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R K Naz

Publications and source records attributed to R K Naz.

At least 91 records · Page 5Linked to original sources
PubMed1987

The fertilization antigen (FA-1) causes a reduction of fertility in actively immunized female rabbits.

Female rabbits were actively immunized against the fertilization antigen (FA-1) isolated from lithium diiodosalicylate (LIS)-solubilized murine testis. Three trials were performed in order to check the effect of immunization on fertility. In all of these trials, there was a significant (P less than 0.001) reduction of fertility as determined by the percentage of 9-day implants/corpora lutea ratio (FA-1, 0-26.3%; adjuvant control, 79.4-100%). A complete block was observed in animals which received intravenous booster immunization with the antigen. Antisera collected from FA-1-immunized rabbits were negative in the agglutination and the immobilization techniques, and demonstrated modal titers of greater than or equal to 1:2560 in the enzyme linked immunosorbent assay (ELISA) using FA-1. Antisera were tissue-specific and showed binding to the specific protein bands of 47,000 and 23,000 Mr, dimeric and monomeric forms of FA-1, respectively, in the Western blot procedure. Ova collected from rabbits inseminated with sperm which had been treated with antiserum from immunized rabbits showed reduced fertilization rates (anti-FA-1, 3.9-27.7%; control rabbit serum, 87.8%). There was again a reduction in percentage of the 9-day implants/corpora lutea ratio in the rabbits inseminated with treated sperm (anti-FA-1, 10.7%; control rabbit serum, 72.7%). It is concluded that active immunization with FA-1 resulted in a tissue-specific immune response which caused a reduction of fertility in rabbits, by a mechanism(s) involving an inhibition of the fertilization process.

Animals↗
PubMed1987

Involvement of fertilization antigen (FA-1) in involuntary immunoinfertility in humans.

Sera from immunoinfertile patients (n = 32) and fertile controls (n = 20) were analyzed for cross-reaction with a purified and characterized sperm-specific glycoprotein, the fertilization antigen (FA-1), employing an enzyme-linked immunosorbent assay. The immunoinfertile sera demonstrated a strong reaction with FA-1 when compared with fertile control sera. There was no correlation between the reaction of sera with FA-1 and the titers obtained through the sperm agglutination technique and the sperm immobilization technique. Immunoinfertile sera showed binding with the protein bands in the regions corresponding to FA-1 on Western blots involving sodium deoxycholate-solubilized human sperm. Antigens isolated with immunoaffinity chromatography involving immunoinfertile sera also demonstrated antigen bands corresponding to FA-1 in sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Of the seven immunoinfertile couples, three that had antibodies to FA-1 in the male as well as female partners demonstrated a block of fertilization (IVF) due to antibodies bound on the sperm surface. The anti-FA-1 antibody activity was detected in serum as well as in follicular fluid and seminal plasma. Immunoinfertile sera that showed an inhibition of human sperm penetration of zona-free hamster ova showed a significant (P less than 0.001) increase in penetration rates after absorption with FA-1. These results indicate that sera from immunoinfertile patients had antibodies reacting with FA-1, and these antibodies are involved in the fertilization process.

Adult↗
PubMed1986

Reduction of fertility in female rabbits and mice actively immunized with a germ cell antigen (GA-1) from the rabbit.

Female rabbits and mice were actively immunized against germ cell antigen (GA-1) of 63 kDa molecular mass isolated from rabbit sperm and testis. There was a significant (P less than 0.05) reduction of fertility in rabbits actively immunized with GA-1 as compared to controls, as seen by the percentage of 9-day implants/corpora lutea ratio (GA-1, 36.3%; controls, 85.7%). In mice, there was again a significant (P less than 0.01) reduction in fertility as seen by mean 7-9 day implants +/- S.D. per mated mouse actively immunized with GA-1 whether through the intraperitoneal route (GA-1, 1.2 +/- 1.6; controls, 8.0 +/- 3.4) or through the subcutaneous/intramuscular route (GA-1, 3.8 +/- 3.4; controls, 10.1 +/- 3.9). The antisera from these actively immunized animals were negative for sperm agglutinating and immobilizing antibodies. In the Western blot enzyme-immunobinding procedure, the antisera showed specific binding to a single protein of 63 kDa. The incidence of fertilization of eggs recovered from rabbits inseminated with anti-GA-1 antibodies-treated sperm was not significantly different from control rabbits. The percentage of fertilized eggs obtained from rabbits inseminated with anti-GA-1 antibodies-treated sperm that reached the blastocyst stage upon in vitro incubation, however, was significantly less than that for embryos obtained from rabbits inseminated with control serum-treated sperm. Incubation of normal fertilized eggs in vitro with the antibodies did not affect development. Neither antiserum nor immune uterine fluid reacted with 4-day blastocysts in the indirect immunofluorescence technique. It is concluded that active immunization with GA-1 results in post-fertilization reduction of fertility in rabbits and mice by inhibiting early embryonic development.

