Persistence of chlamydial antibodies after pelvic inflammatory disease.

The persistence of chlamydial immunoglobulin G (IgG) antibodies and long-term sequelae of pelvic inflammatory disease (PID) were studied in 70 women who had been treated for PID 3 to 6 years previously. Fifty-one women had had PID associated with Chlamydia trachomatis infection (Chlamydia group), and 19 women had had PID not associated with C. trachomatis (non-Chlamydia group). Chlamydial IgG antibodies, as determined by the indirect immunofluorescence test with inclusions of C. trachomatis L2 as antigens, persisted at stable levels in 43% of the women for up to 6 years; 43% of the women showed a decrease in IgG titer, and 13% showed an increase. IgA antibody levels in serum correlated with IgG antibody levels in serum and with the presence of cervical IgA antibodies. Both serum antibodies and cervical IgA antibodies were more often found in the Chlamydia group. Forty-two percent of the women were infertile. Every fifth subsequent pregnancy was ectopic. The presence of cervical IgA antibodies might protect the women from tubal damage.     

Colonization in the rectum and uterine cervix with group B streptococci may induce specific antibody responses in cervical secretions of pregnant women.

We have studied the relationships between genital or rectal carriage of group B streptococci (GBS) with the levels of systemic and mucosal antibodies to GBS in 200 women at about week 17 of pregnancy. Secretions from the uterine cervix were collected with absorbent cylindrical wicks for quantification of antibody levels with whole cell enzyme-linked immunosorbent assay. GBS were cultured from the cervix (with or without concomitant rectal colonization) of 13.5%, from the rectum (with or without concomitant cervical colonization) of 12%, and from both culture sites of 8.5% of the women. Serotypes Ia, II, and III were predominant. Compared with culture-negative women, the group of women colonized rectally had markedly elevated levels of both immunoglobulin A (IgA) and IgG antibodies to GBS in cervical secretions and also had a moderate but significant elevation of IgA antibodies in sera. Women colonized only in the cervix had increases of specific IgA and IgG antibodies in cervical secretions, but their serum antibody levels were not elevated. In cervical secretions, the increase in antibody levels in the groups of colonized women was most pronounced for the IgG isotype, indicating a mucosal immune response involving IgG as well as IgA. A close correlation was found among the levels of antibodies to each of the three GBS serotypes tested. Evidence for such cross-reacting antibodies to different serotypes of GBS, as well as to group A streptococci, was also obtained from absorption experiments. Altogether, our results show that undiluted secretions for antibody determination can be easily collected from the uterine cervix with absorbent wicks and demonstrate that colonization of GBS in the rectum and the uterine cervix may induce a systemic as well as a pronounced local immune response in the female genital tract. The findings may have implications for the development of a mucosal vaccine against GBS disease.     

Rectal Immunization for Induction of Specific Antibody in the Genital Tract of Women

The purpose of the current study was to examine potential routes of vaccine administration for the induction of antigen-specific responses in the genital tract of women. Sixteen women were enrolled in this study, and the level of influenza-specific antibodies induced in the genital tract was measured after rectal or intramuscular immunizations. Both methods of administration induced significant increases in the concentration of flu-specific IgA found in cervical secretions within 28 days after vaccination. Initially flu-specific IgG antibodies were not induced in the genital tract by either route. As expected both IgA and IgG flu-specific antibodies were dramatically increased in serum after intramuscular vaccination. In contrast, rectal administration did not induce significant IgA responses, and only small flu-specific IgG increases in serum. Six months after administration, IgA flu-specific antibody concentrations were significantly higher than baseline levels in vaginal secretions and saliva isolated from both subject groups and flu-specific IgG concentrations in cervical secretions were high in the rectal immunization group. The long-term presence of both IgG and IgA antibody in genital secretions suggests that rectal immunization may be an effective method for induction of immune protection in the genital tract of women.     

