1.Antibody Response of Mouse Splenocytes Using Mixture of Supernatants of Thymocytes, Adherent and Non-adherent Splenocytes: In-vitro Immunization-II.
Dongsoo KIM ; Geun Woong NOH ; Duk Hi KIM ; Oh Hun KWON
Journal of Korean Medical Science 1990;5(1):25-31
We adapted one of the in-vitro immunization methods to induce antibody responses and confirmed the success of the immunization by enzyme-linked immunosorbent assay (ELISA) without hybridization. We have previously reported several methods of in-vitro immunization using different conditioned media. Here we introduce another method of in-vitro immunization using a mixture of three types of supernatant (thymocytes, and adherent and non-adherent splenocytes of mouse). Splenocytes were immunized in-vitro by a recombinant human growth hormone (rhGH) with the above conditioned media, and the results by ELISA showed a much higher optical density than the other in-vitro immunization methods that we had previously reported. Humoral responses as a result of in-vitro immunization to soluble antigens were easily confirmed by ELISA using the above-conditioned media. This finding indicates that any other conditions thought to be critical by other researches may not be essential for in-vitro immunization.
Animals
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*Antibody Formation
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Antigens/immunology
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Cell Adhesion
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Female
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Growth Substances/*immunology
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Immunization
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Lymphocyte Activation/immunology
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Mice
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Mice, Inbred BALB C
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Recombinant Proteins/immunology
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Spleen/cytology/*immunology
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T-Lymphocytes/*immunology
2.Effect of growth factors on the expression of proto-oncogenes c-fos and c-myc in FRTL-5 cell line.
Hwan Young YOON ; Seung Keun OH ; Ka Hee YI ; Bo Youn CHO ; Chang Soon KOH
Journal of Korean Medical Science 1995;10(3):155-163
This study was performed to prove the hypothesis that oncogene expressions would have the same patterns with those of cellular growth to growth factors in FRTL-5 cells. Ribonucleic acids of FRTL-5 were extracted at 15', 30', 60' and 120' after administration of growth factors to quiescent FRTL-5, and blotted to the nitrocellulose membrane. They were hybridized with radiolabelled c-fos, c-myc and beta-actin probes. Hybridized dot blots were autoradiographed and the amount of radioactivity was measured by densitometry. Densitometric readings were used as the indices of oncogene expressions. Expressions of c-fos and c-myc were more prominent in combined administrations of TSH (10 mU/ml) and IGF-I (100 ng/ml) or IgG of Graves' disease (Graves' IgG; 1 mg/ml) and IGF-I than in combined administration of TSH and Graves' IgG. IgG of primary myxedema suppressed oncogene expressions by TSH or Graves' IgG, but not by IGF-I. From the above results, it was suggested that expressions of c-fos and c-myc to growth factors would have similar patterns with those of cell growth to growth factors in FRTL-5, and the actions of TSH and Graves' IgG would be manifested through same signal transduction system, but IGF-I would be manifested by its own.
Animal
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Cell Division/drug effects/genetics
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Cell Line/cytology/physiology
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Gene Expression/drug effects/immunology
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Graves' Disease/immunology
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Growth Substances/genetics/*pharmacology
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Immunoglobulin G/pharmacology
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Insulin-Like Growth Factor I/pharmacology
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Myxedema/immunology
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Proto-Oncogene Proteins c-fos/*genetics
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Proto-Oncogene Proteins c-myc/*genetics
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RNA/analysis
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Rats
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Rats, Inbred F344
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Support, Non-U.S. Gov't
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Thyroid Gland/cytology
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Thyrotropin/pharmacology
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Time Factors