1.The Effect of Recombinant Human Growth Hormone on Neutrophil Activity in Children.
Hun Taeg CHUNG ; Jong Duck KIM ; Hoon RYU ; Du Young CHOI
Korean Journal of Immunology 1997;19(4):629-638
We investigated the acute effect of recombinant human growth hormone (rhGH) on the activity of polymorphoneuclear leukocyte (PMN). We selected 6 patients of growth hormone deficient and 5 normal control children. In both groups, 0.15 IU/kg of rhGH was administered subcutaneously. The plasma growth hormone level were measured by radioimmunoassay on 0, 2, and 6 hours after administration of rhGH. To determined PMN activity, peripheral blood PMN were separated by discontinuous density-gradient centrifigation. Isolated PMN were stimulated hy fMLP and PMA and then respiratory burst activity of PMN was determined. The average growth hormone level of growth horrnone deficient and normal group were increased to the level of 41.6+/-23.7 and 96.3+/-46.5 ng/ml respectively, 2 hours after rhGH injection and decreased to the level of 18.5+/-10.6 and 42.2+/-5.5 ng/ml respectively, 6 hours after rhGH injection. Superoxide (O ) production by PMN which was stimulated by PMA was increased from 9.98+/-5.18 to 38.67+/-19.19 (x 10'cpm) after 6 hours of rhGH injection in control group children. It seemed that administration of the rhGH do not made a any effects acutely on PMN activity in growth hormone deficient group. But in a normal control children, extemal administration of rhGH acutely increased activity of PMN.
Child*
;
Growth Hormone
;
Human Growth Hormone*
;
Humans*
;
Leukocytes
;
Neutrophils*
;
Plasma
;
Radioimmunoassay
;
Respiratory Burst
;
Superoxides
2.Effects of Propolis and Caffeic Acid Phenethyl Ester on Tumorigenesis, Pulmonary Metastases, and Activities of Splenocytes and Macrophages in Mice.
Yo Han SONG ; Hong Yeol HUH ; Chin Soo KIM ; Kang Ju KIM
Korean Journal of Immunology 1997;19(4):617-628
The propolis, honey bee hive product, is a folk medicine for treating various ailrnents and caffeic acid phenethyl ester (CAPE) is an extract of propolis. The purpose of this study was to examine the effects of ethanol extracted propolis (EEP) or CAPE on the tumorigenesis, pulmonary metastases, and proliferation and activity of splenocytes and macrophages in ICR mice. EEP at 0.2, 2 or 20mg/ml applied topically on the back of each mice 30 minutes before application of 7,12-dimethylbenz (a)anthracene and 12-0-tetradecanoylphorbol-13-acetate inhibited the number of tumors per mouse by 61, 75 or 100%, respectively. ...continue...
Animals
;
Bees
;
Carcinogenesis*
;
Ethanol
;
Honey
;
Macrophages*
;
Medicine, Traditional
;
Mice*
;
Mice, Inbred ICR
;
Neoplasm Metastasis*
;
Propolis*
3.Induction of Apoptosis in Synovial Cells from Patients with Rheumatoid Arthritis.
Sung Yeoul CHANG ; Seong Yoon KIM ; Myung Hee KIM ; Young Gyu CHAI ; Jae Chan PARK ; Yong Keel CHOI ; Jae Bum JUN ; Sung Soo JUNG
Korean Journal of Immunology 1997;19(4):609-616
Homeostasis of multicellular organism is controlled by proliferation and differentiation of cells as well as by cell death. The defects in programmed cell death contribute to the pathogenesis of rheumatoid arthritis (RA) and systemic lupus erythematous. RA is considered to be a proliferating disorder of synovial tissue which is accompanied by inflammatory cell infiltration and bone erosion. The aim of the study was to find whether potent inducers of apoptosis could be induced apoptosis in RA synovial cells. We examined the effects of drugs, such as dexamethasone, methotrexate, hydrogen peroxide, and ceramide on induction of apoptosis in cultured RA synovial cells. Used drugs did not induced apoptosis in RA synovial cells. Finally Fas antigen-mediated apoptosis of RA synovial cells was investigated by the addition of anti-Fas antibody. To examine the ICE (interleukin-1p-convertase; caspase-1) expression in synovial cells, RT-PCR of caspase-1 gene was performed. In synovial cells of RA, Fas induces that caspase-1 activation cause apoptosis.
