[Diagnostic values of anti-salivary gland protein-1 antibody combined with anti-parotid secretory protein antibody for Sjögren's syndrome].

OBJECTIVE: To assess the diagnostic value of anti-salivary gland protein-1 (SP1) antibody combined with anti-parotid secretory protein (PSP) antibody for Sjögren's syndrome (SS). METHODS: A total of 60 patients with primary SS (pSS) who were treated in the outpatient and inpatient department of Department of Rheumatology and Immunology of the Second Hospital of Hebei Medical University from January 2020 to December 2022 were collected. Thirty patients with other autoimmune diseases accompanied by dry mouth and/or dry eyes were collected as disease control group. Thirty healthy subjects from the physical examination center were collected for healthy control group, serum samples were obtained from all of them. Their general features and clinical information including clinical manifestations, laboratory examinations and other examinations were recorded. The 2016 American College of Rheumatology (ACR)/European League against Rheumatism (EULAR) classification criteria were adopted as the diagnostic standard of pSS. Immunoglobulin G (IgG) subtype of anti-SP1 antibody and anti-PSP antibody were detected by chemiluminescence immunoassay. The receiver operating characteristic (ROC) curve was used to evaluate the accuracy of anti-SP1 antibody and anti-PSP antibody in diagnosing pSS.The cli-nical characteristics of anti-SP1 antibody and anti-PSP antibody positive patients and negative patients in pSS group were further compared. Independent samples t test, Mann-Whitney U test, variance analysis, Kruskal-Wallis test, Chi-square test or Fisher's exact test and Spearman correlation analysis were used for statistical analysis. RESULTS: There was no significant difference in age (F=1.406, P=0.495) and gender (χ2=2.105, P=0.349) among pSS group, disease control group and healthy control group. The expression levels of anti-SP1 antibody (H=16.73, P < 0.001) and anti-PSP antibody (H=26.09, P < 0.001) were statistically different among the three groups. An intergroup comparison of anti-SP1 antibody expression levels showed that there was a statistically significant difference between pSS and healthy control group (P < 0.001), but no statistically significant difference between the other groups. Comparison of anti-PSP antibody expression levels between the groups showed that there were statistically significant differences between pSS and healthy control group (P < 0.001), and between disease control group and healthy control group (P=0.009), while no statistically significant differences between the other groups. The positive rate of anti-SP1 antibody in pSS group was significantly higher than that in disease control group and healthy control group (58.33% vs. 40.00% vs. 13.33%, P < 0.001). The positive rate of anti-PSP antibody in pSS group was significantly higher than that in disease control group and healthy control group (75.00% vs. 56.17% vs. 16.67%, P < 0.001). The area under the curve for anti-SP1 antibody was 0.688 (P < 0.001). The sensitivity and specificity of anti-SP1 antibody were 58.33% (35/60) and 70.00% (42/60) respectively, the positive predictive value was 66.04% (35/53) and the negative predictive value was 54.55% (42/77) of anti-SP1 antibody.The area under the curve of anti-PSP antibody was 0.720 (P < 0.001), with a sensitivity was 75.00% (45/60), and specificity was 63.33% (38/60).The positive predictive value and negative predictive value of anti-PSP antibody were 67.16% (45/67) and 71.70% (38/53) respectively. All the 13 pSS patients were negative for anti-Sjögren's syndrome A (SSA, including SSA52 and SSA60) antibody and anti- Sjögren's syndrome B (SSB) antibody. Among them, 11 patients were positive for both anti-SP1 antibody and anti-PSP antibody, 1 patient was positive for anti-SP1 antibody and 1 patient was positive for anti-PSP antibody. The clinical features of anti-SP1 antibody and anti-PSP antibody positive and negative groups were compared in pSS patients. The duration of disease in anti-SP1 antibody positive group was shorter (Z=-2.277, P=0.023) when compared with the negative patients. The patients with positive anti-PSP antibody were younger than those in the negative group (t=2.598, P < 0.05), the positive rate of rheumatoid factor (P=0.002) and the serum level of IgG (t=3.806, P=0.003) in anti-PSP antibody positive group were higher than in the negative group. Analysis of the correlation between anti-SP1 antibody and anti-PSP antibody in the pSS patients showed that there was significant correlation between them (r=0.801, P < 0.001). CONCLUSION: Both anti-SP1 antibody and anti-PSP antibody are valuable in the diagnosis of SS, and anti-SP1 antibody is helpful for the early diagnosis of pSS. The combined detection of anti-SP1 antibody and anti-PSP antibody is helpful for the early diagnosis of pSS patients with negative anti-SSA antibody and anti-SSB antibody.

