Antibodies In Vaginal Secretions Research Articles

Removal of the spleen abrogates the long-term hydrosalpinx pathology following Chlamydiagenital infection

Abstract C. trachomatis infections in women may cause pelvic inflammatory disease, chronic pelvic pain, and infertility. In the mouse model, genital Chlamydia disseminates systemically to infect the spleen. Removal of the spleen (splenectomy) abrogates hydrosalpinx pathology in the female reproductive tract (FRT), although it does not affect FRT Chlamydia clearance. Ascension of Chlamydia to the upper FRT (uteri, ovaries, oviducts) is critical for the development of hydrosalpinx. Over the course of genital infection, splenectomized mice exhibit significantly lower Chlamydia burden in the upper FRT and the GI tract compared to controls. Based on the report that Chlamydia-specific IgA secreted in the FRT protects against genital Chlamydia challenge, we hypothesized that splenectomy induces IgA secretion in the FRT. Examination of vaginal washes revealed that unlike controls, splenectomized mice exhibit Chlamydia-specific IgA by 4 weeks post-infection (wpi). However, the presence of IgA at this time did not explain the significant reduction in Chlamydia ascension observed by 2 wpi. Unlike controls, splenectomized mice additionally exhibit Chlamydia-specific IgM antibodies in vaginal secretions as early as 2 wpi. Naïve mice infected with Chlamydia pre-incubated with vaginal washes of splenectomized mice collected at 2 or 6 wpi exhibited reduced Chlamydia burden in the FRT, ILNs, spleen, and the GI tract at 3 and 7 days post-infection compared to controls. Secreted IgM and IgA thus neutralize Chlamydia, diminish its ascension in the upper FRT and thus abrogate the long-term hydrosalpinx pathology. This work reveals a previously unknown role that the spleen plays in the regulation of antibody secretion in the FRT mucosa. Supported by a grant from NIH (1 R21 AI159743-01)

Short Communication: Parallel Analyses of Systemic and Local Vaccinations with Envelope Formulated in Adjuvant for Induction of HIV-Specific Antibodies in the Vaginal Mucosa.

To prevent HIV-1 infections in females, vaccines that elicit antibodies in vaginal secretions are much desired. To induce these antibodies, intravaginal vaccinations are sometimes recommended. However, the benefit of intravaginal vaccination remains a topic of debate, and parallel studies of intravaginal and intramuscular vaccination routes are rarely performed. Here we describe tests of mucosal and systemic antibodies after mouse vaccinations by several routes with HIV-1 envelope protein formulated in adjuvant. Response magnitudes were as follows: intraperitoneal > intramuscular = intravaginal tissue ≥ subcutaneous. We found that the well-accepted and logistically feasible intramuscular immunization was similar to intravaginal tissue immunization for the induction of antibodies in blood and vaginal secretions. Results suggest that a routine comparison of intravaginal and intramuscular immunizations may serve as a beneficial gatekeeper for the development of new intravaginal HIV-1 vaccines.

HIV-1 specific IgA detected in vaginal secretions of HIV uninfected women participating in a microbicide trial in Southern Africa are primarily directed toward gp120 and gp140 specificities.

BackgroundMany participants in microbicide trials remain uninfected despite ongoing exposure to HIV-1. Determining the emergence and nature of mucosal HIV-specific immune responses in such women is important, since these responses may contribute to protection and could provide insight for the rational design of HIV-1 vaccines.Methods and FindingsWe first conducted a pilot study to compare three sampling devices (Dacron swabs, flocked nylon swabs and Merocel sponges) for detection of HIV-1-specific IgG and IgA antibodies in vaginal secretions. IgG antibodies from HIV-1-positive women reacted broadly across the full panel of eight HIV-1 envelope (Env) antigens tested, whereas IgA antibodies only reacted to the gp41 subunit. No Env-reactive antibodies were detected in the HIV-negative women. The three sampling devices yielded equal HIV-1-specific antibody titers, as well as total IgG and IgA concentrations. We then tested vaginal Dacron swabs archived from 57 HIV seronegative women who participated in a microbicide efficacy trial in Southern Africa (HPTN 035). We detected vaginal IgA antibodies directed at HIV-1 Env gp120/gp140 in six of these women, and at gp41 in another three women, but did not detect Env-specific IgG antibodies in any women.ConclusionVaginal secretions of HIV-1 infected women contained IgG reactivity to a broad range of Env antigens and IgA reactivity to gp41. In contrast, Env-binding antibodies in the vaginal secretions of HIV-1 uninfected women participating in the microbicide trial were restricted to the IgA subtype and were mostly directed at HIV-1 gp120/gp140.

