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B S Kwon

Publications and source records attributed to B S Kwon.

At least 109 records · Page 6Linked to original sources
PubMed1989

cDNA sequences of two inducible T-cell genes.

We have previously described a set of human T-lymphocyte-specific cDNA clones isolated by a modified differential screening procedure. Apparent full-length cDNAs containing the sequences of 14 of the 16 initial isolates were sequenced and were found to represent five different species of mRNA; three of the five species were identical to previously reported cDNA sequences of preproenkephalin, T-cell-replacing factor, and a serine esterase, respectively. The other two species, 4-1BB and L2G25B, were inducible sequences found in mRNA from both a cytolytic T-lymphocyte and a helper T-lymphocyte clone and were not previously described in T-cell mRNA; these mRNA sequences encode peptides of 256 and 92 amino acids, respectively. Both peptides contain putative leader sequences. The protein encoded by 4-1BB also has a potential membrane anchor segment and other features also seen in known receptor proteins.

Adjuvants, Immunologic↗
PubMed1989

Isolation, chromosomal mapping, and expression of the mouse tyrosinase gene.

Using a human tyrosinase cDNA probe, we have isolated mouse tyrosinase genomic clones and used them to map the mouse tyrosinase locus and to analyze the promoter sequence of the tyrosinase gene. Southern blot analyses of DNA from somatic cell hybrids, interspecies backcross mice, and albino deletion mice have revealed that the locus for mouse tyrosinase resides at or near the albino locus on mouse chromosome 7. There were three TATA-elements, but only one CAT-element, and the CAT-element appeared to be paired with the third TATA-element, located at the position farthest upstream. Mouse tyrosinase mRNA is approximately 2.4 Kb in size. The amount of tyrosinase mRNA reflects the levels of tyrosinase activity in normal melanocytes and Cloudman S-91 melanoma cell line.

Albinism↗
PubMed1988

Isolation and sequence analysis of serine protease cDNAs from mouse cytolytic T lymphocytes.

Three new cDNA clones (designated MCSP-1, MCSP-2, and MCSP-3) encoding mouse serine proteases were isolated from cloned cytolytic T lymphocytes (CTL) by a modified differential screening procedure. The putative mature proteins of MCSP-2 and MCSP-3 are each composed of 228 amino acids with molecular weights of 25,477 and 25,360, respectively. NH2-terminal amino acids of MCSP-2- and MCSP-3-predicted proteins were identical to those reported for granzyme E and F, respectively. The third species, MCSP-1, was closely related to the two other cDNA species but approximately 30 amino acids equivalents of the NH2-terminal portion of the cDNA were not cloned. The amino acids forming the active sites of serine proteases were well conserved among the three predicted proteins. The active site pocket residue positioned six residues before the active-site Ser184 is alanine in MCSP-1, threonine in MCSP-2, and serine in MCSP-3, indicating that both MCSP-2 and MCSP-3 may have chymotrypsin-like specificity. There are three potential asparagine-linked glycosylation sites in MCSP-1 and MCSP-3, and four in MCSP-2-deduced amino acid sequences. Amino acid comparison of MCSP-1 with four other reported serine proteases whose active site pocket residue is alanine revealed that MCSP-1 was substantially different from the other molecules, indicating that MCSP-1 may be a new member of mouse T cell serine protease family. Antibodies made against a MCSP-1 lacZ gene fusion protein stain granules of CTL and react on immunoblots with two distinct granule protein bands of 29 and 35-40 kD. Only the 35-kD species labels with [3H]DFP. Since a protease cascade may play a key role in cytolytic lymphocyte activation, our isolation of cDNAs representative of unique serine esterases should help to investigate such a cascade process.

Amino Acid Sequence↗
PubMed1988

Sequence analysis of mouse tyrosinase cDNA and the effect of melanotropin on its gene expression.

Using human tyrosinase cDNA as a probe, a mouse tyrosinase cDNA clone representing approximately 75% of the tyrosinase coding region and a mouse genomic clone which includes the tyrosinase 5' coding sequences were isolated: nucleotide and deduced amino acid sequence of the mouse tyrosinase gene were determined from these clones. The predicted amino acid sequence revealed that the mouse tyrosinase is composed of 533 amino acids with a molecular weight of 60,536. The deduced protein contains 6 potential N-glycosylation sites, two cysteine- and two histidine-rich regions which may serve as copper-binding sites, a potential signal and transmembrane sequences. The mouse and human tyrosinase nucleotide and deduced amino acid sequences are approximately 81% homologous. The level of mouse tyrosinase mRNA was elevated after stimulation of Cloudman S-91 melanoma cells with melanotropin and isobutylmethylxanthine and the level of transcript reflected that of tyrosinase activity and melanin content in the cells.

