Formation of different abzymes in autoimmune-prone MRL-lpr/lpr mice is associated with changes in colony formation of haematopoietic progenitors

It was shown that IgGs from the sera of 2-7-month-old control non-autoimmune (CBA x C57BL)F1 and BALB/c mice and 2-3-month-old autoimmune prone MRL-lpr/lpr mice (conditionally healthy mice) are catalytically inactive. During spontaneous development of deep systemic lupus erythematosus (SLE)-like pathology a specific reorganization of immune system of these mice leads to conditions associated with a production of IgGs hydrolyzing DNA, ATP and polysaccharides with low catalytic activities (conditionally pre-diseased mice).A significant increase in DNase, ATPase and amylase IgG relative activities associated with a transition from pre-diseased to deep diseased mice is correlated with additional changes in differentiation and proliferation of mice bone marrow haematopoietic stem cells (HSCs) and lymphocyte proliferation in different organs.The highest increase in all abzyme activities was found in mice immunized with DNA, which in comparison with pre-diseased and diseased mice are characterized by a different profile of HSC differentiation and by a suppression of cell apoptosis. Abzyme activities in the serum of pregnant females were comparable with those for pre-diseased mice, but the profile of HSC differentiation and cell apoptosis levels in pregnant and pre-diseased mice were quite different. Right after the beginning of lactation (4 days after delivery) and in a late time of lactation (14 days after delivery) there was an observed increase in cell apoptosis and two different stages of significant change in the HSC differentiation profiles; the first stage was accompanied with a significant increase and the second with a remarkable decrease in abzyme activities. Overall, all mouse groups investigated are characterized by a specific relationship between abzyme activities, HSC differentiation profiles, levels of lymphocyte proliferation, and cell apoptosis in different organs. From our point of view, the appearance of ATPase, DNase activities may be considered the earliest statistically significant marker of mouse spontaneous SLE and a further significant increase in their activities correlates with the appearance of SLE visible markers and with an increase in concentrations of anti-DNA Abs and urine protein. However, development of autoimmune (AI)-reactions and the increase in the sera anti-DNA antibodies (Abs) and in the abzyme activities in pregnant and lactating mice do not associate with SLE visible markers and proteinuria. The possible differences in immune system reorganizations during pre-disease, disease, pregnancy and lactation leading to production of different auto-antibodies and abzymes are discussed.     

Immunization of rabbits with DNase I produces polyclonal antibodies with DNase and RNase activities

Immunization of rabbits with DNase I leads to the production of antiidiotypic Abs with DNase activity. It is not known at present whether antiidiotypic Abs against DNA-hydrolyzing enzymes can possess RNase activity. Here we show that immunization of healthy rabbits with bovine DNase I produces IgGs with intrinsic DNase and RNase activities. Electrophoretically and immunologically homogeneous polyclonal IgGs were obtained by sequential chromatography of the immune sera on Protein A-Sepharose and gel filtration. Affinity chromatography on DNA cellulose using elution of Abs with different concentrations of NaCl and an acidic buffer separated catalytic IgGs into four Ab subfractions, three of which demonstrated only DNase activity while one subfraction hydrolyzed RNA faster than DNA. The serum of patients with many different autoimmune (AI) diseases contains small fractions of antibodies (Abs) interacting with immobilized DNA, which possess both DNase and RNase activities. Our data suggest that a fraction of abzymes from AI patients hydrolyzing both DNA and RNA can contain a subfraction of Abs against DNase I.     

Comparison of Enzymatic Properties of DNA‐Abzymes and Human DNAses

DNA‐hydrolyzing antibodies (DNA‐abzymes, Abz) were shown to be good biochemical markers of some autoimmune diseases such as systemic lupus erythematosus (SLE) and multiple sclerosis (MS). To better understand mechanisms of abzyme generation, one needs to know optimal conditions for DNA hydrolysis by DNA‐abzymes, as well as their enzymatic properties in comparison with those of enzymes possessing the same activity. In contrast to human urine deoxyribonucleases, DNA‐hydrolyzing antibodies efficiently digested both single‐ and double‐strand DNA. It was shown that polyclonal antibodies (Abs) in MS may contain up to several types of DNase activities, either activated by metal ions or not.     