Passive transfer of antibodies to the linear epitope 60 kD Ro 273-289 induces features of Sjögren's syndrome in naive mice.

Sjögren's syndrome (SS) is an autoimmune inflammatory disease that primarily affects the lacrimal and salivary glands causing dry eyes and mouth. Antibodies to Ro60 are observed frequently in patients with SS; however, the role of these antibodies in SS initiation and progression remains unclear. The sequence Ro60 273-289 (Ro274) is a known B cell epitope of Ro60 and antibodies to this epitope have been observed in a subset of SS patients and in animals immunized with Ro60 protein. Animals immunized with Ro274 linear peptide develop a Sjögren's-like illness. We hypothesized that passive transfer of anti-Ro274-specific immunoglobulin (Ig)G would induce a Sjögren's-like phenotype. To evaluate this hypothesis, we adoptively transferred affinity-purified Ro274 antibodies into naive BALB/c animals, then evaluated salivary gland histology, function and IgG localization 4 days post-transfer. At this time-point, there was no demonstrable mononuclear cell infiltration and salivary glands were histologically normal, but we observed a functional deficit in stimulated salivary flow of animals receiving Ro274 antibodies compared to animals receiving control IgG. Cellular fractionation and enzyme-linked immunosorbent assay revealed Ro274-specific antibodies in the nucleus and cytoplasmic fractions of isolated parotid salivary gland cells that was confirmed by immunohistochemistry. These data support the hypothesis that antibodies to Ro274 deposit in salivary glands can enter intact salivary gland cells and are involved in the dysregulation of salivary flow in SS.

[STUDY OF SAFETY OF PAROTITIS VACCINE].

AIM: Monitoring of post-vaccinal complications in children immunized with a parotitis vaccine. MATERIALS AND METHODS: Observation of 198 945 children, immunized with 16 lots of parotitis vaccine with Leningrad-3 strain (L-3), was carried out for 3 years. Paired samples of sera and saliva were obtained from children, in whom adverse events were registered for 42 days after vaccination. Titers of specific IgM and IgG were determined in blood sera. Analysis of nucleotide sequences of genes F, SH and NH of RNA of parotitis virus was carried out from samples of blood and saliva. RESULTS: Intensive parameter of vaccine-associated aseptic meningitis under the conditions of the experiments was 0 for 100 000 immunized. Frequency of occurrence of post-vaccinal parotitis was 0.06% from the number of vaccinated--18 cases of vaccine-associated parotitis were registered and laboratory confirmed. A significant difference in specific activity was detected for 3 lots of the vaccine, that were associated with cases of development of parotitis, relative to that of 13 lots of vaccine, development of parotitis was not registered after administration of those. CONCLUSION: The study carried out confirmed low neurovirulence of the parotitis vaccine with the L-3 strain of parotitis virus, as well as a low degree of its reactogenicity. A relatively high immunization dose of the used vaccine could be one of the reasons of development of post-vaccinal complications in part of the immunized children.

Immunoglobulin M: local synthesis and selective secretion in patients with immunoglobulin A deficiency.

Local synthesis seems to be decisive for the selective secretion of 19S immunoglobulin M into parotid secretions. The "transport piece" is apparently not involved in the secretion of immunoglobulin M, for there is no association between the two components. The possible significance of the normal association of transport piece with secreted immunoglobulin A remains to be clarified.

Human parotid saliva contains soluble toll-like receptor (TLR) 2 and modulates TLR2-mediated interleukin-8 production by monocytic cells.

