| HOME | HELP | FEEDBACK | SUBSCRIPTIONS | ARCHIVE | SEARCH | TABLE OF CONTENTS |
| ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
From the * Center for Human Reproduction, North
Shore University Hospital, New York University School of Medicine, Manhasset,
New York; the Department of Obstetrics and
Gynecology, Prince of Wales Hospital, the Chinese University of Hong Kong,
Hong Kong, China; the Departments of Urology
and Obstetrics and Gynecology, University of Iowa, Iowa City, Iowa; and the
Department of Urology, Medical College of
Wisconsin, Milwaukee, Wisconsin.
| Correspondence to: Dr Huai L. Feng, Center for Human Reproduction, North Shore University Hospital, NYU School of Medicine, Manhasset, New York 11030 (e-mail: hfeng{at}nshs.edu). |
| Received for publication January 3, 2008; accepted for publication March 10, 2008. |
 |
Abstract |
|---|
 Top Abstract Materials and MethodsResultsDiscussionReferences |
|---|
Key words: Infertility, sperm function
Numerous methods for detection of anti-sperm antibodies are now available. Detection of antibody-bound sperm may be achieved with the immunobead-binding test (Clarke et al, 1985b) or mixed antiglobulin reaction test (Jager et al, 1978), or sperm proteins can be separately identified by two-dimensional gel electrophoresis and transferred to nitrocellulose membranes and incubated with sera from fertile women or immunoinfertile women with anti-sperm antibodies (Bhande and Naz, 2007). The immunobead assay allows for localization of the antigen and identification of specific immunoglobulin (Ig) isotypes. Other assays, such as the tray agglutination and sperm immobilization tests, detect adverse effects of antibodies on sperm agglutination or motility (Isojima et al, 1968; Friberg, 1974). Although these tests can detect anti-sperm antibodies and may yield relatively specific information about the antibodies, the results provide the clinician little indication whether or not sperm exposed to the anti-sperm antibodies are capable of reaching the site of fertilization or binding and penetrating an oocyte.
Treatment options for infertile female patients with anti-sperm antibodies range from timed intercourse to in vitro fertilization with intracytoplasmic sperm injection. Some anti-sperm antibodies, such as tail tip–directed antibodies, do not necessarily preclude couples from achieving pregnancy following intercourse (Bronson et al, 1984). Intrauterine insemination may be effective therapy for overcoming cervical mucus anti-sperm antibodies that interfere with sperm motion and prevent sperm from progressing beyond the cervix. However, intrauterine insemination does not allow sperm to bypass anti-sperm antibodies that dwell in the uterine and tubal fluids. In vitro fertilization may facilitate fertilization for patients who have anti-sperm antibodies in uterine or tubal fluid that cause sperm agglutination or otherwise prevent sperm-oocyte binding (Clarke et al, 1985a, 1986; Mandelbaum et al, 1987). Autosperm antibodies that adversely affect sperm binding and penetration of the oocyte may require intracytoplasmic sperm injection to achieve fertilization in vitro.
The varying degree to which the antibodies impair fertilization suggests that detection of anti-sperm antibodies, identification of relevant sperm surface antigens, and identification of the antigens' role in the fertilization process would facilitate formulation of an effective therapeutic plan to treat patients' infertility. Additionally, identification of sperm antigens that evoke production of anti-sperm antibodies that occlude fertilization may contribute to applications in specific diagnosis, treatment of infertility/immunoinfertility, and development of a new generation of contraceptive modalities (Bhande and Naz, 2007). The objective of this study was to locate antigen-binding sites and determine the mechanism by which anti-sperm antibodies from an infertile female patient's serum interfere with sperm function.
