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Effects of Capsaicin on Production of Cytokines and Nitric Oxide, Salmonella Infection and NF-kappa B Activation.

Tai You HA ; Suhn Young IM ; Jae Seung PARK ; Hyun Ju HA ; Du Young CHOI

Korean Journal of Immunology.1998;20(3):349-363.

"Capsaicin, the pungent principle of hot peppers, is a neurotoxin that depletes primary sensory neurons of neuropeptides like tachykinin. The objectives of these experiment was to examine the effects of capsaicin on Salmonel/a typhimurium-induced production of cytokines such as TNF-a, IL-1B, IL-6, IL-10 and IL-12 and on production of nitric oxide in peritoneal macrophages. In addition, the effects of capsaicin on survival rates of S. typhimurium-infected mice and on nuclear transcription factor (NF-kB) activation were also investigated. Mice were pretreated with a single s.c. injection of 100 ug of capsaicin and were infected i.v. with S. typhimurium (5xO5/mouse) in 0.2 ml volume after capsaicin pretreatment. The serum cytokine levels were measured 30, 60, 120, 180 and 240 min after Salmonella infection, using ELISA kits. The activation of NF-B was also examined by gel shift assay in spleens, thymuses and brains of mice that had been pretreated with a single s.c. injection of 100 ug of capsaicin. It was found that Sa/mone/la infection induced the production of TNF-a, IL-1B, IL-6, IL-10 and IL-12, but capsaicin pretreatment inhibited the production of TNF-a, IL-1B, IL-10 and IL-12, but enhanced IL-6 production 120 min after Salmonella infection. Interestingly, the capsaicin pretreatment inhibited the activation of NF-kB in spleens and thymuses. There were no differences in the numbers of bacteria in livers, brains, spleens, kidneys and lungs between capsaicin- pretreated mice and the control animals in applied experimental conditions. Suprisingly, however, capsaicin pretreatment increased both the survival rates of Sa/mone//a-infected mice and production of nitric oxide by peritoneal macrophages compared with capsaicin-untreated control mice. Taken together, these results indicate that the capsaicin-sensitive primary sensory neurons may play an important modulatory role in the production of cytokine, nitric oxide and NF-B activation and the pathogenesis of salmonellosis."
Animals ; Bacteria ; Brain ; Capsaicin* ; Cytokines* ; Enzyme-Linked Immunosorbent Assay ; Interleukin-10 ; Interleukin-12 ; Interleukin-6 ; Kidney ; Liver ; Lung ; Macrophages, Peritoneal ; Mice ; Neuropeptides ; NF-kappa B* ; Nitric Oxide* ; Salmonella Infections* ; Salmonella typhimurium ; Salmonella* ; Sensory Receptor Cells ; Spleen ; Survival Rate ; Tachykinins ; Thymus Gland ; Transcription Factors

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Effect of Scutellaria baicalensis Extract on the Immune Functions, Microbial Growth and Mutagenicity.

Jeong Ho LEE ; Sook Jeong SHIN ; Yong MOON

Korean Journal of Immunology.1998;20(3):343-348.

Scute/larisa baica/ensis (SB) has been used as a folk medicine for curing ulcer, inflammation and infection. However, surprisingly little has been done to develop and exploit SB's immunomodulating, anti- inflammatory properties. Moreover, the mechanisms of SB-action on immune function had not been elucidated. The present study was undertaken to investigate the effect of 5B on immune functions, microbial growth and bacterial mutagenicity. Boiling water extract of SB was used in this experiment. The proliferation response to PHA-, Con A- or LPS-stimulation and the production of Con A-induced IL-2 and LPS-induced IL-6 of splenocytes from SB-pretreated mice were significantly higher than those of splenocytes from control mice. Daily single injection of 2 mg/mouse SB for 4 days resulted in enhancement of Arthus reaction and DTH to SRBC. And these enhancements were more prominent when SB was treated prior to SRBC- sensitization. SB did inhibit the growth of microorganisms such as C. albicans, C. neoformans, E. coli S. typhimurium and this inhibiting effect was gradually increased in proportion to the increment of SB. SB also remarkably reduced the mutagenicity of mutagens such as sodium azid and benzo[a]pyrene, but this was not certain, because it showed a killing effect on the cell survival test. When SB treated mice were i.p infected with C. albicans, the number of microorganisms in the peritoneal exudates were significantly reduced. Taken together, these results revealed that SB itself has not only multiple effects on events controlling immune responses but also anti-inflammatory properties, which may provide the rational basis for their therapeutic use as one of the biological response modifiers.
Animals ; Arthus Reaction ; Cell Survival ; Exudates and Transudates ; Homicide ; Immunologic Factors ; Inflammation ; Interleukin-2 ; Interleukin-6 ; Medicine, Traditional ; Mice ; Mutagens ; Scutellaria baicalensis* ; Scutellaria* ; Sodium ; Ulcer ; Water

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Generation and Characterization of Alloenic Radiation Bone Marrow Chimera.

