• Animal cells and systems
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• v.8 no.2
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• pp.117-123
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• 2004

The purpose of this study was to analyze inhibitory effects of anti-4-1BB monoclonal antibody on melanoma metastasis The 4-1BB (CD137) T cell molecule is a member of the TNF receptor family and its activation by either 4-1BB ligand or antibody induces T cell activation and growth. In the present study, administration of anti-4-1BB mAb induced inhibition of melanoma metastasis. Agonistic anti-4-1BB mAb induced not only CD$8^+$4-1BBT cells but also CD$8^+$IFN-${\gamma}$$^{+}$ T cell population. In the presence of anti-CD3 antibody, lymphocytes produced high levels of IFN-${\gamma}$ and low levels of IL-4 in anti-4-1BB mAb treated group. Exposure of melanoma cells to IFN-${\gamma}$ induced expression of MHC-I molecules. Thus, the increase in number of CD$8^+$T cells and enhanced MHC-I expression on B16F10 cells by augmented IFN-${\gamma}$ production in response to anti-4-1BB mAb may result in suppression of tumor growth and metastasis.s.

• BMB Reports
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• v.47 no.3
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• pp.122-129
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• 2014

Although considerable progress has been made in understanding how tumors evade immune surveillance, measures to counter the same have not kept pace with the advances made in designing effective strategies. 4-1BB (CD137; TNFRS9), an activation-induced costimulatory molecule, is an important regulator of immune responses. Targeting 4-1BB or its natural ligand 4-1BB ligand (4-1BBL) has important implications in many clinical conditions, including cancer. In-depth analysis revealed that 4-1BB-mediated anti-cancer effects are based on its ability to induce activation of cytotoxic T lymphocytes (CTL), and among others, high amounts of IFN-${\gamma}$. In this review, we will discuss the various aspects of 4-1BB-mediated anti-tumor responses, the basis of such responses, and future directions.

• Journal of Periodontal and Implant Science
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• v.32 no.3
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• pp.457-473
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• 2002

The purposes of this study is to evaluate the combination effects of TGF-${\beta}_1$ and PDGF-BB on the periodontal ligament cells to use as a regeneration promoting agent of periodontal tissue. Human periodontal ligament cells were prepared from the first premolar tooth extracted for the orthodontic treatment and were cultured in DMEM/100% FBS at the $37^{\circ}C$, 5% $CO_2$ incubator. Authors measured the DNA synthesis, total protein, collagen and noncollagenous protein synthesis according to the concentration of TGF-${\beta}_1$,(1,5ng/ml) and PDGF-BB (1,10 ng/ml) in combination. To explore further this delayed effect of TGF-${\beta}_1$, we preincubated human periodontal ligament cells with TGF-${\beta}_1$ for 4 or 24 hours before PDGF-BB stimulation. The results were as follows: The DNA synthetic activity was increased dose dependently by TGF-${\beta}_1$, PDGF-BB. The combination of TGF-${\beta}_1$ and PDGF-BB consistently enhanced the DNA synthetic activity to PDGF-BB alone. The ability of TGF-${\beta}_1$ to enhance DNA synthetic activity in PDGF-BB treated periodontal ligament cells was dose dependent. The maximum mitogenic effect was at the 5ng/ml of TGF-${\beta}_1$ and l0ng/ml of PDGF-BB. Preincubation of cell with TGF-${\beta}_1$ resulted in significantly greater response to PDGF-BB at all TGF-${\beta}_1$ concentration studied, and may be useful for clinical application in periodontal regenerative procedures. The total protein, collagen and noncollagen synthesis was increased dose pendently by TGF-${\beta}_1$, PDGF-BB. The % of collagen was slightly decreased according to the concentration of TGF-${\beta}_1$, PDGF-BB. The effect of TGF-${\beta}_1$, PDGF-BB were not specific for collagen synthesis since it also increased noncollagenous protein synthesis. This study demonstrates that PDGF-BB is major mitogens for human periodontal ligament cells in vitro, and supports a role for TGF-${\beta}_1$ as a regulation of the mitogenic and total protein formation to PDGF-BB in these cells.