|
Materials and Methods |
|---|
|
TopAbstractMaterials and MethodsResultsDiscussionReferences |
|---|
Anti-Sperm Antibody Detection by Indirect Immunobead Assay
Immunobeads—The Ig class of anti-sperm antibodies was
determined by the indirect immunobead-binding test with rabbit anti-human IgA,
IgM, IgG immunobeads (IBT; Irvine Scientific, Irvine, California). The
anti-sperm antibody titer was established using nonspecific rabbit anti-human
Ig (L + H) immunobeads. Complete details of indirect IBT methods have been
reported elsewhere (Clark et al, 1984, 1985b). Lyophilized immunobeads were
washed and resuspended in Dulbecco phosphate-buffered saline (D-PBS; Life
Technologies, Grand Island, New York) supplemented with 5% bovine serum
albumin (BSA; Sigma-Aldrich, St Louis, Missouri).
Sperm Preparation and Passive Antibody Transfer—All assays were performed with freshly ejaculated sperm from a donor who had previously tested negative for anti-sperm antibodies. Following liquefaction, the semen was diluted with pre-equilibrated Biggers, Whitten, and Wittingham (BWW) medium supplemented with 0.4% BSA and washed 3 times by centrifugation at 300 x g for 6 to 8 minutes. After a third wash, the pellet was resuspended in BWW-BSA, and the final concentration of motile sperm was adjusted to 50 to 100 million motile sperm/mL. A 100-µL aliquot of motile sperm (5–10 x 106) was added to 1.6 mL of the patient's serum or anti-sperm antibody–negative serum diluted 1:4 with BWW. The sperm were incubated in the serum-supplemented BWW for 30 minutes, washed 3 times by centrifugation with BWW-BSA, and resuspended at a final concentration of 20 x 106 motile sperm/mL. Incubation periods of 30 minutes and 18 hours were used to determine the anti-sperm antibody titer. An indirect IBT assay was performed with IgG, IgM, IgA, and nonspecific immunobeads immediately after sperm resuspension.
Indirect IBT Assay—Ten microliters of sperm were mixed with 10 µL of immunobeads on a slide and incubated for 10 minutes at room temperature in a humidity chamber. Two sets of 100 progressively motile sperm were scored for total bead binding and regional distribution of immunobead binding. Four replicates of the indirect immunobead assay were performed for IgG, IgM, IgA, and nonspecific Ig (L + H). The anti-sperm antibody titer was determined using 2 additional replicates of the indirect immunobead assay with nonspecific Ig (L + H) beads for sperm incubated for 30 minutes and 18 hours in serum dilutions of 1:4, 1:64, 1:128, 1:512, and 1:1024 in BWW.
Assessment of Sperm Function
Passive Transfer and Ionophore Challenge—Motile donor sperm
(95% ± 0.9% motility) were collected by swim-up into BWW-BSA for 1
hour. Motile sperm (10 x 106) were incubated in 1:4, 1:8,
1:16, and 1:32 dilutions of anti-sperm antibody–positive or anti-sperm
antibody–negative serum in BWW with 0.3% BSA (fraction V) for 3 hours.
At the conclusion of the passive antibody transfer, samples were centrifuged,
the supernatant was removed, and the pellet was resuspended in 2 mL of
BWW-BSA. The final suspensions were aliquoted in 0.5-mL volumes into test
tubes. The acrosome reaction (AR) was induced in 1 aliquot of each treatment
by incubation with 10 µM A23187 calcium ionophore (stock solution in
dimethyl sulfoxide; Sigma) at 37°C under 5% CO2 in air for 1
hour. Sperm motility, agglutination, and acrosomal status of each treatment
group were assessed immediately after the 3-hour passive antibody transfer and
AR induction with the calcium ionophore.
Sperm Viability—Acrosome-intact and acrosome-reacted sperm suspensions were incubated for 15 minutes at room temperature with Hoechst 33258 stain (bis benzimide; Sigma) diluted in PBS at a concentration of 1 µg/mL. The free stain was washed from the sperm suspensions by 2 cycles of centrifugation (450 x g for 8 minutes), and the pellet was resuspended in PBS. Following the second wash, the supernatant was removed and the pellets were resuspended in 200 µL of 95% ethanol and incubated on ice for 30 minutes. Approximately 30 to 40 µL of each sperm suspension was smeared on an ethanol-precleaned glass slide and air-dried. Sperm were examined under epifluorescence. Viability was confirmed by stain exclusion.
