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Biomedical subjects

D L Fulgham

Publications and source records attributed to D L Fulgham.

17 recordsLinked to original sources
PubMed1991

Antisperm antibodies to sperm surface antigens in women with genital tract infection.

Antisperm antibodies to sperm surface antigens in nulligravid women with primary upper genital tract infections were measured by the sperm mixed agglutination reaction assay. As many as 56% of women with a primary episode of pelvic inflammatory disease had antisperm antibodies. In addition, 69% of those women with no history of genital tract infection but with laparoscopic evidence of past pelvic infection had significant levels of circulating antisperm antibodies. Electroimmunoblots of sperm preparations probed with the sera of women who had either known or presumed upper genital tract infection revealed a uniformly recognized 69 kd antigen. In contrast, women with circulating antisperm antibodies before primary upper genital tract infection recognized up to five distinct sperm antigen determinants of 27, 54, 131, 146, and 174 kd. It is a distinct possibility that genital tract infections may lead to immunopotentiation of antisperm antibodies that could affect fertility.

Adolescent↗
PubMed1991

Evaluation of human sperm-zona pellucida tight binding by presence of monoclonal antibodies to sperm antigens.

Characterized WHO monoclonal antibodies (MAbs) to human sperm antigens were evaluated as to whether they inhibited sperm-zona pellucida tight binding as assessed by the hemizona assay (HZA). Of the 26 MAbs tested, only one inhibited zona binding. The whole sperm-specific MAb inhibited zona binding by 70%. The MAb also caused strong agglutination. Two procedures, Sephadex column chromatography and papain digestion, were used to determine whether agglutination or steric hindrance was a factor in the capability of MAb to inhibit zona binding. However, inhibition remained comparable to previous results. The MAb did not prevent capacitation, nor calcium influx and the resulting increase in hyperactivated motility and acrosome reaction. Since its inhibitory influence is not due to agglutination factors, steric hindrance or prevention of normal pre-fertilization maturation, the MAb may be blocking a portion of the zona binding receptor and may be useful in elucidating sperm antigens important to sperm-egg interaction. The approach used in this study allows definition of sperm surface antigens involved in zona pellucida binding.

Antibodies, Monoclonal↗
PubMed1991

Studies on the in vitro spermicidal activity of synthetic magainins.

Two synthetic magainins A and G are shown to have spermicidal activity. Transmission electron microscopic micrographs show that both magainins alter the plasma membranes of sperm and that these actions are rapid. Further studies will better delineate the contraceptive potential of synthetic magainins.

Antimicrobial Cationic Peptides↗
PubMed1990

Sperm preparation affects reactivity of monoclonal antibodies.

The polyacrylamide gel electrophoresis (SDS-PAGE) patterns of extracts prepared from thrice-washed human sperm were compared with extracts of swim-up and non-rise sperm. For sperm extraction sodium deoxycholate (DOC), octylphenoxypolyethoxyethanol (NP-40), dithiothreitol (DTT), lithium 3,5-diiodosalicylate (LIS), or 3-[(3-cholamidopropyl) dimethylammonio]-1-propane sulfonate (CHAPS) were used. A pellet of pooled thrice-washed semen was divided and extracted with these detergents. Similar gel electrophoresis profiles, as revealed by silver staining, were observed with the LIS, DOC, NP-40 and CHAPS extracts, whereas DTT extracts lacked the 150 and 200 kDa bands. Different thrice-washed semen pools varied in their protein pattern, probably due to the presence of sloughed germinal cells, white blood cells and sloughed cells from the reproductive tract. Swim-up sperm extracts had a different protein profile from that of thrice-washed pools but pools of swim-up sperm had a more constant protein pattern than those of thrice-washed semen pellets. Monoclonal antisperm antibodies did not always recognize antigens of the same molecular weights in western blots when extracts of swim-up sperm versus thrice-washed sperm were compared. In immunoblot studies with monoclonals (produced against thrice-washed sperm, cloned five times, reactive by ELISA and sperm function tests), 4/12 reacted with 0.3% NP-40 extracts of thrice-washed sperm and 5/12 with 0.3% NP-40 extract of swim-up sperm. These findings suggest that definition of antigens by monoclonal antisperm antibodies will be influenced by the nature of sperm extract employed.

Antibodies, Monoclonal↗
PubMed1990

Sperm bound to zona pellucida in hemizona assay demonstrate acrosome reaction when stained with T-6 antibody.

