Natural Course of Human T-Cell Leukemia Virus Type 1 Proviral DNA Levels in Carriers During Pregnancy.

The measurement of human T-cell leukemia virus type 1 (HTLV-1) proviral DNA levels by using polymerase chain reaction has been beneficial for confirming HTLV-1 infection during pregnancy. However, the influence of pregnancy on HTLV-1 infection and proviral DNA levels among pregnant women with HTLV-1 has not been clarified. We prospectively gathered blood samples from 36 pregnant women in whom HTLV-1 carriage was previously diagnosed and sequentially measured their proviral DNA levels. The HTLV-1 proviral DNA levels remained at a plateau during pregnancy but were elevated after delivery. Moreover, flow cytometry and serological analyses revealed that the regulatory T-cell population and soluble interleukin 2 receptor levels were similarly elevated after birth in comparison with those in control pregnant women. This study is the first to provide data on sequential changes in HTLV-1 proviral DNA levels during and after pregnancy. These findings will guide the establishment of a better program to prevent mother-to-child transmission of HTLV-1.

HTLV-1 targets human placental trophoblasts in seropositive pregnant women.

Human T cell leukemia virus type 1 (HTLV-1) is mainly transmitted vertically through breast milk. The rate of mother-to-child transmission (MTCT) through formula feeding, although significantly lower than through breastfeeding, is approximately 2.4%-3.6%, suggesting the possibility of alternative transmission routes. MTCT of HTLV-1 might occur through the uterus, birth canal, or placental tissues; the latter is known as transplacental transmission. Here, we found that HTLV-1 proviral DNA was present in the placental villous tissues of the fetuses of nearly half of pregnant carriers and in a small number of cord blood samples. An RNA ISH assay showed that HTLV-1-expressing cells were present in nearly all subjects with HTLV-1-positive placental villous tissues, and their frequency was significantly higher in subjects with HTLV-1-positive cord blood samples. Furthermore, placental villous trophoblasts expressed HTLV-1 receptors and showed increased susceptibility to HTLV-1 infection. In addition, HTLV-1-infected trophoblasts expressed high levels of viral antigens and promoted the de novo infection of target T cells in a humanized mouse model. In summary, during pregnancy of HTLV-1 carriers, HTLV-1 was highly expressed in placental villous tissues, and villous trophoblasts showed high HTLV-1 sensitivity, suggesting that MTCT of HTLV-1 occurs through the placenta.

Cloning and characterization of liver progenitor cells from the scattered cell clusters in primary culture of porcine livers.

The scattered cell clusters that can differentiate into hepatocytes or biliary epithelial cells have been isolated from primary cultures of adult porcine livers. We have generated 11 clonal cell lines from this system and identified liver progenitor cells (LPCs) among the clonal lines. These clonal lines expressed c-kit, HNF-1, HNF-6, and/or CK19 mRNA. An immunocytochemical study of the clonal lines indicated that clonal line CL-11 expressed liver epithelial cell markers CK14, vimentin, CK18, and BD-1. The expression of albumin and alpha1-antitrypsin (alpha1-AT) mRNA was only upregulated in CL-11 among the clonal lines when they were grown as aggregates. Under these conditions, CL-11 also exhibited ammonia metabolic activity and several indicators that suggest hepatocytic differentiation, including the upregulation of liver-specific genes such as dipeptidyl peptidase IV, CYP1A1, and CYP3A4 mRNA, and the downregulation of biliary cell markers such as gamma-glutamyltrans-peptidase (GGT), CK19, and HNF6 mRNA. After culturing CL-11 in Matrigel, the expression of GGT and HNF6 mRNA was upregulated. These results indicate that CL-11 has dual potential: the ability to differentiate as hepatocytes or as bile duct cells. The isolation of scattered cells could provide a simple method to generate LPC lines from adult livers.

Presence of side-population cells in an immortalized nontumorigenic human liver epithelial cell line.

