Electroejaculation and assisted reproductive technologies in the treatment of anejaculatory infertility.

OBJECTIVE: To determine the efficacy of electroejaculation in combination with assisted reproductive technology (ART). DESIGN: Case series. SETTING: University fertility program. PATIENT(S): One hundred twenty-one consecutive couples seeking treatment of anejaculatory infertility. INTERVENTION(S): Electroejaculation with IUI, or gamete intrafallopian transfer or IVF. MAIN OUTCOME MEASURE(S): Pregnancy and pregnancy outcome. RESULT(S): Fifty-two couples became pregnant (43%), 39 by IUI alone (32.2%). Cycle fecundity for IUI was 8.7%. No difference in cycle fecundity was seen among ovarian stimulation protocols (clomiphene citrate, 7.6%, hMG, 13.2%, and natural cycle, 11.2%). Pregnancy was unlikely when the inseminated motile sperm count was <4 million. Female management protocol and etiology of anejaculation did not affect results. Patients undergoing IVF had higher cycle fecundity (37.2%) than did those undergoing IUI. The rates of spontaneous abortion and multiple gestations were 23% and 12%, respectively. CONCLUSION(S): Electroejaculation with stepwise application of ART is effective in treating anejaculatory infertility. Intrauterine insemination with the least expensive monitoring protocol should be used for most couples, because use of more expensive monitoring did not improve results. It is cost-effective to bypass IUI and proceed directly to IVF in men who require anesthesia for electroejaculation and in those with a total inseminated motile sperm count < 4 million.

Canine model of infertility after spinal cord injury: time course of acute changes in semen quality and spermatogenesis.

PURPOSE: We established a canine model of subfertility after spinal cord injury and examined the time course of acute changes in semen quality and spermatogenesis after spinal cord injury. MATERIALS AND METHODS: Seven dogs underwent surgical T7 spinal cord injury. Six dogs were used as controls. Electroejaculation and testicular fine needle aspiration were performed at baseline and twice weekly for 3 weeks after spinal cord injury. Semen quality change was examined by standard semen analysis. Spermatogenesis was assessed by flow cytometry of testicular fine needle aspiration in all dogs as well as by testicular histology at study conclusion in 4 controls and 4 spinal cord injured dogs. RESULTS: No significant changes in spinal cord injured dogs were noted before 3 weeks after injury. From baseline to 3 weeks after injury certain changes were evident in spinal cord injured dogs. Mean antegrade sperm motility decreased from 62.9% to 20.1% (p = 0.008), mean total sperm (antegrade plus retrograde total sperm) decreased from 423 to 294 x 106 which was not statistically significant, and the incidence of testicular haploid cells decreased from 75.6% to 48.3% (p = 0.028). No significant change in any parameter was present in control dogs. The mean number of mature spermatids per cross-sectional tubule on final testicular histology was significantly decreased in spinal cord injured dogs compared with controls (13.6 versus 43.9, p = 0.02). CONCLUSIONS: In the canine model tested the dogs readily survived spinal cord injury, electroejaculation was effective for obtaining ejaculate and fine needle aspiration allowed serial examination of spermatogenesis. Three weeks after spinal cord injury but not before 3 weeks sperm motility and spermatogenesis were significantly decreased. However, at the same point this decrease in spermatogenesis was not yet reflected in the total ejaculated sperm count.

Seminal vesicle aspiration in spinal cord injured men: insight into poor sperm quality.