Apoptosis*
;
Arthritis, Rheumatoid*
;
Cell Death
;
Dexamethasone
;
Homeostasis
;
Humans
;
Hydrogen Peroxide
;
Ice
;
Methotrexate
4.A Study for Germline Mutation of BRCA1 in Early Onset Breast Cancer Patients.
Hyo Joon KIM ; Ji Yeon LEE ; Eun Sook LEE ; Bum Hwan KOO
Korean Journal of Immunology 1997;19(4):601-608
The cumulative incidence of breast cancer in Korea is low, being about one-fifth of that in the United States. This low incidence has been mainly explained by environmental factors, and recently, however, racial variations in the disease-causing genes should also be considered. The BRCA1 is one of the common genes involved in early-onset breast cancer and/or ovarian cancer in the United States and Northern Europe. However, the involvement of BRCA1 in Korean'breast cancer patients are still unclear. We performed germline mutation screening of the BRCA1 gene by DNA single strand conformation polymorphism (SSCP) analysis. We examined 27 breast cancer patients who were diagnosed less than 35 years by age including two cases with family history of breast cancer. Our study showed no germline mutation at the exons 2, 11 and 20, which were known as the supreme susceptible regions of BRCA1 mutations. Even though our cases did not fulfilled the criteria of familial breast/ovarian cancer, the proprotion of families who inherit the mutated BRCA1 allele seems to be very small and might be negligible among Korean population. Therefore, it is considered that the BRCA1 itself cannot be a major susceptibility gene and the contributions of other genes might be important for the breast cancer.
Alleles
;
Breast Neoplasms*
;
Breast*
;
DNA
;
Europe
;
Exons
;
Genes, BRCA1
;
Germ-Line Mutation*
;
Humans
;
Incidence
;
Korea
;
Mass Screening
;
Ovarian Neoplasms
;
United States
5.Effect of Nitric Oxide on the Viability of Bone Marrow - Derived Cultured Mast Cells.
Hun Taeg CHUNG ; Rae Kil PARK ; Chang Duk JUN ; Byung Min CHOI ; Seog Jae LEE
Korean Journal of Immunology 1997;19(4):595-600
It is well established that mast cell proliferation and maturation are regulated by two principle cytokines, IL-3 and the c-kit ligand stem cell factor (SCF). Previous reports have demonstrated that bone marrow-derived IL-3-dependent mast cells exhibit the characteristic apoptosis on removal of IL-3. To know how the number of mast cells is controlled, we observed the effects of nitric oxide (NO) on the murine bone marrow-derived cultured mast cells (BMCMC). Apoptosis was measured by the analysis of flow cytometric data and electrophoretic evidence of DNA fragmentation. Our data showed that sodiurn nitroprusside (SNP)-a NO releasing substance- induced apoptosis in BMCMC. Cell cycle analysis showed that the number of the G,/G, and S phase decreased markedly, while the percentage of cell in G,/M phase was increased. Also, SNP alone induced cell death, whereas SNP in combination with SCF markedly decreased cell death of BMCMC. SNP-induced apoptosis was partially inhibited by the treatment of BMCMC with SCF. Our results suggest that NO might have sorne role in the regulation of the number of mast cells.
Apoptosis
;
Bone Marrow*
;
Cell Cycle
;
Cell Death
;
Cytokines
;
DNA Fragmentation
;
Interleukin-3
;
Mast Cells*
;
Nitric Oxide*
;
Nitroprusside
;
S Phase
;
Stem Cell Factor
6.Effects of Alcohol Administration on Cytokines, Nitric Oxide, Infection Anaphylaxis and Melanoma Growth.