AR—The AR was quantified by rhodamine isothiocyanate (RITC)–labeled Pisum sativum agglutin (PSA; Vector Laboratories, Inc, Burlingame, California) as described by Cross et al (1989). Briefly, sperm smears prepared after viability staining were incubated in a humidity chamber at room temperature with 100 µg/mL RITC-labeled PSA in distilled water for 10 minutes. After incubation, the slides were rinsed 3 times with distilled water and allowed to air-dry, and 200 sperm were examined under epifluorescence. Sperm were scored as acrosome intact when staining was uniform over the entire acrosome region. Sperm devoid of any intense labeling or labeling limited to the equatorial region were considered acrosome reacted. A minimum of 200 viable (Hoechst 33258–excluded) spermatozoa were scored for RITC-PSA binding.
Sperm Agglutination—The sperm agglutination procedure was modified from the tray agglutination test described by Friberg (1974). Sperm were scored for degree of agglutination prior to addition of anti-sperm antibody–positive (1:4, 1:8, 1:16, 1:32) and anti-sperm antibody–negative serum (1:4), following a 3-hour incubation in serum-supplemented BWW and after AR induction by the calcium ionophore. The degree of sperm agglutination was observed at a magnification of 100x on an inverted microscope. Agglutination was scored as a percentage of motile sperm that demonstrated head-to-head, head-to-tail, or tail-to-tail agglutination.
Indirect Immunochemical Technique
Localization of Anti-Sperm Antibodies by Light Microscopy—
The procedure followed that found in previous reports (Feng et al,
1997,
1998,
2004). Swim-up sperm (acrosome
intact) were air-dried onto slides and prepared for immunochemistry staining
in the following manner: 1) cells were fixed with 100% methanol for 15
minutes, permeabilized by a 5-minute exposure to Carnoy solution at
–20°C, and then rinsed twice in PBS (pH 7.4); 2) unfixed specimens
were air-dried on slides. Both fixed and unfixed slides were blocked with PBS
containing 2% BSA (w/v) and 2% normal goat serum (Sigma) for 1 hour followed
by overnight incubation with the infertile patient's serum or anti-sperm
antibody–free, normal human serum (Sigma) at 4°C. After thorough
washing in PBS, immunoperoxidase staining was performed with an
avidin–biotinylated complex kit (Zymed Laboratories, Carlsbad,
California) according to the manufacturer's instructions.
Localization of Anti-Sperm Antibodies by Electron Microscopy—The procedure followed that found in previous reports (Feng et al, 1997, 2004). Swim-up sperm were immersed in 3% paraformaldehyde containing 0.5% glutaraldehyde in PBS for 20 minutes at 4°C. Specimens were washed 3 times with PBS at 4°C, quenched to neutralize any reactive aldehyde groups with 0.1 M glycine for 15 minutes, and washed twice again in PBS. The samples were dehydrated in ethanol and embedded in Spurr resin (Ted Pella, Inc, Redding, California). Ultrathin sections were cut on an ultramicrotome and mounted on nickel grids (300 mesh). The sections were treated with 0.2% Triton X-100 for 10 minutes, rinsed with PBS, and then incubated with 2% BSA and 2.5% normal goat serum for 30 minutes at room temperature to block nonspecific sites. Grids were incubated in either the infertile patient's serum or normal, anti-sperm antibody–free human serum diluted 1:2 with 2% BSA in PBS for 48 hours at 4°C. After rinsing (5 times for 3 minutes each) in PBS, the grids were incubated overnight at 4°C in protein A conjugated to 10-nm colloidal gold (Sigma) diluted 1:25 in 1% BSA in PBS. The grids were washed in PBS, refixed for 20 minutes in 2.5% glutaraldehyde, rinsed in PBS, and stained with 5% uranyl acetate and lead citrate. Specimens were viewed and photographed with a Hitachi 7000 electron microscope.
