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H Taira

Publications and source records attributed to H Taira.

At least 73 records · Page 4Linked to original sources
PubMed1987

Interferon induction by transfection of Sendai virus C gene cDNA.

To elucidate the mechanism of interferon (IFN) induction on virus infection, we constructed two types of plasmids by inserting a part of the cDNA of the Sendai virus into a simian virus 40-derived expression vector (pSV2-0). One, pSV2-PC, contained the P + C gene, which codes for the P and C proteins in overlapping reading frames, and the other, pSV2-C, contained only the C gene. After transfecting the plasmids into mammalian cells, we determined the IFN activity in the culture medium. We found that the level obtained with pSV2-PC was significantly positive but very low, whereas that obtained with pSV2-C was as high as or even higher than that observed in the culture medium after Sendai virus infection. By cleaving pSV2-C between the simian virus 40 promotor and the C gene or by inserting a stop codon within the C gene [pSV2-C(stop)], induction of IFN was greatly diminished. In Northern blot analyses of the transcripts obtained from the cells transfected with the plasmids with cDNA to the P + C gene as a probe, the transcript having the expected size was detected with both pSV2-C and pSV2-C(stop), whereas none was detected with cleaved pSV2-C or pSV2-0. The results indicate that both transcription and translation of the C gene seem to be required for IFN induction after Sendai virus infection.

Animals↗
PubMed1986

Sequence of 2,617 nucleotides from the 3' end of Newcastle disease virus genome RNA and the predicted amino acid sequence of viral NP protein.

DNA fragments complementary to the Newcastle disease virus genome (strain D26) were cloned and sequenced. The sequence of 2,617 nucleotides from the 3' end of the genome was determined and an open reading frame (OP-1) consisting of 1,467 nucleotides, most likely encoding NP protein, was found in this region. This was followed by a second unfinished open reading frame (OP-2) of at least 729 nucleotides which continued beyond the 2,617th nucleotide. Another relatively short (312 nucleotides long) open reading frame (OP-2') was found overlapping with OP-2, but its significance is still unclear. The amino acid sequence deduced from the nucleotide sequence of OP-1 showed a moderate homology to that of the NP protein of Sendai virus in the central portion of the peptide. The leader sequence of 53 nucleotides was also identified. The 5' end of mRNAs synthesized in the infected cells was analyzed and found to be m7GpppA, suggesting that the transcription of viral mRNAs starts with A, but not with G residue.

Amino Acid Sequence↗
PubMed1985

Plasma angiotensin I and serum placental leucine aminopeptidase (P-LAP) in pre-eclampsia.

A study was undertaken on serial measurements of plasma angiotensin I (A-I) and serum placental leucine aminopeptidase (P-LAP) activities in normal and pre-eclamptic pregnancy. There was the significant difference in A-I levels between normal and mild pre-eclamptic pregnancy at weeks 30, 35, 37, between normal and severe pre-eclamptic pregnancy at week 37. There were no differences in serum P-LAP between normal and mild pre-eclamptic pregnancy up to week 33, but thereafter the levels for the mild pre-eclampsia were significantly higher than for the normal pregnancy. The P-LAP activity for the severe pre-eclampsia reached its maximum level at week 31. Around this week, the levels for severe pre-eclampsia were significantly higher than in the normal pregnancy. After week 35, the activities decreased precipitously to week 40; the activities for severe pre-eclampsia in late pregnancy at weeks 39 and 40 were significantly lower than in normal pregnancy. The above data support the idea that P-LAP test is useful for prediction or diagnosis of pre-eclampsia.

Adult↗
PubMed1985

In vitro degradation of oxytocin by pregnancy serum, placental subcellular fractions and purified placental aminopeptidases.