Influence of exogenous reproductive hormones on specific antibody production in genital secretions after vaginal vaccination with recombinant cholera toxin B subunit in humans

The objective of this study was to investigate the influence of exogenous reproductive hormones on the local and systemic production of specific immunoglobulin A (IgA) and IgG antibodies after vaginal vaccination with recombinant cholera toxin subunit B (CTB). Three groups of women using either progesterone-containing intrauterine devices (n=9), oral contraceptives (n=8), or no hormonal contraceptive methods (n=9) were vaginally immunized twice, 2 weeks apart. Cervical secretions, vaginal fluids, and serum were collected before and after vaccination. Total and CTB-specific IgA and IgG antibodies in genital secretions and serum were analyzed by enzyme-linked immunosorbent assay. A majority of the women presented strong CTB-specific IgA and IgG antibody responses in cervicovaginal secretions after vaccination, whereas the antitoxin responses in serum were weaker. Exogenously administered steroid hormones did not seem to have any impact on the production of specific antibodies. Both the frequencies and the magnitudes of IgA and IgG antitoxin responses in genital secretions were comparable among the three immunization groups. An association, in particular for IgA, was found between the magnitudes of the CTB-specific antibody responses in cervical secretions and vaginal fluids after vaccination. The sensitivities and positive predictive values of vaginal antibody analyses to reflect responses in cervical secretions were also high, suggesting that vaginal fluids alone might be used for evaluation of genital immune responses in large-scale vaccination studies in the future.     

Serological responses to papillomavirus group-specific antigens in women with neoplasia of the cervix uteri.

Certain types of human papillomaviruses have been linked to the development of carcinoma of the cervix uteri. We have analyzed 114 serum specimens from women with cervical intraepithelial neoplasia (CIN) or carcinoma of the cervix uteri for the presence of serum antibodies against purified, disrupted bovine papillomavirus (BPV). The titers of immunoglobulin A (IgA) antibodies against BPV were slightly elevated (P less than 0.025) in the sera from CIN or cervical carcinoma patients compared with the titers of 139 serum specimens from sex- and age-matched healthy controls. In contrast, both the IgG and IgM serum antibody titers against BPV were significantly decreased for CIN and cervical carcinoma patients compared with those of healthy controls (P less than 0.001 and P less than 0.005, respectively). These results suggest that the difference between IgA and IgG or IgM antibodies to papillomavirus group-specific antigens may represent interesting serological parameters that could possibly be used in the epidemiologic study of women at risk for CIN.     

Significance of immunoglobulin a titres in the diagnosis of urogenital chlamydial infections

The significance of chlamydial serum IgA compared with IgM and IgG in the diagnosis of urogenital chlamydial infection was evaluated using 120 sera from different categories of patients. In urethritis patients both IgM and IgA antibodies were not found to be present consistently, whereas in patients with deep-seated chlamydial infection, IgA was more often present. Although of limited value in superficial infections, demonstration of IgA antibodies may be of value in the diagnosis of deep-seated chlamydial infections.     

Modified wick method using Weck-Cel sponges for collection of human rectal secretions and analysis of mucosal HIV antibody

Weck-Cel sponges were examined for suitability as an absorbent material for nontraumatic collection of rectal secretions in humans. Sponges were tested in vitro and determined by quantitative enzyme-linked immunosorbent assay (ELISA) to be capable of releasing 100% of absorbed albumin and all immunoglobulin subtypes after treatment with detergent-supplemented buffer. Protein composition in rectal secretions collected from normal women with dry sponges (DS) or with sponges previously softened by moistening with saline (MS) was subsequently compared. DS secretions showed evidence of contamination with blood and interstitial fluid-derived albumin, immunoglobulin G (IgG), and monomeric IgA. MS secretions appeared to represent local mucosal secretions more accurately because they contained negligible blood, a greater percentage of secretory IgA within the total IgA, and both lower albumin/IgG ratios and more dramatic alterations in IgG subclass distribution compared with corresponding serum. Anti-HIV IgG, IgM, IgA, and antibodies with secretory component could be demonstrated by ELISA in rectal secretions collected with moist sponges from 8 of 8, 1 of 8, 5 of 8, and 3 of 8 HIV-infected women, respectively. The data show that Weck-Cel sponges, if premoistened, can be used to collect rectal fluids nontraumatically and to obtain quantitative information about concentrations of immunoglobulins and specific antibodies on rectal mucosal surfaces.     

