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Biomedical subjects

G G Haas

Publications and source records attributed to G G Haas.

At least 37 records · Page 2Linked to original sources
PubMed1990

Evaluation of antisperm complement-dependent immune mediators in human ovarian follicular fluid.

A "sandwich"-type radiolabeled antiglobulin assay using monoclonal anti-C5b-9 neoantigen and polyclonal anti-C5b-9 was used to evaluate the presence of terminal C complexes (SC5b-9 or MC5b-9) in the sera and ovarian follicular fluid (FF) from 45 infertile women. FF SC5b-9 was detectable in all clinical diagnostic categories. The mean SC5b-9 levels in FF and sera were 399 ng/ml (range 75 to 1350 ng/ml) and 798 ng/ml (range 0 to 2700 ng/ml), respectively. Twelve (26.6%) of the 45 FF samples had normal hemolytic C activity, and all FF (n = 44) samples initiated C8/C9-dependent lysis of sensitized sheep E coated with human C1-7. Human plasma IgG antisperm antibodies (ASA) were capable of activating C in 31 (72%) of 43 FF samples as detected by their ability to deposit MC5b-9 on human sperm. Sera from infertile women with ASA in their sera and FF impaired human sperm binding to human zona pellucida and binding and penetration of zona-free hamster oocytes in vitro. The discovery of SC5b-9 and MC5b-9 in ovarian FF implies that the interaction of ASA and C could have a deleterious effect on sperm during in vivo and in vitro sperm-egg interactions in women with antisperm antibodies.

Antigen-Antibody Complex↗
PubMed1990

Assessment by fluorescence-activated cell sorting of whether sperm-associated immunoglobulin (Ig)G and IgA occur on the same sperm population.

Two-color fluorescence-activated cell sorting of antisperm antibody-positive sperm was used to detect simultaneously the presence of immunoglobulin (Ig)A and IgG antisperm antibodies associated in vivo on a man's sperm. Sperm positive for sperm-associated Ig were analyzed using phycoerythrin-conjugated antihuman IgA and fluorescein isothiocyanate-conjugated antihuman IgG; up to 87% of the same spermatozoa were stained with both labels. Sperm positive for only one of the antisperm antibody isotypes stained up to 90% of a man's sperm with only one fluorochrome. Immunocytochemistry studies revealed similar patterns of sperm binding for sperm-associated IgG and IgA. These results suggest that the sperm antigenic determinants reacting with antisperm IgA and IgG are present on the same sperm population at similar locations on the sperm surface.

Cell Separation↗
PubMed1990

Lack of complement activation in the seminal plasma of men with antisperm antibodies associated in vivo on their sperm.

A specific and sensitive "sandwich"-type radiolabeled antiglobulin assay (RAA) using monoclonal anti-human C5b-9 neoantigen and polyclonal anti-human C5b-9 was used to evaluate the presence of the in vivo product of human complement (C) activation (SC5b-9) in the seminal plasma (SP) of 19 fertile and 61 infertile men. SP SC5b-9 was detectable in 7 (8.7%; 1 fertile and 6 infertile men) of the 80 men with a range of 10 to 175 ng/ml. Levels of SP SC5b-9 in other men were below the limit of detection (less than 10 ng/ml). Of the 33 infertile men with sperm-associated immunoglobulin (Ig) G and/or IgA, 27 (82%) had undetectable levels of SP SC5b-9 immunoreactivity. There was no correlation between the SP SC5b-9 levels and the degree of sperm-associated IgG (r = 0.086) or IgA (r = 0.23) activity. However, significant deposition of sperm-bound C5b-9 due to autologous C activation was demonstrated by flow cytometry of donor sperm treated with sera from autoimmune men with ASA in their sera and on their sperm. These findings suggest that sperm-bound Ig cannot activate C in SP.

Autoantibodies↗
PubMed1989

A radiolabeled antiglobulin assay to identify human cervical mucus immunoglobulin (Ig) A and IgG antisperm antibodies.

