Whole mount immunofluorescence analysis of placentas from normotensive versus preeclamptic pregnancies.

INTRODUCTION: Defects in placental angiogenesis and spiral artery remodeling have been proposed to play essential roles in the development of preeclampsia. However, the specific molecular mechanism(s) responsible for aberrant placental angiogenesis in preeclampsia are incompletely understood. The vascular endothelial growth factor receptors (VEGFR1, R2, R3) and STAT3 have critical functions in normal blood vessel development, but their potential roles in preeclampsia are currently unclear. In this study, we utilized a novel whole mount immunofluorescence (WMIF) method to compare expression of VEGFR1, R2, R3 and activated, phosphorylated STAT3 (pSTAT3) in placentas of preeclamptic (PE) versus normotensive (NT) pregnancies. METHODS: Placental biopsies collected from NT and PE pregnant women were fixed and stained with fluorochrome-conjugated antibodies to identify specific cell populations as follows: CD31 for blood vessel endothelial cells, cytokeratin-7 for trophoblast cells, and CD45 for immune cells. Expression of the VEGFRs and pSTAT3 were subsequently characterized by WMIF in conjunction with confocal microscopy. RESULTS: A total of 18 PE and 18 NT placentas were evaluated. No significant differences in the cell type-specific expression patterns or expression levels of VEGFR1, VEGFR2 or VEGFR3 were detected between NT and PE placentas. In contrast, statistically significant increases in pSTAT3 staining were detected in endothelial cells of PE placentas versus NT controls. DISCUSSION: Our study demonstrates that increased pSTAT3 expression in placental endothelial cells is associated with PE. We speculate that elevated pSTAT3 expression in the blood vessels of PE placentas may be due to aberrant angiogenesis, increased pro-inflammatory cytokine expression, and/or placental stress.

Time from hospital admission to CT brain in acute ischaemic stroke.

A retrospective study of all acute ischaemic stroke patients admitted to Midland Regional Hospital Mullingar (MRHM) between January 2004 and September 2009 was undertaken in order to assess the median time from hospital admission to CT brain scan (n = 496). The median time to CT scan ranged from 19-24 hours between 2004-7. In 2008, coinciding with setting up a new Acute Stroke Service (ACSS), the median time to CT scan dropped to 15 hours (n = 130, p =0.03) and decreased further to 3 hours in 2009 (n = 125, p = 0.003). The proportion scanned within 1 hour of admission increased from 7 patients (4.6%) over 2004-7, to 28 patients (21.5%) in 2008 (p = 0) and 44 patients (35%) in 2009 (p = 0.018). This clinically and statistically significant reduction occurred following reorganisation of existing resources on a budget neutral basis at MRHM and was facilitated by the enthusiastic support of a range of disciplines bridging the community and acute hospital interface. Measurement of admission to CT brain scan time is one of several audit parameters which can assess hospitals responsiveness to acute stroke.

Asian women have greater abdominal and visceral adiposity than Caucasian women with similar body mass index.

BACKGROUND: In the Multiethnic Cohort Study, Japanese Americans (JA) have lower mean body mass index (BMI) compared with Caucasians, but show a higher waist-to-hip ratio at similar BMI values and a greater risk of diabetes and obesity-associated cancers. OBJECTIVE: We investigated the abdominal, visceral and hepatic fat distribution in these Asian and Caucasian Americans. DESIGN: A cross-sectional sample of 60 female cohort participants (30 JA and 30 Caucasians), of ages 60-65 years and BMIs 18.5-40 kg m(-2), underwent anthropometric measurements and a whole-body dual energy X-ray absorptiometry (DXA) scan: a subset of 48 women also had abdominal magnetic resonance imaging (MRI). RESULTS: By design, JA women had similar BMIs (mean 26.5 kg m(-2)) to Caucasian women (27.1 kg m(-2)). JA women were found to have a significantly smaller hip circumference (96.9 vs 103.6 cm; P=0.007) but not a significantly lower DXA total fat mass (25.5 vs 28.8 kg; P=0.16). After adjusting for age and DXA total fat mass, JA women had a greater waist-to-hip ratio (0.97 vs 0.89; P<0.0001), DXA trunk fat (15.4 vs 13.9 kg; P=0.0004) and MRI % abdominal visceral fat (23.9 vs 18.5%; P=0.01) and a lower DXA leg fat mass (8.2 vs 10.0 kg; P=<.0001). Their MRI % subcutaneous fat (33.4 vs 30.2%; P=0.21) and % liver fat (5.8 vs 3.8%; P=0.06) did not significantly differ from that of Caucasian women. CONCLUSIONS: Our findings build on limited past evidence, suggesting that Asian women carry greater abdominal and visceral fat when compared with Caucasian women with similar overall adiposity. This may contribute to their elevated metabolic risk for obesity-related diseases.