P.2.044 Dysfunctional supplementary motor area implication during attention and time estimation tasks in neuroleptic naïve schizophrenia

<dm:abstracts xmlns:dm="http://www.elsevier.com/xml/dm/dtd"><ce:abstract xmlns:ce="http://www.elsevier.com/xml/common/dtd" view="all" class="author" id="aep-abstract-id15"><ce:section-title>Publisher Summary</ce:section-title><ce:abstract-sec view="all" id="aep-abstract-sec-id16"><ce:simple-para id="fsabs057" view="all">Many primary simian immunodeficiency virus (SIV) strains use CCR5 to infect simian cells in the absence of CD4. HIV/SIV transmission through an intact mucosal surface requires that the virus bind the epithelial cells and either infect these or be transmitted to underlying lymphoid cells. Langerhans cells and dendritic cells (DCs) also express the major coreceptor CCR5 for SIV/HIV binding and fusion. SIV infection of intact mucosa has been demonstrated by direct application of high doses of SIV into the vagina, rectum, or oral cavity of rhesus monkeys. There is experimental evidence in adult male rhesus macaques that they can be infected with SIV by the placement of cell-free virus onto their foreskins. SIV infected macaques showed predominantly IgG anti-SIV antibodies in vaginal secretions. Findings, both in SIV-infected rhesus macaques and in HIV-infected women, suggest that transudation of serum IgG antibodies is a significant source of antiviral antibodies in vaginal secretion. The development of HIV-SIV hybrid constructs consisting of HIV <ce:italic>env</ce:italic> gene in a SIV mac backbone has extended the use of macaques in immunization with some HIV antigens.</ce:simple-para></ce:abstract-sec></ce:abstract></dm:abstracts>

Induction of an HPV 6bL1-specific mucosal IgA response by DNA immunization

Human papillomavirus (HPV) plays a crucial role in the development of human anogenital dysplasia. To prevent infection, it is important to induce an HPV-specific mucosal immune response. We investigated whether DNA vaccination would induce an intravaginal mucosal antibody response against HPV 6bL1. New Zealand White rabbits were immunized with an HPV 6bL1 DNA vaccine by one of the three routes: muscular, vaginal, or rectal. We found that vaginal immunization of rabbits with HPV 6bL1 DNA induced 6bL1 virus-like particle-specific lgA antibodies in vaginal secretions. They were detectable until at least 14 weeks after the first immunization. The antibodies also showed neutralizing activity in a hemagglutination inhibition assay. No mucosal immune response was detected in vaginal secretions of rabbits immunized intramuscularly or intrarectally. Our data suggest that vaginal immunization with HPV 6bL1 DNA induces long-lasting IgA responses with neutralizing activity in vaginal secretions of rabbits.

Genital and Systemic Immune Responses in a Murine Model of Tritrichomonas foetus Infection

