Gene expression profiles of single human mature oocytes in relation to age.

BACKGROUND: The development competence of human oocytes declines with increasing age. The objective of this study was to investigate the effect of age on gene expression profile in mature human oocytes. METHODS: mRNA was isolated for whole genome gene expression microarray analysis from metaphase II (MII) oocytes donated by IVF or ICSI patients [10 women aged <36 years (younger) and five women aged 37-39 years (both inclusive) (older)] undergoing controlled ovarian stimulation. The oocytes were donated and prepared immediately after recovery from the follicle. RT-PCR on additional four younger and two older oocytes confirmed the array analysis. RESULTS: On the basis of 15 independent replicates of single MII oocytes, 7470 genes (10 428 transcripts) were identified as present in the MII oocytes. Of these, 342 genes showed a significantly different expression level between the two age groups; notably, genes annotated to be involved in cell cycle regulation, chromosome alignment (e.g. MAD2L1 binding protein), sister chromatid separation (e.g. separase), oxidative stress and ubiquitination. The top signaling network affected by age was 'cell cycle and organism development' (e.g. SMAD2 and activin B1 receptor). CONCLUSION: There is a substantial difference between younger and older oocytes in the transcriptional level of genes involved in central biological functions of the oocytes, thus providing information on processes that may be associated with the ageing phenomenon and possibly contributing to decreased fertility.

Recombinant LH supplementation to recombinant FSH during the final days of controlled ovarian stimulation for in vitro fertilization. A multicentre, prospective, randomized, controlled trial.

BACKGROUND: The purpose of this multicentre, multinational trial was to study whether rLH supplementation to recombinant FSH (rFSH) during the late follicular phase increased pregnancy rates. METHODS: After down-regulation with nafarelin, 526 women were randomized on Day 1 of stimulation to use either rFSH (Gonal-F) alone (n = 261) or to continue after Day 6 of stimulation with both rFSH (Gonal-F) and rLH (Luveris) (n = 265) from Day 6. The starting dose of rFSH was 150-225 IU/day according to age below or above 35 years. RESULTS: Ongoing pregnancy rate at week 10-12 was 28.7% after rFSH alone and 27.2% after rFSH + rLH. This showed no evidence of a difference. Administration of rLH significantly (P< 0.001) increased serum LH. Ongoing pregnancy rates in patients with low LH levels (<33 percentile) on Days 1 and 6 of stimulation showed no difference between the group treated with rFSH only (23.9% low Day 1 LH; 22.1% low Day 6 LH) versus rFSH + rLH (25.0% low Day 1 LH; 28.9% low Day 6 LH). CONCLUSIONS: Supplementing rFSH with daily doses of 75-150 IU of rLH during the second half of the follicular phase showed no evidence of increasing the ongoing pregnancy rates in the general population. (ClinicalTrials.gov, trial number: KF02-035/03).

Apolipoprotein E genotypes predict attendance rates at lipid clinic.

Except for the rare epsilon22 genotype it remains largely unsettled whether apolipoprotein E genotype influences an individual's referral to lipid clinics. To test this hypothesis, we compared genotype distributions among 156 hypercholesterolemic and 83 hypertriglyceridemic patients attending a lipid clinic with that among 9241 individuals sampled from the Danish general population. The relative genotype frequencies of epsilon22, epsilon32, epsilon42, epsilon33, epsilon43, and epsilon44 were 0.005, 0.126, 0.026, 0.564, 0.251, and 0.027 in the general population, which differed from genotype frequencies in both hypercholesterolemic (chi2: P = 0.01) and hypertriglyceridemic patients (chi2: P < 0.001). By comparison with epsilon33, epsilon44 predicted a 2-fold increase whereas epsilon32 predicted a 2-fold decrease in the attendance rate at the lipid clinic for hypercholesterolemic patients (95% confidence intervals: 1.1-4.3 and 0.2-0.9). Among hypertriglyceridemic patients, epsilon22, epsilon42, epsilon43, and epsilon44 versus epsilon33 predicted 13-, 3-, 1 1/2-, and 3-fold attendance rates at the lipid clinic, respectively (95% confidence intervals: 4.5-39.9, 1.2-8.4, 1.0-2.8, and 1.1-7.6). These findings are in accordance with the fact that epsilon44 raises cholesterol levels, epsilon32 reduces cholesterol levels, and epsilon22, epsilon42, epsilon43, and epsilon44 raise triglyceride levels in comparison with epsilon33. These data suggest that hypercholesterolemic individuals carrying epsilon44 and hypertriglyceridemic individuals carrying epsilon22, epsilon42, epsilon43, or epsilon44 are relatively more often referred to lipid clinics than carriers of epsilon33.

