No abstract available.

No abstract available.
Luminescence* ; Neutrophils*

To investigate effects of cytokines on rheumatoid synovial cells, proliferation and expression of cytokine and metalloproteinase genes were studied with the primary culture of rheumatoid synovial cells which was treated with TNF-alpha, GM-CSF, TGF-alpha, PDGF and IL-B. By [3H] thymidine incorporation assay, TGF-beta and PDGF increased proliferation of synovial cells by 1.5 and 2.5 folds respectively. Cytokine gene expression was assessed by RT-PCR. Rheumatoid synovial cells expressed constitutively TGF-beta and IL-B at a high level and IL-1B, GM-CSF, and MIP-1a at a relatively low level. TGF-beta, GM-CSF and PDGF increased IL-B expression. Expression of pro-inflammatory cytokines and chemokines was increased by GM-CSF and PDGF. Both GM-CSF and PDGF increased the expression of IL-1B, GM-CSF MIP-la and IL-8. In addition, GM-CSF enhanced expression of TNF-alpha. Stromelysin and collagenase are the major proteinases responsible for destruction ot joints in rheumatoid arthritis (RA). These genes were expressed constitutivefy in rheumatoid synovial cells. In summary, PDGF and GM-CSF may piay an important role by inducing or increasing expression of IL-1B, TGF-beta and PDGF by increasing proliferation of rheumatoid synovial cells.
Tumor Necrosis Factor-alpha

It has been thought that autoimmune diseases like systemic lupus erythematosus and rhumatoid athritis are closely associated with anti-DNA antibodies (Abs). In studies of the control for anti-DNA Ab generation, an understanding of the regulatory mechanisms by anti-idiotypic Abs that influence the production of anti- DNA Abs would be facillitated by the availability of the hybridomas producing the pairs of DNA-specific and anti-DNA's idiotope-specific monoclonal antibodies (mAbs). We have produced a series of anti-DNA mAbs and then monoclonal anti-idiotypic Ab directed against idiotypic determinant of the 3D8 mAb that has the highest affinity to dsDNA and ssDNA among the anti-DNA mABs that we had obtained. The spleen cells of the MRL-Ipr/Ipr, autoimmune prone, mice were fused with P3X63Ag8.653 myeioma cells to obtain anti-DNA Ab secreting hybridomas. Out of the fourteen clones that showed strong binding to ssDNA, four clones had cross-reactivity with dsDNA whereas none of these clones reacted with left-handed z-DNA. The binding activities of the anti-DNA mAbs to various synthetic polynucleotide sequences were different respectively. Anti-idiotypic mAbs were generated by the fusion of myeloma cells and spleen cells from the Balb/c mice immunized with 3DB-Fab. We have produced two anti-idiotypic mAbs, B7 (IgG2a/k) and 02F3 (IgM/k), which were specific to 3DB-Fab and cloned the variable region of the heavy chain from the 02F3 hybridoma.
Mice ; Animals

Transforming growth factor-B1 (TGF-B1) is well known to be one of the most potent Immunosuppressive cytokines. To determine whether TGF-B1 secreted in the latent form can be immunoregulatory, TGF-B1 cDNA driven by the human -actin promotor was transfected into a murine thymoma cell line, EL4 cells. The transfectants (ELJ4) secreted a latent torm of TGF-B1 at a concentration of 5 ng/ml under the influence of TPA. Transfected TGF-b1 transcripts was readily detected by RT-PCR in ELJ4 cells regardless of the presence of TPA, but not in EL4 cells. In addition, we found the degree of Thy-B1 expression, IL-2 secretion and the proliferation rate are not altered by the transfection. Finally, EL4 and ELJ4 cells were injected into C57BU6 mice (syngenic strain), subcutaneously. Tumor cell masses derived from both cell populations survived longer than 1 wk, and the size of tumor derived from ELJ4 was three times larger (2.5 cm of diameter) than that from EL4. Virtually, there was no histopathological difference between two tumors. Taken together, the results from the present study indicates that EL4 thymomas transfected with TGF-1 secretes a latent form of TGF-B1 which may suppress host immune defence system.
Humans ; Mice ; Animals

Human caspase-2, Ich-1 (Ice and Ced-3 homolog), has two different forms of mRNA species derived from alternative splicing, which encodes Ich-1 and Ich-1s. Ich-1v which induces apoptosis is antagonist of Ich-1s which suppresses Rat-1 cell death by serum deprivation. To investigate functions of Ich-1 and Ich-1s in T celi apoptosis, the fusion DNA constructs were made with the ecto and transmembrane of CDB and Ich-lv or Ich-1s and CDS-Ich-1 or CD8-Ich-1s chimeric protein was transiently expressed on Jurkat T cells. Tyrosine phosphorylation of intracellular proteins was induced in these transfectans when activated shortly by anti-CDB Ab. CDB-Ich-li transfectant in serum-rich condition and CDB-Ich-ls transfectant in serum-deprived condition underwent apoptosis when treated with anti-CDS Ab or incubated with NIH3T3 cells expressing stably Fas-L on their surface. We also made six antisense DNA constructs which could specifically inhibit the expression of Ich-1v, Ich- 1s, and then they were transiently transfected into Jurkat T cell. The overexpression of both of the antisese- Ich-1 against N-terminal 42 bp and against C-terminal 366 bp inhibited apoptosis through Fas signalling. But, when three different forms of antisense-Ich-1s were overexpressed in their transfectants, antisense-DNA against N-terminal 197 bp increased knd the one against C-terminal 66 bp inhibited apoptosis, instead the full size of antisense-DNA did not give any effects on apoptosis through Fas pathway.
Humans

T cell activation is a critical event for initiation and regulation of immune responses and inhibitors of such signaling pathways are clinically useful for the treatment of patients received allogratt and autoimmune disease. In the course of screening soil microorganisms from the forest of Cheju island in Korea for new immunosuppressive agent, one of Streptomyces species (CK-95441) was found to produce a new immunosuppressant, tautomycetin which also had antifungal activity. Tautomycetin showed the inhibition of T cell proliferation in murine mixed lymphocyte reaction (MLR) and T cell activation induced by concanavalin A. Tautomycetin also blocked the induction of IL-2 gene expression which was examined in Jurkat TAg cell line in which multiple NFAT-binding sites and minimal IL-2 promoter drive the production of B-galactosidase. Also, the level of inhibition in activation-induced IL-2 receptor expression by tautomycetin was greater than those by cyclosporin A measured by flow cytometry. But, Fas ligand-induced apoptosis in Jurkat cells was unaffected by tautomycetin which was measured by DNA fragmentation assay. These results suggested that tautomycetin will be able to be used as a potent immunosuppressive drug following organ transplantation.

"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

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

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