Ki Young LEE ; Kil Hyoun KIM

Korean Journal of Immunology.1998;20(3):333-341.

Allogenic bone marrow chimera has been used to study the differentiation of donor-derived bone marrow cells under the recipient thymic environment, tolerance generaion between recipient and donor cells. We prepared H-2' to H-2 allogenic chimera by transfering bone marrow cells from H-2dmice to r-irradiated H-2k mice, and examined the differentiation ofthe bone marrow cells under allogenic environment. Complete reconstitution with H-2d+ phenotype cells in the thymus of the chimera mice was observed. However, the number of CD4- CD8+ cells dramatically decreased under the recipient thymic environment, CD8+ cells significantly reduced also in spleen and lymph node, compared with that of normal mice. Interestingly, we also observed coexistence of donor-derived cells (H-2k) and recipient derived cells (H-2d) in lymph node and spleen in the chimera. These results suggested that the decrease of CD4-CD8+ cells could be caused by r-irradiation by affecting the recipient thymic environment, and that in this chimera, tolerance between donor-derived cells and recipient-derived cells was maintained.
Animals ; Bone Marrow Cells ; Bone Marrow* ; Chimera* ; Humans ; Lymph Nodes ; Mice ; Phenotype ; Spleen ; Thymus Gland ; Tissue Donors

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Polymorphism of Antigen Processing ( TAP, HLA-DM, LMP ) Genes in Korean Population.

Tai Gyu KIM ; Hoon HAN ; Hee Baeg CHOI

Korean Journal of Immunology.1998;20(3):325-331.

Antigen processing (TAP, HLA-DM and LMP) genes map within the major histocompatibility complex (MHC) class II region between the HLA-DQB1 and -DPB1 loci, and are involved in the processing of peptides bound to HLA class I or class II molecules. In order to determine the allele frequencies of antigen processing genes and the various linkage disequilibria existing among these genes, we have analyzed TAP1, TAP2, HLA-DMA, and HLA-DMB, LMP2, LMP7 polymorphisms in 184 unrelated healthy Koreans using the rnethod of PCR-SSCP, ARMS-PCR and PCR-RFLP. The frequencies of antigen processing genes were TAP1A (77.7%), TAP1*B (17.1%), TAP1*C (5.2%), TAP2*A (41.6%), TAP2*B (31.3%), TAP2*C (3.3%), TAP2*D (0.8%), TAP2*E (6.5%), TAP2*G (0.8%), HLA-DMA*0101 (81.5%), HLA-DMA*0102 (18.2%), HLA-DMA*0103 (0.3%), HLA-DMB*0101 (42.9%), HLA-DMB*0102 (19.0%), HLA-DMB*0103 (38.0%), LMP2*R (78.8%), LMP2*H (21.2%), LMP7*A (35.3%), LMP7*B (56.0%), LMP7*C (4.9%), and LMP7*D (3.8%). We also analysed two- locus association among each locus. Many significant positive associations were observed between these two loci, such as between HLA-DMB and TAP1, between HLA-DMA and HLA-DMB, between LMP2 and LMP7, and between TAP1 and LMP7. Conversely, any significant linkage disequilibrium was not detected between HLA-DMB and LMP2. These results could be used as control data for disease association and population genetics studies in Korean population.
Antigen Presentation* ; Gene Frequency ; Genetics, Population ; Linkage Disequilibrium ; Major Histocompatibility Complex ; Peptides

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Molecular Cloning of Human 14-3-3 cDNA with CD3 Zeta Peptide Containing Phosphosering Tag.

Jae Seung KANG

Korean Journal of Immunology.1998;20(3):317-323.