Animals↗
PubMed1986

Factors influencing murine embryo bioassay: effects of proteins, aging of medium, and surgical glove coatings.

The 2-cell murine embryo bioassay as quality control for human in vitro fertilization (IVF) was used to evaluate the effects of protein supplements, medium aging, and surgical glove coatings. Ham's F-10 medium (GIBCO, Grand Island, NY) without protein supplementation supported growth of the 2-cell embryos to blastocysts. Addition of bovine serum albumin (BSA), fetal cord serum (FCS), or maternal serum (MS) did not enhance or reduce the blastulation rates (medium alone, 89.4%; BSA, 86.4%; FCS, 90%; MS, 74.7%). Unsupplemented Ham's F-10 medium was found to contain three major peaks of approximately 50,000 daltons and several minor peaks, analyzed on high-performance liquid chromatography (HPLC) and sodium sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), suggesting the presence of protein(s) in the medium itself. The storing of medium up to 425 days at 4 degrees C affected neither the HPLC profile nor its ability to support embryo growth (blastulation rates: fresh, 84%; stored 150 to 425 days, 77.7%). The coating of surgical gloves affected embryo growth. Both talc-coated (TC) and "talc-free," starch-coated (SC) surgical gloves were found to be progressively embryotoxic when they touched the medium for increasing lengths of time, compared with uncoated latex (UL) gloves and untouched control medium. Quality control of medium preparation and handling in murine embryo bioassay is reemphasized, with requirements for protein supplementation, use of fresh medium, and possible contamination with even talc-free, SC surgical gloves reevaluated.

Animals↗
PubMed1986

Characterization of the fertilization antigen 1 for the development of a contraceptive vaccine.

A fertilization antigen, FA-1, was purified from either deoxycholate- or lithium diiodosalicylate-solubilized murine testes by immunoaffinity chromatography using a monoclonal antibody, MA-24, which inhibited fertilization in vitro. The FA-1 was recovered at high (11.4) or low (2.8) pH using stepwise elution procedures of the deoxycholate or lithium diiodosalicylate extracts, respectively. Both of these fractions showed a single band of 47 kDa when analyzed by NaDodSO4/PAGE and silver staining. Following removal of the detergent and extensive dialysis at pH 5.8 or treatment with 0.15 M NaCl, even in the presence of detergent, a monomer of 23 kDa was detected. Two-dimensional PAGE of FA-1 showed, four or five polypeptides in the 47-kDa or 23-kDa range. The dialyzed FA-1 contained a major 23-kDa and a minor 48-kDa band when separated on both sucrose and cesium chloride gradients. High performance size-exclusion chromatography showed a major peak at 23 kDa and a minor peak at 50 kDa. Further analysis of the 23-kDa peak by reverse-phase chromatography resolved the antigen into three peaks, which gave similar two-dimensional gel patterns as the native FA-1. Lectin affinity chromatography on a lens culinaris column demonstrated that a part of the antigen was bound to the lectin while the rest was not. The FA-1 revealed a positive reaction with periodic-Schiff reagent and contained glucose and mannose, which together constituted 18.8% of the total antigen mass. Amino acid analysis showed a high percentage of aspartic acid, glutamic acid, serine, and glycine. As a single injection of MA-24 significantly reduced fertilization rates in vivo, the purified FA-1 is an attractive candidate for the development of contraceptive vaccine.