Urinary phytoestrogen excretion and prostate cancer risk: a nested case-control study in the Multiethnic Cohort.

BACKGROUND: Phytoestrogens are of special interest in prostate cancer research because populations in Asia with a high consumption of phytoestrogens have a lower incidence of the disease than comparable populations in Western countries. METHODS: This case-control study is nested within a large multiethnic cohort in Hawaii and California. Urine samples were analysed for daidzein, genistein, equol, and enterolactone among 249 incident prostate cancer cases and 404 controls matched on age, race/ethnicity, date/time of specimen collection, and fasting status. RESULTS: The median excretion of daidzein was 0.173 nmol mg(-1) creatinine in cases and 0.291 in controls (P=0.01), and the median excretion of genistein was 0.048 in cases and 0.078 in controls (P=0.05). An inverse association was seen for daidzein overall (odds ratio for the highest vs lowest quintile=0.55, 95% confidence interval=0.31-0.98, P(trend)=0.03) and seemed to apply to localized (P(trend)=0.08) as well as advanced or high-grade cancer (P(trend)=0.09). This association was consistent across the four ethnic groups examined. Although the relationship was weaker for genistein, the odds ratios and trends were similarly inverse. Urinary excretion of equol and enterolactone was not significantly related to prostate cancer risk. CONCLUSION: Our findings suggest that high intake of isoflavones, as reflected by urinary excretion of daidzein and genistein, may be protective against prostate cancer.

Cannabinoid activation of PPAR alpha; a novel neuroprotective mechanism.

BACKGROUND AND PURPOSE: Although CB(1) receptor activation evokes neuroprotection in response to cannabinoids, some cannabinoids have been reported to be peroxisome proliferator activated receptor (PPAR) ligands, offering an alternative protective mechanism. We have, therefore, investigated the ability of a range of cannabinoids to activate PPAR alpha and for N-oleoylethanolamine (OEA), an endogenous cannabinoid-like compound (ECL), to evoke neuroprotection. EXPERIMENTAL APPROACH: Assays of PPAR alpha occupancy and gene transactivation potential were conducted in cell-free and transfected HeLa cell preparations, respectively. In vivo estimates of PPAR alpha activation through fat mobilization and gene transcription were conducted in mice. Neuroprotection in vivo was investigated in wild-type and PPAR alpha gene-disrupted mice. KEY RESULTS: The ECLs OEA, anandamide, noladin ether and virodhamine were found to bind to the purified PPAR alpha ligand binding domain and to increase PPAR alpha-driven transcriptional activity. The high affinity synthetic CB(1/2) cannabinoid agonist WIN 55212-2 bound to PPAR alpha equipotently with the PPARalpha agonist fenofibrate, and stimulated PPARalpha-mediated gene transcription. The phytocannabinoid delta 9 tetrahydrocannabinol was without effect. OEA and WIN 55212-2 induced lipolysis in vivo, while OEA pre-treatment reduced infarct volume from middle cerebral artery occlusion in wild-type, but not in PPAR alpha-null mice. OEA treatment also led to increased expression of the NFkappa B-inhibitory protein, Ikappa B, in mouse cerebral cortex, while expression of the NFkappa B-regulated protein COX-2 was inhibited. CONCLUSIONS AND IMPLICATIONS: These data demonstrate the potential for a range of cannabinoid compounds, of diverse structures, to activate PPAR alpha and suggest that at least some of the neuroprotective properties of these agents could be mediated by nuclear receptor activation.