Antisperm immunoglobulin (Ig) A and IgG antibodies in human cervical mucus (CM) were identified by a radiolabeled antiglobulin assay. Cervical mucus samples from fertile and infertile women were exposed to a 1:3,200 dilution of 2-mercaptoethanol (2-ME), and 5 micrograms of the solubilized CM protein were assayed for the presence of IgA and IgG antisperm and anti-Candida activity by the radiolabeled antiglobulin assay. Purified human secretory IgA and IgG exposed to 2-ME retained the molecular integrity and functional activity of the untreated antibody molecules. CM aliquots collected after high-performance liquid chromatography (HPLC) fractionation were assessed for antisperm antibody activity; antisperm antibody activity was retained in the appropriate IgA or IgG CM fractions. The incidence of CM antisperm antibodies was minimally affected when the radiolabeled antiglobulin assay was performed with a motile sperm population. Approximately 70% of the CM IgA antisperm antibodies were of the IgA1 subclass; CM IgG was primarily of the IgG4 subclass. When Candida antigen was substituted for sperm in the radiolabeled antiglobulin assay, the CM antisperm antibodies were found to be exclusively sperm-specific. These data indicate that the radiolabeled antiglobulin assay using 2-ME to extract CM antibodies is a specific method for the assay of antisperm antibodies in CM.

Antibodies↗
PubMed1989

Quantitation of immunoglobulin G on human sperm.

An indirect inhibition assay was devised to quantitate the amount of immunoglobulin molecules on the surface of human sperm. Sperm-associated IgG was used to inhibit competitively 125I-labeled polyclonal anti-human IgG from binding to human IgG affixed to microtissue culture plate wells. Standard curves for the assay were derived by substituting soluble IgG in the supernatant of the tissue culture wells for sperm-associated IgG. The intraassay coefficient of variation was 6.7%; the interassay coefficient of variation was 5.6-21% depending upon the IgG antisperm antibody-positive plasma sample assayed. The assay technique was highly correlated (r = 0.99) with the number of antibody-bound sperm added to the supernatant. When different sperm donors were utilized as an antigen source, the coefficient of variation was 15.6% for positive plasmas and 19.3% for negative plasmas. The indirect inhibition assay showed a strong correlation (r = 0.91) with a previously described radiolabeled antiglobulin assay for sperm-associated IgG.

Humans↗
PubMed1989

Immunoreactive human epidermal growth factor in human seminal plasma.

We measured immunoreactive epidermal growth factor (EGF) by a homologous RIA in seminal plasma (SP) from 31 fertile and 52 infertile men to determine the relationship between SP EGF levels and total sperm count in the ejaculates. The mean SP EGF levels in fertile and infertile men were 41.7 +/- 21.5 (+/- SD) and 53.1 +/- 30.8 micrograms/L, respectively. Infertile men with sperm-associated immunoglobulin G (n = 9), immunoglobulin A (n = 6), or both (n = 8) had mean SP EGF levels of 48.9 +/- 26.1, 47.9 +/- 17.5, and 56.5 +/- 32.1 micrograms/L, respectively. Seven men with severe oligospermia had a mean SP EGF level of 58.5 +/- 35.9 micrograms/L. There was no correlation (r = 0.14; P greater than 0.05) between SP EGF levels and total sperm counts in these men. Fractionation of SP by high performance liquid chromatography on a size exclusion (TSK G2000 SW) column revealed a single immunoreactive peak with an approximate mol wt of 8000, slightly higher than the mol wt of circulating human EGF (6000). We conclude that SP EGF may be distinct from peripheral plasma EGF.

Adult↗
PubMed1988

The effect of fixatives and/or air-drying on the plasma and acrosomal membranes of human sperm.

It has been hypothesized that some fixatives and conditions of slide preparation expose internal sperm antigens and thus are not suitable for demonstrating surface-specific antigens by immunocytochemical assays. This study examined the ability of five fixatives (glutaraldehyde, acetone, methanol, paraformaldehyde, and periodate-lysine-paraformaldehyde [PLP]) and two conditions of slide preparation (air-drying or maintaining sperm in a liquid phase) to maintain the integrity of human spermatozoal membranes at the ultrastructural level as monitored by transmission electron microscopy. Regardless of the fixative employed, air-drying was detrimental to plasma and acrosomal membrane integrity. Acetone and methanol completely disrupted the plasma and acrosomal membranes over the entire sperm surface whether air-drying or liquid phase conditions were employed. Fixation with glutaraldehyde and paraformaldehyde (to a lesser degree) maintained the morphologic integrity of acrosomal and plasma membranes provided the sperm were maintained in a liquid phase. However, glutaraldehyde and paraformaldehyde fixation increased the postfixation binding of immunoglobulins to the sperm surface. We concluded that immunocytochemistry at the light microscopy level may imply erroneous interpretations regarding antigen/antibody interactions on the sperm plasma membrane surface unless care is taken to verify that the antibodies of interest are binding to the antigenically intact plasma or acrosomal membrane.