A reliable laboratory animal model would be useful for the study of immune responses to trichomoniasis, a sexually transmitted disease of human beings and cattle. Murine models are available, but pretreatment with estrogen is used, which may influence immune responses. To evaluate whether vaginal trichomoniasis could be established in nonestrogenized mice and to define the immune responses associated with the infection, CD1 and BALB/c mice were studied with or without estrogen treatment prior to inoculation with Tritrichomonas foetus. Tritrichomonas Foetus was cultured from the vagina and uterus of both estrogen-treated and untreated control mice for up to 26 wk. The infection was sustained better in BALB/c than in CD1 mice, suggesting that the former strain was most susceptible. In CD1 mice, infection was sustained less well in estrogen-treated than in untreated control mice, but there was no difference between treatment groups of BALB/c mice. IgA and IgG antibodies in vaginal secretions, uterine secretions, and serum specific for a surface antigen of T. foetus (TF1.17) were measured by enzyme-linked immunosorbent assay. In infected CD1 mice, vaginal IgA and IgG antibodies were detected by 8 wk postinoculation (PI). In infected BALB/c mice, vaginal IgA and IgG antibodies were detected by 12 wk PI. Uterine IgG responses predominated over IgA in estrogen-treated and untreated CD1 and BALB/c mice. There were high levels of IgG, but relatively no IgA in the sera of CD1 and BALB/c mice. Overall, the highest IgA response was in the vaginal secretions of infected CD1 mice, and some animals of this strain cleared the infection. These results show that a chronic trichomonad infection was established in mice without prior treatment with estrogen. The infection was associated with antibody responses in reproductive secretions and serum. This animal model will be useful in studying immunization to protect against trichomoniasis in mice not immunocompromised by estrogen.

Immunoglobulin G is the main protective antibody in mouse vaginal secretions after vaginal immunization with attenuated herpes simplex virus type 2.

We investigated the protective role of antibodies in vaginal secretions of mice that were immune to vaginal challenge with herpes simplex virus type 2 (HSV-2). Unfractionated vaginal immunoglobulins from immune and nonimmune mice and affinity-purified immunoglobulin G (IgG) and secretory IgA (S-IgA) from immune secretions were adjusted to their concentrations in vivo. Wild-type HSV-2 was incubated in the immunoglobulin preparations for 15 min in vitro, followed by inoculation into vaginae of nonimmune mice. HSV-2 was neutralized by unfractionated antibody and purified IgG from immune secretions but not by unfractionated nonimmune antibody or by purified immune S-IgA. The protective effect of IgG in vivo was investigated by passively transferring purified serum IgG from immune and nonimmune donors to nonimmune recipients before vaginal challenge infection. Immune IgG significantly reduced the percentage of vaginal epithelium infected, concentrations of shed virus protein in the vaginal lumen, and illness scores, even though the viral antibody titers in serum and vaginal secretions of recipient mice at the time of challenge were only 29 and 8%, respectively, of those in actively immunized mice. Additionally, removal of vaginal secretions from immune mice 10 min before vaginal challenge with HSV-2 significantly increased the concentration of shed virus protein in the vaginal lumen after challenge. Collectively, the data indicate that IgG antibody in vaginal secretions of immune mice provides early protection against vaginal challenge infection, probably by neutralizing virus in the vaginal lumen. In contrast, S-IgA antibody contributed relatively little to immune protection of the vagina.

Antibody responses and infertility in mice following oral immunization with attenuated Salmonella typhimurium expressing recombinant murine ZP3.

Ovarian ZP3, the primary sperm receptor, is a major glycoprotein of mouse zona pellucida (ZP). Because antibodies raised against ZP3 block sperm-egg interaction, ZP3 has been considered a candidate immunogen in the development of a contraceptive vaccine. This study explored the possibility of using an attenuated Salmonella typhimurium vaccine strain expressing recombinant ZP3 to elicit an antibody response and infertility in mice. A cDNA sequence generated by the polymerase chain reaction encoding 342 amino acid residues (23-364) of the mouse (m)ZP3 was cloned into an Asd+ vector. An avirulent Salmonella vaccine strain stably expressed the ZP3 polypeptide and colonized the internal organs of mice after oral inoculation. Oral immunization of female BALB/c mice with the recombinant Salmonella vaccine strain expressing mZP3 induced significant levels of anti-native ZP IgG antibodies in serum and IgA antibodies in vaginal secretions. The IgG antibodies thus induced also bound to ZP in vivo. When mated with males, 3 of 6 females immunized with the recombinant Salmonella were infertile. In contrast, none of the mice that received Salmonella containing the vector plasmid produced antibodies to ZP and all were fertile. No ovarian inflammation was observed in the immunized mice at autopsy. The results suggest a potential oral contraceptive vaccine to control populations of rodent vectors of disease and to induce reversible infertility in humans.