Prospective study of grapefruit intake and risk of breast cancer in postmenopausal women: the Multiethnic Cohort Study.

In vitro and in vivo studies have shown that cytochrome P450 3A4 (CYP3A4) is involved in the metabolism of oestrogens. There is evidence that grapefruit, an inhibitor of CYP3A4, increases plasma oestrogen concentrations. Since it is well established that oestrogen is associated with breast cancer risk, it is plausible that regular intake of grapefruit would increase a woman's risk of breast cancer. We investigated the association of grapefruit intake with breast cancer risk in the Hawaii-Los Angeles Multiethnic Cohort Study, a prospective cohort that includes over 50 000 postmenopausal women from five racial/ethnic groups. A total of 1657 incident breast cancer cases were available for analysis. Grapefruit intake was significantly associated with an increased risk of breast cancer (relative risk=1.30, 95% confidence interval 1.06-1.58) for subjects in the highest category of intake, that is, one-quarter grapefruit or more per day, compared to non-consumers (P(trend)=0.015). An increased risk of similar magnitude was seen in users of oestrogen therapy, users of oestrogen+progestin therapy, and among never users of hormone therapy. Grapefruit intake may increase the risk of breast cancer among postmenopausal women.

Nutritional changes among premenopausal women undertaking a soya based dietary intervention study in Hawaii.

BACKGROUND: We conducted a 2-year nutritional intervention among premenopausal women. The goal was to incorporate two daily servings of soya into the regular diet. This report describes the dietary modifications and assesses their nutritional adequacy with regard to major nutrients. METHODS: In this analysis of 100 intervention and 106 control subjects, women completed a validated food-frequency questionnaire at baseline; throughout the study, they participated in at least three unannounced 24-h recalls. RESULTS: At randomization, both groups were similar in age and body weight, reported low soya intake, and did not differ by intake of major nutrients and foods. According to the 24-h recalls, women in the intervention group consumed nearly two servings of soya per day, while the control women remained at 0.2 servings. In comparison with the control group, the intervention group consumed fewer dairy products, primarily milk, but also less meat, nuts and seeds. As a result of the dietary modification, the intervention women consumed less-saturated fat and cholesterol and more protein, dietary fibre, calcium and vitamins than the control group. CONCLUSION: These results suggest that women in the intervention group improved the overall quality of their diet by adding two servings of soya per day.

Plastic bottles as spacers for a pressurized metered-dose inhaler: in vitro characteristics

Homemade spacer devices are commonly used by children with asthma to improve aerosol deposition from pressurized metered dose inhalers (pMDI); however, the efficacy and efficiency of these devices are not fully characterized. We determined the quality of fine particle fraction (< 4.7 microns) and ultrafine particle fraction (< 3.3 microns) of three bottles (from 280 ml to 500 ml) commonly used as spacers in Trinidad and Tobago and compared their performance to the commercially available valved holding chamber (OpT) and pMDI. These data were obtained in vitro using a cascade impactor. All 3 bottles and the OpT were similar (p > 0.05) in reducing the amount of albuterol emitted as large particles (> 4.7 microns) to less than 10 micrograms. The different sized bottles (from 280 ml to 500 ml) produced identical quantities of albuterol in the fine particle and ultrafine particle ranges (p > 0.05). All of the sample bottle spacers emitted a higher amount (p < 0.002) of fine and ultrafine particles than the OpT and pMDI alone. The OpT resulted in a significantly higher fraction of fine particles (p < 0.05) and a greater quantity of drug (p < 0.05) in the ultrafine range as compared to the MDI only. The sizes of particles obtained from the bottle spacers are those that have a high probability of reaching the lower airway; however, the clinical relevance of these findings remains to be determined.