Acetone↗
PubMed1988

ABH blood group antigens in human semen.

Human sperm, erythrocytes, and seminal plasma from the blood and semen of 20 men of known ABH, Lewis, and secretor phenotypes were assayed for ABH blood group antigens. A 1:1 correlation was found between the presence of ABH antigens in seminal plasma and on sperm and if the man had a secretor phenotype. Sperm from nonsecretors or from men of dissimilar ABO blood type could adsorb A antigen when incubated with A antigen-containing seminal plasma. The level of ABH antigens in seminal plasma correlated with the level of ABH antigens on the sperm surface. ABH antigens in semen were present only on a minority of spermatozoa as detected by flow cytometry, and the majority of these sperm were not in the swimup fraction. ABH antigens were not present on sperm within the seminiferous tubules of human testicular material. It was hypothesized that ABH antigens found on human sperm were adsorbed from seminal plasma on a minority of the sperm in an ejaculate.

ABO Blood-Group System↗
PubMed1988

Distribution of human leukocyte antigen-ABC and -D/DR antigens in the unfixed human testis.

The distribution of the major histocompatibility complex (MHC) antigens in the unfixed human testicle was studied by indirect immunofluorescence. Three murine monoclonal antibodies to the common determinants of class I MHC antigens (human leukocyte antigen [HLA]-ABC) and three against class II MHC antigens (HLA-D/DR antigens), respectively, were utilized. No class I MHC antigens were identified on developing testicular germ cells including spermatozoa, but interstitial cells between the seminiferous tubules (including Leydig cells) and blood vessel endothelium expressed the antigen. Class II MHC antigens were not found on any cells within the seminiferous tubules. However, the class II antigen was identified on dendritic-like cells between the seminiferous tubules and on vessel endothelium, although its expression was expectedly limited. These findings indicate that human testicular germ cells express minimal or no MHC antigens.

Adult↗
PubMed1988

Detection and titration of class-specific antisperm antibodies in serum using an enzyme-linked immunosorbent assay.

An antisperm antibody enzyme-linked immunosorbent assay (ELISA) that uses whole unfixed sperm and detects immunoglobulin G (IgG) and IgA antibodies in serum was developed. Donor sperm were washed and plated on poly-L-lysine-treated microtiter plates. The patient's sera were diluted to concentrations of 1:4 to 1:256 and incubated with sperm. Positive and negative sera had been previously tested for IgG antisperm antibody activity with a radiolabeled antiglobulin assay. Samples were considered positive when the mean absorbance of triplicate wells was greater than 2 SD above the pooled negative mean. Intra-assay variation was 7.9 and 9.6% for pooled negative and positive controls, respectively. Identical titers of control positive serum were consistently detected. A correlation of 0.83 was observed between ELISA IgG serum titers and radiolabeled antiglobulin results (N = 12). All negative samples tested negative in both assays (N = 21). Some serum samples showed IgA antisperm antibodies. Determination and titration of class-specific antibodies in serum should facilitate initial screening and follow-up of patients at risk for antisperm antibodies.

Enzyme-Linked Immunosorbent Assay↗
PubMed1987

Influence of estrogens on vascular transudation and mucus production in the rabbit endocervix.

The manner in which estrogens mediate cyclical changes in the viscoelastic properties and volume of cervical mucus is unclear. To identify the response to estrogens of each of the major mucus components (mucin[s], soluble proteins, and small electrolytes and water), the authors quantitated their accumulation rates before and during estrogen stimulation in the rabbit. Circulating radiolabeled markers (35S-sulfate and 131I-albumin) were used to monitor the incorporation of small and large molecular weight intravascular substances into the cervical mucus. The accumulation rate of mucins was unaffected by exogenous estrogen administration, despite significant increases in mucus volume. This increase in mucus volume was attributed to a significantly increased transudation of water and small electrolytes increasing mucus hydration as early as the first 2 hours of estrogen administration. Water, small electrolytes, and soluble proteins significantly increased during the third and fourth hours of estrogen administration, not only when compared with the unstimulated basal levels, but also when compared with the levels found during the first 2 hours of estrogen administration. No significant change occurred in mucin production, while significant changes occurred in the accumulation of proteins and small electrolytes, whether estrogen was given initially or terminally in the experiment protocol.

Animals↗
PubMed1987

How should sperm antibody tests be used clinically?

