DES-induced vascular risk

Immunogenicity of synthetic sex hormones and thrombogenesis, 1985

Study Abstract

The ingestion of synthetic steroidal and non-steroidal estrogens may induce antiestrogen antibodies in women on oral contraceptives, and in prostatic patients treated with diethylstilbestrol (DES). Natural sex hormones have no such effect.

A radioimmunoassay with tritiated ethinylestradiol or DES was applied to study the prevalence of synthetic sex hormone antibodies in 2 populations: 100 women on estroprogestative hormones and 93 cases of DES treated prostatic cancers. Homologous non-treated controls were compared.

Results allowed to identify among treated and asymptomatic subjects an immunoreactive population of 30% women and 47% men. Furthermore, the antibodies were found with a much higher frequency (p less than 0.001) in patients who had experienced a thromboembolic disease while on treatment: 90% of women and 74% of men.

The importance of these antibodies as a risk factor, their possible role in promoting vascular lesions, the interest of their detection for the prevention of the vascular risk induced by synthetic sex hormones, are considered.

References

DES DIETHYLSTILBESTROL RESOURCES

Assessment of myelotoxicity caused by environmental chemicals

The effect of DES on bone marrow, 1982

Study Abstract

Potential antineoplastic agents must be screened for the delayed toxicity that occurs in many cases of drug-induced bone marrow aplasia. In vitro clonal assays for hematopoietic progenitor cells have been developed to assess the degree of myelotoxicity. This adverse side effect is often the limiting factor in the development of new cancer chemotherapeutics. In addition, many environmental chemicals are cytotoxic to rapidly proliferating cells, but a systematic assessment of their myelotoxicity has not been performed.

We have used clonal marrow assays to investigate a panel of chemicals including 2,3,7,8-tetrachlorodibenzo-p-dioxin, polybrominated biphenyls, diethylstilbestrol, benzo(a)pyrene and indomethacin. All were immunotoxic, some to pleuripotent hemopoetic stem cells and other to granulocyte-macrophage progenitors, and at concentrations below those causing other toxic manifestations.

Diethylstilbestrol (DES) was formerly prescribed as a synthetic estrogen and also was used to fatten cattle. Human residues accumulated from ingestion of dairy and meat products have been found. Mice were exposed by subcutaneous injection to 0.2, 2 and 8 mg/kg body weight for 5 consecutive days. There was a dramatic decrease in marrow cellularity, CFU-S and CFU-GM (Table 1). These animals also exhibited highly enhanced sensitivity to Listeria monocytogenes and other generalized immunotoxic responses.

This shows that these bone marrow clonal assays, and hopefully future one for erythroid, B- and T-lymphocytes, and megakaryocytes, will provide the specificity and sensitivity necessary to delineate the myelotoxicity of a broad spectrum of environmental chemicals.

References

  • Full study (free access) : Assessment of myelotoxicity caused by environmental chemicals, Environmental health perspectives, NCBI PubMed, PMC1568900, 1982 Feb.
DES DIETHYLSTILBESTROL RESOURCES

DES immunotoxicity

The comparative immunotoxicity of five selected compounds following developmental or adult exposure, 2006

Study Abstract

It is well established that human diseases associated with abnormal immune function, including some common infectious diseases and asthma, are considerably more prevalent at younger ages. Although not established absolutely, it is generally believed that development constitutes a period of increased immune system susceptibility to xenobiotics, since adverse effects may occur at lower doses and/or immunomodulation may be more persistent, thus increasing the relative risk of xenobiotic exposure to the immunologically immature organism.

To address this issue, a brief overview of immune maturation in humans is provided to demonstrate that functional immaturity alone predisposes the young to infection. Age-dependent differences in the immunotoxic effects of five diverse compounds, diethylstilbestrol (DES), diazepam (DZP), lead (Pb), 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD), and tributyltin oxide (TBTO), which have undergone adult and developmental immunotoxicity testing in rodents, are then reviewed, as are human data when available. For all five chemicals, the developing immune system was found to be at greater risk than that of the adult, either because lower doses produced immunotoxicity, adverse effects were more persistent, or both.