PURPOSE: In men with spinal cord injury poor quality semen is seen when performing electroejaculation and penile vibratory stimulation. We determined whether sperm stasis within the seminal vesicles is a potential cause of this problem. MATERIALS AND METHODS: Seminal vesicle aspiration was performed immediately before electroejaculation or penile vibratory stimulation in men with aspermia secondary to spinal cord injury. Sperm count and quality of seminal vesicle aspiration and subsequent ejaculation were compared with historical ejaculated counts, ultrasound findings and patient characteristics. RESULTS: Mean total number of right plus left seminal vesicle sperm plus or minus standard deviation was 511 +/- 960 x 10(6). Mean total number of sperm obtained by seminal vesicle plus electroejaculation or penile vibratory stimulation was 918 +/- 1,261 x 10(6). Average motility and viability of the seminal vesicle aspirated sperm were 1.3 and 3.2%, respectively. Average motility of the ejaculated sperm was 26.4% after seminal vesicle aspiration versus 16.3% in previous ejaculation induction procedures performed in the same patients. Seminal vesicle aspirated sperm represented 66% of the total number of sperm obtained during the session and was equal to 49% of the sperm obtained at previous electroejaculation or penile vibratory stimulation sessions. The period of abstinence correlated only with ejaculate count (simple regression p = 0.009). No other clinical characteristics had any effect on sperm count or quality. CONCLUSIONS: Large numbers of poor quality sperm are present within the seminal vesicles of spinal cord injured men and these sperm comprise a large portion of the specimens collected by electroejaculation or penile vibratory stimulation. This phenomenon is independent of the period of abstinence, implicating disordered storage of sperm due to spinal cord injury rather than infrequent ejaculation. The large number of senescent sperm within the seminal vesicles appears to be a primary cause of poor sperm quality in spinal cord injured men.

Unilateral obstruction of the vas deferens diagnosed by seminal vesicle aspiration.

Causes of vasal obstruction include vasectomy, inguinal surgery, scrotal surgery, and congenital anomalies. The incidence of unilateral obstruction in various clinical situations is unknown because sperm from the contralateral testicle usually is present in the ejaculate. Vasography is the standard technique used to diagnose a unilateral vas deferens obstruction. We used the technique of seminal vesicle aspiration to accurately diagnose unilateral vas deferens obstruction in a man who had a previous inguinal hernia repaired with mesh. Surgical exploration confirmed the impression of vasal obstruction, and successful vasovasostomy was performed. We believe that seminal vesicle aspiration may be helpful in the diagnosis of unilateral vas deferens obstruction and has potential benefits over vasography.

Fertilization antigen-1 removes antisperm autoantibodies from spermatozoa of infertile men and results in increased rates of acrosome reaction.

OBJECTIVE: To determine if fertilization antigen (FA)-1 will remove autoantibodies from the surface of sperm cells of immunoinfertile men by immune adsorption and permit an increased acrosome reaction (AR). DESIGN: Prospective analytic study. SETTING: University medical center. PATIENT(S): Men from 18 infertile couples with autoantibodies present on their spermatozoa. INTERVENTION(S): Sperm samples after processing were examined for antibody binding and AR before and after adsorption with control medium or FA-1. MAIN OUTCOME MEASURE(S): Sperm-bound antibody was assessed by the immunobead assay (immunoglobulin [Ig] A and IgG) and the AR by induction with ionophore A23187. RESULT(S): Adsorption with FA-1 compared with control medium increased immunobead-free swimming sperm an average of 50% and 76% for IgA and IgG antisperm antibodies, respectively, with 78% and 100% of the 18 semen specimens increasing significantly. The AR rate increased an average of 10.3% compared with control medium and showed improvement in 78% of the sperm samples after FA-1 adsorption. CONCLUSION(S): The FA-1 sperm antigen appears to significantly free sperm cells coated with autoantibodies in the semen of most infertile men examined. Reducing sperm-bound antibodies that inhibited the AR allowed the sperm cells to undergo successful AR induction by calcium ionophore.

Expression of tumor necrosis factor-alpha and its receptors type I and type II in human oocytes.