Amino Acids↗
PubMed1986

Development and application of monoclonal antibodies for specific detection of human enteric adenoviruses.

Monoclonal antibodies were prepared against enteric adenovirus by fusing P3-NS1/-Ag4-1 mouse myeloma cells with lymphocytes from BALB/c mice immunized with enteric adenovirus 40 (Ad40) G2297. Of the several putative clones secreting antibodies to adenovirus, five were found to react specifically to the enteric adenovirus. The specificity of two of these monoclones which recognize a single antigen of a molecular size of 17 kilodaltons was evaluated against 78 clinical isolates. One monoclone (5D8/2C2) reacted with both Ad40 and Ad41, and the other monoclone (2H6/C11) recognized Ad40 only in an enzyme-linked immunosorbent assay (ELISA). These ELISA results correlated well with those of the specific neutralization test or DNA restriction endonuclease analysis or both. The use of this rapid ELISA with these monoclones will find applications in the diagnosis of enteric adenovirus and should facilitate the epidemiologic studies of enteric adenovirus gastroenteritis.

Adenoviridae Infections↗
PubMed1985

Successful human pregnancy following in vitro fertilization using frozen semen.

Frozen-thawed husband's semen was utilized for insemination of spouse's oocytes in vitro. Postthaw semen had a low motility (40%) with a poor forward progression (+2), which subsequently decreased rapidly in the regularly used Ham's F-10 medium (pH 7.42, 280 mosmol/kg) supplemented with 7.5% fetal cord serum. When Ham's F-10 was supplemented with an additional 0.5 mmol each of magnesium sulfate (MgSO4) and potassium bicarbonate (KHCO3) at a higher pH (pH 7.69, 280 mosmol/kg), the sperm maintained their motility and forward progression and fertilized all the oocytes, which subsequently cleaved and developed into normal embryos. Transfer of these embryos resulted in a normal pregnancy. Our results indicate that cryopreserved semen of poor quality can be successfully employed for an in vitro fertilization-embryo transfer (IVF-ET) program, especially when MgSO4 and KHCO3 are used in Ham's F-10 at a higher pH.

Adult↗
PubMed1984

Monoclonal antibody to a human germ cell membrane glycoprotein that inhibits fertilization.

A monoclonal antibody to an antigen in the human germ cell membrane did not agglutinate or immobilize sperm but inhibited binding and penetration of zona-free hamster ova by human sperm and blocked murine fertilization in vitro. The antibody, of the 2a subclass of immunoglobulin G, was germ cell-specific but not species-specific. It recognized a single antigen of 23 kilodaltons that has been isolated from human germ cells. This fertilization antigen, located on the postacrosome , midpiece, and tail of human sperm, is a glycoprotein of testicular origin associated with some types of human involuntary immunoinfertility .

Animals↗
PubMed1984

Characterization of a membrane antigen from rabbit testis and sperm isolated by using monoclonal antibodies and effect of its antiserum on fertility.

An antigen was isolated from deoxycholate-solubilized rabbit testis and sperm by using an immunosorbent column containing IgG from a monoclonal antibody (8C10.5) that inhibits fertility. Elution was by stepwise increases in pH (8.0, 10.0, and 11.4), with the pH 11.4 fraction after recycling through the column showing a single band at 63 kilodaltons in slab NaDodSO4/PAGE with a silver stain. The antigen molecule was composed of two subunits, which on two-dimensional PAGE showed many spots within the same molecular size range (50-70 kilodaltons) but differing in charge. The antigen isolated either from testis or sperm showed mainly the same spots. The antigen is periodic acid/Schiff positive and contained 21% carbohydrate. An asialo-derivative of the antigen did not change its characteristics on NaDodSO4/PAGE. This glycoprotein resolved into two types of polypeptides, those binding and those not binding to a lens culinaris lectin column; some of the polypeptides appeared common to both fractions. Murine antiserum against the antigen neither agglutinated nor immobilized rabbit sperm but in immunofluorescence reacted with the plasma membrane of viable rabbit sperm as well as with murine and human sperm. Fertilization of female rabbits inseminated with treated sperm was not affected, but, by 9 days, fertility was significantly reduced (21% of controls). The postfertilization antifertility effect was not due to parthenogenic activation or to polyspermy. The antiserum reacted with one specific band in the one- and two-dimensional gel electrophoretic transfer blot procedure and was unaffected by absorption with different somatic tissues.