Diethylstilbestrol immunotoxicity

Background

Between 5 and 10 million pregnant women were given diethylstilbestrol (DES), a potent synthetic nonsteroidal estrogen, between 1938 and 1971 to prevent premature delivery or pregnancy loss. Its use was terminated when a rare form of reproductive system cancer was found in female offspring of DES-exposed mothers. Male and female reproductive systems malformations have been reported in children of treated women, as has anecdotal evidence of immune system dysfunction. DES was also used to increase weight gain in livestock, although this use is no longer permitted in most countries.

Effects on the Immune System in Humans

Both female and male children of DES-exposed mothers report a higher incidence of autoimmune diseases and asthma (Baird et al., 1996). In general, these diseases are considered to be the result of inappropriate immune system responses, or possible loss of homeostatic control, instead of immune system suppression.

Effects on the Immune System in Rodents

In utero exposure
Luster et al. (1978b) reported that a single injection of 0.1 mg DES/kg body weight on gestational day (GD) 16 did not affect the antibody response to the T-cell-dependent antigen, sheep red blood cells (SRBC), when evaluated in 7-wk-old male and female offspring of Swiss-Webster mice. The T-independent IgM response of female offspring to bacterial lipopolysaccharide (LPS) was suppressed by DES, but was similar to control responses when females were
immunized for a second time. In marked contrast, the male offspring response to LPS immunization was enhanced after both first and second immunizations, an effect attributed to the stimulating effect of estrogen on the antibody response to LPS. Delayed-type hypersensitivity responses (DTH) were suppressed in female, but not in male, offspring, even though thymus weights and T-cell responses to polyclonal stimulation were suppressed in both genders (Luster et al., 1979). Further studies suggest that DES targets early precursors of T lymphocytes in the fetal liver, accounting for thymic atrophy and suppression of DTH (Holladay et al., 1993), but not for defects in T-independent responses to LPS of female offspring.

Neonatal exposure
Nonspecific T- and B-cell proliferation was reported to be suppressed in 6-wk-old female NMRI mice given 5 μg DES/d (roughly 2.2 mg DES/kg/d) over postnatal days (PND) 1–5 (Kalland et al., 1979); suppression was still evident at 17 mo of age (normal life span ~24 mo). It is noteworthy that neither estradiol nor corticosterone exposure over PND 1–5 produced long-term suppression, and that lymphocyte proliferation was comparable to control values at 6 wk of age in females exposed to DES over PND 6–10. Lower doses (approximately 4.4, 44, or 440 μg/kg/ d) had no effect on proliferative response. The 5-μg DES/d exposure regimen also decreased NK cell activity in 6- to 8-wk-old female inbred C57Bl/6 (75%↓) and BALB/c (53%↓) mice and in outbred NMRI (28%↓) mice (Kalland, 1980a). NMRI or AKR/J female mice, exposed to 5 μg DES/d over PND 1–5, were also more likely to develop tumors after low dose injection of a known carcinogen (Kalland & Forsberg, 1981). A subsequent paper (Kalland, 1984) reported that, on a per cell basis, NK cells from DES mice were as active as cells from the control group, but that exposure reduced the number of NK cell precursors in the bone marrow. In other words, NK cells from experimental animals were as efficient as those from controls, but a deficiency in NK cell precursors produced functional suppression of NK activity at the whole animal level. The same postnatal exposure regimen (Kalland, 1980a) reduced the T-lymphocyte-dependent antibody response to SRBC by ~60%, and the T-independent response to bacterial LPS by ~40% when examined in 16- to 18-wk-old NMRI mice. Suppression of the T-dependent response was reportedly due to a defect in T-helper cells. DTH responses were likewise suppressed in 6- and 9-mo-old NMRI females exposed to approximately 2.2 mg/kg/d over PND 1–5 (Kalland & Forsberg, 1978). Kalland (1980b) also reported a persistent (at least 6.5 mo postpartum) decrease in the proportion of T cells in the spleens of DES-exposed mice.