The present study was conducted to examine the expression of tumor necrosis factor-alpha (TNF-alpha) and its receptors (types I and II, designated TNFR-I and TNFR-II, respectively) in human oocytes and cumulus cells at the mRNA and protein levels. mRNA expression was investigated using a reverse transcriptase-polymerase chain reaction (PCR)/Southern hybridization procedure. DNA-free RNA was isolated from the oocytes/cumulus cells, reverse-transcribed, and PCR-amplified using specific oligonucleotide primers based upon genomic/cDNA sequences. The expected bands of 303 bp and 513 bp were observed in oocytes and cumulus cells using primers based on genomic/cDNA sequences of TNF-alpha and TNFR-II, respectively, that hybridized with specific cDNA probes in Southern blot hybridization procedure. The expected band of 368 bp was not observed in oocytes and cumulus cells using primers based on the TNFR-I cDNA sequence. Similar results were observed for expression at the protein level, as seen by the immunoreactivity of the specific antibodies with the paraformaldehyde-fixed oocytes and cumulus cells in the indirect immunofluorescence technique (IFT). These results indicate that human oocytes and cumulus cells express TNF-alpha and its receptor type II (TNFR-II), and not type I (TNFR-I), both at the mRNA and protein levels. These findings provide further evidence and substantiate the proposed physiologic role of TNF-alpha in ovarian function, and may lead to clinical applications in in vitro fertilization programs and in diagnosis and treatment of infertility in women, especially in cases attributed to ovarian dysfunction.

Electroejaculation versus vibratory stimulation in spinal cord injured men: sperm quality and patient preference.

PURPOSE: We compared semen quality and patient preference between penile vibratory stimulation and electroejaculation in spinal cord injured men. MATERIALS AND METHODS: We treated 11 spinal cord injured men with penile vibratory stimulation and electroejaculation in random order. End points examined were semen analysis, sperm functional assessment, and patient pain scores (1 to 10) and preferred procedure. Differences between the procedures were determined with the paired Student t test. RESULTS: There was no difference in antegrade sperm count but penile vibratory stimulation specimens had greater motility (26.0 versus 10.7%), viability (25.2 versus 9.7%) and motile sperm count (185.0 x 10(6) versus 97.0 x 10(6)). The retrograde sperm count was greater (but not significant) in electroejaculation patients. The total (antegrade plus retrograde) and motile sperm counts were not different. There was no difference in immunobead test (all negative), cervical mucus penetration or sperm penetration assay, although the percent hamster egg penetration approached significance (53.7% for penile vibratory stimulation versus 22.1% for electroejaculation, p = 0.06). There was no difference in the peak blood pressures and no complications were noted. Pain scores were significantly greater for electroejaculation compared to penile vibratory stimulation (5.2 versus 1.7, respectively). All patients preferred penile vibratory stimulation. CONCLUSIONS: There was a slight advantage in sperm quality and a high patient preference in favor of penile vibratory stimulation. Penile vibratory stimulation should be attempted first to induce ejaculation in spinal cord injured men, with electroejaculation reserved for failures.

Procreation after death or mental incompetence: medical advance or technology gone awry?

OBJECTIVE: To review our experience with semen retrieval in men who are incompetent or dead and to formulate general medical, legal, and ethical guidelines for practitioners. DESIGN: Case series and literature review. SETTING: Academic. PATIENT(S): Seven incompetent or neurologically dead individuals in whom sperm retrieval was requested. INTERVENTION(S): Electroejaculation. RESULT(S): Seminal emission was induced in the two men who underwent electroejaculation. Sperm suitable for cryopreservation was obtained in one of these men. Review of the legal and ethical implications of such procedures led to development of general guidelines for determining whether gamete retrieval should be performed when requested. Issues of procreational autonomy, consideration of the decedent's wishes, and assurance of the well-being of any new life created were considered most strongly in the formation of these guidelines. CONCLUSION(S): Although the retrieval of sperm from deceased or incompetent individuals may be achieved readily, it is incumbent upon the practitioner to consider the legal and moral implications of these procedures before proceeding.

Luteal support with both estradiol and progesterone after clomiphene citrate stimulation for in vitro fertilization.