Induction of antigen-specific antibodies in vaginal secretions by using a nontoxic mutant of heat-labile enterotoxin as a mucosal adjuvant.

Immunization of the female reproductive tract is important for protection against sexually transmitted diseases and other pathogens of the reproductive tract. However, intravaginal immunization with soluble antigens generally does not induce high levels of secretory immunoglobulin A (IgA). We recently developed safe mucosal adjuvants by genetically detoxifying Escherichia coli heat-labile enterotoxin, a molecule with a strong mucosal adjuvant activity, and here we describe the use of the nontoxic mutant LTK63 to induce a response in the mouse vagina against ovalbumin (Ova). We compared intravaginal and intranasal routes of immunization for induction of systemic and vaginal responses against LTK63 and Ova. We found that LTK63 is a potent mucosal immunogen when given by either the intravaginal or intranasal route. It induces a strong systemic antibody response and IgG and long-lasting IgA in the vagina. The appearance of vaginal IgA is delayed in the intranasally immunized mice, but the levels of vaginal anti-LTK63 IgA after repeated immunizations are higher in the intranasally immunized mice than in the intravaginally immunized mice. LTK63 also acts as a mucosal adjuvant, inducing a serum response against Ova, when given by both the intravaginal and intranasal routes. However, vaginal IgA against Ova is stimulated more efficiently when LTK63 and antigen are given intranasally. In conclusion, our results demonstrate that LTK63 can be used as a mucosal adjuvant to induce antigen-specific antibodies in vaginal secretions and show that the intranasal route of immunization is the most effective for this purpose.

Oral immunization with attenuated Salmonella expressing human sperm antigen induces antibodies in serum and the reproductive tract.

Induction of immune responses in the reproductive tract will be crucial for a functional gamete antigen-based antifertility vaccine. Here we describe the construction and development of an avirulent Salmonella as an oral vaccine delivery vector to elicit sperm-specific immune responses in reproductive tract secretions. A cDNA sequence encoding the human sperm antigen SP10 was cloned on an asd+vector and expressed to a high level in an avirulent delta cya, delta crp, and delta asd vaccine strain of Salmonella typhimurium. Oral immunization of female BALB/c mice with this recombinant Salmonella elicited high-titer anti-SP10 IgG antibodies in serum and IgA antibodies in vaginal secretions. Anti-SP10 antibody titers could be increased by secondary and tertiary oral administrations of the recombinant Salmonella. Induction of sperm-specific antibodies in the reproductive tract following oral administration of a recombinant Salmonella could lead to the development of a simple, safe, efficient, and easy-to-use antifertility vaccine.

Antichlamydial activity of vaginal secretion

OBJECTIVE: The purpose of this study was to investigate the possible antichlamydial activity of vaginal secretion and to partially characterize the inhibitory principle. STUDY DESIGN: Vaginal secretions obtained from 156 women attending a family planning or gynecologic outpatient clinic for contraceptive advice were studied for the influence on the inclusion formation of Chlamydia trachomatis in cycloheximide-treated McCoy cell cultures. RESULTS: Vaginal secretions from 156 women inhibited the inclusion formation of Chlamydia trachomatis. The inhibitions was concentration dependent and the inhibitory principle had a molecular weight of <10,000 d. It was heat labile. It was not related to antichlamydial antibodies in vaginal secretions. Only three (2%) of the women had a positive culture for Chlamydia trachomatis. Three had immunoglobulin A and three had immunoglobulin G antichlamydial antibodies in vaginal secretions. Secretions of those with a vaginal pH of 3.5 to 4.5 decreased the chlamydial inclusion count by 75% compared with controls. The corresponding percentage for those with a pH of 5.0 to 6.0 was 48% and for those with pH >6 was 33%. Vaginal secretions of oral contraceptive users and nonusers did not differ in the capacity to decrease the chlamydial inclusion count, p > 0.01. CONCLUSIONS: When vaginal secretions were added to McCoy cell cultures infected by Chlamydia trachomatis, the chlamydia inclusion number decreased. There was a correlation between pH of the vaginal secretion and the inhibitory principle. Oral contraceptive use had no influence on the inhibition.