Adult exposure
Luster et al. (1980) reported suppression of the antibody response to SRBC or LPS, and the DTH to keyhole limpet hemocyanin (KLH), in adult female mice exposed to 2 or 8 mg DES/kg/d × 5 d. The DTH was decreased in mice dosed with DES after, but not before, sensitization with KLH, suggesting that the suppressive effects of DES on DTH were not persistent. Using the same exposure regimen, resistance to bacterial or parasite infection was decreased and tumor incidence in animals challenged with tumor cells was increased at ≥2 mg DES/kg/d (Dean et al., 1980). T-cell-mediated resistance to a nematode infection was suppressed by 5 d of exposure to 0.2 mg DES/kg/d if exposure began on the day of infection; if exposure commenced 5 d before or 3 or 8 d after infection, decreased resistance was only observed at the highest dose (8 mg/kg/d) (Luebke et al., 1984).

Mode(s) of Action

DES is a potent estrogen, and likely affects immune function via the estrogen receptor (ER). Evidence includes similar effects of known estrogens (17β-estradiol) on the immune system of adult and neonatal rodents, blockade of certain immunotoxic effects by pharmacologic antagonism of the ER (Luster et al., 1984), and antagonism of estrogen-mediated immune system effects in mice lacking ERα (Staples et al., 1999). DES appears to target precursor cells in the bone marrow (adults and neonates) and fetal liver (neonates), producing a long-lasting or perhaps permanent reduction in numbers of precursor cells. This defect explains a significant portion of long-lived immunosuppressive effects (e.g., Kalland’s 1984 paper on suppressed NK activity), although the effects of adult exposure also includes damage to the thymic epithelium (Luster et al., 1984). The underlying mechanism of long-term suppression following exposure of the developing immune system to DES is not known, but the default assumption is that a critical cell population is lost to developmental exposure; either this purported population is refractory to estrogen-mediated ablation in adults or repair and recovery mechanisms are present in adults that are lacking in the developing immune system.

Data Gaps

There has been no systematic evaluation of persistent DES-mediated immunosuppression in adult animals. Dose-response data are not available for many of the of the developmental exposure studies that revealed persistent effects.

Summary

In utero exposure to 0.1 mg DES/kg during the last trimester of pregnancy suppressed T-cell- and B-cell-mediated responses only in female offspring. The gender dependence of effects was remarkable in that T-independent responses in male offspring were enhanced, yet suppressed in females. Exposure during gestation produced effects that persisted into the equivalent of young adulthood. In neonates there appears to be a critical developmental window during PND 1–5, during which exposure to DES produces persistent immune system defects that last well into adulthood or persist for most of the normal life span of the mouse. These effects are among the most persistent reported for any chemical. In adults, immunosuppression occurs at doses similar to those that produce immunotoxicity in developing animals. However, the immune system-related endpoints that have been evaluated over time in exposed adult animals (bone marrow cellularity, thymus weights) recover relatively quickly (Forsberg, 1984). In adults, recovery may occur so quickly that suppression of cell function or resistance to infection may require ongoing exposure to maintain suppression.

Conclusions

Immunotoxicity has been reported at similar doses when exposure occurs during late gestation, early postpartum, or as adults. However, the distinguishing feature of developmental exposure to DES is the persistence of effects, some of which are still apparent in very old mice. In contrast, immune system-related endpoints that have been evaluated (bone marrow cellularity, thymus, weights) suggest that adults recover relatively quickly (Forsberg, 1984).

References

  • The comparative immunotoxicity of five selected compounds following developmental or adult exposure, Journal of toxicology and environmental health. Part B, Critical reviews, NCBI PubMed, PMID: 16393867, 2006 Jan-Feb.
  • Featured image radicalremission.
DES DIETHYLSTILBESTROL RESOURCES

Alterations in immunological functions

The effect of adult exposure to diethylstilbestrol in the mouse, 1980

Study Abstract

Female adult mice were administered diethylstilbestrol (DES) at dosages of 0.2, 2.0 and 8.0 mg/kg body weight per day for 5 consecutive days.

A dose-related alteration in immunologic function occurred as a consequence of DES exposure and included depression of Ab responses, cutaneous delayed hypersensitivity and in vitro lymphoproliferative responses to mitogens as well as allogeneic leukocytes in one-way mixed leukocyte cultures.