• Molecules and Cells
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• v.24 no.1
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• pp.132-138
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• 2007

4-1BB (CD137), a member of the tumor necrosis factor receptor superfamily, is expressed on activated T-cells, and 4-1BB signaling due to interaction with 4-1BB ligand or ligation with anti-4-1BB monoclonal antibody (mAb) costimulates T cells. It has been shown that administration of anti-4-1BB mAb induces anti-tumor immunity in mice, but the nature of the cellular subsets responsible for this immunity is uncertain. In this study we found that anti-4-1BB mAb administration to B16F10 melanoma-bearing mice induced marked expansion of $CD11c^+CD8^+$ T-cells in parallel with suppression of pulmonary tumors. The mAb-treated mice produced higher levels of $IFN-{\gamma}$ in their tumor tissues, spleen and lymph nodes than mice exposed to control antibody. When the $CD11c^+CD8^+$ T-cells were purified and re-stimulated in vitro, they produced high levels of the Th1 cytokines, $IFN-{\gamma}$ and IL-2, but low levels of the Th2 cytokines, IL-4 and IL-10. Furthermore, they expressed high levels of 4-1BB and CD107a, a marker of activated cytotoxic T-lymphocytes. Our results suggest that $CD11c^+CD8^+$ T-cells play a role in the anti-tumor immunity induced by anti-4-1BB mAb.

• Clinical and Experimental Pediatrics
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• v.54 no.9
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• pp.373-379
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• 2011

Purpose: 4-1BB (CD 137) is a costimulatory molecule expressed on activated T-cells. Repression by 4-1BB is thought to attenuate Th2-mediated allergic reactions. The aim of this study was to investigate the effect of 4-1BB on allergic airway inflammation in a murine asthma model. Methods: BALB/c mice were sensitized to and challenged with ovalbumin (OVA). Hu.4-1BB-Fc was administered 1 day before the first OVA sensitization or 1 day after the second OVA sensitization. Following antigen challenge, airway responsiveness to methacholine was assessed and bronchoalveolar lavage (BAL) fluid was analyzed. Total immunoglobulin (Ig) E, OVA-specific IgE, $IgG_1$, and $IgG_{2a}$ levels in sera were measured by enzyme-linked immunosorbent assay. Lung pathology was also evaluated. Results: In mice treated with Hu.4-1BB-Fc before the first OVA sensitization, there was a marked decrease in airway hyperresponsiveness, total cell count, and eosinophil count in the BAL fluid. In addition, Hu.4-1BB-Fc treatment decreased serum OVA-specific $IgG_1$ levels and increased serum $IgG_{2a}$ level significantly compared with the corresponding levels in mice sensitized to and challenged with OVA. Hu.4-1BB-Fc-treated mice also showed suppressed peribronchial and perivascular inflammatory cell infiltration. In contrast, treatment with Hu.4-1BB-Fc 1 day after sensitization had no effect on airway hyperresponsiveness and showed less suppression of inflammation in lung tissue. Conclusion: Administration of Hu.4-1BB-Fc can attenuate airway inflammation and hyperreactivity in a mouse model of allergic airway inflammation. In addition, administration before sensitization may be more effective. These findings suggest that 4-1BB may be a useful therapeutic molecule against asthma.

• Proceedings of the Korean Society of Applied Pharmacology
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• 1995.04a
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• pp.82-82
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• 1995

4-lBB molecule is expressed on the surface of activated CD4$\^$+/ and CD8$\^$+/ T cells. We generated a panel of anti-4-1 B5 murine mAbs using a fusion protein consisting of the extracellular domain of human 4-1 BB fused to Glutathione S-transferase. The binding activity against cell surface 4-1 BB molecule was assessed by flow cytometry analysis. These studies showed that several anti-4-1 BB mAbs bound to 10-30% of CD4$\^$+/ and CD8$\^$+/T cells in PHA or Con A stimulated PBLs, although these mAbs interacted with only, l-2% of CD4$\^$+/ and CD8$\^$+/ T cells in normal PBLs, indicating the specificity of mAbs to the 4-l BB molecule on activated CD4$\^$+/ and CD8$\^$+/ T cells. Next, we examined the effect of an anti-4-l BB mAb (4B4-1-1) on allogeneic mixed lymphocyte reactions (MLRs). The data indicated that the antibody significantly inhibited the proliferative response at higher concentrations. When tested with several T cell mitogens, the antibody had no stimulatory or inhibitory effects on the mitogen-mediated T cell proliferation. These data suggest that 4-1 BB molecule may play a role in the regulation of antigen-mediated immune response.

• IMMUNE NETWORK
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• v.2 no.3
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• pp.133-136
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• 2002

Background: The costimulatory molecule 4-1BB, a member of nerve growth factor receptor/tumor necrosis factor (NGFR/TNFR) super family, is involved in cell survival and death. Methods: In this study, female C57BL/6 ($H-2^b$) mice were used as a recipient, and DBA/2 ($H-2^d$) as a donor to assess a mixed lymphocyte reaction (MLR) and CTL response in vitro, and skin graft survival. IL-2, IFN level was measured by ELISA. Results: Mixed lymphocyte reaction (MLR) analysis showed that 4-1BB-deficient responder cells showed enhanced cellular proliferation over littermate controls. In contrast, IL-2 production was diminished only in 4-1BB knockout cultures. The IFN expression, on the other hand, was comparable between the groups. When female C57BL/6 ($H-2^b$) mice were grafted with the trunk skin of DBA/2 ($H-2^d$) mice, the in vivo tissue destruction of 4-1BB-deficient mice was not distinct from the normal littermates. Conclusion: These data suggest that 4-1BB is critical for the induction of alloreactive responses in vitro but 4-1BB alone could not change the course of skin rejection in vivo.

