The chemokine, CXCL12, is an independent predictor of poor survival in ovarian cancer.

BACKGROUND: The chemokine CXCL12 and its cognate receptor, CXCR4, have been implicated in numerous tumour types where expression promotes tumour growth, angiogenesis, metastasis and suppresses tumour immunity. METHODS: Using a tissue microarray of 289 primary ovarian cancers coupled to a comprehensive database of clinicopathological variables, the expression of CXCL12 and CXCR4 was assessed by immunohistochemistry and its impact in terms of survival and clinicopathological variables was determined. RESULTS: Patients whose tumours expressed high levels of CXCL12 had significantly poorer survival (P=0.026) than patients whose tumours failed to produce this chemokine. Lack of CXCL12 expression within tumours was associated with a 51-month survival advantage for patients when compared with patients whose tumours expressed high levels of CXCL12. FIGO stage, adjuvant chemotherapy and the absence of macroscopic disease after surgery were all shown to predict prognosis independently of each other in this cohort of patients. CXCL12 was independently predictive of prognosis on multivariate analysis (P=0.016). There was no correlation between CXCL12 and any clinicopathological variable. CONCLUSION: The chemokine CXCL12 is an independent predictor of poor survival in ovarian cancer. High expression of CXCL12 was seen in only 20% of the tumours, suggesting a role for anti-CXCL12/CXCR4 therapy in the management of these patients.

Mutation analysis of the c-mos proto-oncogene in human ovarian teratomas.

Female transgenic mice lacking a functional c-mos proto-oncogene develop ovarian teratomas, indicating that c-mos may behave as a tumour-suppressor gene for this type of tumour. We have analysed the entire coding region of the c-MOS gene in a series of human ovarian teratomas to determine whether there are any cancer-causing alterations. DNA from twenty teratomas was analysed by single-strand conformational analysis (SSCA) and heteroduplex analysis (HA) to screen for somatic and germline mutations. In nine of these tumours the entire gene was also sequenced. A previously reported polymorphism and a single new sequence variant were identified, neither of which we would predict to be disease-causing alterations. These results suggest that mutations in the coding region of the c-MOS gene do not play a significant role in the genesis of human ovarian teratomas.

Somatic and germline mutations of the BRCA2 gene in sporadic ovarian cancer.

The breast and ovarian cancer susceptibility gene BRCA2 has recently been isolated. A role for BRCA2 in sporadic breast and ovarian cancer has been suggested by loss of heterozygosity (LOH) studies which show frequent LOH in the BRCA2 region at chromosome 13q12. In addition, the observation of nonrandom loss of the wild-type chromosome in a breast/ovarian cancer family which shows linkage to BRCA2 suggests it may act as a tumor suppressor gene. To determine the extent of somatic alteration involving BRCA2 in sporadic ovarian cancer, 50 tumors were analyzed for mutations throughout the entire BRCA2 coding region. Mutations predicted to result in truncation of the BRCA2 protein were detected in four tumors. Analysis of germline DNA revealed two of these alterations to be of somatic origin. In addition, all four tumors exhibited loss of the second BRCA2 allele as predicted by Knudson's hypothesis for a tumor suppressor gene. These results suggest that, as is the case with BRCA1, somatic mutations of BRCA2 are infrequent in sporadic ovarian cancer, despite the relatively high frequency of LOH detected around the BRCA2 locus.

The prognostic value of serum CA 125 in patients with advanced ovarian carcinoma: an analysis of 573 patients by the Medical Research Council Working Party on Gynaecological Cancer.

A number of studies have suggested that serum CA 125 levels may be an important prognostic factor for survival of patients with ovarian carcinoma. We investigated, in a large group of patients from 11 UK centers, which combination of CA 125 measurements provided the best prognostic index, and whether the predictive power could be improved by the addition of other factors. Analysis of the data from 248 patients showed that the absolute value of the third CA 125 sample was the single most important factor for predicting progression at 12 months, with the addition of residual bulk only slightly improving the predictive power. Seventy-four patients had CA 125> 70, and of these 57% were correctly predicted to progress or die within 12 months, but 43% remained alive and progression free. The best predictor for progression produced a false positive rate of 19%. We therefore conclude that prognostic information based upon CA 125 measurements up to the start of the third course of initial chemotherapy is not accurate enough to be used to manage individual patients.

Prenatal ultrasound diagnosis of Apert's syndrome.

A mother affected with Apert's syndrome was diagnosed by ultrasound scan at 16-17 weeks to have a fetus similarly affected. The typical features of acrocephaly and symmetrical syndactyly were seen. This is probably the first time that this condition has been diagnosed at such a gestation by ultrasound scan. The patient decided to continue the pregnancy, and intrauterine death occurred at 34 weeks. The diagnosis was confirmed by pathological examination.