Western Blot Analysis
Intact, fresh anti-sperm antibody–free human spermatozoa (10 samples)
were washed twice with PBS (pH 7.4) containing 5 mM EDTA at 4°C. Sperm was
extracted with 1% sodium dodecyl sulfate (SDS) in Tris-HCl containing 1 mM
phenyl-methylsulphonyl fluoride for 2 hours in an ice bath. The cellular
debris was centrifuged at 800 x g for 10 minutes, and the
supernatant removed. Protein concentrations were determined
spectrophotometrically. Samples were diluted in SDS sample buffer, separated
by 12% SDS–polyacrylamide gel electrophoresis, transferred to
nitrocellulose paper, and blocked with 5% powdered milk in TBS buffer (10 mM
Tris-HCl [pH 8.0], 150 mM NaCl) for 1 hour at room temperature. The blots were
postwashed in TBST (TBS containing 0.05% Tween-20) and then incubated
overnight with either the infertile patient's serum or normal human serum
diluted to 1:5 with PBS containing 2.5% BSA. The blots were washed in TBST,
incubated for 1 hour with alkaline phosphatase–conjugated anti-human Ig
(Cappel, Durham, North Carolina) diluted to 1:15 000 in TBST, and then
visualized with freshly prepared 5-bromo-4-chloro-3-indolyl-phosphate/nitro
blue tetrazolium; finally, the reaction was blocked with EDTA, and the
membranes were rinsed with
water.
|
Statistical Analysis
Data were analyzed for significant differences using 
2
analysis. Significance was defined as P < .05.
|
Results |
|---|
|
TopAbstractMaterials and MethodsResultsDiscussionReferences |
|---|
|
|
|
|
Western Blot Analysis
Circulating anti-sperm antibodies reacted with 4 sperm surface antigens, 35
kd, 40 kd, 47 kd, and 65 kd (Figure 5, Lane
A). Control, anti-sperm antibody–negative serum did not
react with alkaline phosphatase–conjugated anti-human Ig second antibody
(Figure 5, Lane B).
|
|
Discussion |
|---|
|
TopAbstractMaterials and MethodsResultsDiscussionReferences |
|---|
Characterization of the infertile woman's serum anti-sperm antibodies has revealed that the antibodies were generated against a heterogeneous population of sperm surface antigens. The dominant Ig class was IgG (79%), with the majority of the IgG immunobeads bound to the sperm head. The prevalence of IgG anti-sperm antibodies in serum has been reported by others and is consistent with a systemic immune response (Clarke et al, 1985b; De Almeida et al, 1991). Large prospective and retrospective studies have failed to provide conclusive evidence of an association between specific serum anti-sperm antibody isotypes or antibody titers and women's fertility (Eggert-Kruse et al, 1989; Check et al, 1995). Lack of correlation between a woman's fertility status and serum anti-sperm antibodies may be partially attributed to the fact that serum anti-sperm antibodies do not necessarily reflect the anti-sperm antibodies secreted in the reproductive tract. Stern et al (1992) could not find direct correlation between anti-sperm antibody titers or Ig isotypes in serum, cervical mucus, and peritoneal fluid. Kutteh et al (1995) reported discrepancies in the IgA subclasses detected in infertile patients' serum and cervical mucus. Despite these discrepancies, some systemic anti-sperm antibodies may be of value for diagnosis of infertility and for treatment. Investigators have suggested that systemic and local immune responses may be triggered by a common antigen and that IgG and IgA isotypes may recognize the same antigen (Marthur et al, 1987; Haas et al, 1990; Snow and Ball, 1992).