Immunoglobulin G antibodies in human vaginal secretions after parenteral vaccination

The induction of antibodies in vaginal secretions by systemic (intramuscular) immunization in humans was investigated by using the tetanus toxoid vaccine. Five women, 30 to 40 years old, were injected with a currently used dose of toxoid (40 IU), and serum, saliva, and vaginal secretion samples were collected on day 0 and on day 6 or day 10. All of these subjects had been previously vaccinated at least 5 years before; four were in good health, whereas one suffered from AIDS in clinical category B3. In most cases, analysis of specific antibodies in the vaginal wash showed a dramatic rise after boosting. These antibodies were primarily of the immunoglobulin G (IgG) isotype. The specific activity (ratio of antibody titer to IgG concentration) was shown to increase after the booster injection, irrespective of variations in the IgG level during the menstrual cycle. Comparison between serum and genital antibodies showed no difference in terms of both specific activity and level of avidity. These results demonstrate that parenteral injections can induce a systemic-derived antibody release in the vaginal fluid. Hence, systemic vaccinations can be efficient at the genital level and thus could reinforce or even replace a local vaccine.

Cell mediated responses of immunised vervet monkeys to defined T cell epitopes

<dm:abstracts xmlns:dm="http://www.elsevier.com/xml/dm/dtd"><ce:abstract xmlns:ce="http://www.elsevier.com/xml/common/dtd" view="all" class="author" id="aep-abstract-id15"><ce:section-title>Publisher Summary</ce:section-title><ce:abstract-sec view="all" id="aep-abstract-sec-id16"><ce:simple-para id="fsabs057" view="all">Many primary simian immunodeficiency virus (SIV) strains use CCR5 to infect simian cells in the absence of CD4. HIV/SIV transmission through an intact mucosal surface requires that the virus bind the epithelial cells and either infect these or be transmitted to underlying lymphoid cells. Langerhans cells and dendritic cells (DCs) also express the major coreceptor CCR5 for SIV/HIV binding and fusion. SIV infection of intact mucosa has been demonstrated by direct application of high doses of SIV into the vagina, rectum, or oral cavity of rhesus monkeys. There is experimental evidence in adult male rhesus macaques that they can be infected with SIV by the placement of cell-free virus onto their foreskins. SIV infected macaques showed predominantly IgG anti-SIV antibodies in vaginal secretions. Findings, both in SIV-infected rhesus macaques and in HIV-infected women, suggest that transudation of serum IgG antibodies is a significant source of antiviral antibodies in vaginal secretion. The development of HIV-SIV hybrid constructs consisting of HIV <ce:italic>env</ce:italic> gene in a SIV mac backbone has extended the use of macaques in immunization with some HIV antigens.</ce:simple-para></ce:abstract-sec></ce:abstract></dm:abstracts>

Antigenic specificity of antibodies in vaginal secretions during infection with Neisseria gonorrhoeae.

Antibodies in genital secretions of patients with gonorrhea have been shown to inhibit the attachment of gonococci to epithelial cells. The gonococcal antigens for which these antibodies are specific were studied by adsorption of the genital secretions from a patient infected with gonorrhea with purified lipopolysaccharide, outer membrane complex, or purified pili of homologous Neisseria gonorrhoeae and measurement of the reduction of inhibition of attachment of the gonococci to epithelial cells. The removal of antibodies was documented with the use of a solid-phase radioimmunoassay in which the amount of antibody in the adsorbed secretions that bound to a specific gonococcal antigen was shown to be reduced as compared with the amount of antibody in unadsorbed secretions. The antibody in the secretions that inhibited attachment was removed primarily by adsorption with the homologous pili, not with homologous lipopolysaccharide. A preparation of the homologous outer membrane complex that contained pili, cell-wall proteins, and lipopolysaccharide also blocked the inhibitory antibody.