Tai You HA ; Hyun Ju HA ; Young Hyun KIM
Korean Journal of Immunology 1997;19(4):583-594
The present study was undertaken to investigate the effect of ethanol administration on the resistance of mice to Cryptococcus neoformans, IL-2 production of murine splenocytes, active systemic anaphylaxis induced by ovalbumin (OVA), serum TNF-alpha production, nitric oxide (NO) production by peritoneal machrophages and B16F10 melanoma colonization in lungs in mice. It was found that ethanol administration significantly inhibited the resistance of mice to C. neoformans infection, IL-2 production, active systemic anaphylaxis induction, serum TNF- alpha production and NO production. Ethanol administration significantly enhanced lung colonization when it was administered before i.v. melanoma inoculation. These results demonstrate that ethanol may play a critical role in tumorigenesis and immunoregulation as an immunomodulator.
Anaphylaxis*
;
Animals
;
Carcinogenesis
;
Colon
;
Cryptococcus neoformans
;
Cytokines*
;
Ethanol
;
Interleukin-2
;
Lung
;
Melanoma*
;
Mice
;
Nitric Oxide*
;
Ovalbumin
;
Tumor Necrosis Factor-alpha
7.Immunological Activities of Korean Mistletoe Extract ( Viscum album coloratum ; KM - 110 ).
Jong Bae KIM ; Taek Joon YOON ; Yung Choon YOO ; Tae Bong KANG ; Myoung Sool DO ; Ichiro AZUMA
Korean Journal of Immunology 1997;19(4):571-582
Immunological effect of the extract (KM-110) from Korean mistletoe (Viscum album coloratum) was examined. Lymphocytes obtained from KM-110-administrated mice showed increased responsiveness to mitogens, concanavalin-A (Con.A) and lipopolysaccharide (LPS). In order to study cytokine induct ability of the KM-110, macrophages from the Balb/c mice were cultivated in the medium containing the extract. the macrophages were shown to induce secretion of tumor necrosis factor-alpha (TNF-alpha), interleukin 1 and 6 (IL-1, IL-6) and interferon-gamma (INF-gamma). We then tested antitumor activity of the macrophages activated by the KM-110. Peritoneal macrophages harvested from the KM110-treated Balb/c mice exhibited cytotoxicity against the syngeneic 3LL carcinoma cells. ...continue...
Animals
;
Interferon-gamma
;
Interleukin-1
;
Lymphocytes
;
Macrophages
;
Macrophages, Peritoneal
;
Mice
;
Mistletoe*
;
Mitogens
;
Tumor Necrosis Factor-alpha
;
Viscum album*
;
Viscum*
8.Immunomodulators Extracted from Korean - style Fermented Soybean Paste and Their Function . 1 . Isolation of B Cell Mitogen from Korean - style Fermented Soybean Paste.
Bong Ki LEE ; Yun Soo JANG ; Sook Yi YI ; Kun Sub CHUNG ; Shin Yang CHOI
Korean Journal of Immunology 1997;19(4):559-570
Responses of mouse lymphocytes to the soybean paste fermented by Korean traditional fashion was examined to clarify its effects in cytokine production in vitro. A fraction of the soybean paste (KFSP-100) was prepared by precipitation with ammonium sulfate and by filtration through ultrafiltration membrane. KFSP-100 were added into cultures of fresh mouse splenic cells in vitro. KFSP-100 significantly enhanced the amount of IL-6 and TNF-a produced by macrophages and IL-6 and IFN-r produced by lymphocytes. Production of IL-12 by macrophages was not much affected by KFSP-100 treatments. The most noticeable finding was the fact that lymphocytes treated with KFSP-100 proliferated to an exceeding numbers (more than 10 times to the control) in 72 hours. The KFSP-100-induced proliferative response was specific to B cells since almost all of the KFSP-100-induced cells in the cultures of splenic cells were B cells. Furthermore, such a proliferative responses were equally observed only in cultures of purified B cells but not in cultures of T cells. In thermostability test, the biologically active components of the KFSP-100 is assumed to be either linear protein or glycoprotein. KFSP-100 did not induce agglutination of lymphocytes demonstrated by lectins in the same cells. These observations suggest that KFSP-100 may be a novel mitogen for B lymphocytes. The component (s) responsible for the B cell proliferation in KFSP-100 might be a factor gained by natural fermentation. None of the fractions of not fermented soybean paste prepared by the same methods demonstrate the same effect.