Toll-like receptor (TLR) family members are pattern-recognition receptors and very important molecules in innate immunity. Although TLRs are originally type I transmembrane receptors, soluble forms of TLRs are detected in human plasma and milk. This study showed that soluble TLR2 (sTLR2) is detected in human parotid saliva. Western blotting with anti-TLR2 antibodies (Abs) showed that three polypeptides are detected as sTLR2 with molecular weights of 55, 40 and 27kDa, respectively. Parotid saliva neutralized the binding of anti-TLR2 polyclonal Ab to cell-surface TLR2 on THP-1, a human monocytic cell line. Immunohistochemical analysis revealed that TLR2 is expressed in serous and interlobular ductal cells of human salivary gland. Human salivary gland cell lines, AZA3 and HSY, constitutively expressed TLR2. Parotid saliva augmented IL-8 production of THP-1 cells stimulated with a synthetic TLR2 ligand, Pam(3)Cys-Ser-(Lys)(4) (Pam(3)CSK(4)). Depletion of sCD14 from parotid saliva by immunoprecipitation eliminated the augmentation of IL-8 production, indicating that the augmentable effects depended on sCD14 in parotid saliva. On the other hand, preincubation of Pam(3)CSK(4) with parotid saliva abrogated the augmentation of IL-8 production, indicating that sTLR2 in saliva bound to Pam(3)CSK(4) and neutralized its function. These results suggest that parotid saliva modulates the TLR2-mediated immune responses with binary mechanisms via sTLR2 and sCD14 in the oral cavity.

Age-dependent deficiency in saliva and salivary antibodies secretion in Down's syndrome.

OBJECTIVE: Down's syndrome (DS) individuals suffer from an increased susceptibility to infections. Here, we assessed age-related changes in the salivary-specific humoral immunity of DS subjects. DESIGN: Parotid and whole saliva were collected from a young group of DS (YDS, n=30, 23.3+/-4 years), an older group of DS individuals (ODS, n=10, 51.9+/-8 years) and compared to two age-matched groups of healthy volunteers--a young group (YC, n=29, 22.8+/-5 years) and an older group (OC, n=10, 48.4+/-9 years). The levels of total IgA, and specific antibodies to three common oral pathogens (Porphyromonas gingivalis, Actinobacillus (Aggregatibacter) actinomycetemcomitans and Streptococcus mutans) were analysed. RESULTS: The limited increases in IgA concentrations could not compensate the dramatic reduction in the salivary flow rate observed in DS individuals. Therefore, the median secretion rates of the specific antibodies in whole and parotid saliva were 70-77% and 34-60% (respectively) lower in YDS individuals as compared to YC and farther 77-100% and 75-88% (respectively) lower in ODS compared to YDS. In contrast, the antibody secretion rates were similar for parotid saliva, or even increased for whole saliva of OC, compared with YC. Consequently, a dramatic cumulative extreme reduction (>92%) in the bacterial specific salivary antibodies differentiated the adult DS individuals from to their age-matched controls. CONCLUSIONS: Our results indicate a severe immunodeficiency in the secretion rate of the specific salivary IgA response of in DS individuals which intensifies with age.

Nonspecific oral immunity in individuals with HIV infection.

Lactoferrin, lysozyme, interferon, and neopterin levels were determined in parotid saliva from 44 individuals with different clinical stages of human immunodeficiency virus (HIV) infection and 19 HIV-seronegative controls. The secretory output of individual components was calculated according to the fluid flow rate. No parotid interferon activity was found in any of the HIV-infected subjects or controls, and no significant differences in parotid lysozyme or neopterin outputs were observed. The lactoferrin output was significantly decreased in HIV-seropositive subjects in parallel with their markedly reduced parotid secretory IgA output. This combined deficiency of parotid lactoferrin and secretory IgA may well contribute to the frequent oral infections seen in subjects with HIV infection.

Immune-reactive cells in multiple sclerosis mucosal secretions.