Spectrum of LDL receptor gene mutations in Denmark: implications for molecular diagnostic strategy in heterozygous familial hypercholesterolemia.

Heterozygous familial hypercholesterolemia (FH) is one of the most common potentially fatal single-gene diseases leading to premature coronary artery disease, but the majority of heterozygous FH patients have not been diagnosed. FH is due to mutations in the gene coding for the low-density lipoprotein (LDL) receptor, and molecular genetic diagnosis may facilitate identification of more FH subjects. The Danish spectrum of 29 different mutations, five of which account for almost half of heterozygous FH, is intermediate between that of countries such as South Africa, where three mutations cause 95% of heterozygous FH in the Afrikaners, and Germany or England, where there are many more mutations. In clinical practice, a strategy for the genetic diagnosis of heterozygous FH, tailored to the mutational spectrum of patients likely to be seen at the particular hospital/region of the country, will be more efficient than screening of the whole LDL receptor gene by techniques such as single-strand conformation polymorphism (SSCP) analysis in every heterozygous FH candidate. In Aarhus, Denmark, we have chosen to examine all heterozygous FH candidates for the five most common LDL receptor gene mutations (W23X, W66G, W556S, 313 + 1G --> A, 1846 - 1G --> A) and the apoB-3500 mutation by rapid restriction fragment analysis. Negative samples are examined for other mutations by SSCP analysis followed by DNA sequencing of the exon indicated by SSCP to contain a mutation. If no point mutation or small insertion/deletion is detected, Southern blot or Long PCR analysis is performed to look for the presence of large gene rearrangements. In conclusion, our data suggest that an efficient molecular diagnostic strategy depends on the composition of common and rare mutations in a population.

Lipoprotein(a) and dietary proteins: casein lowers lipoprotein(a) concentrations as compared with soy protein.

BACKGROUND: Substitution of soy protein for casein in the diet decreases LDL cholesterol and increases HDL cholesterol. How the 2 proteins affect lipoprotein(a) [Lp(a)], an independent risk factor for coronary artery disease, is unknown. OBJECTIVE: We compared the effects of dietary soy protein and casein on plasma Lp(a) concentrations. DESIGN: Nine normolipidemic men were studied initially while consuming their habitual, self-selected diets, and then, in a crossover design, while consuming 2 liquid-formula diets containing either casein or soy protein. The dietary periods lasted 45 d (n = 7) or 33 d (n = 2). Fasting total cholesterol, LDL-cholesterol, HDL-cholesterol, triacylglycerol, and Lp(a) concentrations were measured throughout. RESULTS: After 30 d of each diet, the mean concentration of Lp(a) was not significantly different after the soy-protein and self-selected diets. However, Lp(a) decreased by an average of 50% (P < 0.001) after the casein diet as compared with concentrations after both the soy-protein and self-selected diets. Two weeks after subjects switched from the self-selected to the soy-protein diet, Lp(a) increased by 20% (P = 0.065), but subsequently decreased to baseline. In contrast, the switch to the casein diet did not cause an increase in Lp(a), but instead a continuing decrease in mean concentrations to 65% below baseline (P < 0.0002). Total cholesterol, LDL cholesterol, and HDL cholesterol were significantly lower > or =30 d after both the casein and soy-protein diets than after the self-selected diet (P < 0.001). HDL cholesterol was 11% higher after the soy-protein diet than after the casein diet (P < 0.002), but LDL cholesterol, total cholesterol, and triacylglycerol were not significantly different after the casein and soy-protein diets. CONCLUSION: These findings indicate that soy protein may have an Lp(a)-raising effect, potentially detrimental to its use in antiatherogenic diets.

Variable lipemic response to dietary soy protein in healthy, normolipemic men.