Fertilization in vivo requires sperm capacitation, AR, and sperm binding and penetration of the zona pellucida. Previous studies using the hamster oocyte penetration assay or the hemi–zona pellucida–binding assay have demonstrated that sperm function is impaired when these assays are performed in the presence of anti-sperm antibody sera from infertile women (Tsukui et al, 1988; Shibahara et al, 1993; Hall et al, 1994; Shibahara et al, 1996). Additionally, supplementation of culture media with anti-sperm antibody–positive sera has been shown to reduce fertilization rates in vitro (Clarke et al, 1985a; Mandelbaum et al, 1987). Results reported in these articles clearly show that not all patients' sera block sperm-zona binding or penetration of the oocytes to the same degree. This response supports the hypothesis that antibodies are generated in response to a diverse population of antigens.
It has been well documented that the ability of anti-sperm antibodies to bind to sperm surfaces may change after sperm undergo capacitation or the AR (Fusi and Bronson, 1990; Monroe et al, 1990; Margalioth et al, 1992; Bohring and Krause, 2003). We did not see a change in the degree of immunobead binding after 18 hours of incubation; however, the initial 79% binding for nonspecific Ig immunobeads may have masked an increase in immunobead binding after capacitation. We observed similar immunolabeling patterns and intensity for both capacitated and noncapacitated spermatozoa (data not shown). Furthermore, capacitated and preacrosome-reacted sperm still remained immunogold particles (Figure 4C).
The AR induction by ionophore challenge test was performed to evaluate the effect of the anti-sperm antibodies on the AR. The percentage of sperm undergoing spontaneous AR after 3 hours of incubation in 1:4, 1:8, 1:16, or 1:32 dilutions of anti-sperm antibody–positive serum or 1:4 dilutions of anti-sperm antibody–negative serum did not differ. The frequency of AR following exposure to the calcium ionophore was significantly reduced when sperm were incubated in anti-sperm antibody–positive serum. Our results were similar to those of previous studies that reported suppression of spontaneous and/or calcium ionophore–induced ARs when sperm are incubated in anti-sperm receptor antibodies (Feng et al, 1997, 1998, 2004) and anti-sperm antibody–positive serum (Tsukui et al, 1988; Benhoff et al, 1993; Tasdemir et al, 1995). Benhoff et al (1993) suggested that the changes in the phospholipid membrane sterol content associated with capacitation may be inhibited by anti-sperm Ig. This hypothesis was based on demonstration of anti-sperm Ig's ability to inhibit capacitation-related decreases of plasma membrane cholesterol content and increases of mannose receptor surface expression. Also, anti-sperm antibodies may block calcium influx by passing calcium channel or calcium-ATPase activities, resulting in reduced calcium ionophore–induced ARs (Feng et al, 2006, 2007).
Sperm agglutination was observed during the ionophore challenge test. Agglutination was enhanced by exposure to increasing concentrations of anti-sperm antibody–positive serum. There was no evidence of sperm agglutination after induction of the AR with calcium ionophore A23187. Sperm agglutination after induction of the AR was reduced to less than 10% in all treatment groups, whereas the proportion of acrosome-reacted sperm was suppressed in a dose-response fashion (Figure 3). These results suggest that the calcium ionophore caused a change in antigen expression in the plasma membrane that reduced sperm agglutination but did not overcome the antibodies' ability to block capacitation and/or the AR.
Acrosome-intact sperm proteins at 35 kd, 40 kd, 47 kd, and 65 kd reacted
with the anti-sperm antibodies. Sperm surface proteins of similar molecular
weights previously were recognized by infertile patients' circulating
antibodies (Snow and Ball,
1992; Benhoff et al,
1993; Domagala et al,
2007). Snow and Ball
(1992) reported
immunoreactivity of sera from male and female infertility patients toward the
35-kd, 40- to 45-kd, and 66-kd sperm proteins correlated with detection of
anti-sperm antibodies by the immunobead assay. Their Western blot analysis of
proteins isolated from subcellular sperm fractions suggested that the 66-kd
and 34- to 35-kd proteins recognized by circulating antibodies were located on
the inner acrosomal membrane. The serum from infertile men indicated that the
anti-sperm antibody directed against purified 57-kd protein inhibited binding
of human sperm to zona-free hamster oocytes in a dose-dependent manner. This
protein (57 kd) was localized to the head of nonacrosome-reacted spermatozoa
and shifted to the equatorial segment in acrosome-reacted spermatozoa
(Rajeev and Reddy, 2004;
Paradowska et al, 2006).