The degradation of oxytocin (formula; see text) by human placental particulate and soluble fractions, pregnant and non-pregnant sera, and purified enzymes such as microsomal placental leucine aminopeptidase (P-LAP), retroplacental serum P-LAP and placental post-proline endopeptidase, was studied by measuring liberated amino acids with a high performance liquid chromatograph (HPLC). While the placental particulate fraction degraded oxytocin almost completely into single amino acid and amide, the placental soluble fraction did not liberate any amino acid and amide. Purified retroplacental P-LAP liberated Tyr2, Ile3, Gln4 and Asn5 from the cyclic structure of oxytocin actively. Pregnancy serum containing the retroplacental P-LAP liberated also these amino acids and amides. Purified microsomal P-LAP liberated Leu8 in addition to these amino acids and amides. Although purified placental post-proline endopeptidase or porcine kidney leucine aminopeptidase (LAP) could not liberate any amino acid and amide from oxytocin, the combination of the post-proline endopeptidase with porcine kidney LAP or with placental microsomal P-LAP actively liberated all amino acids and amides detectable by HPLC. When the ratio of amino acid liberation velocity to LAP activity measured with leu-p-nitroanilide was calculated, the ratios for cyclic amino acids such as Tyr2 and Ile3 were high with the placental particulate fraction, the mixture of post-proline endopeptidase and microsomal P-LAP, retroplacental P-LAP, and pregnancy serum. The ratio for Leu8 was high with the placental particulate fraction and the mixture of post-proline endopeptidase and microsomal P-LAP.(ABSTRACT TRUNCATED AT 250 WORDS)

Amino Acids↗
PubMed1985

Comparative studies on (2'-5')oligoadenylate-related enzyme systems and the antiviral effect of interferon in two mouse cell lines which differ in (2'-5')oligoadenylate sensitivity of their protein synthesizing system.

Effect of (2'-5')oligoadenylate (2-5A) on cellular and viral protein and RNA syntheses was investigated with two mouse cell lines, L929 and Lz (a subclone of L929). The oligonucleotide was introduced into the cells either by using calcium phosphate coprecipitation technique or by microinjection method. In L929 cells protein and viral RNA syntheses were severely inhibited by 2-5A, whereas in Lz cells, both were only slightly inhibited. The activities of 2-5A synthetase and double-stranded (ds)RNA-dependent protein kinase were enhanced by interferon (IFN) treatment roughly to the same extent and there was no significant difference in the level of 2'-5' phosphodiesterase activity either. On the other hand, 2-5A-dependent RNase (RNase L) activity in Lz cells was low, being about 10-20% of that of L929 cells. It was increased twofold after IFN treatment, but protein synthesis of Lz cells was not as sensitive to 2-5A as that of L929 cells even after IFN treatment. L929 and Lz cells were sensitive to the antiviral effect of mouse IFN against vesicular stomatitis virus (VSV) and Mengovirus. In contrast, however, Lz cells were relatively insensitive to the antiviral effect of IFN on vaccinia virus, whereas L929 cells were sensitive.

2',5'-Oligoadenylate Synthetase↗
PubMed1985

In vitro degradation of angiotensin II (A-II) by human placental subcellular fractions, pregnancy sera and purified placental aminopeptidases.

The degradation of angiotensin II (Asp1-Arg2-Val3-Tyr4-Ile5-His6-Pro7-Phe8: A-II) by human placental particulate and soluble fractions, pregnant and non-pregnant sera, and highly purified placental enzymes such as placental leucine aminopeptidase P-LAP (microsomal), retroplacental serum P-LAP (soluble), aminopeptidase A and post-proline endopeptidase, was studied by measuring liberated amino acids by high performance liquid chromatography. Placental particulate and soluble fractions degraded A-II almost completely into single amino acids. The purified P-LAP (microsomal) actively liberated five amino acids from the N-terminal. The placental particulate fraction containing P-LAP (microsomal) also actively liberated these amino acids. The purified aminopeptidase A liberated Asp1 very actively as expected. When the ratio of the velocity of liberation of each amino acid to P-LAP activity measured with leu-p-nitroanilide as a substrate was calculated, placental soluble fraction liberated Asp1 very actively, but the liberation rate of Asp1 with the purified P-LAP (soluble) was very low. Therefore it seems that the enzyme in the placental soluble fraction and pregnancy serum responsible for the Asp1 liberation is not P-LAP (soluble), but aminopeptidase A. The mixture of purified P-LAP (soluble) and aminopeptidase A showed higher liberation rate of Arg2 and Val3 than that with purified aminopeptidase A alone, demonstrating that once the N-terminal Asp1 was liberated, the P-LAP (soluble) attacks the shorter peptide (angiotension III) very actively. It was concluded that P-LAP (microsomal) together with aminopeptidase A seem to contribute greatly to the degradation of A-II in pregnant women.