We found previously that dietary soy protein, compared with casein, reduced plasma LDL cholesterol and increased HDL cholesterol concentrations in healthy women and men. However, there was considerable variation among individuals. The aim of this study was to characterize the lipoprotein responsiveness of individuals to examine whether different response patterns could be identified. Nine normolipemic men consumed 2 liquid-formula diets of identical composition except that the protein component was either soy protein or casein. After 1 mo of consuming each diet, the subjects' plasma HDL cholesterol (P < 0.01) and apolipoprotein (apo) A-I (P < 0.05) concentrations were increased by the soy-protein diet whereas the ratio of LDL cholesterol to HDL cholesterol was decreased (P < 0.01); total cholesterol, triacylglycerol, LDL cholesterol, apo B and apo A-II were insignificantly affected. In 5 individuals, however, soy protein reduced mean LDL cholesterol, LDL2 cholesterol, and LDL2 apo B concentrations by 26% and plasma apo B by 16%, whereas HDL cholesterol increased by 11%. In 3 other individuals, soy protein increased mean HDL cholesterol by 17% and plasma apo A-I by 12%, but did not lower LDL. In 1 subject, soy protein decreased LDL2 cholesterol by 11% and increased plasma triacylglycerol by 40%, but neither HDL cholesterol nor apo A-I increased. We identified 3 types of lipemic responses to dietary soy protein involving a reduction in atherogenic LDL and increase in antiatherogenic HDL. In most subjects, the effects on both LDL and HDL were favorable, although fewer experienced either an increase in HDL or a decrease in LDL2.

Association of mutations in the apolipoprotein B gene with hypercholesterolemia and the risk of ischemic heart disease.

BACKGROUND: Familial hypercholesterolemia leads to premature ischemic heart disease and is often caused by mutations in the gene for the low-density lipoprotein receptor. Mutations in the apolipoprotein B gene, which encodes a ligand for this receptor, may also result in this phenotype. METHODS: We studied the genotypes of 9255 women and men from the general population, 948 patients with ischemic heart disease, and 36 patients with familial hypercholesterolemia, all from Denmark, for three mutations in the apolipoprotein B gene: Arg3500Gln, Arg3531Cys, and Arg3500Trp. RESULTS: The prevalence of heterozygotes in the general population was 0.08 percent (95 percent confidence interval, 0.03 to 0.16 percent) for both the Arg3500Gln and the Arg3531Cys mutations, and 0.00 percent (95 percent confidence interval, 0.00 to 0.18 percent) for the Arg3500Trp mutation. Among carriers of the Arg3500Gln mutation, cholesterol levels were significantly higher than among noncarriers in the general population - by 100 mg per deciliter (2.6 mmol per liter) among carriers in the general population, 154 mg per deciliter (4.0 mmol per liter) among patients with ischemic heart disease, and 172 mg per deciliter (4.5 mmol per liter) among patients with familial hypercholesterolemia. Heterozygous carriers of the Arg3500Gln mutation were significantly more common among patients with ischemic heart disease (odds ratio, 7.0; 95 percent confidence interval, 2.2 to 22; P=0.003) and patients with familial hypercholesterolemia (odds ratio, 78; 95 percent confidence interval, 16 to 388; P=0.001) than in the general population. Heterzygous carriers of the Arg3531Cys mutation in the general population did not have higher-than-normal plasma cholesterol levels or an increased risk of ischemic heart disease (odds ratio; 1.4; 95 percent confidence interval, 0.2 to 11; P=0.54). CONCLUSIONS: The Arg3500Gln mutation in the apolipoprotein B gene, which is responsible for familial defective apolipoprotein B-100 and is present in approximately 1 in 1000 persons in Denmark, causes severe hypercholesterolemia and increases the risk of ischemic heart disease.

Phenotypic characterization of a patient homozygous for the D558N LDL receptor gene mutation.

We describe the clinical, biochemical, and genetic features of a patient with true homozygous familial hypercholesterolemia due to the D558N low-density lipoprotein receptor gene mutation, previously designated FH Cincinnati-4. Functional flow-cytometric analysis of the LDL receptorR protein on upregulated EBV-transformed lymphocytes indicated reduction of the number of receptors on the cell surface by 87% and reduction of receptor activity by 89% compared to control cells. With drugs and a portacaval shunt operation, performed when the patient was 15 years old, serum cholesterol was reduced from about 28 to about 15 mmol/l. He died at the age of 32 of a myocardial infarction. The autopsy showed generalized atherosclerosis, especially in the coronary arteries, which were severely stenosed proximally. A rare finding was a large intracranial xanthoma that apparently had been asymptomatic.

Treatment of patients with familial defective apolipoprotein B-100 with pravastatin and gemfibrozil: a two-period cross-over study.