Amino Acid Sequence↗
PubMed1988

Human tyrosinase gene, mapped to chromosome 11 (q14----q21), defines second region of homology with mouse chromosome 7.

The enzyme tyrosinase (monophenol,L-dopa:oxygen oxidoreductase; EC 1.14.18.1) catalyzes the first two steps in the conversion of tyrosine to melanin, the major pigment found in melanocytes. Some forms of oculocutaneous albinism, characterized by the absence of melanin in skin and eyes and by a deficiency of tyrosinase activity, may result from mutations in the tyrosinase structural gene. A recently isolated human tyrosinase cDNA was used to map the human tyrosinase locus (TYR) to chromosome 11, region q14----q21, by Southern blot analysis of somatic cell hybrid DNA and by in situ chromosomal hybridization. A second site of tyrosinase-related sequences was detected on the short arm of chromosome 11 near the centromere (p11.2----cen). Furthermore, we have confirmed the localization of the tyrosinase gene in the mouse at or near the c locus on chromosome 7. Comparison of the genetic maps of human chromosome 11 and mouse chromosome 7 leads to hypotheses regarding the evolution of human chromosome 11.

Animals↗
PubMed1988

Tyrosinases of murine melanocytes with mutations at the albino locus.

Tyrosinase (monophenol monooxygenase; monophenol, L-dopa:oxygen oxidoreductase, EC 1.14.18.1) is a key enzyme in the synthesis of melanin. Reduced levels of tyrosinase play an important role in albinism. The data described here show differences in the expression and characteristics of tyrosinase in cutaneous murine melanocytes grown in culture from normal wild-type strains (C/C); from three albino locus mutants: himalayan (ch/ch), chinchilla (cch/cch), and albino (c/c); and from the double-mutant heterozygous pink-eyed chinchilla (cchp/cp). Our results suggest that the diminished pigmentation in all mutants is due to abnormal posttranslational modification of the enzyme: the levels of mRNA for tyrosinase in wild-type, himalayan, and pink-eyed chinchilla melanocytes are similar; the himalayan mutation confers a deficiency in N-linked glycosylation, which results in an extremely unstable enzyme that is also temperature sensitive; the chinchilla and albino mutations confer susceptibility to proteolytic cleavage; the pink-eye dilution confers a reduction in the levels of immunoprecipitable tyrosinase, and what little enzyme there is fails to be translocated from the trans-Golgi network to melanosomes. The kinetics of activation and inhibition of the enzyme by the cofactor dopa are unique for the mutants tested and differ from those of tyrosinase from wild-type melanocytes. The findings support the conclusion that the albino locus in mice encodes the structural gene of tyrosinase.

Animals↗
PubMed1988

bFGF as an autocrine growth factor for human melanomas.

Normal human melanocytes in culture require specific additives such as basic fibroblast growth factor (bFGF) and dibutyryl cyclic adenosine monophosphate (dbcAMP) in order to proliferate in defined or serum-containing medium (Halaban et al., 1987). This stringent requirement is absent in cells derived from metastatic melanomas which not only proliferate in regular culture medium, but also produce a substance immunologically related to bFGF (Halaban et al., 1987). We show here that the mitogenic activity necessary for normal human melanocytes is constitutively present in several lines of human metastatic melanomas and that this activity is inactivated by anti-bFGF antibodies. Melanoma cells, but not normal melanocytes, express bFGF gene transcripts. Although the molecular mechanism underlying the abnormal expression of bFGF in melanomas is not known, the results suggest that bFGF acts as an autocrine growth factor in melanomas.

Cell Transformation, Neoplastic↗
PubMed1987

Effects of acyclovir therapy during simultaneous reactivation of latent HSV-1 in rabbits.