Amino Acids↗
PubMed1983

Structure and expression of cloned murine IFN-alpha genes.

The mouse has an interferon-alpha (MuIFN-alpha) gene family containing at least four, and likely more than ten members. A segment of mouse chromosomal DNA and cDNAs encoding murine alpha IFNs have been cloned, and the sequence of two MuIFN-alpha DNAs determined. No intron was found in the chromosomal gene. The two coding sequences produced biologically active IFN when expressed in monkey cells under the control of an SV40 promoter, and in E.coli under the control of the ampicillinase promoter. MuIFN-alpha 1 had no detectable activity on human cells, while MuIFN-alpha 2 was 20% as active on human as on mouse cells.

Animals↗
PubMed1982

Structure and expression of human IFN-alpha genes.

Copy DNA (cDNA) was prepared from induced leucocyte poly(A) RNA and cloned in Escherichia coli. IFN-alpha cDNA clones were isolated by subculture cloning with the use of a translation hybridization assay. Definitive identification of the clones was based on the production of an interferon-like protein by the transformed bacteria. Different IFN-alpha cDNAs, with characteristic target cell specificities, were identified. The cloned cDNAs typically encode a mature polypeptide of 166 (or, for IFN-alpha 2, 165) amino acids and a signal sequence of 23 amino acids. A human chromosomal library was screened with IFN cDNA and 17 distinct IFN-alpha-related sequences were isolated and identified, of which 7 proved to be nonallelic authentic genes and 4 pseudogenes; 6 sequences remain to be elucidated. Taking into account the work of Goeddel and his colleagues, 13 non-allelic authentic genes and 6 pseudogenes can be distinguished. In addition, 9 genes believed to be allelic to the 13 authentic genes have been sequenced. The IFN-alpha genes may be classified into two major subfamilies, which diverged at least 33 Ma ago, but perhaps much earlier, if sequence rectification occurred. At least one IFN-alpha gene appears to have resulted by a recombinational event between members of the subfamily I and II. IFN-beta is distantly related to IFN-alpha's and may have diverged from a common ancestor at least 500 Ma ago. Both IFN-alpha and IFN-beta genes differ from most other genes of higher organisms by being devoid of introns. The mouse was found to possess an IFN-alpha gene family of a size similar to that of man; the murine genes also do not have introns. IFN-alpha genes devoid of their signal sequence were joined to prokaryotic promoters to produce the mature interferons in E. coli in high yield. IFN-alpha 2, purified to homogeneity, has been crystallized by T. Unge and B. Strandberg (Uppsala). Hybrid genes consisting of IFN-alpha 1 and IFN-alpha 2 segments were constructed and expressed in E. coli; the target cell specificities of such hybrids were dependent on the arrangement of the segments and were different from those of either parent. The chromosomal gene for HuIFN-alpha 1 was introduced into mouse L cells to study the mechanism of its expression. Correct transcription was only detected after induction (with Newcastle disease virus); expression was transient, with the same kinetics as those of the endogenous mouse IFN mRNA. Natural murine IFNs and human IFN-beta and IFN-gamma are glycosylated. Because E. coli cells transformed with the genes of eukaryotic glycoproteins are not expected to yield correctly glycosylated polypeptides, we prepared lines of hamster cells permanently transformed with hybrid plasmids, which contained an IFN gene linked to the SV40 early promoter, as well as dihydrofolate reductase as a selective marker. After intracellular amplification of the introduced genes, cell lines were obtained which constitutively produced IFN at about 40 000 units ml-1 and could be propagated for at least several months.

Base Sequence↗
PubMed1980

Effect of interferon-alpha 1 from E. coli on some cell functions.