OBJECTIVE: To determine if pregnancy rates (PRs) for clomiphene citrate (CC)-stimulated IVF-ET can be increased by luteal support with E2 and P. DESIGN: Prospective randomized crossover clinical study. SETTING: Infertile women volunteers in an academic research environment. PATIENTS: Ninety-three infertile women underwent a total of 143 IVF-ET cycle using CC for ovulation induction. INTERVENTIONS: Each woman received either no luteal support (control group) or luteal support with both oral E2 (2 mg three times daily) starting on the day of retrieval and vaginal P suppositories (100 mg twice daily) starting on the day of ET. MAIN OUTCOME MEASURE: Clinical PR. RESULTS: In 79 of 143 (55%) of the cycles, at least one embryo was transferred. Compared with the control group (n = 35 cycles), the luteal support group (n = 44 cycles) had a significantly higher PR per retrieval (control: 2% versus luteal support: 16%) and were older (control: 33 +/- 4 versus luteal support: 35 +/- 4 years; mean +/- SEM). They did not differ in terms of E2 or P levels, endometrial thickness on the day of hCG, number of follicles > 16 mm in diameter, mature oocytes retrieved, or embryos transferred. CONCLUSIONS: Luteal support with both E2 and P significantly increase the clinical PRs for CC-stimulated IVF-ET.

Penile vibratory stimulation in spinal cord injured men: optimized vibration parameters and prognostic factors.

OBJECTIVE: To study the efficacy of penile vibratory stimulation (PVS) with optimized vibration parameters in spinal cord injured (SCI) men and to examine prognostic factors for success. DESIGN: Case series. SETTING: University hospital outpatient clinic. PATIENTS: Thirty-four consecutive SCI men seeking fertility treatment. INTERVENTION: PVS with optimized vibration parameters to induce reflex ejaculation. MAIN OUTCOME MEASURES: Ejaculatory response; semen analysis. RESULTS: Antegrade ejaculation was seen in 65% of patients. High rates were seen in lesions above T10 (81%) and in presence of hip flexion and bulbocavernosus reflexes (77%). Of men with lesions above T10, those with a penile prosthesis had lower ejaculation rates (40% vs 90%). Average total sperm counts were 968 million, with 26% motility. CONCLUSIONS: High rates of ejaculation are seen with optimized vibration parameters, especially in men with lesions above T10 and intact lower spinal reflexes. A penile prosthesis may impair success with PVS.

Assessment of sperm function and clinical aspects of impaired sperm function.

Fertility is dependent on a complex set of events, involving both male and female components. Normal sperm function involves many steps, including motility, capacitation, acrosome reactivity and, ultimately, fertilization of the oocyte. While male fertility is most often assessed by means of gross semen parameters, infertility may also be caused by abnormal sperm function, and only by performing specific tests of this function, may the reasons for infertility become evident. Specific tests which may be helpful include semen analysis, detailed sperm motility assessment, motility longevity, hypo-osmotic swelling test, mucus penetration assay, acrosome reactivity, antisperm antibody tests, sperm penetration assay and in vitro fertilization. Relatively well-defined syndromes of abnormal sperm function include immunologic infertility, immotile-cilia syndrome, anejaculation and nifedpine-associated infertility.

Treatment with thymosin alpha-1 increases fertilizing capacity of sperm of infertile men: a multicenter trial.

Thymosin alpha-1 (T alpha 1) significantly enhances the fertilizability of sperm from fertile men by increasing capacitation and/or acrosome reaction. The present study was conducted to investigate the effect of T alpha 1 on fertilizing capacity of sperm of infertile men using the sperm penetration assay (SPA) in a multicenter clinical trial. T alpha 1 significantly (p = .0006 to < .0001) increased (31 to 45%) fertilizing capacity of sperm of 76% (50/68) of infertile men as defined by the sperm penetration index in SPA. The enhancing effect of T alpha 1 and the degree of enhancement depended on the T alpha 1 concentration in the seminal plasma, since these parameters showed a significant intercorrelation (r = .65 to .74, p = .039 to .01). It would appear that T alpha 1 may have clinical application in the diagnosis and treatment of male infertility especially mediated through defective sperm function.

Levels of interferon-gamma and tumor necrosis factor-alpha in sera and cervical mucus of fertile and infertile women: implication in infertility.