14-3-3 proteins are highly conserved proteins of about 29 kDa and have a minimum of seven isoforms. 14- 3-3 proteins interact with many signalling proteins by the recognition of phosphoserine. For the identification of proteins which react with ITAM (immunoreceptor tyrosine-based activation motif) of CD3 zeta chain, labeled synthetic peptides representing the CD3 zeta chain structual motifs (ITAMs) with a tag of PKC substrate sequence were used for western blotting. One major protein band of approximately 29 kDa was identified in lysate of Jurkat T cell, B cells and HeLa cells. Screening of lamda gt 11 library derived from HeLa cell gave two clones of 14-3-3 protein cDNA. Inspection of their nucleotide sequences identified these two full length cDNA clones as the 29 kDa human homologue of rat 14-3-3 gamma and the human 14-3-3 zeta protein. The human 14-3-3 gamma isoform also showed high homology with other species in amino acid and nucleotide sequence. Although 14-3-3 proteins are phosphoserine-binding proteins, there may be another way of interaction between ITAMs of CD3 and 14-3-3 proteins.
14-3-3 Proteins ; Animals ; B-Lymphocytes ; Base Sequence ; Blotting, Western ; Clone Cells ; Cloning, Molecular* ; DNA, Complementary* ; HeLa Cells ; Humans* ; Mass Screening ; Peptides ; Phosphoserine ; Protein Isoforms ; Rats

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Length diversity in CDR3 Domain of Immunoglobulin Kappa Chain during the Human Deelopment.

Ji Soo LEE ; Soo Kon LEE ; Chan Hee LEE ; Chang Ho SONG

Korean Journal of Immunology.1998;20(3):309-316.

The third complementarity determining region (CDR3) of the immunoglobulin (Ig) kappa () chain is known to be located at the center of antigen binding groove and critical for antibody specificity. Ig chain has been characterized by limited junctional diversity due to the absence of N-region addition resulting in relative conservation of CDR3 lengths with 9 or 10 amino acids. CDR3 region of 11 amino acids is only possible with N-region addition. Recently, x transcripts with 11 amino acids CDR3 was found to be expressed in normal individuals, and in autoimrnune disease such as rheumatoid arthritis, the fraction of 11 amino acids CDR3 of humkv325-derived chains was overexpressed compared to conventional adult peripheral B cells. However, the significance of this bias is difficult to interpret without a clear understanding of normal repertoire of CDR3 length during development. The purpose of this study is to determine whether developmental regulation of CDR3 amino acids codon lengths exists in chains expressed in the fetal liver, cord blood, and adult peripheral blood lymphocytes (PBL). Lymphocytes were seperated from fetal liver, cord blood and adult PBL and cDNA was generated from extracted mRNA. PCR-based CDR3 finger- printing assay was performed with VI-IV family specific primers. CDR3 length diversity of Ig x chain increases as the development proceeds. The length diversity most frequently occured in Vlll family derived transcripts including 11 amino acids CDR3. transcripts with 11 amino acids CDR3 were consitently expressed in both fetal and adult Ig repertoire. These results support the hypothesis that v chain CDR3 length is developmentally regulated and implicates the diversity of antigen-antibody specificity generation.
Adult ; Amino Acids ; Antibody Specificity ; Arthritis, Rheumatoid ; B-Lymphocytes ; Bias (Epidemiology) ; Codon ; Complementarity Determining Regions ; DNA, Complementary ; Fetal Blood ; Humans* ; Immunoglobulin kappa-Chains* ; Immunoglobulins* ; Liver ; Lymphocytes ; RNA, Messenger ; Sensitivity and Specificity

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Production and Characterization of Murine Monoclonal Antibodies ( MAbs ) which Specifically Recognize B-Subunit of Human Chorionic Gonadotropin ( HCG ).

Heui Keun KANG ; Jin Dong CHANG ; Hyung Jun LEE ; Jung Hak CHA ; Moo Young SONG ; In Young KO

Korean Journal of Immunology.1998;20(3):303-308.

We have constructed several panels of MAbs which specifically recognize B-subunit of HCG (BHCG). Splenocytes from Balb/c mice immunized with B-subunit of HCG were fused with SP2/o-Ag14 myeloma cells by PEG method. Fifteen different hybridorna clones (individually named as mG10.127, mG10.61, mG9.5, mG9.18, rnG9.20, mG6.3, mG6.36, mG6.8, mG7.31, mG7.79, mG9.11, mG9.51.6, mG9.51.12, mH4.17, and mH4.4) were obtained by indirect ELISA screening and three to five successive cloning procedures. The distinct features of these MAbs were determined by specificity, western blot, isotyping, and isoelectrofocusing. All of the MAbs except mG9.20 and mG6.8 specifically bind to BHCG without cross- reaction with B-subunit of LH (BLH). In western blot analysis, all of the MAbs bind to non-denatured form of BHCG suggesting that the MAbs recognize conformation-dependent epitope of BHCG. This new panels of MAbs to BHCG should be useful for developing diagnostic reagent such as pregnancy, choriocarcinoma, Down's syndrome as well as for the fine quantitation of serum or urinary HCG.
Animals ; Antibodies, Monoclonal* ; Blotting, Western ; Choriocarcinoma ; Chorionic Gonadotropin* ; Clone Cells ; Cloning, Organism ; Down Syndrome ; Enzyme-Linked Immunosorbent Assay ; Female ; Humans* ; Mass Screening ; Mice ; Pregnancy ; Sensitivity and Specificity