Another report distinguished a panel of antigens localized exclusively on the
surface of sperm cells and reacting with anti-sperm antibody–positive
sera (in the molecular weight range of 15 to 115 kd); some of them seem to
present novel entities as judged by the molecular weights
(Domaga
a and Kurpisz,
2004). These results are in contrast to our findings, which
indicate that the antigens are located on the plasma membrane over the
acrosomal region.
In summary, our data demonstrated that IgG, head-directed anti-sperm antibodies isolated from this particular patient interfere with fertilization through mechanisms that increase sperm agglutination. Antigen localization efforts revealed that our patient's anti-sperm antibodies are directed toward antigens that are present on the plasma membrane of noncapacitated, and perhaps, capacitated sperm. These findings indicate that evaluation of sperm function in the presence of the anti-sperm antibodies may provide valuable information when a course of infertility therapy is selected. Further characterization of the antigens recognized by these Ig may be useful for applications in specific diagnosis and treatment of infertility/immunoinfertility.
|
Footnotes |
|---|
|
References |
|---|
|
TopAbstractMaterials and MethodsResultsDiscussionReferences |
|---|
Benhoff S, Cooper G, Hurley I, Mandel FS, Rosenfeld DL. Antisperm antibody binding to human sperm inhibits capacitation induced changes in the level of plasma membrane sterols. Am J Reprod Immunol. 1993; 30: 113 –130.[Medline]
Bhande S, Naz RK. Molecular identities of human sperm proteins reactive with antibodies in sera of immunoinfertile women. Mol Reprod Dev. 2007;74: 332 –340.[CrossRef][Medline]
Bohring C, Krause W. Characterization of spermatozoa surface antigens by antisperm antibodies and its influence on acrosomal exocytosis. Am J Reprod Immunol. 2003; 50: 411 –419.[Medline]
Bronson R, Cooper G, Rosenfeld D. Sperm antibodies: their role in infertility. Fertil Steril. 1984; 42: 171 –183.[Medline]
Check JH, Katsoff D, Bollendorf A, Callan C. The effect of sera antisperm antibodies in the female partner on in vivo and in vitro pregnancy and spontaneous abortion rates. Am J Reprod Immunol. 1995; 33: 131 –133.[Medline]
Clarke GN, Hsieh C, Koh SH, Cauchi MN. Sperm antibodies, immunoglobulins, and complement in human follicular fluid. Am J Reprod Immunol. 1984;5: 179 –181.[Medline]
Clarke GN, Lopata A, Johnston WIH. Effect of sperm antibodies in females on human in vitro fertilization. Fertil Steril. 1986;46: 435 –441.[Medline]
Clarke GN, McBain JC, Lopata A, Johnston WI. In vitro fertilization results for women with sperm antibodies in plasma and follicular fluid. Am J Reprod Immunol Microbiol. 1985a; 8: 130 –131.[Medline]
Clarke GN, Stojanoff A, Cauchi MN, Johnston WI. The immunoglobulin class of antispermatozoal antibodies in serum. Am J Reprod Immunol. 1985b;7: 143 –147.
Cross NL, Meizel S. Methods for evaluating the acrosomal status of mammalian sperm. Biol Reprod. 1989; 41: 635 –641.[Abstract]
De Almeida M, Zouari R, Jouannet P, Feneux D. In-vitro effects of
anti-sperm antibodies on human sperm movement. Hum
Reprod. 1991;6: 405
–410.