Concentrations of two immune cytokines, namely interferon-gamma (INF-gamma) and tumor necrosis factor-alpha (TNF-alpha), were determined in the sera and cervical mucus samples of fertile (n = 16), idiopathic infertile (n = 44), and immunoinfertile women (n = 45) to investigate their role, if any, in female infertility. Sera of idiopathic infertile women demonstrated significantly (P < 0.0001) higher levels of INF-gamma compared to those in fertile controls, whether expressed as pg/ml or pg/mg serum protein. Similarly, sera of immunoinfertile women demonstrated significantly (P = 0.0008) higher levels of INF-gamma compared to fertile controls and idiopathic infertile women. Cervical mucus of idiopathic infertile women also demonstrated significantly (P < 0.0001) higher concentrations of INF-gamma compared to those in fertile controls. Cervical mucus of immunoinfertile women demonstrated significantly (P < 0.0001) higher concentrations of INF-gamma compared to those in fertile controls and idiopathic infertile women. INF-gamma levels in serum did not significantly (P > 0.05) correlate (r = 0.12-0.43) with the concentrations in cervical mucus, when all the three groups were compared together. However, when the serum levels were compared with the cervical mucus concentrations by condition, only the idiopathic infertile group showed a significant (P = 0.005) correlation (r = 0.70). Serum levels of TNF-alpha did not differ significantly (P > 0.05) among three groups of women. Cervical mucus concentrations of TNF-alpha, however, varied among groups with levels being significantly (P = 0.04) higher-in idiopathic infertile women compared with fertile controls and in immunoinfertile women significantly (P = 0.0007) higher than in fertile controls as well as idiopathic infertile women. TNF-alpha levels in serum correlated (r = 0.65) significantly (P < 0.001) with the concentrations in cervical mucus when all the three groups were compared together or individually by infertility condition. These findings suggest the involvement of cytokines in infertility, and thus may have potential applications in diagnosis and treatment of female infertility.

Electroejaculation does not impair sperm motility in the beagle dog: a comparative study of electroejaculation and collection by artificial vagina.

Sperm samples obtained by electroejaculation (EEJ) in men with anejaculatory infertility have a markedly lower quality than those obtained by normal ejaculation. An electrical effect of the EEJ procedure has been implicated by some investigators as a direct cause of low sperm quality. To determine whether the EEJ procedure causes direct sperm damage, we compared ejaculates obtained from dogs by EEJ and by artificial vagina (AV). In seven adult beagle dogs, semen was collected weekly, alternating between the two procedures. The average (antegrade) sample from AV had a volume of 2.6 ml., sperm concentration of 150.1 x 10(6) per ml., total sperm count of 381.7 x 10(6) and motility of of 26.3%. The average antegrade sample from EEJ had a volume of 1.8 ml., a concentration of 129.6 x 10(6) per ml., a total sperm count of 166.8 x 10(6) and a motility of 30.1%. Of these differences only the total sperm counts and the total motile sperm counts were statistically significant (p < 0.05). There were no significant differences between the antegrade motilities, total sperm counts (antegrade plus retrograde-381.7 versus 243.4 x 10(6), for AV and EEJ, respectively) or the total motile sperm counts from the two procedures (103.9 versus 78.0 x 10(6)). There were no differences in the average curvilinear velocity (VCL) measured by computer-assisted sperm motion analysis (56.9 mu. per second for AV, 47.4 mu. per second for antegrade EEJ specimens and 41.7 mu. per second for retrograde EEJ specimens). Since routine semen parameters between artificial vagina and electroejaculation did not differ in dogs, we conclude that the electroejaculation procedure is not responsible for the gross semen abnormalities reported in electroejaculation of anejaculatory men.

Antisperm antibodies: origin, regulation, and sperm reactivity in human infertility.