Acyclovir (ACV) therapy with a simultaneous reactivation of latent HSV-1 was evaluated in HSV-1 infected rabbit eyes. When the latently infected rabbits received epinephrine iontophoresis into corneas without ACV therapy 100% of eyes shed virus into tear film. The shedding was initiated on the second day of the epinephrine iontophoresis and lasted for an average of 4.6 days. When the rabbits received ACV (60 mg/kg body weight) intravenously once daily and topically (5% ACV ointment) twice daily for 6 consecutive days while a 0.01% epinephrine solution was iontophoresed into cornea for the first 3 consecutive days, 33% (2/6) of eyes showed detectable HSV-1 in the tear film only after terminating the ACV therapy, and the duration of shedding was for only one day. The average quantity of virus detectable in the tear film was decreased 14-fold in the latter group compared to the epinephrine iontophoresis group without ACV therapy. Four days after the last ACV therapy the titer of HSV-1 in the cell-free homogenates of the trigeminal and superior cervical ganglia was determined. For the epinephrine iontophoresed group, 67% of ganglia (8/12) were HSV-1 positive, while only 33% (4/12) of the ganglia from the combined treatment group were HSV-1 positive. The difference was statistically significant (p = 0.034). Furthermore, the titer of virus detectable in the cell-free homogenates of the virus-positive ganglia from the combined treatment group was less than that from the ganglia of the epinephrine iontophoresed group or untreated group. This suggests a reduction in the total number of latent foci for the combined treatment group.

Acyclovir↗
PubMed1987

Isolation and sequence of a cDNA clone for human tyrosinase that maps at the mouse c-albino locus.

Screening of a lambda gt11 human melanocyte cDNA library with antibodies against hamster tyrosinase (monophenol, L-dopa:oxygen oxidoreductase, EC 1.14.18.1) resulted in the isolation of 16 clones. The cDNA inserts from 13 of the 16 clones cross-hybridized with each other, indicating that they were from related mRNA species. One of the cDNA clones, Pmel34, detected one mRNA species with an approximate length of 2.4 kilobases that was expressed preferentially in normal and malignant melanocytes but not in other cell types. The amino acid sequence deduced from the nucleotide sequence showed that the putative human tyrosinase is composed of 548 amino acids with a molecular weight of 62,610. The deduced protein contains glycosylation sites and histidine-rich sites that could be used for copper binding. Southern blot analysis of DNA derived from newborn mice carrying lethal albino deletion mutations revealed that Pmel34 maps near or at the c-albino locus, the position of the structural gene for tyrosinase.

Albinism↗
PubMed1987

Isolation and initial characterization of multiple species of T-lymphocyte subset cDNA clones.

A modified differential screening procedure was applied to analyze cDNA libraries of cloned helper T lymphocytes (Th) and cytolytic T lymphocytes (CTL). Negative and positive differential screening and RNA blot analysis were used to identify cDNA clones that were expressed preferentially in Th or CTL. Seven clones corresponded to previously described T-cell genes, and 16 additional types of cDNA clones were isolated, 9 from Th and 7 from CTL. Of these, 3 were expressed in both Th and CTL, 7 were expressed in only Th, and 6 only in CTL. These clones were analyzed for induction after stimulation by interleukin 2 or Con A or after stimulation of the T-cell antigen receptor (TCR). Three different patterns of expression were seen: induction only by Con A, induction by Con A and interleukin 2, and induction by Con A and TCR stimulation. The approach is potentially useful for analyzing paths of T-cell differentiation and detecting cDNA clones encoding unrecognized cytokines.

Animals↗
PubMed1987

A melanocyte-specific complementary DNA clone whose expression is inducible by melanotropin and isobutylmethyl xanthine.

Two groups of cDNA clones were isolated by screening a lambda gt11 cDNA library of normal human melanocytes with antityrosinase antibodies: one group of 13 was related to the human tyrosinase gene. The properties of the other group of three cDNA clones was investigated by the use of a representative clone, Pmel 17-1. The cDNA hybridized to an mRNA species of approximately 2600 bases from human and murine melanocytes. The transcript of Pmel 17-1 (17-1 mRNA) was expressed preferentially in melanocytes and its abundance paralleled the melanin content. The expression of Pmel 17-1 mRNA increased after stimulation of human and murine melanoma cells with agents that increase the levels of melanization. Immunocompetition assays with monoclonal antibodies to gp75, a known pigmentation-associated antigen of melanocytes, suggested that Pmel 17-1 encodes a 75,000 Mr glycoprotein that is highly abundant in melanotic cells and shares some immunological homology with tyrosinase. The gene for Pmel 17-1 did not map at or near the c-albino locus in mice. The cDNA of Pmel 17-1 detected a single hybridizing restriction fragment in both human and murine DNA, indicating that the gene has been conserved between these two species and exists as a single gene in each.