DNase,RNase, and phosphatase activities of antibodies formed upon immunization by DNA,DNase I,and DNase II

Relative DNase, RNase (efficiency of hydrolysis of ribo- and deoxyribooligonucleotides (ON)), and phosphatase (removal of the ON 5′ terminal phosphate) catalytic activities of antibodies (AB) obtained after rabbit immunization by DNA, DNase I, and DNase II were compared. It is shown that electrophoretically homogeneous preparations of polyclonal AB from non-immunized rabbits did not exhibit such activities. Immunization of rabbits by DNA, DNase I, and DNase II results in generation of IgG abzymes that exhibit high activity in the ON hydrolysis reaction and even higher activity in cleavage of 5′ terminal phosphate of ON. In this case K _m values for supercoiled plasmid DNA and ON found in reactions of their AB-dependent nuclease hydrolysis and phosphatase cleavage of 5′ terminal phosphate differ by 2–4 orders of magnitude. This shows that nuclease and phosphatase activities belong to different abzyme fractions within polyclonal AB. Thus, in this work data indicative of the possibility of a formation of antibodies exhibiting phosphatase activity after immunization of animals with DNA, DNase I, and DNase II, were obtained for the first time. Possible reasons for production of AB with phosphatase activity after immunization of rabbits with these immunogens are discussed.     

Comparison of enzymatic properties of DNA-abzymes and human DNAses

DNA-hydrolyzing antibodies (DNA-abzymes, Abz) were shown to be good biochemical markers of some autoimmune diseases such as systemic lupus erythematosus (SLE) and multiple sclerosis (MS). To better understand mechanisms of abzyme generation, one needs to know optimal conditions for DNA hydrolysis by DNA-abzymes, as well as their enzymatic properties in comparison with those of enzymes possessing the same activity. In contrast to human urine deoxyribonucleases, DNA-hydrolyzing antibodies efficiently digested both single- and double-strand DNA. It was shown that polyclonal antibodies (Abs) in MS may contain up to several types of DNase activities, either activated by metal ions or not.     

Induction of glutathione S-transferases A, B and C in rat liver cytosol by trans-stilbene oxide

RNAase H, which catalyzes the hydrolysis of the RNA moiety of an RNA-DNA hybrid, was measured in the mammary gland of virgin, pregnant, lactating, and weaning Fischer rats and in the R3230AC mammary tumor grown in the same animals. In the normal mammary gland when DNA levels were low, as in the virgin state or during involution, RNAase H activity was also low. During pregnancy and lactogenesis when DNA levels increased, RNAase H activity, either on the basis of mammary gland weight or DNA content, also increased. During lactation when cellular proliferation ceases but rates of RNA and protein synthesis continue to reach peak values, RNAase H activity decreased. Compared to the corresponding enzyme from host glands, RNAase H from the R3230AC mammary tumor grown in pregnant and lactating hosts changes similarly, but to a lesser extent. The RNAase H activity which, ona tissue weight basis, was higher than in normal tissue also increased during pregnancy and directly after parturition, but decreased during lactation. During pregnancy these changes were accompanied by an increase in tumor DNA values. During lactation the tumor DNA values returned to the level seen in virgin hosts. These results are consistent with a role for RNAase H in DNA replication in rat mammary gland and in R3230Ac mammary tumor.     

Prevalence and avidity of human herpesvirus-6 specific IgG antibodies in pregnant women in Hungary

The prevalence, the level and the avidity of human herpesvirus-6 (HHV-6) specific IgG were examined in pregnant women and age-matched female blood donors. The study group consisted of 180 women (age 14-45); 60 women with normal pregnancy, 60 pregnant women with fetuses suspected of having any viral infection and 60 healthy blood donors with no history of pregnancy. Plasma or serum samples were tested for HHV-6 IgG antibodies by an immunofluorescence assay. Ninety-eight percent of blood donors and 97% of 120 pregnant women had IgG antibodies to HHV-6. The rate of seropositivity in women with normal pregnancies and women with fetuses suspected to have viral infection was the same. Pregnant women (n = 120) had significantly lower antibody titer than blood donors. No significant differences were found in the same respect between the two groups of pregnant women. Low avidity of IgG antibodies to HHV-6 was detected in 5% of pregnant women.     

Alkaline ribonuclease and ribonuclease inhibitor in mammary gland during the lactation cycle and in the R3230AC mammary tumour.