Thirty patients with familial defective apolipoprotein B-100 were treated in a two-period (8 weeks each) cross-over study with pravastatin and gemfibrozil. Cholesterol, LDL cholesterol, and apo B were reduced by 20-25% (P < 10(-4)) by pravastatin and by 4-6% by gemfibrozil (pravastatin vs. gemfibrozil: P < 10(-4)). Response to pravastatin was variable and not correlated to gender, age, or apo E genotype. Gemfibrozil lowered triglycerides by 25% (P < 10(-4)) and raised HDL cholesterol by 11%. The effects of pravastatin on these two interrelated variables were significantly smaller. Both drugs increased Lp(a) significantly by about 10%. The LDL cholesterol lowering effect of pravastatin in patients with FDB is similar to that observed in patients with familial hypercholesterolemia.

Incidence of the apolipoprotein B-3500 mutation in Denmark.

A total of 5000 consecutively samples newborn screening cards were anonymously selected for screening for the apolipoprotein B-3500 (apo B-3500) mutation, which causes familial defective apolipoprotein B-100 (FDB). The mutation was found in 5 of 5000 Danish children, of whom 2 were twins. This indicates a lower prevalence of this mutation in Danes than that reported in the UK, Germany, USA, Austria, Canada and especially Switzerland. Haplotype studies suggest that Caucasian subjects with the apo B-3500 mutation have a common founder. The apparently lower prevalence in Denmark than in Switzerland and Central Europe may indicate that the mutation was brought from these areas to Denmark after the initial settling of Denmark. In 101 unrelated Danish subjects with familial hypercholesterolemia, diagnosed on clinical and biochemical criteria including tendon xanthomata, 2 were heterozygous for the apo B-3500 mutation (2%).

Continuous combined and sequential estradiol and norethindrone acetate treatment of postmenopausal women: effect of plasma lipoproteins in a two-year placebo-controlled trial.

OBJECTIVE: Our purpose was to examine the effects of postmenopausal estrogen therapy supplemented with progestogen on plasma lipoprotein levels. STUDY DESIGN: One hundred thirteen women were randomized to receive either placebo or a combination of 17 beta-estradiol and norethindrone acetate administered continuously (Kliogest) or sequentially (Trisequens). Plasma lipoprotein levels were measured at baseline and after 2 years of treatment and compared by analysis of variance. RESULTS: Hormone therapy lowered plasma cholesterol levels (p < 0.001) and low-density lipoprotein cholesterol (Kiogest, p < 0.001; Trisequens, p < 0.01), whereas high-density lipoprotein cholesterol levels were unchanged (Trisequens) or reduced (Kliogest, p < 0.01), primarily because of a decrease in the high-density lipoprotein-2 subfraction (p < 0.05). Low-density lipoprotein/high-density lipoprotein cholesterol ratios remained unchanged. CONCLUSIONS: Although hormonal replacement therapy with estradiol combined with norethindrone acetate eliminated the increase in high-density lipoprotein cholesterol levels observed with estrogen monotherapy, the reductions in low-density lipoprotein cholesterol concentrations still suggest reduced cardiovascular risk, according to the National Cholesterol Education Program and to recent observations indicating that risk is not necessarily inversely proportional to high-density lipoprotein cholesterol levels.

Characteristics of 46 heterozygous carriers and 57 unaffected relatives in five Danish families with familial defective apolipoprotein B-100.

Plasma concentrations of cholesterol, high-density lipoprotein (HDL) cholesterol, low-density lipoprotein (LDL) cholesterol, apolipoprotein (apo) B, and lipoprotein(a) (Lp[a]) in 46 persons heterozygous for the apo B-3500 mutation causing familial defective apo B-100 (FDB) were compared with those in 57 non-FDB relatives. FDB patients had 50% to 70% higher mean concentrations of cholesterol, LDL cholesterol, and apo B than non-FDB relatives (P < 10(-4) for all three variables). Triglycerides were higher (P = .016) and HDL cholesterol was lower (P = .021) in FDB patients. The concentration ranges of these variables were broad in each family, and there was no between-family difference in means for cholesterol and LDL cholesterol. There was no phenotype-specific difference in Lp(a) concentrations between FDB patients and non-FDB relatives. Apo E4 is normally associated with higher concentrations of LDL and apo E2 with lower concentrations. This relation was partly reversed in FDB patients: apo E4 was associated with lower apo B concentrations and apo E2 with higher apo B concentrations. Tendon xanthomata were found in members of two of the five families. Six of 12 FDB patients > 50 years old had atherosclerotic disease. In contrast, all 18 non-FDB relatives > 50 years old were apparently healthy. A total of 8 FDB patients with atherosclerotic disease had 36% higher cholesterol concentrations, 28% higher apo B concentrations, 50% higher triglyceride concentrations, and 120% higher Lp(a) concentrations than FDB patients without clinical atherosclerosis.(ABSTRACT TRUNCATED AT 250 WORDS)

Characterization of a disease-causing Glu119-Lys mutation in the low-density lipoprotein receptor gene in two Danish families with heterozygous familial hypercholesterolemia.