A monoclonal antibody, T-6, useful for detecting acrosome-reacted sperm based on an immunofluorescent assay, was employed to evaluate acrosomal status of human sperm that were tightly bound to hemisected human zonae pellucidae (hemizona assay). Over 90% of the bound sperm evaluated exhibited immunofluorescent patterns indicative of acrosome reaction. This staining method for evaluating the acrosomal status of sperm bound to the zona pellucida may enable definition of a group of male infertility patients heretofore not recognized.

Acrosome↗
PubMed1990

Serum hormone levels affect sperm function.

If hormone levels affect sperm motility during capacitation, then the serum added to samples prepared for artificial insemination could affect sperm fertilizability. We investigated various sperm functional movement characteristics (percent motile, progressive velocity, linearity, beat cross frequency, lateral head displacement, longevity, and hyperactivation) in specimens incubated with women's sera, as well as exogenous hormone preparations of estradiol (E2) and progesterone (P). Early follicular phase serum (low E2 and low P) maintained motility and longevity. Sperm in E2-treated medium exhibited higher progressive velocity, linear motility, and longevity. In contrast, the percent motility decreased as sperm exhibited hyperactivated motility with P.

Estradiol↗
PubMed1990

Characteristics of monoclonal antibodies reactive with human sperm and seminal plasma.

The characteristics of monoclonal antibodies developed against human spermatozoa are described. Out of 10 monoclonal antibodies 9 did not react in ELISA with human RBC, WBC, platelets, Raji cells nor mouse sperm. Four monoclonal antibodies reacted with monkey sperm and all 10 reacted with human seminal plasma. Monoclonal antibodies showed differential reactivity with pre- and post-capacitated sperm. Four monoclonal antibodies were able to agglutinate sperm whereas none of these were positive in sperm-immobilization assay. Interestingly, two monoclonal antibodies (MA-46 and MA-50) were able to block the attachment of pre-capacitated sperm to zona denuded hamster oocytes. MA-46 and MA-50 recognized in immunoblot spermatozoa antigens having apparent molecular weights of 14 and 20 K Da and greater than 200 K Da respectively. The monoclonal antibodies reported in this study will be useful in further delineating the spermatozoa antigens involved in regulation of fertility.

Animals↗
PubMed1988

Production of large amounts of mouse polyclonal antisera.

A simple and inexpensive method for producing larger volumes of mouse polyclonal antisera is described. Groups of BALB/c mice were immunized with different human sperm antigens classified according to molecular weight. Mice were immunized twice with the antigen, twice pristane pretreated, and then injected with a non-antibody-secreting myeloma cell line to induce ascites formation. Antibody activity of the ascites fluid approximated that of the serum. Thus, one mouse can provide about six times the amount of antibody-containing fluid usually obtained after conventional immunization. This method also may be applicable for ascites fluid production in nude mice.

Animals↗
PubMed1988

Distribution of T cell subsets in follicular fluid.

Examination of follicular fluid (FF) from in vitro fertilization patients revealed a significant difference in concentrations of lymphocytes and T cell subpopulations with increased oocyte maturation. A total of 111 follicles containing 82 oocytes were aspirated from 10 patients undergoing laparoscopic oocyte retrieval. FF from 61.3% of the follicles was classified as clear and 38.7% as bloody, based on gross and microscopic appearance. A mean of 1.78 X 10(6) lymphocytes/ml was obtained from peripheral blood (PB) as compared to 2.14 X 10(5) and 2.79 X 10(5) lymphocytes/ml for clear and bloody FF, respectively. There were 6.3 X 10(5) T4 and 3.7 X 10(5) T8 lymphocytes in PB, resulting in a T4/T8 ratio of 1.72, which is not significantly different from that of the general population. The mean concentration of FF T4 and T8 lymphocytes decreased with increased oocyte maturation; the T8 reduction was statistically significant (P less than 0.05). The proportion of T4 to T8 lymphocytes in FF remained unchanged and was unaffected by maturity of the oocyte. Although estradiol (E2) did not vary with oocyte maturity, progesterone (P) increased and E2/P decreased. There was no correlation between E2 or P levels and distribution of T cells. Fertilization rates were higher in more mature oocytes, but there was no correlation between fertilization and E2, P, E2/P, or T cell subpopulations. It remains to be determined what factors result in the decrease in lymphocytes with increased oocyte maturity and the observed difference in FF T4/T8 compared to PB.

Female↗
PubMed1987

In vitro immune absorption of antisperm antibodies with immunobead-rise, immunomagnetic, and immunocolumn separation techniques.

