated with Bacteroides gingivalis cells or protease isolated from the culture supernatant of B. gingivalis; the digestion of IgA was determined by immunoelectrophoresis and sodium dodecyl sulphate-polyacrylamide gel electrophoresis. B. gingivalis cells almost completely degraded HC-IgA; protease isolated from the culture supernatant cleaved both HC-IgA and secretory IgA in human parotid saliva. Thus by degradation of IgA, the protease may mediate in part the periodontopathogenic role of B. gingivalis by decreasing the oral defence mechanism.