IgG, IgA, and IgM antibodies to mite in sera and sputa from asthmatic patients

In order to examine roles of antibodies to allergens in bronchial secretions, IgG, IgA, and IgM antibodies to mite in sputa from mite-sensitive asthmatics were measured by ELISA and compared with antibodies in sera. IgA antibodies to mite in sputa were significantly higher in mite-sensitive patients than in normal controls or mite-unsensitive asthmatic patients (P less than .01), whereas IgG and IgA antibodies in sera were significantly higher in mite-sensitive patients than in the other two groups (P less than .01). There were no significant differences of serum or sputum IgM antibodies among the three groups. The relative ratio of the level of IgA antibodies: the level of IgG antibodies was higher in sputum than in sera. IgA antibodies in bronchial secretions may play a protective role for asthma when allergens enter into the bronchial trees.     

Serological responses to human papillomavirus type 6 and 16 virus-like particles in patients with cervical neoplastic lesions.

Serum samples from 36 cervical carcinoma patients, 33 patients with high-grade squamous intraepithelial lesions, and 31 cytologically normal women were tested by enzyme-linked immunosorbent assay (ELISA) using human papilloma virus type 6 (HPV 6) and HPV 16 virus-like particles as antigens. Forty serum specimens from 1-year-old children were used to assign cutoff points. When serum samples from the subjects infected with HPV 16 were tested in an HPV 16 ELISA detecting immunoglobulin A (IgA), IgG, and IgM binding, 61% showed IgA, 44% showed IgG, and 39% showed IgM reactivity. Of HPV 6- or 11- or HPV 18-infected subjects. fewer than 17% showed IgA or IgG responses and 33% showed IgM reactivity. In contrast, 13% showed IgA, 10% showed IgG, and 16% showed IgM reactivity in the HPV DNA-negative controls. The results suggest that the IgA and IgG responses are HPV 16 specific and the IgM response is cross-reactive to different HPV types. On the other hand, the serological responses to HPV 6 did not differ in the patient and control groups. The percentages of patients positive for both IgA and IgG antibodies were significantly higher in the groups with high-grade squamous intraepithelial lesions (12% [4 of 33]; P = 0.04) and cancer (17% [6 of 36]; P = 0.02) than in the healty women (0% [0 of 31]), and the percentages for either IgA or IgG were higher for the cancer group (47% [17 of 36]; P = 0.01) than in the normal group (19% [6 of 31]). Most sera positive for IgA and IgG in the patient groups showed higher titers than those in the normal group. All these results suggest that high IgA and IgG responses are good indicators for estimating HPV 16 infection.     

Mucosal and peripheral immune responses to chlamydial heat shock proteins in women infected with Chlamydia trachomatis

Most of the studies on 60-kDa and 10-kDa chlamydial heat shock proteins (HSPs) to date have been carried out with blood lymphocytes or serum antibody responses, which do not provide a clear picture of the actual pathogenesis as they do not differentiate primary infection from recurrent infection. Thus, in the present study induction of the immune response was evaluated by studying lymphoproliferation of both cervical and peripheral lymphocytes to synthetic peptides of cHSP60, cHSP10 and major outer membrane protein (MOMP) antigen. In addition, cervical antibody prevalence to MOMP antigen, cHSP60 and cHSP10 and cytokine levels in cervical washes was also determined. Positive proliferative responses of cervical lymphocytes to cHSP10 peptide were significantly higher (P < 0.05) in women with recurrent infections and that to MOMP antigen were significantly higher in primary infection. On proliferation of PBMCs with the above antigens, no significant difference was observed between primary and recurrent infection. Prevalence of cervical IgG and IgA antibodies to Chlamydia trachomatis was significantly higher (P < 0.05) during primary infection than recurrent infections. In contrast, prevalence of IgG and IgA antibodies to cHSP10 and IgG antibodies to cHSP60 was higher during recurrent infections than primary infections. Interferon (IFN)-gamma levels were significantly higher in cervical washes of women with recurrent infection and correlated strongly with cHSP60 antibody titres. Our data thus suggest that mucosal responses are more appropriate in understanding the pathogenesis of chlamydial infection and IFN-gamma could be involved in the modulation of immune responses towards chlamydial infection directly, by causing acute inflammation, or indirectly through modulation of HSP expression.     