OBJECTIVE: To follow-up and expand discussion on the action mechanisms of antisperm antibodies in human infertility, the etiology and control of antisperm antibody induction, sperm antigens involved in immunoinfertility, and strategies for therapy. DESIGN: A review of the recent literature with an emphasis on female immunoinfertility. RESULTS: The role of antisperm antibodies in clinical infertility continues to be defined. Through assisted reproductive technologies, antisperm antibodies were shown to exert detrimental effects on different prefertilization and possibly postfertilization events. The female reproductive tract is part of the common mucosal immune system and is able to mount effective immune responses against infectious agents, foreign antigens, and, occasionally, sperm cells. Sperm membranes and constituents contain numerous antigenic components foreign to the human body, and yet antisperm antibodies become problematic in few women exposed to semen. Semen and sperm cells contain immunosuppressive factors capable of inhibiting different immune cells. Fertile women apparently produce antisperm antibodies but also possess neutralizing serum anti-idiotypic antibodies that are lacking in virgin and immunoinfertile women. CONCLUSIONS: Antisperm antibodies can affect adversely human fertility but normally may be controlled by anti-idiotypic antibodies, which along with immunosuppressor factors in semen prevent their induction to a significant degree. This balance between detrimental and "beneficial" immune response to sperm may be shifted toward an antisperm antibody response by stimulatory factors such as infection. Therapies may be devised to stimulate the anti-idiotypic antibody system, to induce immune tolerance to sperm antigens, and to use antigens to adsorb antisperm antibodies from spermatozoa.

Antiidiotypic antibodies to sperm in sera of fertile women that neutralize antisperm antibodies.

The presence of antiidiotypic antibodies (ab-2) to sperm was investigated in the sera of fertile, infertile, and virgin women using sperm-specific anti-FA-1 monoclonal antibody Fab'.ab-2 were detected in 71% (17/24) of sera from fertile women and in none (0/12) of the sera from virgin females by the enzyme-linked immunosorbent assay, Western blot procedure, and immunoprecipitation procedure. Sera from infertile women that had antisperm antibodies showed a minimal presence of ab-2, with only three sera (13%, 3/23) demonstrating the presence of low levels of ab-2. The ab-2 present in fertile women were capable of neutralizing the fertilization-inhibitory activity of anti-FA-1 antibody in a concentration-dependent manner in a human sperm penetration assay (SPA) of zona-free hamster oocytes. ab-2 were also capable of inhibiting the binding of antisperm antibodies to the sperm surface as determined by the immunobead binding technique. This is the first report demonstrating the presence of ab-2 in the sera of fertile women that are capable of neutralizing antisperm antibodies present in sera of infertile women. These findings suggest that the inability to detect antisperm antibody activity in the sera of fertile women may be due to higher levels of ab-2 present in these sera than levels found in sera of infertile women, although both groups may be producing antisperm antibody response after sexual exposure to sperm.

Immunoglobulin (Ig) G, IgA, and IgA subclass antibodies against fertilization antigen-1 in cervical secretions and sera of women of infertile couples.

OBJECTIVES: To assess the occurrence of immunoglobulin (Ig) G, IgA, and IgA subclass antibodies against human sperm fertilization antigen-1 (FA-1) in cervical mucus (CM) and serum of women of infertile couples. DESIGN: Enzyme-linked immunosorbent assay methodology was used to detect anti-FA-1 antibodies. Antisperm antibodies were detected by agglutinating, immobilizing, and indirect immunobead (IB) methods. Control samples for the ELISA were from 10 women negative in the antisperm antibody assays. PARTICIPANTS: Samples were from women of 32 infertile couples undergoing antisperm antibody analysis. RESULTS: One of 10 control CM samples was slightly positive for IgG anti-FA-1 and none for IgA. Of the 22 CM samples from antisperm antibody-positive women, 9 were positive for IgG antibodies, 9 for IgA, 7 for IgA1, and 6 for IgA2. Cervical mucus samples from eight women were positive for both IgA and IgG antibodies. Assay of 19 serum samples, including 8 controls, by ELISA, indicated 9 of 11 from antisperm antibody-positive women and none from controls were positive for IgA and IgG (7 of 9 identical women). In addition, of the nine IgA-positive sera, seven were of the A1 subclass and five were of the A2 subclass. Positive IB assays occurred more frequently in CM and serum samples positive for anti-FA-1 antibodies than in negative samples. CONCLUSION: The results suggest that cervical secretions and sera of antisperm antibody-positive women contain IgA and IgG antibodies against sperm antigen FA-1 that may be involved in antifertility effects.