Alkaline RNAase (ribonuclease) and RNAase inhibitor were assayed to determine the potential role of the degradative process in regulating the amount of RNA in the mammary gland and mammary tumour. Very little free alkaline RNAase activity was found in the cytosol fraction of the mammary gland of virgin, pregnant, lactating or involuting Fischer rats. However, addition of p-chloromercuribenzoate to the assay medium revealed latent RNAase which, when expressed on a DNA basis, decreased during pregnancy and lactation. The cytosol latent RNAase is stable in 0.125 M-H2SO4. The non-cytosol RNAase activity also decreased during pregnancy and lactation. Addition of Triton X-100 produced slightly higher activity at all stages tested. The inhibitor activity in rat mammary gland was very low before pregnancy, increased gradually during pregnancy and more dramatically at parturition, continued to increase throughout lactation and returned to resting-gland values by the sixth day of involution. The increase during pregnancy may be due to the increased cellularity of the gland, whereas the gain during lactation was more than could be accounted for by increases in cell number. The R3230AC transplantable mammary tumour resembles the normal gland in early lactation with respect to both its cytosol and non-cytosol alkaline RNAase activities and its moderately high content of RNAase inhibitor. The relatively high inhibitor and low RNAase activities in both the gland of the lactating rat and in the tumour are of potential significance in maintaining high amounts of RNA and increased rates of protein synthesis in these tissues.     

Antibodies against RNA hydrolyze RNA and DNA

Immunization of animals with DNA leads to the production of anti-DNA antibodies (Abs) demonstrating both DNase and RNase activities. It is currently not known whether anti-RNA Abs can possess nuclease activities. In an attempt to address this question, we have shown that immunization of three rabbits with complex of RNA with methylated BSA (mBSA) stimulates production of IgGs with RNase and DNase activities belonging to IgGs, while polyclonal Abs from three non-immunized rabbits and three animals immunized with mBSA are catalytically inactive. Affinity chromatography of IgGs from the sera of autoimmune (AI) patients on DNA-cellulose usually demonstrates a number of fractions, all of which effectively hydrolyze both DNA and RNA, while rabbit catalytic IgGs were separated into Ab subfractions, some of which demonstrated only DNase activity, while others hydrolyzed RNA faster than DNA. The enzymic properties of the RNase and DNase IgGs from rabbits immunized with RNA distinguish them from all known canonical RNases and DNases and DNA- and RNA-hydrolyzing abzymes (Abzs) from patients with different AI diseases. In contrast to RNases and AI RNA-hydrolyzing Abs, rabbit RNase IgGs catalyze only the first step of the hydrolysis reaction but cannot hydrolyze the formed terminal 2',3'-cyclophosphate. The data indicate that Abzs of AI patients hydrolyzing nucleic acids in part may be Abs against RNA and its complexes with proteins. Copyright (c) 2008 John Wiley & Sons, Ltd.     

Peculiarities of abzymes from sera and milk of healthy donors and patients with autoimmune and viral diseases

The detection of catalytic activity of antibodies is the earliest indicator of development of autoimmune diseases (AID). In early stages of AID, the repertoire of abzymes with various properties is relatively small, but it is greatly increased during their development. Catalytic diversity of the abzymes includes DNase, RNase, ATPase, and oxidoreductase activities; there are antibodies phosphorylating proteins, lipids, and polysaccharides. This review summarizes new data on abzyme heterogeneity and possible reasons for this phenomenon. A possible role of abzymes and their exceptional multiplicity in the pathogenesis of different AID is discussed.     

Natural antibodies to nucleic acids

Blood of healthy donors contains low concentrations of autoantibodies to its own components, including DNA and RNA. Increased concentrations of antibodies to DNA and RNA have been found in blood of people and animals with autoimmune diseases and viral and bacterial infections. Detection of different antibodies with catalytic activities, including abzymes with DNase and RNase activities, is the earliest indicator of the development of some autoimmune diseases. This review reveals possible mechanisms of generation of anti-DNA and anti-RNA antibodies without catalytic activities and abzymes in normal organisms and in organisms with different pathologies. A possible role of these autoantibodies and the reasons of their exceptional diversity in normal organisms and in organisms with different autoimmune diseases are discussed.     