Plastic bottles as spacers for a pressurized metered-dose inhaler: In vitro characteristics

Homemade spacer devices are commonly used by children with asthma to improve aerosol deposition from pressurized metered dose inhalers (pMDI); however, the efficacy and efficiency of these devices are not fully characterized. We determined the quality of fine particle fraction (<4.7 um) and ultrafine particle fraction (<3.3 um) of three bottles (from 280 ml to 500 ml) commonly used as spacers in Trinidad and Tobago and compared their performance to the commercially available valved holding chamber (OpT) and pMDI. These data were obtained in vitro using a cascade impactor. All 3 bottles and OpT were similar (p>0.05) in reducing the amount of albuterol emitted as large particles (>4.7 um) to less than 10 ug. The diffreent sized bottles (from 280 ml to 500 ml) produced identical quantities of albuterol in the fine particle and ultrafine particle ranges (p>0.05). All of the sample bottle spacers emitted a higher amount (p<0.002) of fine and ultrafine particles than the OpT and pMDI alone. The OpT resulted in a significantly higher fraction of fine particles (p<0.05) and greater quantity of drug (<0.05) in the ultrafine range as compared to MDI only. The sizes of particles obtained from the bottle spacers are those that have a high probability of reaching the lower airway; however, the clinical relevance of these findings remains to be determined. (AU)

Evaluation of validity of items for a food behavior checklist.

OBJECTIVES: To evaluate the validity of food behavior items, using a biological measure (serum carotenoids) as the criterion for validity of fruit and vegetable intake, and the results from multiple 24-hour recalls to test convergent validity with nutrient intake. DESIGN: Participants responded to 39 food-behavior questions and later completed three 1-day dietary recalls. Serum carotenoid levels were determined for a 59% randomly selected subsample. SUBJECTS/SETTING: A convenience sample of 100 English-speaking, low-income women participating in a Food Stamp Nutrition Education Program in 8 California counties. Statistical analyses Spearman correlation coefficients were calculated between responses to the food behavior items and (a) serum carotenoid levels and (b) mean nutrient intakes from the 24-hour recalls. Cronbach's coefficient alpha was determined for items within broad food behavior topics. RESULTS: Responses to 10 food behavior items were significantly correlated with serum carotenoid levels (correlations greater than 0.45 were found for choosing low-fat foods and a self-evaluation of overall dietary quality). An additional 12 items showed hypothesized associations with the 24-hour recall data (with a maximum correlation 0.50 for number of eggs per week and dietary cholesterol). Cronbach's coefficient alpha ranged from 0.28 (for 5 fat and cholesterol items) to 0.79 (for 9 fruit and vegetable items). APPLICATIONS/CONCLUSIONS: Nutrition professionals can use these methods to validate items for food behavior checklists for specific populations. The items described here may be useful when designing instruments to administer to low-income women in a community setting.

Food insecurity is positively related to overweight in women.

Although individuals with poor food security might be expected to have reduced food intake, and thus reduced body fat and less likelihood of being overweight, these associations have not been adequately studied. The purpose of the current study was to examine the relationship between food insecurity and overweight as measured by body mass index (BMI) using data from the nationally representative 1994-1996 Continuing Survey of Food Intakes by Individuals (CSFII). Overweight was defined as BMI >27.3 kg/m(2) for women and 27.8 kg/m(2) for men. Food insecurity was related to overweight status for women (n = 4509, P < 0.0001), but not for men (n = 4970, P = 0.44). Excluding the 11 severely insecure women, the prevalence of overweight among women increased as food insecurity increased, from 34% for those who were food secure (n = 3447), to 41% for those who were mildly food insecure (n = 966) and to 52% for those who were moderately food insecure (n = 86). Food insecurity remained a significant predictor of overweight status, after adjustment for potentially confounding demographic and lifestyle variables (P < 0.01). In a logistic regression analysis, mildly insecure women were 30% more likely to be overweight than those who were food secure [odds ratio (OR) 1.3, P = 0.005]. Thus, food insecurity had an unexpected and paradoxical association with overweight status among women with a higher prevalence of overweight among the food insecure, and a resulting potential for increased incidence of obesity-related chronic diseases. Given that the rates of both overweight and food insecurity are on the rise, this research area warrants further investigation.