Domagaa A, Kurpisz M. Immunoprecipitation of sperm and
somatic antigens with antibodies from sera of sperm-sensitized and anti-sperm
antibody-free individuals. Am J Reprod Immunol. 2004; 51: 226
–234.[Medline]
Domagaa A, Pulido S, Kurpisz M, Herr JC. Application of
proteomic methods for identification of sperm immunogenic antigens.
Mol Hum Reprod. 2007; 13: 437
–444.
Eggert-Kruse W, Christmann M, Gerhard I, Pohl S, Klinga K,
Runnebaum B. Circulating antisperm antibodies and fertility prognosis: a
prospective study. Hum Reprod. 1989; 4: 513
–520.
Feng HL, Han YB, Hershlag A, Zheng LJ. The impact of
Ca2+ flux inhibitors on acrosome reaction of hamster spermatozoa.
J Androl. 2007;28: 561
–564.
Feng HL, Hershlag A, Han YB, Zheng LJ. Localizations of intracellular calcium and Ca2+-ATPase in hamster spermatogenic cells and spermatozoa. Microsc Res Tech. 2006; 69: 618 –623.[CrossRef][Medline]
Feng HL, Sandlow JI, Sandra A. Expression and function of the c-kit proto-oncogene protein in mouse sperm. Biol Reprod. 1997; 57: 194 –203.[Abstract]
Feng HL, Sandlow JI, Sandra A. Presence of c-kit receptor and its possible signaling pathway in mouse sperm. Mol Reprod Dev. 1998; 49: 317 –326.[CrossRef][Medline]
Feng HL, Sandlow JI, Zheng LJ. c-kit receptor and its possible function in human spermatozoa. Mol Reprod Dev. 2004; 70: 103 –110.
Francavilla F, Romano R, Santucci R, Marrone V, Properzi G, Ruvolo G. Interference of antisperm antibodies with the induction of the acrosome reaction by zona pellucida (ZP) and its relationship with the inhibition of ZP binding. Fertil Steril. 1997; 67: 1128 –1133.[CrossRef][Medline]
Friberg J. A simple and sensitive micromethod for demonstration of sperm agglutinating activity in serum from infertile men and women. Acta Obstet Gynecol Scand. 1974; 36: 21 –29.
Fusi F, Bronson RA. Effects of incubation time in serum and capacitation on spermatozoal reactivity with antisperm antibodies. Fertil Steril. 1990; 54: 887 –893.[Medline]
Haas GG Jr, D'Cruz OJ, DeBault LE. Assessment by fluorescence-activated cell sorting whether sperm-associated immunoglobulin (Ig)G and IgA occur on the same sperm population. Fertil Steril. 1990;54: 127 –132.[Medline]
Hall JL, Engel D, Naz RK. Significance of antibodies against human sperm FA-1 antigen in immunoinfertility. Arch Androl. 1994; 32: 25 –30.[Medline]
Isojima S, Li TS, Ashitaka Y. Immunological analysis of sperm immobilizing factor found in sera of women with unexplained sterility. Am J Obstet Gynecol. 1968; 101: 677 –683.
Jager S, Kremer J, Van Slochteren-Draaisma T. A simple method of screening for antisperm antibodies in the human male. Detection of spermatozoal surface IgG with the direct mixed antiglobulin reaction carried out on untreated fresh human semen. Int J Fertil. 1978; 23: 12 –21.[Medline]
Kohl B, Kohl H, Krause W, Deichert U. The clinical significance of
antisperm antibodies in infertile couples. Hum Reprod. 1992; 7: 1384
–1387.
Kremer J, Jager S. The significance of antisperm antibodies for
sperm-cervical mucus interaction. Hum Reprod. 1992; 7: 781
–784.