Ceruloplasmin decreases respiratory burst reaction during pregnancy

Testing of pregnant women reveals weakening of neutrophil-mediated effector functions, such as reactive oxygen species generation. This study provides data confirming the phenomenon, gained through application of the flow cytometry technique. Key factors influencing neutrophil functional activity in blood plasma of pregnant women have not been detected so far. At the same time, concentration of ceruloplasmin – a copper-containing glycoprotein – is known to increase in blood significantly during pregnancy. We observed the negative correlation between ceruloplasmin concentration in blood plasma of pregnant women and the intensity of respiratory burst of neutrophils. Fractionation of plasma using gel-filtration revealed that ceruloplasmin-containing fraction demonstrated suppression of the respiratory burst reaction. Partial elimination of ceruloplasmin from the blood of pregnant women, performed with the help of specific antibodies and followed by immunoprecipitation, leads to an increased respiratory burst reaction. On the contrary, addition of ceruloplasmin to blood samples of healthy donors noticeably decreases the respiratory burst reaction. The results presented prove that change in ceruloplasmin level in plasma is necessary and sufficient for modulating the ability of neutrophils to produce reactive oxygen species during pregnancy.     

Antibodies to rubellavirus, herpes simplex virus and Toxoplasma gondii in serial serum samples of pregnant and non-pregnant women.

By testing serial serum samples of 213 pregnant women for rubellavirus, of 196 for herpes simplex virus and of 134 for Toxoplasma gondii, it was found that during pregnancy there was a fall in the humoral antibody level. Presence and titre of antibodies were lower in sera of pregnant than of non-pregnant women. Alteration of the humoral antibody level during pregnancy may influence serological studies aimed at clarifying the role of infections in fetal malformations. Serial serum samples (4 samples from each pregnant woman involved) should be tested for obtaining reliable data regarding the frequency of infections during pregnancy.     

Detection of circulating mammary mucin (Muc1) and MUC1 immune complexes (Muc1-CIC) in healthy women

There is convincing epidemiological evidence that multiparity provides protection against the development of breast cancer. In the present study we evaluated the levels of MUC1 and MUC1 circulating immune complexes (MUC1-CIC) in 135 serum samples obtained from healthy women. The study population included 13 women who had never been pregnant, 31 primiparous pregnant women, 36 multiparous pregnant women who had lactated, 5 multiparous pregnant women who had never lactated, 24 multiparous non-pregnant women who were lactating at the time of the study, 24 multiparous non-pregnant women who had lactated, and 2 multiparous non-pregnant women who had never lactated. The purpose of this work was to detect MUC1 variations during pregnancy and lactation as well as to study the possible induction of a humoral immune response against MUC1 in these conditions. We employed ELISA techniques to measure MUC1 (CASA test) and MUC1-CIC (IgM and IgG) using two anti-MUC1 monoclonal antibodies (MAbs): C595 and SM3. Statistical analysis was performed using the ANOVA test. The pooled results pertaining to pregnant versus non-pregnant women were compared and significant differences were observed in MUC1 and MUC1-CIC-lgM levels detected with both MAbs; the MUC1-CIC-lgG levels detected with C595 were increased in the pregnant group while the MUC1-CIC-lgG levels detected with SM3 did not show any significant differences. When the results were compared between lactating and non-lactating women, no significant differences were found. In conclusion, MUC1 and MUC1-CIC-lgM, detected with both MAbs, and MUC1-CIC-4gG levels detected with the MAb C595 are apparently induced by pregnancy.     

Metabolic adaptations during lactogenesis. Lactose synthesis in rabbit mammary tissue during pregnancy and lactation.