Plastic bottles as spacers for a pressurized metered-dose inhaler: in vitro characteristics.

Homemade spacer devices are commonly used by children with asthma to improve aerosol deposition from pressurized metered dose inhalers (pMDI); however, the efficacy and efficiency of these devices are not fully characterized. We determined the quality of fine particle fraction (< 4.7 microns) and ultrafine particle fraction (< 3.3 microns) of three bottles (from 280 ml to 500 ml) commonly used as spacers in Trinidad and Tobago and compared their performance to the commercially available valved holding chamber (OpT) and pMDI. These data were obtained in vitro using a cascade impactor. All 3 bottles and the OpT were similar (p > 0.05) in reducing the amount of albuterol emitted as large particles (> 4.7 microns) to less than 10 micrograms. The different sized bottles (from 280 ml to 500 ml) produced identical quantities of albuterol in the fine particle and ultrafine particle ranges (p > 0.05). All of the sample bottle spacers emitted a higher amount (p < 0.002) of fine and ultrafine particles than the OpT and pMDI alone. The OpT resulted in a significantly higher fraction of fine particles (p < 0.05) and a greater quantity of drug (p < 0.05) in the ultrafine range as compared to the MDI only. The sizes of particles obtained from the bottle spacers are those that have a high probability of reaching the lower airway; however, the clinical relevance of these findings remains to be determined.

Living arrangements affect dietary quality for U.S. adults aged 50 years and older: NHANES III 1988-1994.

The number and proportion of older U.S. adults who live alone have increased dramatically in the past three decades, and there is concern that these individuals may have particularly poor dietary quality. We examined the association of four living arrangements (living with a spouse only, with a spouse plus someone else, with someone other than a spouse or living alone) with dietary quality (the number of low nutrients out of a possible 15, with low defined as <67% of the recommended dietary allowance) among 6525 U.S. adults aged 50-64 y and those >/=65 y in the third National Health and Nutrition Examination Survey (NHANES III 1988-1994). Among non-Hispanic Caucasian adults, those who lived with a spouse only had better dietary quality, with significant differences ranging from 0.8 to 1.5 fewer low nutrients compared with those with other living arrangements. Effects of living arrangements on dietary quality were also seen among non-Hispanic African-Americans, Mexican-Americans, and those of "other" races, but differences were significant only for African-American men aged >65 y living with a spouse plus others (1.6 additional low nutrients compared with those living with a spouse only). Energy intake was strongly associated with dietary quality, but did not account for the associations between living arrangements and dietary quality. Although middle-aged and older adults with living arrangements other than living with a spouse only (including those living alone) tended to have poorer dietary quality, the effects varied substantially across age, gender and ethnic categories.

FE(NO): relationship to exhalation rates and online versus bag collection in healthy adolescents.

Measurement of exhaled nitric oxide (FE(NO)) is a noninvasive and practical method for assessing airway inflammation. We conducted this investigation to determine the most appropriate flow rate for FE(NO) measurement and to obtain normal values for FE(NO). We determined which expiratory flow was easy to sustain, generated reproducible values, and provided good correlation between offline and online measurements. Thirty-two healthy subjects (15- 18 yr old) underwent spirometry and FE(NO) measurements, using a chemiluminescent NO analyzer at expiratory flow rates of 46, 31, 23, 15, 10, 7, 5, and 4 ml/s. The major findings were as follows: (1) FE(NO) increased as flow rates decreased, with strong correlation between FE(NO) values and flow rates at the four highest flows (0. 85- 0.93, p < 0.001); (2) there were no significant differences and good agreement between offline bag and online FE(NO) values for the four highest flows (p < 0.09-0.83); (3) online FE(NO) values increased with age 15-17 yr at all flow rates, but decreased at age 18 yr; and (4) using multiple regression, significant predictors of FE(NO) were flow, body surface area, age, and FEF(25-75). On the basis of these results, we provide FE(NO) values for healthy adolescents and propose that the ideal flow rate for children is between 30 and 50 ml/s.