External secretions from multiple sclerosis (MS) patients show immunoglobulin abnormalities consistent with mucosal inflammation. In this study we collected tears and parotid saliva from ten normal subjects and ten MS patients to examine for free-floating inflammatory cells. We found lymphocytes, macrophages, and plasma cells at low numbers in normal secretions, but at much higher numbers in MS secretions. Using an immunobead rosette technique, most of the lymphocytes were null cells. However, we found increased T lymphocytes in the secretions of clinically active MS patients. The extensive mucosal surfaces of the MS patient could provide a peripheral source for activated lymphocytes that subsequently enter brain.

Radioimmunoassay of class-specific antibodies to Streptococcus mutans in monkey serum and saliva.

A radioimmunoassay (RIA) has been developed to measure class-specific antibodies to Steptococcus mutans in the serum and saliva of monkeys (Macaca fascicularis). Anti-human immunoglobulin antibodies purified by affinity chromatography on immobilised monkey immunoglobulins and labelled with 125I were employed. Formolised cells of S. mutans and an extract of culture supernatant adsorbed to polystyrene wells were used as solid-phase antigens. The coefficients of variation for IgG, IgA and IgM assays were less than or equal to 10% for both antigen systems. Two monkeys were immunised with formolised cells of S. mutans by subcutaneous injection and subsequent instillation of bacterial cells into their right parotid ducts. IgG, IgA and IgM antibody responses to S. mutans in samples of serum and saliva were quantitated by RIA. Immobilisation of purified components of S. mutans on polystyrene wells enabled the measurement of antibody response to a number of antigens to be made. The RIA is a sensitive, reproducible and quantitative method of measuring serum and salivary antibody responses in monkeys.

Improved method for post-embedding cytochemistry using reduced osmium and LR white resin.

We established an improved method for post-embedding cytochemistry by which highly specific cytochemical reactions on excellent cellular ultrastructures are possible. The method is a combination of post-fixation in potassium ferrocyanide-reduced OsO4 and embedment in acrylic-based LR White resin. It permits both immuno- and lectin-gold cytochemistry with fine ultrastructures comparable to those obtained by conventionally osmicated and epoxy-embedded tissues. Fixation with reduced osmium appeared to contribute to the preservation of immunoreactivity and membranous structures. By this method, the immunocytochemical localization of secretory proteins (amylase and chymotrypsinogen), actin filaments by polyclonal antibodies, 105 KD Golgi-associated protein by a monoclonal antibody (GF-1), and binding sites for gold-labeled lectin could be demonstrated. Also possible was multiple staining with enzyme cytochemistry of thiamine pyrophosphatase and immunocytochemistry of GF-1 and anti-amylase antibodies. This multiple staining made possible partial characterization of the trans-Golgi network in parotid acinar cells.

Tropism-restricted neutralization by secretory IgA from parotid saliva of HIV type 1-infected individuals.

In this study we examined secretory IgA, isolated from the parotid saliva of 10 HIV-1-infected subjects, for its ability to influence HIV-1 infection of peripheral blood mononuclear cells with two R5 and two X4 primary isolates. Salivary IgA from four subjects was found to inhibit both R5 viruses but not the X4 viruses. In another subject, salivary IgA inhibited both X4 viruses but not the R5 viruses. The specificity of these antibodies seemed to be directed against, but not restricted to, gp160 and gp120. Compared with subjects whose salivary IgA did not inhibit HIV-1 infection, subjects who displayed neutralizing activity were in relatively early stages of disease and had CD4(+) T cell counts greater than 200 cells/microl. Our data indicate the presence of tropism-specific (more frequently R5-specific) neutralizing antibodies in HIV-1-infected subjects. Because mucosal transmission of HIV-1 occurs exclusively in R5 viruses, and X4 viruses often emerge in established infection and account for viral persistence later in disease, our data suggest a potential role for secretory IgA in preventing viral transmission, but a less likely effect on chronic infection.

Variation of secretory antibodies in parotid saliva to human immunodeficiency virus type 1 with HIV-1 disease stage.