Fourteen men with a mean duration of infertility greater than 3 years who had significant sperm immobilizing or sperm-agglutinating antibodies were studied. All patients had greater than 20% IgG or IgA immunobinding to sperm in their seminal plasma and 7 had immunobinding levels of greater than 50%. Sperm from these men were less able to penetrate an overlaying buffer layer than sperm from a fertile control. Addition of immunobeads to the specimen was of little use, because few motile sperm could swim into the overlaying buffer; retained immunobeads were noted in the buffer layer of 18-hour capacitated specimens. Magnetic isolation of antibody-coated sperm from antibody-free sperm avoids potential damage to fragile sperm through centrifugation. Viable spermatozoa were isolated from magnetite-complexed spermatozoa, but the motility of the isolated spermatozoa deteriorated rapidly during the subsequent capacitation period. Passage of diluted ejaculate through a column of dextran beads for antisperm antibody processing (ASAP) was associated with superior sperm quality and fertilizing potential. The use of ASAP resulted in good sperm velocity and linearity and improved sperm function, as measured with the hamster egg penetration test. Sperm from men with immunologically mediated infertility can be processed through the ASAP and used for artificial insemination of their partners or in an in vitro fertilization program.

Autoantibodies↗
PubMed1987

Rectal infusion of semen results in transient elevation of blood prostaglandins.

Repeated semen deposition in the gut may be linked to the development of viral infections in homosexual men. Other investigators have suggested that rectal insemination may diminish immune responsiveness. We approximated conditions of human insemination by infusing 2 ml of pooled human seminal plasma (SP) into the rectum and/or vagina of rhesus monkeys. This resulted in increased blood plasma concentrations of the bicycloderivative of prostaglandin E (PGEM-II) which reached peak concentrations 2 h after rectal SP instillation in seven of eight test monkeys, but not the controls. The rate of PGE2 diffusion appeared to occur more rapidly across vaginal than rectal mucosa. Suppression of peripheral cellular immune functions was not demonstrated after the single exposure of this study, although persistent and repeat exposures could lead to local or generalized suppression of host defense mechanisms. Absorption of PGE's from the gut may be a cofactor in the development of sexually transmitted viral diseases.

Animals↗
PubMed1986

Antisperm antibodies and circulating immune complexes of vasectomized men with and without coronary events.

We compared antisperm antibody and circulating immune complex (CIC) levels in serum samples from 101 vasectomized and 101 normal age-matched nonvasectomized men; 31 of each group had histories of coronary heart disease (CHD). Vasectomy and CHD status were treated as categorical independent variables in the two-way analysis of variance. Elevations of both systolic and diastolic blood pressures were significantly associated with age and body mass index but not vasectomy. Antisperm antibodies (immobilizing and agglutinating) were significantly associated with vasectomy (P less than or equal to .001); the incidences were similar in men with and without CHD. The CICs were significantly associated with vasectomy in a Staphylococcus aureus (FcSa) CIC assay (P less than or equal to .001) and a Raji cell CIC assay (P less than or equal to .05). A third CIC assay, the Clq binding assay, did not reveal a difference between any subgroups. Generally, CICs occurred more frequently in the CHD group by the FcSa assay and particularly the Raji cell assay (P less than or equal to .001). In summary, vasectomized men had a higher incidence and higher levels of circulating antisperm autoantibodies and CICs than did age-matched controls.

Antigen-Antibody Complex↗
PubMed1985

Circulating immune complexes in cynomolgus macaques.

Nonhuman primates can be used as models for the study of immune-complex-associated diseases. Recognizing that very little is known about the levels of circulating immune complexes (CICs) in normal monkeys, we have used three assays to measure the levels in serum collected from 313 adult and 106 juvenile cynomolgus macaques (Macaca fascicularis). The prevalence was higher than expected. There was a strong statistical association between CIC levels and country of origin. Monkeys from Indonesia were more likely to have elevated CICs than those from Malaya or the Philippines. This relationship was observed with all three assays. Furthermore, juvenile macaques tended to have lower levels than did adults. This study indicates that it may be important to consider genetic factors, the country of origin, or both when selecting cynomolgus macaques for research on immune-complex-associated diseases.

Age Factors↗
PubMed1977

Cell-mediated immunity in vasectomized rhesus monkeys.

Cell-mediated immunity in rhesus monkeys that had been vasectomized for 2, 4, 7, or 11 years was measured by lymphocyte blastogenesis following stimulation with concanavalin A, phytohemagglutinin (PHA), and pokeweed mitogens. Several of the 7- and 11-year vasectomized animals had significantly reduced PHA reactivity when compared with control animals, and the percentage of animals with reduced PHA reactivity increased with time after vasectomy.

Alpha-Globulins↗