1-Methyl-3-isobutylxanthine↗
PubMed1987

Timolol induces HSV-1 ocular shedding in the latently infected rabbit.

Timolol iontophoresis into the eye can induce herpes simplex virus type 1 (HSV-1) shedding in rabbits latently infected with HSV-1 strain McKrae. Anodal iontophoresis of 0.01% timolol was done at 0.8 mAmp for 8 min once a day for 3 consecutive days. Viral shedding was determined by the presence of HSV-1 in the preocular tear film obtained by eye swabs. In two experiments, iontophoresis of 0.01% timolol resulted in all eyes (18/18) shedding HSV-1 for an average duration of 4.3 days. When 5.0% timolol was applied topically to rabbit eyes supersensitized by iontophoresis of 6-hydroxydopamine (6-HD), all eyes (10/10) shed virus for an average duration of 2.9 days. All eyes (12/12) receiving iontophoresis of 6-HD, pre- and posttreatment with topical application of 5.0% timolol, and posttreatment with topical application of 1.0% epinephrine shed virus for an average duration of 3.6 days. Eyes treated with topical application of 5.0% timolol alone showed no difference in HSV-1 ocular shedding, compared with untreated eyes. We concluded that both iontophoresis of 0.01% timolol and topical application of 5.0% timolol to adrenergically supersensitized eyes induced HSV-1 shedding reliably and with a high frequency, and that topically applied timolol does not block the HSV-1 ocular shedding induced by epinephrine in adrenergically supersensitized eyes.

Administration, Topical↗
PubMed1985

Iontophoresis of epinephrine isomers to rabbit eyes induced HSV-1 ocular shedding.

Iontophoresis of 0.01% levo(-) epinephrine for 8 min at 0.8 mAmp once daily for 3 consecutive days induces ocular shedding of herpes simplex virus type 1 (HSV-1) in latently infected rabbits. In the present experiment, we tested dextro(+) and levo(-) epinephrine for their comparative effects on induced HSV-1 ocular shedding. One hundred percent of the eyes shed virus after either 0.01% dextro(+) or levo(-) epinephrine iontophoresis (8 min, 0.8 mAmp). However, the shedding frequency caused by 0.005% levo(-) epinephrine was significantly higher (P less than 0.05) than that by 0.005% dextro(+) epinephrine when the iontophoresis was conducted at 0.4 mAmp for 4 min. Iontophoresis of 0.001% levo(-) epinephrine for 8 min at 0.8 mAmp once daily for 3 consecutive days and iontophoresis of 0.001% dextro(+) epinephrine for 8 min at 0.8 mAmp once daily for 3 consecutive days did not induce HSV-1 ocular shedding in latently infected rabbits. The data suggest that the mechanism of induction of HSV-1 ocular shedding by epinephrine is correlated to the receptor potency of levo(-) epinephrine.

Animals↗
PubMed1984

Mouse mammary tumor virus-related sequences in mouse lymphocytes are inducible by 12-O-tetradecanoyl phorbol-13-acetate.

cDNA libraries from EL-4 cells treated with 12-O-tetradecanoyl phorbol-13-acetate (TPA) were screened for TPA-inducible sequences by differential hybridization. The most abundant inducible species was a sequence similar to that of mouse mammary tumor virus (MMTV). Induction of the mRNA corresponding to the MMTV-related sequences was already evident 30 min after TPA treatment, whereas the maximum accumulation occurred after 20 h of exposure to TPA. TPA also increased levels of MMTV-related RNA in the normal spleen cells of BALB/c and C57BL/6 mice. The level of RNA expression corresponding to MMTV-related sequences, however, was markedly elevated in EL-4 cells as compared with normal spleen cells. Southern blots of EL-4 cell DNA showed that the MMTV-related sequences were inserted into multiple locations of the EL-4 genome. Sequence analysis revealed that the MMTV-related cDNA clones included a part of the env gene and the right long terminal repeat of MMTV. However, the cDNA sequences were substantially different from published MMTV proviral sequences, most notably because of a contiguous deletion of 491 base pairs in the open reading frame within the U3 region.

Amino Acid Sequence↗
PubMed1984

Antibody-dependent cellular cytotoxicity mediated by polymorphonuclear leukocytes and mononuclear cells against HSV-1 infected primary cultures of rabbit corneal epithelium.