The secretory immune response to pathogens of the gut-associated lymphoid tissue is often independent of the systemic response. We investigated and compared the presence of antibodies to human immunodeficiency virus type 1 (HIV-1) antigens in parotid saliva and serum by Western blotting in 22 HIV-1-infected individuals. Antibodies to the HIV-1 envelope antigen gp160 were detected in saliva samples from 21 of 22 individuals and in serum from all individuals who were classified as CDC Group II, III, or IV. Antibody titers to gp160 were approximately 3000 times higher in serum than in saliva. Antibodies to viral core antigen p24 were detected in 6 of 7 Group II individuals in saliva and in 7 of 7 in serum. Antibodies to p24 were not found in the parotid saliva, but were detected in the sera of 3 of 3 Group III and 11 of 12 Group IV patients. The absence of secretory antibodies to HIV-1 core antigen p24 was correlated with CD4+ cell counts of less than 200/mm3. The results suggest that loss of secretory anti-p24 antibodies may be an early sign of progression to higher CDC clinical stages in HIV-1-infected individuals.

Plasma and salivary IgG subclasses in HIV type 1 infection: evidence of both transudation and local synthesis of IgG in parotid saliva.

In this study we measured the levels of plasma and salivary IgG subclasses in 81 HIV-1-infected individuals and 30 uninfected controls. Salivary IgG1 was increased in HIV-1-infected patients, while salivary IgG2 and IgG4 were decreased. Patients with high levels of plasma anti-HIV-1 IgG antibodies presented a higher CD4+ cell count and lower viral load. High levels of anti-HIV-1 IgG antibodies in plasma were also associated with high levels of anti-HIV-1 IgG antibodies in parotid saliva. In comparing the HIV-1 recognition patterns of salivary IgG with plasma IgG, we determined that plasma and salivary IgG antibodies had similarities as well as differences in their reactivity to HIV-1 antigens. The present study demonstrates that HIV-1 infection affects both plasma and salivary IgG and provides evidence that the origin of HIV-1-specific IgG antibodies in parotid saliva is primarily transudated from plasma; however, some local synthesis of IgG in parotid saliva also occurs.

Immunoglobulin isotypes in human minor gland saliva.

Lower labial, upper labial, palatine (minor), and parotid (major) gland saliva samples from 18 young adult males were quantitatively assayed for the presence of IgA1, IgA2, IgM, and IgG. The mean (+/- standard deviation) concentrations of IgA (sum of IgA1 + IgA2) were 79 +/- 42 micrograms/mL (parotid), 111 +/- 42 micrograms/mL (lower labial), 69 +/- 72 micrograms/mL (upper labial), and 88 +/- 68 micrograms/mL (palatine). Total IgA concentrations were positively correlated among different minor-gland samples from the same subject, although these correlations did not reach significance. Upper-labial-gland saliva samples contained significantly (at least p less than 0.05) lower concentrations of IgA1 than those found in parotid or lower-labial minor-gland secretions. All three minor-gland sources of saliva contained significantly (p less than 0.002) higher levels of IgG than did parotid saliva. Upper-labial fluids had significantly (p less than 0.02) higher IgG concentrations than lower-labial saliva. IgM could be detected in 89% of parotid saliva samples and 75% of the palatine saliva samples. Palatine IgM concentrations (8.2 +/- 17.8 micrograms/mL) were significantly (p less than 0.05) higher than parotid IgM concentrations (0.6 +/- 0.4 micrograms/mL). IgM was detected much less frequently and at lower concentrations in lower- and upper-labial-gland saliva. These data reveal that minor-gland saliva from different oral sites may contain distinctive immunoglobulin isotype patterns, and expressions of host defense may vary within each micro-environment.

Synthesis and secretion of secretory immunoglobulins: with special reference to dental diseases.