Mutations in the gene for the low-density lipoprotein receptor (LDL receptor) cause the autosomal dominant inherited disease familial hypercholesterolemia (FH). In 15 Danish patients with heterozygous FH we have screened exon 4 of the LDL receptor gene for point mutations and small rearrangements employing genomic DNA amplification and bidirectional solid-phase sequencing. Two subjects were found to be heterozygous for a guanine to adenine base substitution at nucleotide position 418 of the LDL receptor cDNA. This point mutation results in an amino acid change from glutamic acid to lysine at amino acid residue 119 in the third repeat of the cysteine-rich ligand binding domain of the mature LDL receptor. Disruption of LDL receptor function by the Glu119-Lys mutation was confirmed by site-directed mutagenesis and expression in COS-7 cells. By Western blotting the mutation was found to affect the processing of the LDL receptor protein. Using flow cytometric analysis of the transfected cells a decreased binding and internalization of LDL by the mutant receptor was documented. By means of a mutation-specific PCR-based assay the Glu119-Lys mutation was not detected in another 85 apparently unrelated Danish heterozygous FH patients. We identified six persons in the index families with the Glu119-Lys mutation cosegregating with the clinical syndrome of FH in these families. Furthermore, haplotype analysis revealed that the haplotype [SfaNI+, StuI+, AvaII-, (dTA)7] of the mutation carrying allele was the same in the two apparently unrelated patients. This indicates that the mutation has been inherited from a common ancestor.

Cholesteryl esters in lymph chylomicrons: contribution from high density lipoprotein transferred from plasma into intestinal lymph.

Most of the cholesterol in intestinal chyle and chylomicrons is derived from plasma. Our aim was to determine how much plasma low density (LDL) and high density (HDL) lipoproteins contribute to the cholesterol in chyle and chylomicrons, and to examine how plasma cholesterol becomes associated with lymph chylomicrons. Intravenous injection of radioiodinated plasma lipoproteins into two chyluric patients showed that 82% of the HDL plasma pool transferred daily to intestinal chyle, corresponding to 58% of lymph cholesterol; LDL contributed 18% of its plasma pool, corresponding to 18% of lymph cholesterol. When plasma HDL radiolabeled in both the protein and cholesteryl ester moieties was injected, the isotope ratios of plasma HDL and lymph lipoproteins were identical; 85% of the HDL cholesteryl esters transferred to triglyceride-rich lipoproteins, while the apolipoproteins remained largely (70%) in the higher density lipoproteins of the chyle. Incubations of similarly labeled plasma HDL showed preferential transfer of cholesteryl esters to artificial chylomicrons mediated by a factor present in lipoprotein-free plasma. Thus, a sizable portion of plasma HDL enters intestinal lymphatics probably as intact HDL, and then transfers part of their cholesteryl esters to chylomicrons, possibly mediated by transfer proteins. Reverse cholesterol transport may therefore include an extravascular loop via lymph chylomicrons and chylomicron remnants to the liver.

Apolipoprotein(a) polymorphism predicts the increase of Lp(a) by pravastatin in patients with familial hypercholesterolaemia treated with bile acid sequestration.

HMG-CoA reductase inhibitors effectively reduce the concentration of low density lipoproteins (LDL) in plasma. Lipoprotein(a) [Lp(a)] may be as atherogenic as LDL. A few studies, only one of which was placebo controlled, suggest that the HMG CoA reductase inhibitors either do not affect Lp(a) or they increase Lp(a). The response of Lp(a) to HMG-CoA reductase inhibition has not been related to apolipoprotein(a) phenotypes in previous studies. We conducted a double-blind, placebo controlled study of pravastatin in 51 patients with familial hypercholesterolemia (FH) (n = 43) or probable FH (n = 8). All patients had LDL-cholesterol concentration above 4.1 mmol l-1 despite treatment with diet and bile acid sequestration. In patients assigned to pravastatin (n = 34), the mean concentrations of total cholesterol and LDL cholesterol fell significantly (P < 0.01) when compared to placebo. Lp(a) increased (P < 0.01) from a mean (+/- SD) of 33.6 +/- 40.8 mg dl-1 to 41.1 +/- 46.1 mg dl-1 on pravastatin but was unchanged during placebo treatment. The percentage increase in Lp(a) was the same in patients with different apo(a) phenotypes, and hence the absolute increase in Lp(a) was greatest in patients with the low molecular weight apo(a) phenotypes.