Surface expression of secretory component and HLA class II DR antigen on glandular epithelial cells from human endometrium and two endometrial adenocarcinoma cell lines.

The expression of secretory component (SC) by human glandular endometrial cells cultured in vitro was significantly increased by estradiol in the medium. Interferon-gamma and interleukin-4 stimulated the expression of SC only in the presence of estrogen. Tumor necrosis factor-alpha plus estrogen also caused a significant increase in the number of cells expressing SC. HLA class II antigen DR was detected on few glandular epithelial cells of human endometrium cultured in control medium, whereas interferon-gamma and interleukin-4, but not tumor necrosis factor-alpha, caused significant increases in the expression of DR. Estrogen in the culture medium did not significantly affect DR expression. The human endometrial adenocarcinoma cell lines, HEC and RL-95, expressed SC in approximately 50 and 20% of the cells. Also, approximately 20% of the RL-95 cells stained for DR antigen. Interferon-gamma did not influence the degree of expression of either surface marker of the two cell lines. Cells of both lines bound polymeric IgA and IgM but showed little to no binding of monomeric IgA, IgG, or an IgM previously shown not to bind SC.

Immune response of the female rat genital tract after oral and local immunization with keyhole limpet hemocyanin conjugated to the cholera toxin B subunit.

The immune response of the female rat genital tract was evaluated with Lewis rats given primary and secondary immunizations with keyhole limpet hemocyanin (KLH) alone or coupled to the cholera toxin (CT) B subunit (CTB) by the oral or intravaginal-uterine route or a combination of routes. CT (2 to 5 micrograms) was administered as an adjuvant with the KLH-CTB conjugate. While a significant mucosal immunoglobulin A (IgA) response was induced by KLH, there were no significant differences among the immunized groups in the levels of IgA antibodies in salivary gland, gut, vaginal, and uterine secretions, with the exception that rats immunized only orally with the KLH-CTB conjugate lacked a detectable vaginal response. Levels of IgA antibodies to CT, however, were significantly increased in genital tract secretions of rats immunized locally versus orally with the KLH-CTB conjugate. Antibody activity of the IgG isotype against both KLH and CT was significantly elevated in genital tract secretions of rats immunized with KLH-CTB by the oral or intravaginal-uterine route and given genital tract boosters, in comparison with the results for the other groups. IgM antibody titers were generally negligible in the different secretions. An enzyme-linked spot-forming assay revealed IgA and IgG antibody-secreting cells in salivary gland and uterine tissues. A highly significant correlation between the numbers of antibody-secreting cells and antibody titers existed for uterine IgG but not IgA responses to KLH among the different groups of rats. In conclusion, a vigorous local immune response was induced after immunization of the female rat reproductive tract alone or in combination with peroral challenge with the KLH-CTB conjugate.

Comparison of intracervical, intrauterine, and intratubal techniques for donor insemination.

OBJECTIVE: To compare the efficacy of intracervical insemination (ICI), intrauterine insemination (IUI), and a combination of intratubal and intrauterine insemination (ITI/IUI) for donor insemination. DESIGN: Prospective randomized clinical trial. SETTING: The University of Michigan donor insemination program. PATIENTS, PARTICIPANTS: Forty-one women undergoing donor insemination with cryopreserved sperm for either isolated male factor or male factor plus ovulatory dysfunction corrected by clomiphene citrate. INTERVENTION: Each patient was randomly assigned to receive each of the three insemination techniques in consecutive cycles until pregnancy occurred or the patient dropped from the study. MAIN OUTCOME MEASURES: Cycle fecundity rates were compared using the chi 2 test, and cumulative pregnancy rates (PRs) determined by life table analysis were compared using a log-rank test. RESULTS: Cycle fecundity rate was significantly higher for IUI (18.3%) than for ICI (3.9%) or ITI/IUI (7.3%). By life table analysis, the cumulative PR for IUI was significantly higher than for ICI, but the PR for ITI/IUI was not. CONCLUSION: For donor insemination with cryopreserved sperm, IUI increases cycle fecundity compared with ICI. The addition of ITI to IUI, however, interferes with the apparent beneficial effect of IUI alone.