A comparison of the growth promoting properties of ascitic fluids, cyst fluids and peritoneal fluids from patients with ovarian tumours.

The growth promoting properties of ascitic fluids, cyst fluids and peritoneal fluids from patients with ovarian malignancy, benign ovarian tumours and non-tumour related gynaecological conditions have been investigated using an ovarian carcinoma cell line (OAW 42), mesothelial cells (58MC) and rat kidney cells (NRK-49F). Colony stimulating activity (CSA) for tumour cells and transforming activity (TA) for mesothelial cells were weakly correlated, but whereas elevated TA was tumour-associated, CSA was not. However, TA was not cancer-associated and, although the difference between the mean TA values of benign and malignant cyst fluids was of borderline significance, some benign cyst fluids from cystadenomas showed high TA values. Higher levels of TA in the cystadenomas showed a significant correlation with the menopausal status of the patient and higher levels of TA in the malignant cyst fluid/peritoneal fluid groups were associated with more advanced disease. Results indicated that some fluids contained TGF-beta-like activity, but there was no direct evidence for the presence of TGF-alpha/EGF-like activity in the fluids. Heparin inhibited clonogenic growth of tumour cells but not mesothelial cells. The reduced CSA which was observed after treatment of fluids with both heparin and thrombin implicated coagulation factors in the manifestation of CSA. It was concluded that CSA in the fluids was due, at least partly, to fibrin coagulation, and TA was due to unknown growth factor(s) which may include TGF-beta-like activity. The results are discussed in the context of the aetiology of ovarian carcinoma, and the possible clinical significance of TA.

A prospective evaluation of OC125 and magnetic resonance imaging in patients with ovarian carcinoma.

Eighteen patients with suspected primary or recurrent ovarian carcinoma have been investigated in each case by the assay of serum levels of the antigen CA125, immunoscintigraphy using 131I-OC125 antibody and magnetic resonance imaging using a 0.15 Tesla system. The final diagnosis was confirmed by laparotomy or laparoscopy. Serum levels of CA125 ranged between 5 and 780 units/ml (normal range less than 35). Antibody images and MRI were truly positive in 11 patients, 2 of whom were subsequently found to have bowel tumours. MRI showed greater detail of smaller lesions whilst immunoscintigraphy was more suited to the detection of distant metastases. In 7 patients the antibody images were positive whilst the serum marker levels were normal. This pilot study provides a preliminary comparison of the more recent techniques currently being evaluated for the detection of ovarian carcinoma.

Cytotoxic activity and phenotypic analysis of natural killer cells in early normal human pregnancy.

Peripheral blood lymphocytes taken from healthy women planning a pregnancy and then at various intervals up to the 16th week of pregnancy were assayed for natural killer (NK) cell activity against K562 target cells both in a 51Cr-release assay and in a single cell cytotoxicity assay. Results indicated a depression in NK activity from the earliest stages of pregnancy. The target binding capacity of the effector cells remained unimpaired up to 16 weeks, but a significant reduction in the post-binding lytic potential was observed, which parallelled the drop in cytotoxicity as assayed by the 51Cr-release method. The ability of individual effector cells to recycle and kill multiple targets remained essentially unimpaired. Analysis of lymphocyte subpopulations using the monoclonal antibodies anti-Leu-7 and anti-Leu-11b, which recognize NK cell-associated antigens, showed a significant reduction in the proportion of the mature, lytically active Leu-7-11+ cells capable of both binding and lysing K562 target cells. The suggestion that the depression in cytotoxicity may be associated with the reduction in the Leu-7-11+ subpopulation is supported by the high correlation levels observed between the proportion of Leu-7-11+ cells and target cell lysis.

Phenotypic heterogeneity and recycling capacity of natural killer cells in normal human pregnancy.

Natural killer (NK) cells have the ability to kill a variety of target cell types and the possibility that such cells could mount an effective attack on the developing fetus has not been discounted. The present study extends previous work showing that maternal NK reactivity against K562 target cells (TC) is reduced during pregnancy. Here we demonstrate using cytotoxicity assays at both the population and single cell level that, although depressed in number, maternal NK cells exhibiting the capacity to kill K562 TC are as lytically active in their ability to recycle and destroy multiple TC as NK cells from non-pregnant females. Moreover, two colour immunofluorescence analysis of the NK cell-associated markers Leu-7 and Leu-11b indicates that, in addition to a reduction in the absolute number of TC conjugate-forming cells, pregnant females present in their peripheral blood a larger proportion of TC-binding Leu-7+11- cells. These cells may be lytically immature. Small changes in NK cell profile and activity in maternal peripheral blood may be indicative of much more significant changes at the feto-maternal interface. It is, however, clear that pregnant females retain a population of highly active NK cells, thus minimising the possibility of immunocompromise.

