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402 Virulence Factors of Pyelonephritis Associated E. Coli from Pregnant Women
386
400
SPO Abstracts
PRENATAL DIAGNOSIS OF HIV INFECTION: THE USE OF CORDOCENTESIS, POLYMERASE CHAIN REACTION, AND P24 ANTIGEN ASSAY Mark T Cullen MP Richard R Viscarello MO, Sharon Paryani MO', L Sanchez-Ramos M.O, University of Florida Health Science Center, Jacksonville, FL. and Yale University School of Medicine, New Haven, CT.
Perinatal transmission of the Human Immunodeficiency Virus is thought to be between 25 to 30%. Neonatal infection is devastating with a 50% mortality in the first year. Antenatal knowledge of fetal infection would allow the option of pregnancy termination or fetal treatment with antiretroviral agents. Thirteen HIV-infected, pregnant women underwent cordocentesis for c~nical indications. Two had AIDS (18%) and 11 were seropositive asymptomatic. A single insertion into an anterior cord was performed in all cases. Blood was sent for HCT, PCR, p24 antigen, T-cell phenotyping, and a Kleihauer-Betke. Gestational age ranged from 22 to 34 weeks (mean of 28.2 ± 5.4). Fetal CD4 counts were available in nine cases. There was no difference between the infected and uninfected groups. There was 1 fetal demise secondary to an abruption at term in a PCR-negative fetus. Three cordocentesis samples were positive for HIV-1 by PCR, 2 were indeterminate, and 7 negative. Of the three fetuses who had positive tests by PCR, 2 have developed AIDS, and one is asymptomatic and <15 months (PO). Both of these fetuses also had negative p24 antigen tests. One false negative PCR test was observed. Conclusion: HIV detected by PCR at cordocentesis appears to predict which fetus will develop HIV infection. A negative test is reassuring, but not absolute.
401
HIV IN PREGNANCY: FACTORS PREDICTIVE OF MATERNAL AND FETAL OUTCOME. Mark T Cyllen MIl. Issac Oelke MO, Joseph Greenhaw MO', Richard R. Viscarello MO, Sharon Paryani MO', Luis Sanchez-Ramos MO, University of Florida, Jacksonville, FL. Yale University, New Haven, CT
In a 3 year study period, 7,596 pregnant women were screened for HIV-1. 82 (1.07%) were found to be HIVseropositive and followed prospectively. The mean age at entry was 24.7 yrs. 23 were IVDAs, 10 admitted to prostitution, 25 had multiple heterosexual partners, 2 h.ad received infected blood products, and 28 had no known nsk factors. 71 were asymptomatic HIV-positive, 7 had AIDS, and 1 had ARC. 12 of 82 (14%) chose termination of pregnancy for the following reasons: 2 had hysterectomy for CIN3, 1 had new onset PCP pneumonia, and the rest were elective. To date, 64 deliveries have occurred. There were 56 vaginal deliveries and 8 cesarean sections. There was 1 third trimester stillbirth secondary to an abruption. 2 maternal deaths occurred in the peripartum period, which were HIVrelated. 8 mothers experienced infectious morbidity. Mean CD4 nadir, while lower for this group did not reach statistical significance (314 vs 550 p=.06). Of the 63 infants, 1~ are infected, 15 are seronegative, and 35 are PO. The pennatal transmission rate is currently 22.4%. Mean birthweight was 2,739g (936-4225) and gestation age was 37.7 weeks. 20:'0 were premature deliveries. There were no differences In demographics. mode of delivery. length of ROM. or the use of a scalp clip between the infected and uninfected groups. Transmission rates were influenced by clinical status in the mother. Eight mothers were placed on AZT for CD4 counts<500/cc. There were no ill effects of the drug on either mother or fetus. Conclusion: Clinical status appears to be the best predictor of transmission and pregnancy course.
January 1992 Am J Obstet Gynecol
402
VIRUlENCE FACfORS OF PYELONEPHRmS ASSOCIATED E. COll FROM PREGNANT WOMEN. M. Martens', A. Hart', B. Nowicki', S. Nowicki·, C. Peyton·, L. Schaiter·, G. Anderson, Department of Obstetrics and Gynecology, University of Texas Medical Branch, Galveston, Texas Pyelonephritis is a serious complication in pregnancy. E. coli is the most frequent organism associated with pyelonephritis. Virulence determinants such as P funbriae have been determined to be associated with the pathogenesis of pyelonephritis in non-pregnant women. However, virulence factors have not been extensively characterized in pregnant women with pyelonephritis. Different flmbrial types on E. coli including P, Dr and type I were detected on E. coli isolates from IS pregnant and 12 non-pregnant patients with pyelonephritis and 31 non infected individuals. E. coli strains were tested for the presence of P, Dr, and type 1 fimbriae (colonization factors) by agglutination with human pp, Dr (a-) erythrocytes or yeast cells (Table 1).