The distribution of immunoglobulin-containing cells in human tonsils, salivary glands, and inflamed gingiva is described. The cellular localization of J chain indicates that this peptide is a basic gene product of B cells that is expressed only in the early phase of clonal differentiation. Gland-associated immunocytes apparently are derived from this phase, which may be relevant to their local homing mechanism. The selective glandular transport of dimeric or polymeric IgA and 19S IgM may be determined by the content of J chain in these immunoglobulins. A J-chain-dependent configuration seems to be responsible for their noncovalent affinity for SC, and this may explain their specific reception at the epithelial cell membranes. Subsequent stabilization of the Ig-SC complexes takes place during their external transport, probably because of disulfide exchange. The latter process is more efficient for secretory IgA than for secretory IgM. The secretory dynamics of parotid IgA differs from that of other parotid proteins and is highly dependent on the degree of secretory stimulation. The secretion rate (mug/min/gland) seems to be a better measure of an individual's parotid IgA output than the absolute concentration of IgA in the secretion. A low parotid IgA secretion rate is associated with high susceptibility to dental caries, perhaps reflecting inferior resistance to dental plaque formation. It is not known whether such resistance, in part, is determined by specific antibodies to certain bacterial antigens. A potential candidate for an efficient preventive action of salivary antibodies could be the GTF enzyme system of S mutans. However, we were able to demonstrate only extremely rare immunocytes producing antibodies to GTF in human tonsils and gingiva, and none at all in salivary glands.

Effect of age on immunoglobulin content and volume of human labial gland saliva.

Stimulated lower labial (LLGF) and parotid salivary volumes and IgG, IgA, and IgM concentrations were measured in 264 subjects whose ages ranged from 17 to 76 years. A significant (p < 0.001) age-related decline in LLGF output was observed for subjects over this age range. Sixty-three percent of the subjects in the 18-20-year-old group (n = 46) secreted at least 10 microL of labial saliva in a 7-10-minute period, while approximately 70% of the subjects in the two oldest groups (61-70 and 71-76 years old) secreted less than 1 microL of LLGF during this time period (n = 64). No significant gender-based differences occurred in the volumes of labial saliva secreted. Stimulated parotid salivary flow showed no age-related trend in these subjects. Lower labial gland salivary IgA concentrations in an older population (mean age +/- SD = 55.6 yr +/- 1.3) were significantly lower (p < 0.025) than IgA concentrations in a younger population (20.7 yr +/- 0.8), when IgA was expressed as microgram/mL LLGF collected. Immunoglobulin A concentrations in parotid saliva and IgG and IgM concentrations in labial and parotid saliva were not significantly different when the two age populations were compared. These data suggest that the physiological and immunological potential of labial gland saliva may decrease with age.

Secretory immunoglobulin A heavy chain presents Galbeta1-3GalNAc binding structures for Actinomyces naeslundii genospecies 1.

Adherence of Actinomyces naeslundii ATCC 12104 to hydroxyapatite beads coated with protein fractions of parotid saliva, obtained by gel filtration on S-200 HR columns, showed GalNAcbeta1-3Galalpha-O-ethyl-inhibitable binding to high-molecular-weight proteins (Strömberg et al., 1992). The present study investigates the nature of these high-molecular-weight binding proteins and determines their specific ability to mediate adherence to representative strains of Actinomyces species. Strain ATCC 12104 bound specifically in a lactose-inhibitable manner to the heavy chain of secretory immunoglobulin A (S-IgA), contained within a high-molecular-weight parotid protein fraction separated on SDS-PAGE and transferred to a solid membrane support. Lactose-inhibitable binding to the heavy chain of S-IgA from human colostrum was also demonstrated. Peanut agglutinin bound to the heavy chain of parotid and colostrum S-IgAs contained on solid support membranes, confirming the presence of Galbeta1-3GalNAc residues on these molecules. Both salivary and colostrum S-IgA aggregated with strain ATCC 12104 in a GalNAcbeta1-3Galalpha-O-ethyl-inhibitable fashion. Further separation of high-molecular-weight salivary proteins on S-500 HR columns showed GalNAcbeta1-3Galalpha-O-ethyl-inhibitable binding to both mucin- and S-IgA-containing fractions. The presence of S-IgA in salivary pellicles formed in vivo on teeth was demonstrated by Western blot analysis of pellicle extracts with anti-IgA antibodies. Among strains representing A. naeslundii genospecies 1 and 2 and A. odontolyticus, only those of genospecies 1 with a particular adherence profile showed efficient GalNAcbeta1-3Galalpha-O-ethyl-inhibitable binding to S-IgA. Thus, oligosaccharides on S-IgA may promote bacterial aggregation (or adherence) and provide a mechanism by which S-IgA can interact with bacteria without prior immunological challenge.