Animals↗
PubMed1984

Isoantigenicity of rabbit sperm, testis, and their extracts as demonstrated by Western blot enzyme immunobinding procedure.

The isoantigenicity of rabbit germ cells and their extracts was studied by using a Western blot enzyme immunobinding procedure after systemic and local intrauterine immunization. In the deoxycholate (DOC) sperm extract, the antisperm isoantiserum (ASA) recognized proteins of 100, 96, 92, 82, 53, 47, 43, and 29 kD, and the antitestis isoantiserum (ATA) recognized proteins of 100, 96, 92, 82, 43, and 29 kD. In the lithium diiodosalicylate (LIS) sperm extract, ASA reacted with four antigen bands of 92, 30, 14, and 6 kD, and ATA reacted with proteins of 94, 92, 91, 23, 17, 7, and 6 kD. In the reduced sodium dodecyl sulfate (SDS) sperm extract, ASA recognized proteins of 95, 76, 51, 41, 33, 30, 28, 26, and 22 kD; ATA showed a reaction only with proteins of 100, 98, and 95 kD, and anti-LIS/sperm isoantiserum (ALA) recognized three proteins of 98, 81, and 65 kD. The IgA of the immune uterine fluid (IUF) from the LIS/sperm-immunized rabbits recognized a 14-kD antigen in the DOC conceptus extract, a 51-kD antigen in the DOC testis extract, and 53-, 49-, 31-, and 28-kD proteins in the reduced SDS-sperm pellet extract. The IgA of the IUF from the NP-40/pellet-immunized rabbits showed two proteins of about 100 kD, plus ones of 56, 36, 31, 29, 19, 17, and 14 kD in the DOC conceptus extract; proteins of 92, 70, 51, 29, and 14 kD in the DOC testis extract; and of 61, 57, 53, 49, 35, 31, and 28 kD in the reduced SDS-sperm pellet extract.

Animals↗
PubMed1983

Inhibition of fertility in rabbits by monoclonal antibodies against sperm.

Monoclonal antibodies were prepared against rabbit sperm antigens by fusing P3-X63-Ag8-653 mouse myeloma cells with lymphocytes from Balb/c mice immunized with Tergitol NP-40 detergent-solubilized rabbit epididymal sperm. Ascites fluid from mice injected with two of these hybridomas (8C4.1 and 8C10.5) was negative in immobilization and agglutination methods, however, acrosome positive on methanol fixed sperm and plasma membrane positive on unfixed sperm in indirect immunofluorescence. Insemination of female rabbits with the sperm treated with either of these monoclonal antibodies resulted in significant reduction in fertility as seen by the percentage of 9-day implants/corpora lutea ratio (8C4.1, 25.7%; 8C10.5, 1.9%; and control, 64.7%). Though the antibodies inhibited in vitro binding of the rabbit sperm with zonae pellucidae of rat ova, fertilization in vivo was not affected significantly. The antibodies did not demonstrate antiblastocyst activity by immunofluorescence. Both of these monoclonal antibodies appeared to recognize the same antigen by the SDS-PAGE/Protein Blot enzyme immunobinding procedure. The antigen was of testicular origin and had a molecular weight of approximately 63,000 daltons. It is concluded that these monoclonal antibodies which were organ specific, block post-fertilization fertility by inhibiting some step necessary for viable embryo formation.

Animals↗
PubMed1981

Immunological sterilization of male dogs by BCG.