The roles of rabbit polymorphonuclear leukocytes (PMNL) and mononuclear cells (MC) for the regulation of ocular herpes simplex virus-1 (HSV-1) infection were studied. The antibody-dependent cellular cytotoxicity (ADCC) mediated by PMNL and MC from normal rabbit peripheral blood was assessed kinetically employing a specific 51Cr release assay. The HSV-1 infected primary cultures of rabbit corneal epithelium (PRCE) were used as the target cells to obtain a homologous assay system. The PRCE was prepared by an epithelium outgrowth technique and identified by electron microscopy. The expression of the surface HSV-1 antigens on PRCE was examined by indirect immunofluorescent staining; the cell population stained by fluorescein increased from 40% at 3 hr postinfection (PI) to 100% at 8 hr PI. To determine how early the cytotoxicity occurs, PRCE were infected with HSV-1 for 2 hrs. After 2 hrs, the ADCC was checked every 10 min for the first 40 min and then at 1, 2 and 4 hr of incubation. The cytotoxicity was apparent at 10 min postincubation and reached 46% by PMNL and 40% by MC at 4 hr postincubation (6 hr PI). Significant cytotoxic effect (26% by PMNL and 16% by MC) occurred as early as 3 hr PI. When the one-step growth cycle of HSV-1 was studied in the PRCE, HSV-1 had an eclipse period of 4 hr and a rise period of 8 hr. This suggests that rabbit PMNL and MC have the potential to eliminate the HSV-1 infected rabbit corneal epithelium before HSV matures in the cells.

Animals↗
PubMed1984

Thymine arabinoside (Ara-T) topical and iontophoretic application for herpes simplex virus type 1 and type 2 skin infections in hairless mice.

The antiviral agent thymine arabinoside (9-beta-D-arabinofuranosyl thymine, Ara-T) was applied topically and by anodal (+) iontophoresis in a NaCl solution to Herpes simplex virus type 1 (HSV-1), and Herpes simplex virus type 2 (HSV-2) skin infections in hairless mice. The chemotherapeutic efficacy of Ara-T was evaluated employing the parameters of mean lesion score, number of mice with signs of neurological involvement (paralysis), mortality and mean survival time. Anodal (+) iontophoresis of 0.4% Ara-T in a 0.1 M NaCl solution significantly suppressed HSV-1 skin infection, but the infection was not altered by topical application of 3% Ara-T. HSV-2 infection was refractory to both topical and iontophoretic application of Ara-T. This is the first report of the in vivo efficacy of Ara-T for HSV-1 skin infections.

Administration, Topical↗
PubMed1983

Herpes simplex virus recovery in neural tissues after ocular HSV shedding induced by epinephrine iontophoresis to the rabbit cornea.

Ocular HSV-1 shedding from latently infected rabbits was induced by iontophoresis of 0.01% epinephrine into the eye. Anodal Iontophoresis of epinephrine was performed at 0.8 mAmps for 8 min once a day for 3 consecutive days. Shedding was determined by the presence of HSV-1 in the preocular tear film obtained via eye swabs. Bilateral epinephrine iontophoresis performed on selected days during 220-280 days after inoculation resulted in HSV-1 shedding in 75% of the eyes (30/40) and 100% of the rabbits (20/20). Following the induction of ocular HSV-1 shedding, rabbits were killed and selected neural tissues were homogenized. Cell-free preparations were assayed for the presence of infectious virions using primary rabbit kidney cell monolayers. When the tissues were homogenized immediately after death, virus was detected in only one neural tissue, the trigeminal ganglia. However, when the tissues were incubated in vitro for 18-24 hours prior to the homogenization, infectious HSV-1 was recovered from homogenates of the trigeminal ganglion, superior cervical ganglion, the ophthalmic branch of the trigeminal nerve, and the root entry zone of the trigeminal nerve. A relationship was noted between the time of the last ocular shedding and recovery of infectious HSV from the tissue homogenates. Furthermore, a positive correlation in 11 eyes between the recovery of HSV-1 from the perocular tear film and HSV-1 recovery from one or more corresponding neural tissues was found. These results suggested that epinephrine iontophoresis to the cornea triggered an "alteration" in the state of the virus in the neural tissues of the latently infected rabbits and that the change can be related to the induced ocular shedding.

Animals↗