No./Patient Pregnant (IS) <.002· with pyelonephritis Non-pregnant(12) <.01· with pyelonephritis Healthy women
NS
No. of E.coli 16
TYPE! 3 (19%)'
FIMBRIAE P Dr 11 value* 12 (75%)* 3 (19%)·
11
5 (45%)
9(81%)'
3 (27%)*
15
9 (60%)
8(53%)
4 (27%)
The results indicate that P funlriae are expressed with higher frequency in pyelonenephritis - associated E. coli. Among cervical isolates of healthy non-infected funbrial types are more distributed among all strains with type 1 funbriae being the most frequent. Therefore, it appears that P funbriae may be virulence factors associated with the development of pyelonephritis with the greatest risk associated with infection in pregnancy.
403 HORMONAL RECEPTORS AND ADHERENCE OF
PYELONEPHRlTIS ASSOCIATED ESCHERICHIA COLI IN PREGNANCY. M. Martens', B. Nowicki', A. Hart', B. Taylor*, S. Nowicki', G.D. Anderson, Department of Obstetrics and Gynecology, University of Texas Medical Branch, Galveston, Texas The incidence of pyelonephritis is increased in pregnancy, and has been attributed to various alterations of the human host. However an investigation of the changes which pregnancy has on bacteria has not been undertaken. Kinetics of various hormones in pregnancy and incidence of pyelonephritis was analyzed, and a correlation was noted for HCG. HCG and a control growth hormone (GH), were tested for possible receptor binding sites on pathogenic E. coli strains. GH or HCG were coated onto microtitre trays. E. coli, (type I pili positive and negative), and guinea pig RBC were added. Erythrocytes not trapped by bacteria-hormone complexes were removed by washing, and 0.001 % Tryton X was added to lyse bound red cells. The amount of released hemoglobin was measured in an ELISA reader. Results indicate that pathogenic type I funbriated E. coli bind to HCG in a dose dependent fashion. Saturation of E. coli-HCG binding occur at 4 ug (approximately 65,000 mIU) of HCG per well. Attachment of E. coli cells to HeG was inhibited by a1pha- methyl-mannose, which is an inhibitor and receptor analog for type I pili adhesin. The alpha subunit ofHCG, but not beta or GH. demoDStrated dose dependent binding of E. coli. We conclude that E. coli via type I pili can bind to the mannose receptor on the alpha subunit of HeG. This phenomenon may be important in the pathogenesis of pyelonephritis in the early trimesters of pregnancy due to an increase in adherence to receptors found in genito-urinary tract tissues.
400
SPO Abstracts
PRENATAL DIAGNOSIS OF HIV INFECTION: THE USE OF CORDOCENTESIS, POLYMERASE CHAIN REACTION, AND P24 ANTIGEN ASSAY Mark T Cullen MP Richard R Viscarello MO, Sharon Paryani MO', L Sanchez-Ramos M.O, University of Florida Health Science Center, Jacksonville, FL. and Yale University School of Medicine, New Haven, CT.
Perinatal transmission of the Human Immunodeficiency Virus is thought to be between 25 to 30%. Neonatal infection is devastating with a 50% mortality in the first year. Antenatal knowledge of fetal infection would allow the option of pregnancy termination or fetal treatment with antiretroviral agents. Thirteen HIV-infected, pregnant women underwent cordocentesis for c~nical indications. Two had AIDS (18%) and 11 were seropositive asymptomatic. A single insertion into an anterior cord was performed in all cases. Blood was sent for HCT, PCR, p24 antigen, T-cell phenotyping, and a Kleihauer-Betke. Gestational age ranged from 22 to 34 weeks (mean of 28.2 ± 5.4). Fetal CD4 counts were available in nine cases. There was no difference between the infected and uninfected groups. There was 1 fetal demise secondary to an abruption at term in a PCR-negative fetus. Three cordocentesis samples were positive for HIV-1 by PCR, 2 were indeterminate, and 7 negative. Of the three fetuses who had positive tests by PCR, 2 have developed AIDS, and one is asymptomatic and <15 months (PO). Both of these fetuses also had negative p24 antigen tests. One false negative PCR test was observed. Conclusion: HIV detected by PCR at cordocentesis appears to predict which fetus will develop HIV infection. A negative test is reassuring, but not absolute.