The effect of a single intratesticular injection of Freund's complete adjuvant (FCA), incomplete Freund's adjuvant (IFA) and various components has been investigated in dogs. FCA caused severe granulomatous reaction with wide-spread degeneration and vacuolation of the tubules. IFA and Arlacel A induced oligo - but not azoospermia, which was of short duration (6 weeks). Bacillus Calmette Guerin (BCG) suspended in saline given with uniform distribution at doses of 10 units or higher was effective in inducing aspermatogenesis. The sperm count declined to near zero level within 3 to 6 weeks after injection of BCG, and sperm suppression was maintained for 6 to 11 months of the observation period. At moderate doses of 10 to 25 units partial to complete inhibition of spermatogenesis was seen in different tubules with leukocyte infiltration in the interstitium. The basement membrane of the tubules, the peritubular cells as well as the Leydig cells were intact. At high doses (75-110 units) and at sites where bacillary deposits were high even in moderate dose animals, there was massive leukocyte infiltration with nonspecific damage to the tubules. The animals, on low or high dose, retained libido and copulated with bitches in oestrus. The mating was, however, infertile. The treatment, especially with low and moderate doses, was largely free of significant side effects. At higher doses, scrotal pain and swelling lasting for 7-15 days was observed; a small decrease in the size of the testes was also noted.

Animals↗
PubMed1981

Immunological control of male fertility.

A notable feature of the male gametes is the presence in them of proteins that are "foreign" to the immune system of both male and the female. It is there that are considered responsible for the elicitation of auto- and iso-antibodies in certain natural infertility cases. By virtue of their dual application in both sexes the sperm antigens have interesting potential for exploration as possible agents for control of fertility.

Animals↗
PubMed1979

A practicable immunological approach to block spermatogenesis without loss of androgens.

The intrinsic capacity of the immune system to elicit immune response selectively against late developing sperm proteins has been mobilized to intercept spermatogenesis. Bacillus Calmette-Guérin given in appropriate doses intratesticularly is effective in bringing about this effect. In dogs and rhesus monkeys, the sperm count in the semen declined precipitously, and almost complete azospermia was attained in 4-6 weeks after immunization. The few sperm cells that were present were immotile. Examination of serial sections of testes in immunized rats showed about 98% of the tubules to be devoid of sperm. The tubules were partially or fully atrophied. The basement membrane was, however, intact and the pertubular cell layer was normal. Sertoli cell nuclei were apparently normal but the cytoplasm was vacuolated and, in most cells, partially disintegrated. The lumen of the tubules was exhausted of formed elements and at times filled with eosinophilic debris. Leydig cells were present and hyperplasia of interstitial cells was seen, with massive infiltration of leukocytes. Blood testosterone levels were in the normal range and Leydig cells were responsive to gonadotropins. Libido was intact. The method was applicable to a variety of mammalian species. The implications of the results are discussed.

Adjuvants, Immunologic↗
PubMedDate not supplied

Reversibility of azoospermia induced by bacillus Calmette-Guerin.

A single intratesticular injection of bacillus Calmette-Guerin induces azoospermia within 3 to 6 weeks in a variety of animals without loss of androgens. After a period of azoospermia, the return of spermatogenesis was observed in dogs and monkeys. Seven dogs that showed aspermatogenesis after a local instillation of bacillus Calmette-Guerin (20-110 units per testis) again demonstrated spermatogenesis after a period of 153 to 325 days of azoospermia. Sperm counts and motility were restored to preimmunization levels within 6 weeks after initiation of spermatogenesis. In one dog, systemic treatment with antimycobacterial drugs helped to clear the bacillus Calmette-Guerin from the testis, thus hastening the reinitiation of spermatogenesis. In two monkeys injected with 110 and 160 units of bacillus Calmette-Guerin, azoospermia lasted up to 150 days. Subsequently, fertility was restored and the treated animals sired healthy offspring. A second injection of bacillus Calmette-Guerin of a lesser dose (5-20 units per testis) in these fertile dogs and monkeys again produced azoospermia. Histologically, the monkeys having a second period of azoospermia demonstrated aspermatogenic orchitis in the testis and leukocyte infiltration in the interstitium. A majority of the tubules had intact basement membranes, with a block of spermatogenesis at the spermatocyte stage. In dogs, the second azoospermia lasted for 113 to 277 days with a subsequent restoration of sperm counts and sperm motility to preimmunization levels. There were no circulating antisperm antibodies detected in sera of these animals during the azoospermic period or during its reversal. These observations suggest that the bacillus Calmette-Guerin-induced azoospermia is reversible, both in terms of sperm count and fertility.

Animals↗