Exhaled nitric oxide as an indicator of severity of asthmatic inflammation.

Traditional assessment of severity of asthma relies on an evaluation of signs and symptoms and pulmonary function tests. These pulmonary function tests, such as peak expiratory flow rates, forced vital capacity, and forced expiratory flow rates, are indirect measures of airway caliber only, and not inflammation. Since asthma is an inflammatory disease, a measure of the degree of inflammation would be helpful in quantitating severity and titrating of anti-inflammatory therapy. A noninvasive method for measuring pulmonary inflammation would therefore be helpful to assist the emergency physician in initial treatment and assist in titration of anti-inflammatory therapy during repeat visits. Exhaled nitric oxide (NO) assays are convenient and practical and may fulfill this role. In this review, we discuss the role of NO in asthmatic inflammation and the role that exhaled NO values may play in the emergency management of asthma.

Intake of flavonoids and lung cancer.

BACKGROUND: To investigate the possible relationship between intake of flavonoids-powerful dietary antioxidants that may also inhibit P450 enzymes-and lung cancer risk, we conducted a population-based, case-control study in Hawaii. METHODS: An in-person interview assessed smoking history and usual intake of 242 food items for 582 patients with incident lung cancer and 582 age-, sex-, and ethnicity-matched control subjects. Subjects who donated a blood sample were genotyped for the P450 enzyme variant allele CYP1A1*2 by use of a polymerase chain reaction-based method. Logistic regression analysis was used to compute odds ratios (ORs) and 95% confidence intervals (CIs). All P values are two-sided. RESULTS: After adjusting for smoking and intakes of saturated fat and beta-carotene, we found statistically significant inverse associations between lung cancer risk and the main food sources of the flavonoids quercetin (onions and apples) and naringin (white grapefruit). The lung cancer OR for the highest compared with the lowest quartile of intake was 0.5 (95% CI = 0.3-0.9) for onions (P for trend =.001) and 0.6 (95% CI = 0.4-1.0) for apples (P for trend =.03). The OR for the highest compared with the lowest tertile of intake for white grapefruit was 0.5 (95% CI = 0.2-0.9) (P for trend =.02). No association was found for important food sources of other flavonoids. Using published food-composition data for flavonoids, we found an inverse association between intake of quercetin and risk of lung cancer (P for trend =.07) that appears consistent with associations for its food sources. The effect of onions was particularly strong against squamous cell carcinoma (a cell type specifically associated with CYP1A1*2 in our study) and was modified by the CYP1A1 genotype, suggesting that CYP1A1 may play a role in this association. CONCLUSION: If replicated, particularly in prospective studies, these findings would suggest that foods rich in certain flavonoids may protect against certain forms of lung cancer and that decreased bioactivation of carcinogens by inhibition of CYP1A1 should be explored as underlying mechanisms.

Apoptosis induced by DNA uptake limits transfection efficiency.