• Korean Journal of Veterinary Research
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• v.29 no.4
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• pp.457-460
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• 1989

This work was carried out to get some informations about blood types and their researches, involved blood stock and genetic identification. Horses examined were total 55 heads of sire, mare and their progeny in Korean Horse Affairs Association. 1. Albumin phenotypes of 26 mare were examined. The appearance of phenotype AA, BB, AB, was 1, 18, 7 respectively. The gene frequency of albumin A was 0.17 and albumin B was 0.76. 2. The appearance of phenotype AA, BB, AB in 29 progeny was 1, 16, 12 respectively. The gene frequency of albumin A was 0.24 and albumin B was 0.76. The gene frequency of gene A was higher than their parents. 3. Identification of the relationship between parents and their progeny was also examined. 4 of type AB between AA & BB, 4 of type BB between BB & BB, 13 of type AB between BB & AB were borned. In third case, all of progeny was type AB. This results suggest positive relationship between them.

• Journal of Veterinary Clinics
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• v.9 no.2
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• pp.433-449
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• 1992

Total CPK activities and CPK isoenzymes fractions of the sera and tissues were examined to obtain the physiological basic data of ruminant available in veterinary clinical practice. For the sera total CPK activities and CPK isoenzymes fractions, total 39 clinically healthy Korean native goats (3 to 10 months old, IS of female and 18 of male) and 6 of Korean native goats (1 to 2 years old, 3 of female and 3 of male) were used. Seventeen Korean native cattle (3 to 6 years old, 10 of female and 7 of male) and 27 Holstein-Friesian cattle (2 to 8 months old, 7 of female and 3 to 12 years old, 20 of female) were also examined for the sera total CPK activities and CPK isoenzymes fractions. For the total CPK activities and CPK isoenzyme fractions, 3 of female Korean native goats (7 months old), 3 of female Korean native cattle (2 years old) and 3 of dairy cattle (2 years old, 2 of female and 1 of male) were used. The tissues examined were the cerebrum (2 of Korean native cattle), spinal cord (1 of Korean native cattle), heart, lung, diaphragm, reticulum, liver, spleen, kidney, jejunum. colon and femoral muscle. The results obtained were as follows : 1. In Korean native goats less than 1-year-old. serum total CPK activities were 67.8${\pm}$17.7(39.0~96.5) IU/$\ell$ in female and 63.4${\pm}$19.0(28.7~94.4) IU/$\ell$ in male. Further they were 67.0${\pm}$5.3(59.5~70.7) IU/$\ell$ and 54.5${\pm}$11.1(39.1~69.4) IU/$\ell$ in female and male Korean native goats over 1-year-old, respectively. Serum total CPK activities of female were slightly higher than those of male. Significance between age and sex was not found. 2. Serum total CPK activities were 56.8${\pm}$19.7(27.6~90.5) IU/$\ell$ and 65.6${\pm}$10.8(52.8~78.0) IU/$\ell$ in female and male Korean native adult cattle, respectively, Serum total CPK activities of male were slightly higher than those of female, but they were not significant 3. Serum total CPK activities we,e 72.5${\pm}$8.2(57.2~83.2) IU/$\ell$ and 60.8${\pm}$12.5(42.7~80.6) IU/$\ell$ in calves and adult of dairy acttle, respectively. Serum total CPK activities of calves were significantly higher than those of adult(p<0.05). 4. In Korean native goats less than 1-year-old, serum CPK isoenzymes fractions were high with decreasing order of MM>MB>BB and MM>BB>MB in female and male, respectively. Further they were high with decreasing order of MM>MB>BB and MM>B8>MB in female and male Korean native goats over 1-year-old, respectively. The main fractions of CPK isoenzymes were MM in sera of Korean native goats. 5. Serum CPK Isoenzyme fractions were high with decreasing order of MM>MB>BB In both female and male of Korean native cattle. The main fraction among them was MM. 6. Serum CPK isoenzymes fractions were high with decreasing order of MM>BB>MB in both calves and adult of dairy cattle. The main fraction among them was MM. 7. Total CPK activities were high with decreasing order of the femoral muscle>kidney>reticulum>diaphragm>liver>spleen>heart>colon>lung>jejunum in Korean native goats. 8. Total CPK activities were high with decreasing order of the spinal cord >cerebrum>femoral muscle>reticulum>kidney>liver>spleen>diaphragm>lung>colon>heart>jejunum in Korean native cattle. 9. Total CPK activities were high with decreasing order. of the femoral muscle >liver>retoculum>kidney>heart>colon>lung>spleen>jejunum>diaphrasm in dairy cattle. 10. The pattern of the cardiac CPK isoenzymes fractions was identical in Korean native goats, Korean native cattle and dairy cattle. They were high in the order of MM>MB without BB fractions and the main fraction was MM. 11. The pattern of the pulmonary CPK isoenzymes fractions was the same Korean native goats, Korean native cattle and dairy cattle. They were high with decreasing order of MM>MB>BB and the main fraction among them was MM. 12. The pattern of CPK isoenzymes fractions of the diaphragm was Identical in Korean native goats and Korean native cattle. They were high with decreasing order of MM >BB >MB except dairy cattle (MM>MB>BB) but the main fraction among them was MM. 13. The pattern of the reticular CPK isoenzymes fractions was identical in Korean native cattle and dairy cattle. They were high with decreasing order of BB >MM >MB except Korean native goats(BB>MB>MM) but the main, fraction among them was BB 14. The pattern of the hepatic CPK isoenzymrs fractions was identical in Korean native cattle and dairy cattle. They were high with decreasing order of MB >BB >MM except Korean native goats(MB>MM>BB)but the main fraction was MB. 15. The splenic CPK isoenzymes fractions showed different pattern. They were high with decreasing order of MB>BB>MM, MM>BB>MB and BB>MB>MM in Korean native goats, Korean native cattle and dairy cattle, respectively. The main fraction among them was different from each other. 16. The pattern of the renal CPK isoenzymes fractions was identical in Korean native cattle and dairy cattle. They were high with decreasing order of MM >MB>BB except Korean native goats(BB>MB>MM). 17. The CPK isoenzymes fractions of the Jejunums showed different pattern. They were high with decreasing order MM>MB>BB, MM>BB>MB and BB>MM>MB in Korean native goats, Korean native cattle and dairy cattle, respectively. The main fractions were MM In Korean native goats and Korean native cattle, and BB in dairy cattle. 18. The colonic CPK isoenzymes fractions showed different pattern. They were high with decreasing order of MM>MB>BB, MM>BB>MB and BB>rrfB>MM in Korean native goats, Korean native cattle and dairy cattle, respectively. The main fractions were MM in Korean native goats and Korean . native cattle, and BB in dairy cattle. 19. The cerebral CPK isoenzymes fractions were high with decreasing order of BB >MM without MB detected in Korean native cattle and those of spinal cord were high with decreasing order of BB >MM >MB. The main fractions in both cerebrum and spinal cord were BB.