Chewing stimulates secretion of human salivary secretory immunoglobulin A.

Immunoglobulin A (IgA) is the most abundant immunoglobulin in saliva and other mucosal secretions and plays an important role in mucosal immunity. The present study examined whether secretion of IgA, like other salivary proteins, is increased by reflex stimulation. Parotid saliva was collected from subjects into separate vials under resting conditions and during chewing-stimulated secretion over 45 min. Enzyme-linked immunosorbent assay (ELISA) indicated that chewing increased IgA secretion. The extent and pattern of the increase were similar to those of total protein and acinar cell amylase. SDS gel electrophoresis and Western blotting showed that high-molecular-weight forms of IgA-containing secretory component predominated in all saliva samples. Secretory component, the cleaved epithelial receptor for polymeric IgA, was secreted in a pattern very similar to that of IgA. It is concluded that chewing stimulates epithelial cell transcytosis of IgA and increases secretion of secretory IgA into saliva.

Generation of specific antibody-secreting cells in salivary glands of BALB/c mice following parenteral or oral immunization with Porphyromonas gingivalis fimbriae.

The humoral immune response and induction of antigen-specific antibody-secreting cells in mucosal lymphoid tissues were examined in mice immunized subcutaneously or orally with fimbrial protein purified from Porphyromonas gingivalis strain 381. A group of BALB/c mice was immunized subcutaneously with P. gingivalis fimbriae and semisynthetic adjuvant GM-53 in Freund's incomplete adjuvant on days 0 and 28. Another group of mice was immunized perorally with liposome containing fimbriae and GM-53 on days 0, 1, 27 and 28. In the mice immunized subcutaneously, salivary anti-fimbrial IgM antibodies were detected transiently on day 5, followed by the appearance of specific IgG and IgA antibodies on day 14. Higher concentrations of salivary IgG and IgA anti-fimbrial antibodies were found after the second immunization. Fimbria-specific IgM and IgG spot-forming cells (SFC) were detected in cervical lymph nodes of the immunized mice by the ELISPOT method. Fimbria-specific IgA SFC but not IgM and IgG appeared in the parotid and submandibular glands of subcutaneously immunized mice. On the other hand, mice immunized by gastric intubation generated almost exclusively salivary anti-fimbrial IgA antibodies. In agreement with this finding, increased numbers of antigen-specific IgA but not IgM and IgG SFC were seen in parotid and submandibular glands, but not in cervical lymph nodes of orally immunized mice. It can be concluded that systemic or oral immunization with fimbrial antigen induces distinct immune responses in specific lymphoid tissues or in the salivary glands in respect of their temporal sequences and the numbers of plasma cells secreting antigen-specific and non-specific immunoglobulins.

Keratoconjunctivitis sicca: immunological evaluation of 62 canine cases.

The etiology of keratoconjunctivitis sicca (KCS) in 62 dogs was evaluated, using immunologic techniques. Using direct fluorescent antibody testing, autoantibodies within the lacrimal, salivary, or pancreatic glands were detected in 5 of 8 dogs tested. Circulating antibodies to the nictitating membrane gland, main lacrimal gland, parotid salivary gland, or mandibular salivary gland were detected using indirect fluorescent antibodies in 9 of 31, 3 of 31, 5 of 31, and 5 of 31 sera, respectively. Using radial immunodiffusion, hyper-gamma-globulinemia was detected in 21 of 30 dogs with KCS. Antinuclear antibodies, primarily in a nucleolar pattern, were demonstrated in 20 of 50 dogs with KCS. Lymphocytic infiltrates were evident in 5 of 9 labial salivary biopsies, 2 of 4 parotid gland specimens, 2 of 4 mandibular gland specimens, and 2 of 3 thyroid gland specimens taken from dogs with KCS. Autoimmune diseases had been previously documented in 4 of 62 dogs. Twenty-five of the 62 dogs (40%) had concurrent problems indicative of an underlying immunologic disorder.