Electrotransfection is an effective method for transfecting lymphoid cells. However, the transfection efficiency of certain lymphoid cells is low. L1210 subclones and NFS-70 pro-B cells, which are highly refractory to various transfection methods, were used to identify the limiting factors. Cells were electrotransfected with plasmids coding for green fluorescence protein or luciferase. The luciferase expression of L1210 subclone 3-3 was found to increase 6-12 h after electroporation, but decreased significantly from 12 to 48 h. The lower level of luciferase activity at later time periods correlated with decreases in cell viability, which was shown to be due to apoptosis, as determined by propidium iodide/acrindine orange staining, DNA laddering, and prevention of cell death by addition of caspase inhibitors. Similar results were observed with NFS-70 pro-B cells and select L1210 subclones. In contrast, L1210 parental and L1210 subclone 7-15.6 cells undergo only low levels of apoptosis (< or = 5%). Apoptosis occurred only when DNA (plasmids or salmon sperm DNA) was present during electroporation, but was not dependent on the conformation of the DNA used or the expression of transgenes. Cells pulsed in the presence of dextran sulfate (MW 500,000) did not apoptose. Similar results were observed when L1210 subclone 3-3 was transfected using the cationic lipid 1, 2-dioleoyl-3-trimethylammonium propane, although the transfection efficiency and corresponding rate of apoptosis were significantly lower. Applying the caspase inhibitor fluoromethyl ketone (Boc-ASP-FMK) dramatically improved cell viability and transgene expression of select L1210 subclones and NFS-70 pro-B cells.

Absence of MHC class II antigen expression in trophoblast cells results from a lack of class II transactivator (CIITA) gene expression.

Although the mechanism(s) underlying the failure of the maternal immune system to reject the semiallogeneic fetus have not been clearly defined, the absence of MHC class II antigen expression by fetal trophoblast cells very likely plays a critical role in the maintenance of normal pregnancy. However, the regulation of class II antigen expression in trophoblast cells is poorly understood. Class II transactivator (CIITA) is a transacting factor that is required for both constitutive and IFN-gamma-inducible class II gene transcription. In this report we demonstrate that the inability of trophoblast cells to express class II antigens is due to a lack of CIITA gene expression. Trophoblast cell lines derived from human, mouse, and rat do not express CIITA, and expression is not inducible by IFN-gamma. The absence of CIITA gene expression in trophoblasts treated with IFN-gamma does not result from a defect in the IFN-gamma receptor or the JAK/STAT pathway, because the classical IFN-gamma inducible gene encoding the guanylate-binding protein is expressed. Transfection of CIITA expression vectors into trophoblast cells results in activation of class II promoters, endogenous class II mRNA expression, and subsequent expression of class II antigens on the cell surface. In contrast, class I mRNA is not expressed in human trophoblast cells transfected with CIITA expression vectors. Thus, trophoblast cells contain all of the DNA binding factors necessary for class II transcription, and ectopic expression of CIITA is sufficient to activate class II, but not class I expression. The failure of trophoblast cells to express CIITA, and therefore class II antigens, provides a potential mechanism by which the fetus is protected from the maternal immune system during pregnancy.

Repression of MHC class II gene transcription in trophoblast cells by novel single-stranded DNA binding proteins.

The maintenance of the fetus during pregnancy has been attributed to the absence of major histocompatibility complex (MHC) class II antigens on fetal trophoblastic cells that make contact with the maternal immune system. However, the mechanism(s) by which class II genes are regulated in trophoblast cells is unclear. We have identified a negative regulatory element (IA alpha NRE) in the promoter of the mouse class II gene IA alpha that represses IA alpha transcription in trophoblast cells. IA alpha NRE, located from-839 to -828, binds transacting factors from rat, mouse and human trophoblast cells, but not from 18 other cell lines tested. These results indicate that IA alpha NRE binding proteins (IA alpha NRE BPs) are conserved in species with hemochordial placentas, and suggest that IA alpha NRE binding activity is restricted primarily to trophoblast cells. Interestingly, the IA alpha NRE BPs bind to the IA alpha NRE antisense strand in a sequence-specific manner. IA alpha NRE represses transcription from the IA alpha promoter in a position-dependent manner, and has a minor down-regulatory effect on the activity of the SV40 promoter/enhancer. Our results demonstrate that MHC class II gene transcription is repressed in fetal trophoblast cells by sequence-specific, single-stranded DNA binding proteins, and suggest a possible mechanism by which the conceptus is protected from immune rejection during pregnancy.