Lactose biosynthesis and relevant enzymatic activity in rabbit mamma ry tissue during various stages of pregnancy and lactation are investigated by using a tissue-slice incubation method in order to understand the temporal relationships. Ovulation was induced in 27 New Zealand white rabbits and they were bred by artificial insemination. Sacrifice occurred on days 15, 24, and 29 of pregnancy, and day 2, 5, 8, 15, and 22 post partum. Nucleic acids were extracted and concentratons of DNA determined spectrophotometrically at 600 nm with diphenylamine reagent and RNA determined with orcinal reagent. The tissue incubations were made with (U-14C) glucose. (14C) lactose was then separated by paper chromatography from unchanged radioactive glucose. Enzyme analysis including determining the activities of phosphoglucomutase, UDP-glucose pyrophosphorylase, and UDP-glucose 4-epimerase. Lactose synthase was determined, as well as, hexokinase. A biphasic adaptation in the rate of lactose synthesis and in the RNA concentration was noted during lactogenesis. The 1st increase in the rate of lactose biosynthes is occurred between days 15 and 24 of pregnancy. A 2nd substantial increase was noted immediately post partum. The overall rate of lactose biosynthesis increased 12-fold from day 24 of pregnancy to day 15 of lactation post partum, and then decreased from 15 to 22 days post partum. The RNA concentration/g wet weight of tissue and the ratio of RNA/DNA closely represented the biphasic ability of the mammary-tissue slice to synthesize lactose. Increases in the activities of UDP-glucose 4-epimerase and lactose synthase were most closely correlated with increases in the rate of lactose biosynthesis. UDP-glucose pyrophosphor ylase activity was unrelated with the ability to synthesize lactose, and hexokinase and phosphoglucomutase activities were variable during pregnancy and lactation. Lactose synthase activity was present by day 15 of pregnancy, but the ability to synthesize lactose was undetected until day 24 of pregnancy.     

Multidimensional analysis of the essential elements in pregnant women's whole blood and characterization of maternal status by elemental pattern

BackgroundDuring pregnancy, the fetus needs to obtain a lot of nutrients from the mother, but the micronutrient deficiencies in pregnancy are not clear at present, and there is no reliable basis for nutrient intake and supplement. The purpose of this study was to understand the levels of essential elements in whole blood of pregnant women during various pregnancy stages at different ages and in different regions, to evaluate the deficiency of essential elements in Chinese pregnant women, and to explore the feasibility of using the elemental pattern to characterize maternal status.MethodsWhole blood samples of 11222 healthy pregnant women enrolled in different areas of China from Jan–Dec 2019, were analyzed for concentrations of six essential elements including Mn, Cu, Zn, Ca, Mg, and Fe, using the inductively coupled plasma mass spectrometer. A retrospective comparative study during different pregnancy periods at different ages and in different regions in whole blood essential elements content from non-pregnant normal women and pregnant normal women was developed using multivariate statistical analysis. Principal component analysis evaluation elemental pattern was used to characterize pregnancy status of pregnant women.ResultsIn general, the levels of six essential elements in whole blood of pregnant women can satisfy the needs of normal physiological activities. With the development of pregnancy, the contents of Cu and Mn increased, while the contents of Fe and Mg decreased, and the contents of Zn and Ca have no noteworthy change. At the same gestation stage, the Cu content in whole blood of elderly pregnant women was higher. There were some differences in whole blood essential elements content of pregnant women in different regions. Principal component analysis and heat map analysis showed the feasibility of using bioinformatics research strategies to identify different pregnancies.ConclusionsThere are differences in the content of whole blood essential elements of women at different stages of pregnancy in different regions. It was found that there was no obvious deficiency in whole blood essential elements levels of pregnant women in recent years. The pattern of essential elements has a certain application potential in the evaluation of pregnancy and pregnant women's health status.     

A noninvasive approach to studying fetal cells for prenatal diagnosis of chromosomal aneuploidies

Fetal cells isolated from maternal peripheral blood during the second trimester of pregnancy were analyzed. Blood samples were centrifuged in a Ficoll-Paque gradient, the mononuclear cell fraction was isolated and stained with fluorescent monoclonal antibodies against glycophorine A (GPA + PE), transferrin (CD71 + FITC), and Hoechst 33342. Fluorescence-activated cell sorting (FACS) was conducted on a Vantage flow cytofluorimeter (Becton Dickinson). Fluorescence in situ hybridization (FISH) with Y chromosome-specific DNA probe revealed fetal cells that exhibited Y signal in all 20 blood samples obtained from women pregnant with healthy male fetuses. The concentration of these fetal cells averaged about 1.34% and ranged from 0.1 to 4.2% in different blood samples. In six cases, blood samples were obtained from pregnant women, in which prenatal cytogenetic analysis revealed various fetal aneuploidies. Using FISH with DNA probes specific for chromosomes X, 18, and 13/21, Fetal cells with chromosomal aberrations were detected in these six maternal blood samples at a concentration from 1.5 to 5.6% (on average 3.7%). These results indicate the possibility of a new noninvasive approach, which is safe for both mother and fetus when used for isolation of fetal cells from pregnant women's blood samples and prenatal diagnosis of a broad spectrum of fetal cell chromosomal aberrations.     