Side-population (SP) cells have been shown to be highly enriched stem cells. We investigated whether an immortalized, nontumorigenic human liver cell line, THLE-5b, contains SP cells. Flow cytometry analysis after Hoechst 33342 staining demonstrated that the THLE-5b line contained a small component of SP cells. These SP cells were essentially eliminated by treatment with verapamil and expressed higher levels of ABCG2 mRNA than non-SP cells. In addition, the level of these SP cells detected by Hoechst 33342 staining was affected by the experimental conditions including the incubation medium. This is the first report of the presence of SP cells in the immortalized, nontumorigenic human liver cell line.

A very rare case of an ectopic cervical intramural pregnancy.

OBJECTIVE: To report a case of a very rare ectopic cervical intramural pregnancy. DESIGN: Case report. SETTING: Prefectural hospital. PATIENT(S): A 22-year-old woman, gravida 1, para 0, was referred to our hospital with the suspicion of a cervical ectopic pregnancy (EP). Pelvic examination revealed an enlarged uterine cervix with no genital bleeding. We found a clear gestational sac (GS) and fetal heart beat in the anterior muscular layer of the uterine cervix by ultrasonography, and confirmed these findings by magnetic resonance imaging (MRI). INTERVENTION(S): We injected methotrexate (MTX) into the GS cavity and around the GS. One week later, the GS was removed surgically without massive bleeding. MAIN OUTCOME MEASURE(S): On the 11th postoperative day, she recovered and was discharged from our hospital. Her menstruation restarted on the 35th postoperative day. RESULT(S): We have shown a case of a very rare ectopic cervical intramural pregnancy with successful treatment. CONCLUSION(S): We have explained a case and successful treatment of a very rare ectopic cervical intramural pregnancy with clear GS and fetal heart beat. Our strategy was injecting MTX into the GS cavity and around the GS, then performing an operation to remove the GS.

A novel positive regulatory element for exfoliative toxin A gene expression in Staphylococcus aureus.

A 1.4 kb positive regulatory element (ETA(exp)) that controls staphylococcal exfoliative toxin A (sETA) transcription was cloned from Staphylococcus aureus. ETA(exp) is located upstream of the cloned 5.8 kb eta gene (etaJ1) obtained from the chomosomal DNA of S. aureus ZM, the standard ETA-producing strain. The cETA prepared from an Escherichia coli transformant into which the recombinant plasmid petaJ1 (5.8 kb eta/pUC9) had been introduced was expressed at high levels in the culture supernatant and the ammonium-sulfate-precipitated culture supernatant fraction as shown by immunoblotting and the single radial immunodiffusion test. However, cETA produced by the recombinant plasmid petaJ3 containing the 1.7 kb eta sequence (etaJ3) with a 1.45 kb ETA(exp)-deficient eta fragment (1.7 kb eta/pUC9) obtained from the 5.8 kb eta sequence by subcloning was not detected in either the culture supernatant or the ammonium-sulfate-precipitated culture supernatant fraction (167-fold concentrate of the culture supernatant) by immunoblotting or the single radial immunodiffusion test. A large amount of cETA was produced by the 1.7 kb eta sequence when it was linked to ETA(exp) amplified by PCR (1.7 kb eta-ETA(exp)/pUC9), regardless of the orientation of ETA(exp) insertion. Northern blot hybridization showed lower levels of the transcripts of the 1.7 kb eta sequence than of the 5.8 kb eta sequence. The rsETA prepared from an S. aureus transformant into which the recombinant plasmid 3.4 kb eta-ETA(exp)/pYT3 (pYT3-etaJ6) had been introduced was expressed at high levels in the culture supernatant fraction as shown by the latex agglutination test. However, the agglutination titre in the culture supernatant fraction of rsETA produced by the recombinant plasmid (1.7 kb eta/pYT3) containing the 1.7 kb eta sequence carrying the 1.4 kb ETA(exp)-deficient eta fragment (pYT3-etaJ3) was 2500-4000 times lower than that of pYT3-etaJ6.