The percentage of splenic leukocytes possessing a T cell membrane marker was decreased following DES exposure although the percentage of cells with B cell markers was normal.

Co-culture studies indicated that immune depression is due, at least in part, to induction of suppressor cells that reside in the adherent cell population although an additional effect of DES on T lymphocytes is also probable.

References

  • The effect of adult exposure to diethylstilbestrol in the mouse: Alterations in immunological functions, Journal of the Reticuloendothelial Society, NCBI PubMed, PMID: 6450833, 1980 Dec.
  • Featured image animalsake.
DES DIETHYLSTILBESTROL RESOURCES

Suppression of antibody response of female mice exposed to DES

Suppression of cell-mediated immunocompetence after subchronic exposure to diethylstilbestrol in female B6C3F1 mice

1983 Study Abstract

The effect of the nonsteroidal estrogenic compound, diethylstilbestrol (DES) on cell-mediated immunocompetence in the mouse has been assessed.

Adult female B6C3F1 mice were injected (s.c.) with 0.2, 1.0 and 4.0 mg/kg of DES daily for 14 days. All immunological tests and toxicological parameters were determined at least 24 hr after the last exposure.

Thymic involution and hepatomegaly were noted at the lowest dose of DES. The most sensitive indicator of immunosuppression by DES was the delayed hypersensitivity response (DHR) to keyhole limpet hemocyanin in mice sensitized without adjuvant. The lowest dose of DES produced a 93% decrease, compared with a 39% decrease for the same DHR model in mice sensitized to keyhole limpet hemocyanin plus adjuvant and a 63% decrease for the DHR to sheep erythrocytes. A 35% decrease of the acute inflammatory response to carrageenin was produced by the lowest dose of DES. Day 2 proliferative responses to both concanavalin A and phytohemagglutinin (T-cell mitogens) were depressed by the lowest dose of DES, whereas significant suppression of the response to lipopolysaccharide (B-cell mitogen) was only noted at the highest dose of DES. The effects of DES on Day 3 proliferative responses were less dramatic. The mixed lymphocyte response was significantly suppressed by 1.0 and 4.0 mg/kg of DES on all days in culture. The reversibility of the DES effects was studied by resting the mice for 30 days between exposure and measuring a given parameter. All effects on organ weights and the depression of both the sheep erythrocytes DHR and the carrageenin inflammatory response were reversed.

References

  • Suppression of cell-mediated immunocompetence after subchronic exposure to diethylstilbestrol in female B6C3F1 mice, The Journal of pharmacology and experimental therapeutics, NCBI PubMed, PMID: 6225865, 1983.
  • Featured image my.vanderbilt.edu.
DES DIETHYLSTILBESTROL RESOURCES

Effect of DES on the immune responsiveness

Effects of single administration of diethylstilbestrol on murine langerhans cells

2005 Study Abstract

Diethylstilbestrol (DES) is a synthetic compound with potent estrogenic actions useful in the treatment of prostate carcinoma despite the fact that it can also induce some forms of neoplasia.

Both effects are thought to be related to its estrogenic actions and very little attention has been focused on the possible effect of DES on the immune response.

Skin is the largest organ of the body and constitutes the first line of defense against xenobiotics. The Skin Immune System (SIS) has become the center of attention of research for the development of new therapeutic approaches for neoplasic diseases. Langerhans cells (LC), as an element of SIS, are “professional” antigen presenting cells resident in the skin that participate in the immune response associated with tolerance and acquired immunity to antigens.

Hence in this work we studied the effect of DES on LC of murine skin as a model to analyze the possible effect of DES on the immune response.

Male CD-1 mice (20 to 35 g body weight) were treated topically (TO) or subcutaneously (SC) with DES (10 and 100 mg/kg, dissolved in ethanol) and sacrificed at 12, 84 and 228 hr. LC were quantified in the ear skin of mice using both an enzymatic histochemical technique to demonstrate ATP-ase activity; and an indirect immunohistochemical assay for detecting class II molecules of the major histocompatibility complex (MHC-II).