Agglutination
;
Ammonium Sulfate
;
Animals
;
B-Lymphocytes
;
Cell Proliferation
;
Fermentation
;
Filtration
;
Glycoproteins
;
Immunologic Factors*
;
Interleukin-12
;
Interleukin-6
;
Lectins
;
Lymphocytes
;
Macrophages
;
Membranes
;
Mice
;
Soybeans*
;
T-Lymphocytes
;
Ultrafiltration
9.Production of Human Fab Monoclonal Antibody to Surface Protein, preS1, of Hepatitis B Virus using Antibody Phage Display Library.
Hee Sun KIM ; In Hak CHOI ; Ik Jung KIM ; Jun Ho CHUNG ; Se Kwang PARK
Korean Journal of Immunology 1997;19(4):549-558
Human monoclonal antibodies have considerable potential in the prophylaxis and treatment of viral disease. By cloning human Ig gene segments from the B cells of volunteer into pComb3 phagemid vector, antibody library was created of filamentous phage particles displaying Fab fragments on their surface after being rescued with M13KO7 helper phages. The size of library was 7x10' pfu. Phage antibodies (phabs) were panned against biotinylated preS1 using streptavidine coated Dynabead. The soluble Fab antibodies were prepared from phagemid colonies and assayed directly for the ability to bind preS1 by ELISA. And then 3DW and SGW specific to preS1 which have both heavy and light chain to form Fab fragment, were selected. The soluble Fab antibody from 3DW was expressed highly at the concentration of 0.1 - 1.0 mM of IPTG, and 5 hours postinduction. The soluble antibodies from 3DW and SGW showed their relative affinities of 2x10' M ', and Sx10 M ', respectively, and the specificities to preS1 on ELISA. Our results suggest that antibody phage display library is very useful method to generate the human monoclonal antibody and that the human Fab monoclonal antibodies specific to preS1 selected in this study open the way to treat hepatitis B as a component of passive irnmunotherapeutics.
Antibodies
;
Antibodies, Monoclonal
;
B-Lymphocytes
;
Bacteriophages*
;
Clone Cells
;
Cloning, Organism
;
Enzyme-Linked Immunosorbent Assay
;
Genes, Immunoglobulin
;
Hepatitis B virus*
;
Hepatitis B*
;
Hepatitis*
;
Humans*
;
Immunoglobulin Fab Fragments
;
Isopropyl Thiogalactoside
;
Streptavidin
;
Virus Diseases
;
Volunteers
10.Identification of the CDR3 Gene Sequence on the beta Chain of the T Cell Receptor in T Cell Leukemia Cell Line.
In Hong CHOI ; Se Jong KIM ; Joo Deuk KIM ; Yong Joon CHWAE
Korean Journal of Immunology 1997;19(4):541-548
In order to develop a method for the detection of minimal residual leukemic disease (MRD) in T cell acute lymphocytic leukemia (T cell ALL), T cell leukemia cell line was used to detect clonal TCR p chain mRNA and to synthesize CDR3 specific oligonucleotide probe. For the Jurkat cell line clonal TCR p chain cDNA was amplified by using RT-PCR with oligonucleotide primer, Vp universal primer. As the result of RT-PCR an approximate 300 bp fragment of the TCR chain was obtained, and the partial identification of the TCR p chain gene and the amino acid sequence of the fragment were done by gene cloning and sequencing. The gene sequence of TCR p obtained was identified as Vp8-Jp1.2-Cp2. Diversity gene segment could not be found. Within the p chain, the CDR3 region was identified as 12 amino acids (SFSTCSANYGYT). It is kown that TCR is expressed in about 40% of the all T cell ALL. However it is not kown what percentage of the TCR p chain mRNA expression translates into the actual TCR molecule. It is not certain how many patients with MRD can be detected by this method used in this study, but this technique might be useful to detect MRD in at least 40% of the patients with T-cell ALL.
Amino Acid Sequence
;
Amino Acids
;
Cell Line*
;
Clone Cells
;
Cloning, Organism
;
DNA, Complementary
;
Humans
;
Jurkat Cells
;
Leukemia, T-Cell*
;
Precursor Cell Lymphoblastic Leukemia-Lymphoma
;
Receptors, Antigen, T-Cell*
;
RNA, Messenger
;
T-Lymphocytes