Antisperm antibodies in split ejaculates.

The split ejaculation technique concentrates the most motile and viable spermatozoa in the first part of the ejaculate. Several clinicians use this fraction for insemination and in vitro fertilization. Eleven vasovasostomized men with antisperm antibodies participated in this study. The mixed antiglobulin reaction (MAR) for IgG, IgA, and secretory component was carried out on the whole ejaculate and the fractions of the split ejaculate. The isotype and concentration of free antisperm antibodies were determined in serum, the whole ejaculate and the fractions of the split ejaculate by the indirect MAR and the tray agglutination test. The detection of bound antisperm antibodies revealed almost identical reactions in fractions one and two and in the whole ejaculate. The results suggested that the majority of antisperm antibodies in the ejaculate from vasovasostomized men are transuded from serum not only at the epididymal and/or the prostatic level but also in the seminal vesicles.

Antisperm antibodies on epididymal spermatozoa.

Still under debate is the location in the genital tract where antisperm antibodies transuded from serum and locally produced antibodies, respectively, become attached to the surface of the spermatozoa. The mixed antiglobulin reaction (MAR) for immunoglobulin (Ig) G, IgA, and locally produced secretory IgA (sIgA) could be carried out on motile spermatozoa from epididymis from 13 of 34 men undergoing vasovasostomy. Four months and one year after the operation the MAR was repeated on a fresh semen sample from all patients. It was found that transuded as well as locally produced antisperm antibodies were bound to spermatozoa at the epididymal level of the genital tract. The follow-up results on semen were almost identical to the results obtained with epididymal spermatozoa.

Effects of dietary proteins on plasma lipoprotein levels in normal subjects: interaction with dietary cholesterol.

We have compared the effects of dietary soy protein and casein in diets low in cholesterol (less than 100 mg/d) and in diets enriched in cholesterol (500 mg/d) to examine whether the level of cholesterol intake affects the response of plasma lipoproteins to dietary proteins of plant and animal origin. Normal men and women consumed formula diets containing 20% of calories as soy protein or casein, 27% as fat and 53% as carbohydrate in 2 crossover studies. The dietary periods lasted for 31 days and were separated by a month-long interim period on self-chosen food. Following an initial reduction of plasma total cholesterol and low-density lipoprotein-cholesterol (LDL-C) levels on all diets, the plasma lipid and lipoprotein concentrations stabilized. On low-cholesterol diets the concentration of each of the major lipoprotein classes were similar during the soy and the casein dietary periods. On cholesterol-enriched diets, the concentration of LDL-C stabilized at a 16% lower level on soy protein than on the casein diet (p less than 0.02), while the concentration of high-density lipoprotein-cholesterol (HDL-C) was 16% higher (p less than 0.01). Since the difference in LDL-C (p less than 0.05) and in HDL-C (p less than 0.025) levels on casein and on soy protein diets were significantly greater on the high than on the low cholesterol intake, the findings indicate that the level of dietary cholesterol may determine whether plant and animal dietary proteins have similar or different effects on plasma LDL-C and HDL-C concentrations.

Antisperm antibodies and fertility after vasovasostomy: a follow-up study of 216 men.

A group of 216 vasovasostomized men were tested with the mixed antiglobulin reaction for immunoglobulin (Ig)G, IgA, and secretory IgA antisperm antibodies bound to the sperm membrane. Free antisperm antibodies in serum and seminal plasma were detected with the gelatin agglutination test and the tray agglutination test. The results were related to the conception rates. In a subgroup with a pure IgG response, the conception rate reached 85.7%, whereas only 42.9% of the men who also had IgA on the sperm induced pregnancy. When 100% of the spermatozoa were covered with IgA, the conception rate was reduced to 21.7%. The combination of IgA on all sperm and a strong immune response (titer in serum greater than or equal to 256) was associated with a conception rate of zero.