Immunoglobulin-specific assays are currently available for detection of patients with antibody-mediated infertility. Care must be taken in insuring that the correct immunoglobulin class is assayed, the assay is directed to the possible interaction of antibodies and sperm in the male or female genital tract, and the antigen employed will not detect antibodies against antigens not found on the plasma membrane of living sperm.

Antibodies↗
PubMed1987

Antibody-mediated causes of male infertility.

It is probable that antisperm antibodies identified directly on a man's sperm's surface correlate best with the presence of antibody-mediated infertility in males. Care must be taken in reviewing the literature regarding antisperm antibodies in men, since many studies are based on measurement of antibodies in the circulation or have utilized antigens (such as solubilized or fixed sperm) that are not on the sperm surface. The results of these assays that are not measuring antibodies against the sperm plasma membrane may not be clinically significant, since the sperm plasma membrane remains intact in sperm participating in in-vivo fertilization until after the acrosome reaction. Treatment of antibody-mediated infertility in the male is difficult at the present time, although newer techniques are promising.

Adrenal Cortex Hormones↗
PubMed1987

Immunologic infertility.

In general, the management of antibody-mediated infertility has been plagued by misdiagnosis due to the choice of assay system, therapies that may be associated with major side effects, and a failure to learn from the models of other antibody-mediated diseases. An important consideration in diagnosing antibody-mediated infertility is to use an immunoglobulin-specific technique that employs intact or living spermatozoa. Once this is done, a search should be made for what functional deficits in reproduction are associated with the presence of the antisperm antibodies. This would include postcoital testing, tests for ovum penetration (human sperm/hamster egg penetration assay), and tests for the ability of the sperm in the presence of antibody to undergo the acrosome reaction. Once the extent of the reproductive deficit is known, appropriate therapy can then be suggested. I believe that a "case" must be built to prove that antibody-mediated fertility actually exists in a particular couple, even in the presence of a positive test that is known to be highly specific. Sperm antibodies can attach to the sperm's surface without detrimentally affecting reproductive function. In a similar vein, low levels of sperm antibodies could give a positive test result, but the level may be insufficient to be a major detriment to fertility. It is hoped that in the near future isolation of specific sperm antigens will be used to identify antibodies against antigens critical to reproductive function. It may be possible to determine the minimal amount of antisperm antibody that is necessary to disturb each individual step in reproductive function. If a patient is found to have sperm antibodies whose quantity exceeds this amount, then the patient can be appropriately labeled as having antibody-mediated infertility. The array of therapies available for the couple's unique fertility problem(s) can then be described to the patient, and an appropriate therapeutic choice made.

Antibodies↗
PubMed1987

A double-blind, placebo-controlled study of the use of methylprednisolone in infertile men with sperm-associated immunoglobulins.

Forty-three men with antibody-mediated infertility identified by measuring sperm-associated immunoglobulin with a direct radiolabeled antiglobulin assay were enrolled in a double-blind, placebo-controlled protocol to judge the efficacy of corticosteroids in this condition. In the active drug group, men were given 96 mg methylprednisolone in three divided doses for 7 days followed by a 2-day tapering of the drug. The control group was given placebo in the same manner. Both regimens were repeated three times approximately 1 month apart on the basis of their female partner's menstrual cycle. There was not a favorable effect of corticosteroids over placebo on the men's semen parameters, the results of assays for sperm-associated IgA and plasma IgG antisperm antibodies, or the men's subsequent fertility. There was a statistically significant effect on sperm-associated IgG in men given methylprednisolone compared with men given placebo.

Antibodies↗
PubMed1986

Circulating antisperm antibodies in recurrently aborting women.

One hundred seventy-three women with a history of three or more recurrent consecutive abortions were analyzed for circulating antisperm antibodies with a radiolabeled antiglobulin assay (RAA), a modified enzyme-linked immunosorbent assay (ELISA), a tray agglutination test (TAT), and a sperm immobilization test (SIT). No pregnancies were subsequently gestated to term in women who were antisperm antibody-positive unless they were inoculated with their husband's leukocytes as treatment for an immune basis (not related to antisperm antibodies) for their recurrent abortions. In women with an immune basis for their recurrent abortions, immunization with leukocytes from their male partners increased the ability of these women previously aborting their fetuses to carry their fetuses to term, even if they had positive results in the ELISA, TAT, and SIT; women with positive results in the RAA continued to abort subsequent pregnancies, despite leukocyte immunization. Immunization of antisperm antibody-positive women with their partner's leukocytes did not incite or increase the antisperm antibody titer, with any of the assay techniques.

Abortion, Spontaneous↗