Interferon-alpha 1 from Escherichia coli transformed with a hybrid plasmid containing a human leukocyte complementary DNA insert, induces resistance to virus in appropriate target cells. It also shares the following properties with natural leukocyte interferon (IFN). (i) It enhances natural killing activity of human lymphocytes, (ii) it enhances antibody-dependent cell-mediated cytotoxicity, (iii) it suppresses antigen- and mitogen-induced leukocyte migration inhibition, (iv) it inhibits growth of IFN-sensitive Burkitt lymphoma cells. Since these activities are exhibited by a cloned protein species, they are due to IFN itself and not to other human proteins.

Antibody-Dependent Cell Cytotoxicity↗
PubMed1980

Synthesis in E. coli of a polypeptide with human leukocyte interferon activity.

Double-stranded cDNA prepared from the 12S fraction of poly(A) RNA from interferon (IF)-producing human leukocytes was cloned in Escherichia coli using the pBR322 vector. One of the resulting clones had a 910-base pair insert which could hybridise to IF mRNA and was responsible for the production of a polypeptide with biological IF activity. Up to 10,000 units IF activity per g of cells was obtained from some clones.

Base Sequence↗
PubMed1980

Mouse interferons: amino terminal amino acid sequences of various species.

Mouse interferons of three size classes (A, 35,000 to 40,000 daltons; B, 26,000 to 33,000 daltons; and C, 20,000 daltons) were purified from Ehrlich ascites tumor cells infected with Newcastle disease virus. The sequences of the first 24 amino acids (No. 17 has not been identified) of interferons A and B are identical. The sequence of the first 20 amino acids of interferon C differs from that of A and B in 18 positions. There is partial homology in amino terminal sequence between mouse interferons A (or B) and a human fibroblast interferon and between mouse interferon C and a human lymphoblastoid interferon.

Amino Acid Sequence↗
PubMed1980

Comparisons of several biological and physicochemical properties of human leukocyte interferons produced my human leukocytes and by E. coli.

Human interferon (IFN) prepared from virus-induced human leukocyte suspensions (leukocyte-derived interferon) was compared to the IFN extracted from Escherichia coli harboring a human interferon-alpha cDNA hybrid plasmid (Hif-SN35-AH-L6). E coli-derived IFN was 20 to 50 times more active than leukocyte-derived IFN on heterologous bovine, feline, murine and guinea pig cells, relative to the activity on human cells. After partial purification by affinity chromatography on an anti-human lymphoblastoid IFN antibody column, the IFN was analyzed by SDS-polyacrylamide gel electrophoresis. While leukocyte-derived IFN gave a heterogeneous pattern with major peaks of activity of 24000 and 19000 daltons, E. coli-derived IFN gave a heterogeneous peak of activity at about 17-18000 daltons. The leading edge of leukocyte-derived IFN in SDS-polyacrylamide gels was significantly more active on bovine cells than on human cells and coincided in mobility with E. coli-derived IFN, which was also much more active on bone than on human cells. After reduction with mercaptoethanol in SDS, the E. coli-derived IFN lost no activity, whereas the leukocyte-derived IFN lost about 90% of its activity. After reduction, E. coli-derived IFN migrated in SDS-polyacrylamide gels as a single peak at 24000 daltons, as did the residual activity of reduced leukocyte-derived interferon. Out data suggest that the interferon produced by the E. coli harboring the clone Hif-SN35-AH-L6 is analogous in size and cross-species activity to one of the molecular species of leukocyte-derived interferon.

Chromatography, Affinity↗
PubMed1980

Large scale production of mouse interferons from monolayers of Ehrlich ascites tumour cells.

Conditions are described for the large scale production of mouse interferons from Ehrlich ascites tumour cells cultured as monolayers in roller bottles. With the procedure reported here, we have used 50 to 65 600 cm(2) roller bottles to produce routinely 2 X 10(9) to 3 X 10(9) International units of crude mouse interferon/week with a specific activity of 1 X 10(6) to 1.5 X 10(6) units/mg protein.

Animals↗