Cytotoxic reactivity of human natural killer (NK) cells during normal pregnancy: a longitudinal study.

A longitudinal analysis of natural killer (NK) cell-mediated cytotoxicity of maternal peripheral blood lymphocytes was carried out at various stages (16-36 weeks) of normal human pregnancy, within 1 week following delivery and up to 40 weeks post-partum. NK cell-mediated lysis of K562 target cells (TC) in short term 51Cr-release assays was significantly depressed throughout pregnancy, returning to control levels 9-40 weeks post-partum. Natural cytotoxicity of unseparated maternal peripheral blood was also substantially depressed at all stages of pregnancy and, in addition, remained impaired in the late post-partum period. Longitudinal enumeration of Leu 3a+ and Leu 2a+ lymphocytes indicated an absence of helper/suppressor T-cell imbalance during pregnancy, and analysis of Leu 7+ cells showed no difference in population sizes between control and pregnancy groups. Comparison of blood and lymphocyte cytotoxicity in control and pregnancy samples suggested a complex regulation of NK reactivity during pregnancy. The potential role in vivo of both plasma-associated and cellular regulatory elements is discussed.

Natural killer cells in normal pregnancy: analysis using monoclonal antibodies and single-cell cytotoxicity assays.

Peripheral blood lymphocytes (nylon wool non-adherent) from healthy pregnant women and normal non-pregnant females were tested for natural killer (NK) cell-mediated cytotoxicity against K562 target cells both by 51Cr-release assay and single-cell cytotoxicity assay in agarose. The results indicated depression of NK cytotoxicity in pregnancy due to a decrease in the proportion of target-binding lymphocytes as well as a reduction in the lytic capacity of target-bound cells. The ability of active pregnancy-associated NK lymphocytes to recycle appeared to be unimpaired. Analysis of lymphocyte populations with monoclonal antibodies recognizing NK cell-associated antigens showed that the number of Leu-11+ lymphocytes was reduced in pregnancy. Enumeration of Leu-7+ cells and correlation of NK cell subpopulation data with cytotoxicity assay data suggest that pregnancy is associated with a reduction in the number of mature NK cells and probably also an inhibition of post-binding lytic activity.

Placental steroid sulphatase deficiency.

Low maternal plasma and urinary oestrogen concentrations in pregnancy are usually indicative of fetal problems, either placental insufficiency or fetal adrenal hypoplasia. Paediatricians, however, should follow obstetricians in becoming increasingly aware that deficiency of placental steroid sulphatase activity, a condition related to X linked ichthyosis, may produce the same abnormalities.

Serum free thyroxine concentration and free thyroid hormone indices in normal pregnancy.

Serum free thyroxine (FT4) concentrations were shown to be significantly reduced at 36-38 weeks normal pregnancy, both as measured FT4 by the Amerlex method (P less than 0.001), and as calculated FT4 (P less than 0.001) using accepted molecular weight and affinity constant data for the binding proteins. Serum FT4 concentrations as determined by the Immophase method were normal at 36-38 weeks normal pregnancy. All methods gave normal serum FT4 concentrations in subjects taking the oral contraceptive pill. FT4I and FT3I, derived using the MAA T3 uptake-value, were higher than normal at 36-38 weeks pregnancy (P less than 0.001), whereas T4/TBG and T3/TBG were both reduced (P less than 0.001). The observation that serum FT4 concentrations may fall in late pregnancy, as demonstrated both by the Amerlex radioimmunoassay technique and by calculation, suggests that circulating FT4 may not be the sole determinant of thyroid status at this time. From a practical viewpoint, it is important to note that currently available direct and indirect methods of assessing serum FT4 concentrations produce different patterns of change in pregnancy.

Intensive plasma exchange in rhesus isoimmunisation.

Intensive plasma exchanges were carried out in a 34-year-old pregnant woman to lower the level of circulating anti-D. The patient proved interesting from a number of serological aspects. The possible protective effect of coexistent ABO incompatibility is discussed. The effect of plasma exchanges on cytotoxic HLA antibodies is also demonstrated.

The amine oxidases of human placenta and pregnancy plasma.

1. The purification of monoamine oxidase and diamine oxidase from normal human term placental tissue is described. 2. The properties of these enzymes are reported and compared with the properties of unpurified human pregnancy plasma. 3. This comparison shows that the amine oxidase of pregnancy plasma has properties corresponding to purified placental diamine oxidase, suggesting a placental origin for the plasma enzyme system. 4. Detailed kinetic study of the purified placental diamine oxidase suggests that it has a Ping Pong sequence, a mechanism of action and rate-limiting step similar to the diamine oxidase of pig kidney. 5. It is suggested that the enzyme system is important in protecting the foeto-placental unit from excesses of biogenic amines.