DES induced a significant time- and dose- dependent reduction in the number of LC (P < 0.05). Data presented here suggest that estrogens may exert a modulatory action on LC.

References

  • Effects of single administration of diethylstilbestrol on murine langerhans cells, Proceedings of the Western Pharmacology Society, NCBI PubMed, PMID: 16416678, 2005.
  • Featured image maths.ox.ac.uk.
DES DIETHYLSTILBESTROL RESOURCES

Prenatal DES exposure linked to Hashimotos thyroiditis

Drug exposure, pregnancy outcome and fetal and childhood development occurring in the offspring of mothers with systemic lupus erythematosus and other chronic autoimmune diseases

2006 Study Abstract

Most autoimmune diseases occur more commonly in females and many of these young women wish to become mothers. For pregnancy to proceed successfully immunomodulation and physiological changes preparing the reproductive system need to occur.

Pregnancy occurring in a chronically ill mother who requires medications in order to maintain her own health and who may have already incurred significant organ pathology gives rise to several problems and so four questions arise:

  1. What will be the effect of the pregnancy on the underlying disease?
  2. What will be the effect of the disease on the outcome of pregnancy?
  3. How to manage the disease, just prior to, throughout and immediately after the pregnancy?
  4. The long term fetal and childhood effects of maternal disease and its management.

This paper reviews the current literature pertaining to these questions in patients with systemic lupus erythematosus (SLE) and other chronic rheumatic and autoimmune diseases.

Diethylstilbestrol

… “Other evidence suggests that the risk of autoimmune disease, particularly Hashimotos thyroiditis, is also increased in individuals exposed to DES in utero.” …

References

  • Drug exposure, pregnancy outcome and fetal and childhood development occurring in the offspring of mothers with systemic lupus erythematosus and other chronic autoimmune diseases, Lupus, NCBI PubMed, PMID: 17153855, 2006.
  • Featured image cleveland clinic.
DES DIETHYLSTILBESTROL RESOURCES

How DES exposure leads to reduced fertility in males

Diethylstilbestrol induces morphological changes in the spermatogonia, Sertoli cells and Leydig cells of adult rat

2019 Study Abstract

It is now established that diethylstilbestrol (DES) has damaging effects on the male reproductive system. However, to date there have been no studies morphological analysis of adult rat testes upon treatment with DES.

Here, we examined whether DES has any significant morphological effect on steroidogenesis and spermatogenesis.

DES was injected subcutaneously at 3 μg/day and 30 μg/day in adult male Sprague-Dawley (SD) rats for two different treatment lengths (1 or 3 weeks), after which rats were necropsied. TUNEL labeling, cell counting, and morphological analysis were used to evaluate the effects of DES.

A high dose of DES and longer exposure severely affected the cellular development of the testis. Specifically, DES treatment disrupted both steroidogenesis and spermatogenesis by decreasing the number of spermatogonia, Sertoli cells, and Leydig cells in a dose- and time-dependent manner.

Thus, DES may account for decreases in the number of spermatogenic cells, Sertoli cells and Leydig cells, which in turn may lead to reduced fertility in males.

References

  • Diethylstilbestrol induces morphological changes in the spermatogonia, Sertoli cells and Leydig cells of adult rat, Research in veterinary science, NCBI PubMed, PMID: 31082573, 2019 Apr.
  • Featured image Bart Plantenga.
DES DIETHYLSTILBESTROL RESOURCES

Immunosuppressive effects of DES exposure

Immunotoxic effects of diethylstilbestrol on host resistance: comparison with cyclophosphamide, 1984

Abstract

To evaluate the usefulness of host resistance assays for measurement of immunotoxicologic effects of chemicals, the immunosuppressive effects of exposure to diethylstilbestrol (DES) were compared with the effects of treatment with the known immunosuppressive drug cyclophosphamide (CPS).

A panel of six host resistance models was evaluated, including

  • infection with the bacterium Listeria monocytogenes,
  • herpes simplex virus type 2 (HSV-2),
  • and encephalomyocarditis virus (EMC),
  • the yeast Cryptococcus neoformans,
  • the parasite Naegleria fowleri,
  • and transplantation of the B16F10 melanoma tumor.

