Immunoprecipitation of Sperm and Somatic Antigens with Antibodies from Sera of Sperm‐Sensitized and Anti‐sperm Antibody‐ Free Individuals

Domagala, Alina; Kurpisz, Maciej

doi:10.1111/j.1600-0897.2004.00151.x

Cited by 10 publications

(5 citation statements)

References 11 publications

(14 reference statements)

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“…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 (Domagała 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.…”

Section: Discussionmentioning

confidence: 99%

Characterization of Human Sperm Antigens Reacting With Anti‐Sperm Antibodies From an Infertile Female Patient's Serum

Feng

Han

Sandlow

2008

Journal of Andrology

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Identification of sperm antigens that elicit immunoglobulin (Ig) production and knowledge of their roles in sperm transport and fertilization may enhance diagnosis and treatment of immunologic infertility. Sperm antigens recognized by a female patient's serum anti-sperm antibodies were characterized using an indirect immunobead-binding test, immunoblot analysis, and immunochemical labeling. The anti-sperm antibodies' effect on sperm function was evaluated by acrosome induction by calcium ionophore. Immunobeads specific for IgG were bound to the head of 79% of motile donor sperm. Immunochemical labeling of antibody-binding sites was restricted to the plasma membrane over the acrosomal crescent. No labeling was observed on the inner acrosomal membrane of acrosome-reacted sperm. The antibodies reacted with 35-, 40-, 47-, and 65-kd proteins extracted from acrosome-intact donor sperm. Sperm incubated in 1:4, 1:8, 1:16, and 1:32 dilutions of anti-sperm antibody-positive serum had similar rates of spontaneous acrosome reaction and significantly lower rates of ionophore-induced acrosome reaction compared with sperm incubated in control serum. These results suggest that sperm antigens recognized by the patient's serum anti-sperm antibodies are restricted to the acrosomal region of the plasma membrane. The antibodies may impair fertility by compromising the sperm's ability to undergo capacitation and/or acrosome reaction.

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Section: Discussionmentioning

confidence: 99%

Characterization of Human Sperm Antigens Reacting With Anti‐Sperm Antibodies From an Infertile Female Patient's Serum

Feng

Han

Sandlow

2008

Journal of Andrology

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“…Antibodies directed toward specific antigens are thought to disrupt the fertilization process in some way [56,57]. Unfortunately, the number and types of antibodies required to induce infertility are still unclear [58,59]. One aspect of this mystery may be resolved by our demonstration of a coating protein on the sperm surface that protects the sperm from immune recognition and antibody attack.…”

mentioning

confidence: 93%

Beta-Defensin 126 on the Cell Surface Protects Sperm from Immunorecognition and Binding of Anti-Sperm Antibodies1

Yudin

Generao

Tollner

et al. 2005

Biology of Reproduction

118

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Beta-defensin 126 (DEFB126), formerly known as epididymal secretory protein 13.2 (ESP13.2), coats the entire primate sperm surface until completion of capacitation, and it is a candidate for providing immune protection in the female reproductive tract. To further examine the potential role of DEFB126 as a means of protection from immune recognition, cynomolgus macaque sperm were exposed to a number of treatments that are known to alter sperm surface coats, including capacitation. We used a novel in vivo assay to determine immune recognition: aldehyde-fixed whole sperm injections into rabbits. Following booster injections, immunoblot analyses of whole sperm prepared in various manners was conducted. On Days 60 and 80 post-initial immunization, the antisera showed a remarkably strong reaction to a single 34-36 kDa protein, which was shown to be DEFB126. Sera from rabbits that were immunized with sperm washed more rigorously using Percoll gradients showed an increase in the number and intensity of proteins recognized on whole sperm Western blots, although DEFB126 was still the major immune response. When capacitated sperm, from which most DEFB126 had been released, were used as the immunogen, there was a dramatic increase in the immune recognition to a variety of protein bands. Sperm treated with neuraminidase to remove sialic acid on DEFB126 before fixation were shown to still possess DEFB126, but lacked the sialic acid component of the glycoprotein. These sperm were as immunogenic as capacitated sperm even though the desialylated DEFB126 still covered the entire cell surface. These sperm lost their highly negative charge (the isoelectric point of DEFB126 shifted from pI 3.0 to pI 6.4). Experiments using different sperm plasma membrane protein-specific Igs showed that recognition did not occur when DEFB126 was present, but following capacitation these Igs readily recognized the exposed sperm membrane. Our data suggest that DEFB126 protects the entire primate sperm surface from immune recognition and that the sialic acid moieties are responsible for the cloaking characteristic of this unique glycoprotein.

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“…Proteins were transferred to nitrocellulose membranes as previously described (Domagała & Kurpisz, ). After the electrotransfer, the membranes were incubated with serum samples at 1 : 2000 dilution overnight at 4 °C.…”

Section: Methodsmentioning

confidence: 99%

“…Such identification is difficult because of insufficient information on sperm proteome and due to a great heterogeneity of immunological reactions among individuals. Despite of these obstacles, many experiments identifying sperm antigens have been performed by proteomic methodology (Shetty et al ., , , ; Shibahara et al ., ; Bohring & Krause, ; Domagała & Kurpisz, ; Bhande & Naz, ; Domagała et al ., ; Feng et al ., ).…”

Section: Introductionmentioning

confidence: 99%

Proteomic identification of sperm antigens using serum samples from individuals with and without antisperm antibodies

et al. 2015

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The aim of the study was to identify human sperm antigens reacting with polyclonal antisperm antibodies. Protein sperm extracts were subjected to electrofocusing, and next immune reactions (immunoblotting) were carried out with positive for antisperm antibodies and control (not containing antisperm antibodies) serum samples. Proteomic analysis of human sperm proteins resulted in identification of 80 sperm antigens that could be divided into three groups: antigens specific for patients with antisperm antibodies (32), antigens recognised by both infertile patients and control sera (35) and antigens detected by control serum samples only (13). Among antigens specific for infertile patients, there were 12 sperm entities known to be involved in fertilisation process. We have also characterised three protein entities identified only by sera of infertile women. Altogether, the proteomic analysis resulted in identification of 27 sperm entities not reported previously in human sperm proteome. Identified proteins are sperm antigens that could be potentially responsible for immunological infertility. The study also sheds new light on the sperm antigens in aspect of gender specificity. The investigation of human sperm proteome by the use of antisperm antibodies-containing sera of infertile individuals not only may indicate new proteins but also can draft their immunological nature.

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Immunoprecipitation of Sperm and Somatic Antigens with Antibodies from Sera of Sperm‐Sensitized and Anti‐sperm Antibody‐ Free Individuals

Cited by 10 publications

References 11 publications

Characterization of Human Sperm Antigens Reacting With Anti‐Sperm Antibodies From an Infertile Female Patient's Serum

Characterization of Human Sperm Antigens Reacting With Anti‐Sperm Antibodies From an Infertile Female Patient's Serum

Beta-Defensin 126 on the Cell Surface Protects Sperm from Immunorecognition and Binding of Anti-Sperm Antibodies1

Proteomic identification of sperm antigens using serum samples from individuals with and without antisperm antibodies

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