Detection of Antispermatozoal Antibodies of IgG,IgA, and IgM Immunoglobulin Classes in Cervical Mucus

ABSTRACT: The immunobead test (IBT) was applied to bromelin-treated cervical mucus (CM) samples from 78 infertile women. Seven (8.9%) of the patients were positive for IgG and/or IgA class antibodies, whilst none (0/35) were found positive for IgM class antibodies. We found that 57% of the positive samples contained IgG and IgA classes, whilst the remaining samples contained IgA alone. The implications of these results were discussed.     

Dog immunoglobulins. II. The antibacterial properties of dog IgA, IgM and IgG antibodies to Vibrio cholerae.

Secretory IgA antibodies to Vibrio cholerae were purified from the parotid saliva and mammary secretions of locally and orally immunized dogs using gel filtration, ion-exchange chromatography and anti-immunoglobulin immuno-absorbents. IgM and IgG antibodies were isolated from serum by gel filtration and ion-exchange chromatography. IgA antibodies proved to have minimal, if any, activity in direct killing of bacteria in the presence of complement or in the promotion of phagocytosis. The minimal activity which IgA had in these assays could be accounted for by extremely small quantities of IgM antibody. The same IgA antibodies, mixed with the challenge innoculum of Vibrio cholerae and fed to infant mice, protected these mice as efficiently as IgG or IgM antibodies.     

Antibody responses to Toxoplasma gondii in sera, intestinal secretions, and milk from orally infected mice and characterization of target antigens.

Toxoplasma gondii-specific antibody responses in serum, intestinal secretions, and milk were identified with an enzyme-linked immunosorbent assay following a single oral infection of mice with strain 76K cysts of T. gondii. Immunoglobulin A (IgA) production began during week 2 of infection in serum and milk and during week 3 of infection in intestinal secretions and persisted in all three throughout the experiment (17 weeks). IgG but not IgM antibodies were detected in intestinal secretions later in the infection. Serum and milk IgG and IgM production began at the same time after infection as did the IgA response. In Western blotting (immunoblotting), intestinal IgA antibodies were shown to react with antigens comigrating with the T. gondii proteins p22, p23, p30, and p43, the 28-kilodalton antigen, and the 55- and 60-kilodalton rhoptry proteins, as recognized by specific monoclonal antibodies. Milk IgA antibodies reacted with antigens comigrating with p30 and p43. Most of the antigens recognized by IgA antibodies were also detected by IgG antibodies. IgA antibodies from all three biological samples detected the same major T. gondii antigens; thus, there was apparently no specific antibody production unique to one locality.     

Comparison between cell culture and serology for detecting Chlamydia trachomatis in women seeking abortion.

The efficiency of an immunoperoxidase serological assay and culture of Chlamydia trachomatis were compared in 127 women seeking first trimester abortion. Serum IgG and IgA antibodies specific for C trachomatis were detected by a single serovar (L2) inclusion immunoperoxidase assay (IPA). Eighty (63%) women were seropositive for chlamydial IgG and 31 (24%) for IgA antibodies. C trachomatis was isolated from 21 of 127 (17%) women. Twenty of the 80 women (25%) seropositive for specific IgG antibodies and one of 47 (2%) patients without these antibodies were culture positive (p less than 0.001). Compared with isolation, chlamydial antibodies at a titre of greater than or equal to 16 showed high sensitivity and negative predictive value (95% and 98%, respectively), but low specificity and efficiency (43% and 52%, respectively). Chlamydial IgA antibodies at a titre of greater than or equal to 8 showed low sensitivity (52%), but a higher specificity, negative predictive value, and efficiency of 81%, 90%, and 76%, respectively. C trachomatis IgG antibodies at a titre of 16 as determined by IPA can be used as an efficient negative exclusion marker for active chlamydial infection in screening women seeking abortion.     