401
HIV IN PREGNANCY: FACTORS PREDICTIVE OF MATERNAL AND FETAL OUTCOME. Mark T Cyllen MIl. Issac Oelke MO, Joseph Greenhaw MO', Richard R. Viscarello MO, Sharon Paryani MO', Luis Sanchez-Ramos MO, University of Florida, Jacksonville, FL. Yale University, New Haven, CT
In a 3 year study period, 7,596 pregnant women were screened for HIV-1. 82 (1.07%) were found to be HIVseropositive and followed prospectively. The mean age at entry was 24.7 yrs. 23 were IVDAs, 10 admitted to prostitution, 25 had multiple heterosexual partners, 2 h.ad received infected blood products, and 28 had no known nsk factors. 71 were asymptomatic HIV-positive, 7 had AIDS, and 1 had ARC. 12 of 82 (14%) chose termination of pregnancy for the following reasons: 2 had hysterectomy for CIN3, 1 had new onset PCP pneumonia, and the rest were elective. To date, 64 deliveries have occurred. There were 56 vaginal deliveries and 8 cesarean sections. There was 1 third trimester stillbirth secondary to an abruption. 2 maternal deaths occurred in the peripartum period, which were HIVrelated. 8 mothers experienced infectious morbidity. Mean CD4 nadir, while lower for this group did not reach statistical significance (314 vs 550 p=.06). Of the 63 infants, 1~ are infected, 15 are seronegative, and 35 are PO. The pennatal transmission rate is currently 22.4%. Mean birthweight was 2,739g (936-4225) and gestation age was 37.7 weeks. 20:'0 were premature deliveries. There were no differences In demographics. mode of delivery. length of ROM. or the use of a scalp clip between the infected and uninfected groups. Transmission rates were influenced by clinical status in the mother. Eight mothers were placed on AZT for CD4 counts<500/cc. There were no ill effects of the drug on either mother or fetus. Conclusion: Clinical status appears to be the best predictor of transmission and pregnancy course.
January 1992 Am J Obstet Gynecol
402
VIRUlENCE FACfORS OF PYELONEPHRmS ASSOCIATED E. COll FROM PREGNANT WOMEN. M. Martens', A. Hart', B. Nowicki', S. Nowicki·, C. Peyton·, L. Schaiter·, G. Anderson, Department of Obstetrics and Gynecology, University of Texas Medical Branch, Galveston, Texas Pyelonephritis is a serious complication in pregnancy. E. coli is the most frequent organism associated with pyelonephritis. Virulence determinants such as P funbriae have been determined to be associated with the pathogenesis of pyelonephritis in non-pregnant women. However, virulence factors have not been extensively characterized in pregnant women with pyelonephritis. Different flmbrial types on E. coli including P, Dr and type I were detected on E. coli isolates from IS pregnant and 12 non-pregnant patients with pyelonephritis and 31 non infected individuals. E. coli strains were tested for the presence of P, Dr, and type 1 fimbriae (colonization factors) by agglutination with human pp, Dr (a-) erythrocytes or yeast cells (Table 1).
No./Patient Pregnant (IS) <.002· with pyelonephritis Non-pregnant(12) <.01· with pyelonephritis Healthy women
NS
No. of E.coli 16
TYPE! 3 (19%)'
FIMBRIAE P Dr 11 value* 12 (75%)* 3 (19%)·
11
5 (45%)
9(81%)'
3 (27%)*
15
9 (60%)
8(53%)
4 (27%)
The results indicate that P funlriae are expressed with higher frequency in pyelonenephritis - associated E. coli. Among cervical isolates of healthy non-infected funbrial types are more distributed among all strains with type 1 funbriae being the most frequent. Therefore, it appears that P funbriae may be virulence factors associated with the development of pyelonephritis with the greatest risk associated with infection in pregnancy.
403 HORMONAL RECEPTORS AND ADHERENCE OF
PYELONEPHRlTIS ASSOCIATED ESCHERICHIA COLI IN PREGNANCY. M. Martens', B. Nowicki', A. Hart', B. Taylor*, S. Nowicki', G.D. Anderson, Department of Obstetrics and Gynecology, University of Texas Medical Branch, Galveston, Texas The incidence of pyelonephritis is increased in pregnancy, and has been attributed to various alterations of the human host. However an investigation of the changes which pregnancy has on bacteria has not been undertaken. Kinetics of various hormones in pregnancy and incidence of pyelonephritis was analyzed, and a correlation was noted for HCG. HCG and a control growth hormone (GH), were tested for possible receptor binding sites on pathogenic E. coli strains. GH or HCG were coated onto microtitre trays. E. coli, (type I pili positive and negative), and guinea pig RBC were added. Erythrocytes not trapped by bacteria-hormone complexes were removed by washing, and 0.001 % Tryton X was added to lyse bound red cells. The amount of released hemoglobin was measured in an ELISA reader. Results indicate that pathogenic type I funbriated E. coli bind to HCG in a dose dependent fashion. Saturation of E. coli-HCG binding occur at 4 ug (approximately 65,000 mIU) of HCG per well. Attachment of E. coli cells to HeG was inhibited by a1pha- methyl-mannose, which is an inhibitor and receptor analog for type I pili adhesin. The alpha subunit ofHCG, but not beta or GH. demoDStrated dose dependent binding of E. coli. We conclude that E. coli via type I pili can bind to the mannose receptor on the alpha subunit of HeG. This phenomenon may be important in the pathogenesis of pyelonephritis in the early trimesters of pregnancy due to an increase in adherence to receptors found in genito-urinary tract tissues.