Selenium and malondialdehyde content and glutathione peroxidase activity in maternal and umbilical cord blood and amniotic fluid

Placenta tissue may be a major source of lipid peroxidation products in pregnancy. It was proven that placental peroxidation activity increases with gestation. Selenium (Se), as an essential constituent of glutathione peroxidase (GSH-Px), takes part in the reduction of hydrogen peroxides and lipid peroxides. Malondialdehyde (MDA) is a major breakdown product split off from lipid peroxides. In this study, Se and MDA content and GSH-Px activity were measured in blood and plasma taken from 20 apparently healthy nonpregnant women between 19 and 38 yr of age and from 115 unselected pregnant women between 17 and 45 yr of age (35 in the first trimester, 22 in the second trimester, 38 in the third trimester, and 20 within 2 d of delivery). Samples of umbilical cord blood and amniotic fluid were taken from women in the second and third trimesters and at delivery. The Se content was measured by atomic absorption spectrometry (AAS), plasma MDA concentration by thiobarbituric acid reaction, and Se-dependent GSH-Px spectrometrically. Blood and plasma Se contents of nonpregnant women were below those considered adequate, indicating low selenium intake. In comparison to nonpregnant women, pregnant women had significantly decreased whole-blood and plasma Se levels in the second and third trimesters and at delivery. The significant drop of whole-blood SeGSH-Px activity was observed in the first trimester of pregnancy and its lower activity was maintained until delivery. A significant drop in plasma SeGSH-Px activity occurred in the second trimester and attained the minimal level at delivery. The Se level and SeGSH-Px activity in maternal and umbilical cord blood were at similar levels. Amniotic-fluid SeGSH-Px activity was nondetectable or exceptionally low and its Se content remained unchanged during pregnancy. Plasma levels of MDA were significantly decreased in the second and third trimesters and at delivery. The fetal blood plasma at birth had a lower MDA level compared to the levels of MDA of their mothers at delivery. A low, but significant inverse correlation existed between blood SeGSH-Px activity and plasma MDA content and between plasma Se and plasma MDA contents during pregnancy. A significant decrease of Se and SeGSH-Px activities (antioxidant enzyme) in both blood and plasma suggests a possible drop in total antioxidant status during pregnancy. Elevated MDA plasma levels might be the result of increased lipid peroxidation in placental tissue during pregnancy.     

Recognition of nonclassical HLA class I antigens by gamma delta T cells during pregnancy

The healthy trophoblast does not express classical HLA-A and HLA-B products; therefore, an MHC-restricted recognition of trophoblast-presented Ags is unlikely. In the decidua and also in peripheral blood of healthy pregnant women, gammadelta T cells significantly increase in number. We investigated the possible role of gammadelta T cells in recognition of trophoblast-presented Ags. PBL and isolated gammadelta T cells from healthy pregnant women as well as from those at risk for premature pregnancy termination were conjugated to choriocarcinoma cells (JAR) transfected with nonclassical HLA Ags (HLA-E, HLA-G). To investigate the involvement of killer-inhibitory/killer-activatory receptors in trophoblast recognition, we tested the effect of CD94 block on cytotoxic activity of Vdelta2(+) enriched gammadelta T cells to HLA-E- and/or HLA-G-transfected targets. Lymphocytes from healthy pregnant women preferentially recognized HLA(-) choriocarcinoma cells, whereas those from pathologically pregnant patients did not discriminate between HLA(+) and HLA(-) cells. Normal pregnancy Vdelta2(+) T cells conjugated at a significantly increased rate to HLA-E transfectants, whereas Vdelta2(+) lymphocytes from pathologically pregnant women did not show a difference between those and HLA(-) cells. Blocking of the CD94 molecule of Vdelta2(+) lymphocytes from healthy pregnant women resulted in an increased cytotoxic activity to HLA-E-transfected target cells. These data indicate that Vdelta2(+) lymphocytes of healthy pregnant women recognize HLA-E on the trophoblast, whereas Vdelta1 cells react with other than HLA Ags. In contrast to Vdelta2(+) lymphocytes from healthy pregnant women, those from women with pathological pregnancies do not recognize HLA-E via their killer-inhibitory receptors and this might account for their high cytotoxic activity.