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Stimulatory Effect of IL-10 on Antitumor Cytolytic Activity of Murine Spleen Cells.

Hong Gu JOO

Korean Journal of Immunology.1998;20(3):295-301.

L-10 has been recognized as an irnmune suppressive cytokine which inhibits Ag-specific activation and proliferation of T cells. It also inhibits Ag presenting capacity of monocyte/macrophage and down-regulates monokine production. However it has also shown that IL-10 has stimulatory effect on immune effector cells in recent studies. This report shows that IL-10 has direct stimulatory effect on antitumor cytolytic activity suppressed by TGF-B. To assess the effect of IL-10 on cytolytic activity against tumor, spleen cells prepared from tumor-bearing mice were cultured with mitomycin C-treated MOPC-315 cells in the presence of IL-10. Unexpectedly, IL-10 was able to reverse the cytolytic activity suppressed with TGF-B. The stimulatory effect of IL-10 was dependent on the addition time of IL-10. At day 0, 4, those effects were shown higher than those of the other days. Also, the stimulatory effect of IL-10 showed specificity against MOPC-315 tumor cells. To elucidate the role of endogenous IL-10, TGF-B in MLTC cultures, anti-IL-10 and anti-TGF-B mAb were used. The inhibition of IL-10 release in MLTC cultures by using anti-IL-10 mAb resulted in the suppression of cytolytic activity against MOPC-315 tumor cells. Taken together, although IL- 10 has been recognized as a strong immunosuppressive cytokine derived of tumor cells, IL-10 showed the direct stimulatory effect on the antitumor cytolytic activity of spleen cells.
Animals ; Interleukin-10* ; Mice ; Mitomycin ; Sensitivity and Specificity ; Spleen* ; T-Lymphocytes ; Transforming Growth Factor beta

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Effective Production and Clinical Application of Anti-Interleukin-6 Monoclonal Antibodies.

Kyung Soo NAM ; In Pyo CHOI ; Cheorl Ho KIM ; Mi Jung CHOI ; Jung Hwa YANG ; Jeon Ok MOON

Korean Journal of Immunology.1998;20(3):289-294.

Highly specific and sensitive immunoassay method for soluble human recombinant interleukin-6 (hu rlL-6) was established by two different immunization methods. One is conventional method by Freund's adjuvant method and the other is special method which is directly injected to mouse spleen. Among seven established monoclonal antibodies (mAbs), two typical monoclonal antibodies, designated YB3 (IgG1) and NY2 (IgM), were further characterized. These mAbs highly bound to IL-6, however did not show cross reactivity with IL-1B and IL-2. As the results of ELISA inhibition assay and western blotting method, it was further identified that YB3 and NY2 had high binding specificity with IL-6. And the limiting detection amount of rlL-6 for YB3 was 5 ng/ml and for NY2 was 0.5 ng/ml. Furthermore, N-glycosylated human rlL-6 was also bound to YB3 on ELISA. On the other hand YB-3 furtherly recognized N-glycosylated human rlL-6 by sandwich ELISA method. These mAbs may be of use to diagnose the gynecopathy which contains abortion and preterm labor.
Animals ; Antibodies, Monoclonal* ; Blotting, Western ; Enzyme-Linked Immunosorbent Assay ; Female ; Freund's Adjuvant ; Hand ; Humans ; Immunization ; Immunoassay ; Interleukin-2 ; Interleukin-6 ; Mice ; Obstetric Labor, Premature ; Pregnancy ; Sensitivity and Specificity ; Spleen

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Molecular Cloning of Mouse Interleukin-2 Receptor bata-Chain Gene Promoter.

Chang Kwon KO ; Sang Kyu YE ; Bon Hong MIN ; Kwang Ho LEE

Korean Journal of Immunology.1998;20(3):285-288.

No abstract available.
Animals ; Cloning, Molecular* ; Interleukin-2* ; Mice*

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