• Journal of Periodontal and Implant Science
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• v.26 no.4
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• pp.873-887
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• 1996

Platelet-derived growth factor(PDGF) is one of the polypeptide growth fators. PDGF has been reported as a biological mediator which regulates activities of wound healing process including the cell proliferation, migration and metabolism. Recent studies indicated that demineralized root surface as the primary site for growth factor application has advantages over other application method, especially due to binding capacity of growth factor for exposed matrix component of deminera1ized dentin surface. The purpose of this study is to evaluate optimal application time of PDGF-BB on proliferation of human gingival fibroblasts using deminera1ized dentin surface as primary application site. Human gingival fibroblasts and dentin slabs were prepared from the first premolar tooth extracted for the orthodontic treatment, cells were cultured in DMEM/I0% FBS at the $37^{\circ}C$, 5% CO2 incubator. All of the dentin slabs were preconditioned with Tetracycline HCI(100mg/ml) solution and rinsed in PBS. In the cell proliferation experiment, experimental group was immersed in DMEM containing 10% FBS, 50ng/rnl PDGF-BB during different time(30sec, 1, 2, 4, 8 minutes) and dried. Cells at concentration of $1{\times}10^5$cells/ml were seeded in each culture well which contained dentin slabs and incubated for 6 hours. Then, all of the dentin slabs were moved into new 24 well culture dish and incubated for 24, 48, 72 hours. The cell counting was done by hemocytometer with inverted phase contrast microscope after trypsinization. The results were as follows : The application of PDGF-BB for 1, 2 min slightly increased the number of gingival fibroblasts, and the application of PDGF-BB for 4, 8 min prominently increased the number of gingival fibroblasts. The application of PDGF-BB for 4 min showed maximum proliferation rate of gingival fibroblasts at 24, 48, 72 hours, and the application of PDGF-BB for 8 min showed less proliferation rate of gingival fibroblasts compared to the application of PDGF-BB for 4 min at 24, 48, 72 hours. In conclusion, the application of PDGF-BB for 4 min appeared to be optimal to obtain maximum proliferation of gingival fibroblasts using demineralized dentin surface as primary applicaton site of PDGF-BB.