Boosting of Secretory IgA Antibody Responses in Man by Parenteral Cholera Vaccination

The occurrence of specific antibodies to Vibrio cholerae lipopolysaccharide in serum, milk, and saliva of Pakistani women from a very low socioeconomic group was studied before and after a single subcutaneous cholera vaccination. Before immunization all women had low levels of specific antibodies in serum, primarily of IgM class, and in many cases cholera IgA angibodies were found in milk and saliva as well, indicating earlier natural exposure. The vaccination consistently induced a marked rise in serum antibody titer, and notably also produced significant titer increases in 70% of the milk and in 45% of the saliva samples. Whereas the serum antibodies induced were predominantly of the IgG class, secretory IgA was responsible for most of the titer increase in the secretions. The results indicate that parenteral cholera vaccination can boost local secretory IgA antibody responses in intestinally primed individuals.     

Protective local and systemic antibody responses of swine exposed to an aerosol of Actinobacillus pleuropneumoniae serotype 1.

The isotype-specific antibody responses in serum and in nasal and pulmonary lavage fluids of swine following aerosol immunization with an attenuated strain of Actinobacillus pleuropneumoniae serotype 1, strain CM5A, was investigated. The presence of immunoglobulin G (IgG), IgA, and IgM with specificities for capsular polysaccharide, lipopolysaccharide, and hemolysin was determined by enzyme-linked immunosorbent assay by using purified antigens. Strain CM5A induced serum antibodies of each isotype to the three antigens. The serum antibody response was sustained and typical of persistent antigenic stimulation. The specific IgM response decreased and the specific IgG response increased after challenge with strain CM5. IgA specific for the three antigens was detected in nasal secretions from all immune pigs, whereas specific IgG could only be detected in samples contaminated with blood. Both IgA and IgG specific for each of the antigens were detected in pulmonary lavage samples. There was no significant increase in specific IgA in nasal secretions; however, levels of lipopolysaccharide-specific and hemolysin-specific IgG and IgA in pulmonary secretions rose after aerosol challenge with strain CM5. Passive transfer of immune swine serum resulted in protection against pleuropneumonia and in levels of specific serum IgG which were similar to those in actively immunized pigs. It is concluded that specific serum IgG antibodies are important in protection from porcine pleuropneumonia.     

Immunoglobulins in human cervico-vaginal secretions

The concentration of IgG, IgA and IgM has been measured in the cervico-vaginal secretions of 8 women with a normal menstrual cycle, 52 pregnant women, 6 post-menopausal women and 12 women with total hysterectomy. No significant difference in immunoglobulin levels was found in the cervico-vaginal secretions of women with a normal cycle as compared to those of post-menopausal or pregnant women. A significant decrease of the IgG/IgA ratio was noticed during ovulation as a consequence of increased IgA secretion. In patients with hysterectomy, the secretions are of vaginal origin only and contain negligible quantities of IgA. Secretory IgA is found essentially in the superior genital tract. IgM is present in trace amounts in all secretions and does not vary considerably. The secretion of immunoglobulins may be under hormonal control: in addition to ovarian hormones, corticosteroids seem also to be involved. The local application of a fluorinated corticosteroid into the vagina has produced a significant decrease of secretory IgA production. The therapeutic possibilities of administering a fluorinated corticosteroid in cases of sterility which are due to the production of anti-spermatozoal antibodies are discussed.     