Kutteh WH, Kilian M, Ermel LD, Mestecky J. Antisperm antibodies in infertile women: subclass distribution of immunoglobulin (Ig)A antibodies and removal of IgA sperm-bound antibodies with a specific IgA1 protease. Fertil Steril. 1995; 63: 63 –70.[Medline]
Mandelbaum SL, Diamond MP, DeCherney AH. Relationship of antisperm antibodies to oocyte fertilization in vitro fertilization-embryo transfer. Fertil Steril. 1987; 47: 644 –651.[Medline]
Margalioth EJ, Cooper GW, Taney FH, Scholl GM, Rosenfeld DL. Capacitated sperm cells react with different types of antisperm antibodies than fresh ejaculated sperm. Fertil Steril. 1992; 57: 393 –398.[Medline]
Marshburn PB, Kutteh W. The role of antisperm antibodies in infertility. Fertil Steril. 1994; 61: 799 –811.[Medline]
Marthur S, Chao L, Schulte BA. Sperm and seminal plasma antigens from autoimmune men induce immunological infertility. Arch Androl. 1987;19: 161 –175.[CrossRef][Medline]
Monroe JR, Altenbern DC, Mathur S. Changes in sperm antibody test results when spermatozoa are subjected to capacitating conditions. Fertil Steril. 1990; 54: 1114 –1120.[Medline]
Naz RK, Menge AC. Antisperm antibodies: origin, regulation, and sperm reactivity in human infertility. Fertil Steril. 1994; 61: 1001 –1013.[Medline]
Paradowska A, Bohring C, Krause E, Krause W. Identification of evolutionary conserved mouse sperm surface antigens by human antisperm antibodies (ASA) from infertile patients. Am J Reprod Immunol. 2006;55: 321 –330.[Medline]
Rajeev SK, Reddy KVR. Sperm membrane protein profiles of fertile
and infertile men: identification and characterization of fertility associated
sperm antigen. Hum Reprod. 2004; 19: 234
–242.
Shibahara H, Burkman LJ, Isojima S, Alexander NJ. Effects of sperm-immobilizing antibodies on sperm-zona pellucida tight binding. Fertil Steril. 1993; 60: 533 –539.[Medline]
Shibahara H, Shigeta M, Inoue M, Hasegawa A, Koyama K, Alexander
NJ, Isojima S. Diversity of the blocking effect of antisperm antibodies on
fertilization in human and mouse. Hum Reprod. 1996; 11: 2595
–2599.
Shibahara H, Shigeta M, Toji H, Koyama K. Sperm immobilizing antibodies interfere with sperm migration from the uterine cavity through the Fallopian tubes. Am J Reprod Immunol. 1995; 34: 120 –124.[Medline]
Snow K, Ball DG. Characterization of human sperm antigens and anti-sperm antibodies in infertile patients. Fertil Steril. 1992;58: 1011 –1019.[Medline]
Stern JE, Dixon PM, Manganiello PD, Brinck-Johnsen T. Antisperm antibodies in women: variability in antibody levels in serum, mucus, and peritoneal fluid. Fertil Steril. 1992; 58: 950 –958.[Medline]
Tasdemir I, Tasdemir M, Fukuda J, Kodama H, Matsui T, Tanaka T. Effect of sperm-immobilizing antibodies on the spontaneous and calcium-ionophore (A23187)-induced acrosome reaction. Int J Fertil. 1995;40: 192 –195.
Tsukui S, Noda Y, Fukuda A, Matsumoto H, Tatsumi K, Mori T. Blocking effect of sperm immobilizing antibodies on sperm penetration of human zonae pellucidae. J In Vitro Fert Embryo Transf. 1988; 5: 123 –128.[CrossRef][Medline]
Witkin SS, David SS. Effect of sperm antibodies on pregnancy outcome in a subfertile population. Am J Obstet Gynecol. 1988; 158: 59 –62.[Medline]
| ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
| HOME | HELP | FEEDBACK | SUBSCRIPTIONS | ARCHIVE | SEARCH | TABLE OF CONTENTS |