The results demonstrate a general correlation between the effects of CPS and DES on host resistance. Acute treatment with CPS (200 mg/kg) markedly depressed resistance to the microbial infections with L. monocytogenes and HSV, and exposure to DES usually also decreased resistance in a dose dependent manner. Moreover, CPS had no marked effect on resistance to N. fowleri and EMC virus, and exposure to DES also had a neglible or slight effect. There were, however, two model systems in which the effects of CPS and DES diverged. Whereas treatment with DES produced no significant effect on resistance to C. neoformans, acute treatment with CPS prior to the fungal infection produced a marked increase in resistance. Also, while treatment with CPS markedly increased B16F10 lung metastases, treatment with DES significantly decreased the incidence and number of lung metastases.

The data support the general validity of host resistance assays, particularly with models of short disease course, for measuring immunosuppression. However, the results also emphasize the complexity of interpreting effects of environmental chemicals on host resistance, because of the interplay of such factors as relative times of exposure to the chemical in relation to pathogenesis of infection, the length of the disease course, the nature of the operative host defense mechanisms, and the compensatory recovery of these mechanisms.

References

  • Immunotoxic effects of diethylstilbestrol on host resistance: comparison with cyclophosphamide, Journal of leukocyte biology, NCBI PubMed, PMID: 6323603, 1984.
  • Featured image vanderbilt edu.
DES DIETHYLSTILBESTROL RESOURCES

DES exposure linked to laterality process disruption

Obstetrical complications and subsequent schizophrenia in adolescent and young adult offsprings: is there a relationship?, 2004

Abstract

Schizophrenia is a psychiatric disease affecting around 1% of the population, the negative signs of which are correlated with inactivity of the prefrontal dorsolateral cortex, while an increased, more deeply localized, activity in the mesolimbic pathway may explain the positive signs. Several events occurring during pregnancy are likely to be involved in its genesis: hormonal supplementation by diethylstilbestrol, severe maternal denutrition, exposure to influenza virus, repeated psychological stress.

From multicentric studies and meta-analyses in the psychiatric literature, the risk of schizophrenia appears to be multiplied by two if pregnancy is complicated, mainly by diabetes, Rhesus incompatibility, bleeding, preeclampsia, premature rupture of membranes and preterm birth. When delivery is linked to an abnormal presentation or happens via a caesarean birth for acute foetal distress, the time when the first signs of psychosis appear seems to be earlier in adolescence or in early adulthood.

Cerebral imaging of schizophrenic patients shows ventriculomegaly and gray matter reduction, mainly in hippocampal volumes and in the dorsolateral prefrontal cortex. Similar alterations in the neuronal pathways have been experimentally reproduced in rats after repeated prenatal stress and perinatal hypoxia. A region on the distal portion of chromosome 1 has shown evidence for linkage to schizophrenia.

Exposure to diethylstilbestrol (DES) in the second trimester of pregnancy is likely to disrupt the laterality process, leading to an outstanding number of left-handed individuals, be they boys or girls. In the same way data from the literature suggest that people exposed to DES have a higher risk of presenting depressive signs. Nevertheless it may not be the only cause, since it is obvious that the gynecological troubles which required hormonal prescription may have affected the course of pregnancy. Katz et al. reported a higher risk of psychosis after the mother took DES about four cases: DES doses varied from 7 to 12.8 g, but two pregnant women out of four also received progestatives (total dose of 1.950 mg of medroxyprogesterone to 3.600 mg of progesterone).

Therefore, a two factor model seems to be able to explain the onset of schizophrenia in which obstetrical complications may interact with a genetic liability and in which the consequences of hypoxic events may lie on a continuum ranging from cerebral palsy in some children to subtle cognitive and behavioural disturbances in others.

References

  • Obstetrical complications and subsequent schizophrenia in adolescent and young adult offsprings: is there a relationship?, European journal of obstetrics, gynecology, and reproductive biology, NCBI PubMed, PMID: 15140504, 2004.
  • Featured image wallpapercave.
DES DIETHYLSTILBESTROL RESOURCES