The embryotoxicity of sera from patients with autoimmune diseases on post-implantation rat embryos in culture persists during remission and is not related to oxidative stress

Abstract

We evaluated the embryolethality and embryotoxicity of sera from patients suffering from autoimmune diseases during remission on post-implantation rat embryos cultured on these sera and determined the association between the patients' clinical history, high blood levels of specific antibodies, medications, and oxidative stress parameters. One hundred and eighty, 10.5-day-old rat embryos were cultured in their yolk sacs in 33 sera of systemic lupus erythematosus (SLE)/antiphospholipid syndrome (APS) patients, and compared with 84 embryos cultured in rat sera and 88 embryos cultured in control human sera. The sera proved to be lethal and embryotoxic but not teratogenic resulting in smaller yolk sacs and embryos, lower protein level and lower developmental score. Significantly less embryos cultured in 'toxic' SLE/APS sera had peak 2 of low molecular weight antioxidants (LMWA) wave, implying a delayed maturation of the antioxidant defense. Lower peak 1 of LMWA correlated with a history of recurrent abortions. Embryonic levels of superoxide dismutase (SOD) and catalase (CAT) did not correlate with sera toxicity, patients' clinical history or specific antibodies. We conclude that SLE/APS patients' clinical remission did not prevent death or developmental delay accompanied by later appearance of peak 2 of LMWA in post-implantation rat embryo cultures. The normal levels of the antioxidant enzymes evaluated may indicate that sera toxicity is not related to oxidative stress.

PreImplantation factor (PIF) protects cultured embryos against oxidative stress: relevance for recurrent pregnancy loss (RPL) therapy

“…Thus, in RPL PIF can negate disparate toxic elements and NK cells cytotoxicity supporting therapeutic potential [34]. RPL serum contains several embryo-toxic compounds and antibodies [36, 61]. Consequently, they are used in patients’ workup and possibly as targets for therapy.…”

“…Matured oocytes were transferred to a modified IVF medium (Fert-TALP; [61]) supplemented with 0.5 mM fructose, 0.2 mM non-essential amino acids, 6 mg/ml BSAFFA Fraction V, 30 μM penicilamine, 15 μM hypotaurine, 1.5 μM epinephrine (PHE), 22 μg/ml heparin, 20 μg/ml gentamicin, covered with light mineral oil in a humidified atmosphere at 38.5°C with 5 % CO 2 in air for 18 h (pH of 7.38 ± 0.01, 285 ± 1 mOsm). The IVF procedure for bovine species was recently reported.…”

“…Recovered COCs were matured in groups of 10-30 for 24 hrs in TCM-199 (with Earle's Salts) enriched with 10 % FCS, 0.2 mM sodium pyruvate, 1 mM alanyl-glutamine, 0.1 mM taurine, 0.1 mM cysteamine, 1 μg/ml estradiol, 85 mU/ml bovine follicle stimulating hormone (FSH, SIOUX Biochemical, Inc., Sioux Center, IA), 0.1 μg/ml gentamicin, and 10 ng/ml epidermal growth factor (EGF) at pH 7.35 ± 0.02 and osmolarity of 300 ± 2 mOsm and covered with light mineral oil in a humidified atmosphere at 38.5°C with 5 % CO 2 in air. Matured oocytes were transferred to a modified IVF medium (Fert-TALP; [ 61 ]) supplemented with 0.5 mM fructose, 0.2 mM non-essential amino acids, 6 mg/ml BSAFFA Fraction V, 30 μM penicilamine, 15 μM hypotaurine, 1.5 μM epinephrine (PHE), 22 μg/ml heparin, 20 μg/ml gentamicin, covered with light mineral oil in a humidified atmosphere at 38.5°C with 5 % CO 2 in air for 18 h (pH of 7.38 ± 0.01, 285 ± 1 mOsm). The IVF procedure for bovine species was recently reported.…”

Embryonic and Placental Damage Induced by Maternal Autoimmune Diseases - What Can We Learn from Experimental Models

“…Additionally a significant positive correlation between the growth parameters was demonstrated among the embryos cultured in sera from patients and controls: the larger embryos had larger yolk sacs; had higher general and neurologic developmental score and more somites, implying more mature embryos. (Ergaz et al, 2010). A high rate of anomalies in embryos cultured in sera from women with recurrent spontaneous abortions (Abir et al, 1994) or following the addition to the culture medium of purified IgG from SLE and APLs patients (Abir & Ornoy, 1996), differed from the study using sera obtained from non-pregnant patients in remission, where there was no increase in the rate of anomalies compared to controls.…”