The Induction and Expression of IgA Anti-DNP Antibodies in Rat Bile and Secretions

Pregnant rats were immunized with dinitrophenylated type III-pneumococcal vaccine by the intravenous, gastric, or intramammary routes. Anti-DNP antibody responses in the IgA. IgG and IgM isotypes were measured in serum, secretions and bile. Gastric intubation was most effective at eliciting IgA antibody responses in bile and secretion, whereas the other routes were more effective at inducing IgG responses in serum and bile. IgM antibody responses were infrequent and were found in fluids most closely associated with the immunization route. Isoelectric focusing studies of IgA antibodies appearing in secretions and bile revealed that the gastric route consistently elicited antibody spectrotypes with shared components. Intravenous and intramammary immunization resulted in IgA spectrotypes possessing less homology, suggesting that these protocols lead to independent antibody responses triggered in spleen, draining lymph nodes, or secretory sites. After gastric stimulation, the appearance of IgA antibodies with identical spectral components in secretions and bile favours the concept that IgA precursor cells with identical clonotype potential migrate from the gastrointestinal area to secretory sites. Antibody expression in bile appears to result from the selective transfer of IgA populations gaining access to serum after synthesis at a secretory site.     

Kinetics of local and systemic immune responses after vaginal immunization with recombinant cholera toxin B subunit in humans

Vaginal vaccination seems to be the best strategy for inducing specific immunoglobulin A (IgA) and IgG antibody responses in the female genital tract. The relative efficiencies of one, two, and three vaginal doses of recombinant cholera toxin B subunit (CTB) in generating mucosal and systemic immune responses in healthy women were evaluated, and the kinetics of the immune responses were monitored for responding volunteers for up to 12 months after the last vaccination. A single dose of CTB failed to generate CTB-specific IgA antibody responses in cervical secretions. Two vaccinations induced significant increases in IgA antitoxin titers in seven of nine volunteers, and four volunteers also developed IgG antitoxin responses. The magnitudes of the responses were 20-fold for IgA antitoxin and 7.1-fold for IgG antitoxin. A third vaccination did not significantly increase the antitoxin responses, although the frequency of IgG responses was slightly higher than that after the second vaccination. In serum, CTB-specific antibodies were observed already after a single vaccination. However, two vaccinations were required to induce marked IgA as well as IgG antitoxin titer increases in the majority of volunteers. The postvaccination levels of antitoxin antibodies in serum were comparable after two and three vaccinations. At 12 months after vaccination, significantly elevated IgA and IgG antitoxin levels in cervical secretions could still be detected in approximately half of the volunteers who had initially responded to the vaccine. Antitoxin titer increases in serum were found in most of the vaccinees at follow-up.     

Kinetics of Local and Systemic Immune Responses after Vaginal Immunization with Recombinant Cholera Toxin B Subunit in Humans

Vaginal vaccination seems to be the best strategy for inducing specific immunoglobulin A (IgA) and IgG antibody responses in the female genital tract. The relative efficiencies of one, two, and three vaginal doses of recombinant cholera toxin B subunit (CTB) in generating mucosal and systemic immune responses in healthy women were evaluated, and the kinetics of the immune responses were monitored for responding volunteers for up to 12 months after the last vaccination. A single dose of CTB failed to generate CTB-specific IgA antibody responses in cervical secretions. Two vaccinations induced significant increases in IgA antitoxin titers in seven of nine volunteers, and four volunteers also developed IgG antitoxin responses. The magnitudes of the responses were 20-fold for IgA antitoxin and 7.1-fold for IgG antitoxin. A third vaccination did not significantly increase the antitoxin responses, although the frequency of IgG responses was slightly higher than that after the second vaccination. In serum, CTB-specific antibodies were observed already after a single vaccination. However, two vaccinations were required to induce marked IgA as well as IgG antitoxin titer increases in the majority of volunteers. The postvaccination levels of antitoxin antibodies in serum were comparable after two and three vaccinations. At 12 months after vaccination, significantly elevated IgA and IgG antitoxin levels in cervical secretions could still be detected in approximately half of the volunteers who had initially responded to the vaccine. Antitoxin titer increases in serum were found in most of the vaccinees at follow-up.