Diagnostic assay for the detection of preeclampsia

Chemistry: molecular biology and microbiology – Measuring or testing process involving enzymes or... – Involving viable micro-organism

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436 63, 436 86, 436 65, C12Q 102

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052388190

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BRIEF SUMMARY
BACKGROUND OF THE INVENTION



Field of the Invention

This invention relates to the diagnosis of preeclampsia using an assay to measure a mitogenic factor in blood. Preeclampsia is a serious problem in pregnant women. It is an idiopathic life threatening hypertensive condition. The condition of preeclamptic women can often deteriorate to a point where serious injury will occur to either the mother, child or both. Preeclampsia is a leading cause of death both maternal and infant.


Information Disclosure

Preeclampsia occurs in 7-10% of pregnancies and is responsible for significant maternal and fetal morbidity (Roberts, J. M., Pregnancy-related hypertension. In: Creasy, R. K.; Resnick R., eds., Maternal-Fetal Medicine-Principles and Practice, Philadelphia: W. B. Saunders, 1984, 703-752). Despite decades of interest and research, the pathogenesis of this disease is still poorly understood. Recent evidence, however, suggests that endothelial cell injury may play an important role in the preeclamptic syndrome. The histopathological findings of endothelial lesions in renal and umbilical vessels obtained from preeclamptic patients have been recognized (Spargo, B. H.; McCartney, C. P.; Winemiller, R., Glomerular capillary endotheliosis in toxemia of pregnancy. AMA Arch. Pathol., 1959, 68:593-497; Dadak C., Ulrich W., Sinzinger H. Morphological changes in the umbilical arteries of babies born to preeclamptic mothers: an ultrastructural study. Placenta 1984, 5:419-426). Lately, several reports have documented biochemical abnormalities in serum from preeclamptic patients which support the concept that endothelial cell perturbation and sublethal injury of these cells may contribute to the pathogenesis of preeclampsia. Most of these studies have been indirect, showing elevated levels of endothelial cell products in serum (e.g., fibronectin (Stubbs, T. M.; Lazarchick, J.; Horger, E. O., III, Plasma fibronectin levels in preeclampsia: a possible biochemical marker for vascular endothelial damage. Am. J. Obstet. Gynecol, 1984, 150:885-887), factor VIII antigen (Redman, C. W. G.; Beilin, L. J.; Stirrat, G. M., et al., Factor VIII consumption in preeclampisa. Lancet 1977, II:1249-1252), or platelet and coagulation abnormalities (Roberts, J.M., Pregnancy-related hypertension, In: Creasy, R. K.; Resnick, R., eds., Maternal-Fetal Medicine-Principles and Practice. Philadelphia: W. B. Saunders, 1984, 703-752). We have recently demonstrated directly that serum from preeclamptic women injures endothelial cells in vitro (Rodgers, G. M.; Taylor, R. N.; Roberts, J. M., Preeclampsia is associated with a serum factor cytotoxic to human endothelial cells, Am. J. Obstet. Gynecol, 159:908-914, 1988).


BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1
Mitogenic dose-response of cultured fibroblasts exposed to serum from normal and preeclamptic patients. Quiescent fibroblasts were incubated with paired pre- and postpartum sera from normal and preeclamptic patients as described in Materials and Methods. Incorporation of .sup.3 H-thymidine into fibroblast DNA increased in a dose-dependent, logarithmic fashion. Shown are mitogenic activities of prepartum (open box) and postpartum (closed box) sera from normal (A) and preeclamptic (B) patients. There was a significant left-shift of the curve for prepartum as compared to postpartum preeclamptic serum but not in paired sera from normal pregnancy (P<0.01).
FIG. 2
Mitogenic activity of sera from normal and preeclamptic patients. (A) Direct comparison of prepartum sera from preeclamptic and normal patients diluted to a final concentration of 2% demonstrated a 46% increase in mitogenic activity in preeclamptic sera as compared to normal (P<0.01). (B) Serum collected at 24-48 hours postpartum revealed no difference between the two groups (P>0.6).
FIG. 3
No indirect effect of magnesium sulfate (MgSO.sub.4) treatment on the mitogenic activity of sera from preeclamptic patients. Two groups of preeclamptic patients were compared. One group (n=4) had received intravenous MgSO.sub.4 prior to the prepar

REFERENCES:
Spargo, et al., "Glomelular capillary endotheliosis in toxemia of pregnancy," AMA Arch. Pathol. 68:593-597 (1959).
Redman, et al., "Factor VIII consumption in preeclamsia," Lancet 2:1249-1252 (1977).
Dadak, et al., "Morphological changes in the umbilical arteries of babies born to preeclamptic mothers: an ultrastructural study," Placenta 5:419-426 (1984).
Roberts, J. M., "Pregnancy-related hypertension," in Maternal-Fetal Medicine-Principles and Practice, Creasy, R. K. and Resnick, R., eds., W. B. Saunders, Philadelphia, (1984) pp. 703-752.
Stubbs, et al., "Plasma fibronectin levels in preeclampsia: a possible biochemical marker for vascular endothelial damage," Am. J. Obstet. Gynecol. 150:885-887 (1984).
Rodgers, et al., "Preeclampsia is associated with a serum factor cytotoxic to human endothelial cells," Am. J. Obstet. Gynecol. 159:908-914 (1988).
Musci, T. J., et al., "Mitogenic activity is increased in the sera of preeclamptic women before delivery," Am. J. Obstet. Gynecol. 159(6):1446-1451 (1988).
Keski-Oja, J., et al., "Transforming Growth Factors and Control of Neoplastic Cell Growth," J. Cell. Biochem. 33:95-107 (1987).
Cagnoli, L., et al., "Lymphocyte Hyporesponsiveness During Edema, Proteinuria and Hypertension (EPH) Gestosis," La Ricerca Clin. Lab, 11:229-238 (1981).
Gaugas, J. M., et al., "Complement Fixing Antibody Against Solubilized Placental Microsomal Fraction in Pre-Eclampsia Sera," Br. J. exp. Path. 55:570-574 (1974).
Redman, C. W. G., et al., "Plasma Urat and Serum Deoxycytidylate Deaminase Measurements for the Early Diagnosis of Pre-Eclampsia," Br. J. Obstet. Gynecol. 84:904-908 (Dec. 1977).
Chemical Abstracts General Subject Index--11th Collective (1982-1986) p. 25973GS.
Musci et al., --Am. J. Obstet. & Gynecology vol. 159 (Dec. 1988) pp. 1446-1451.
Westley et al.,--J. Biological Chemistry vol. 259(16) Aug. 1984 pp. 10030-10035.

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