The Expression and Biologic Effects of Ovine Interleukin-4 on T and B Cell Proliferation
| Autor: | Gerard Casey, Jean-Pierre Y. Scheerlinck, Robert De Rose, Paul R. Wood, Glen S. Buchan, Paul Chaplin |
|---|---|
| Rok vydání: | 2000 |
| Předmět: |
medicine.drug_class
T-Lymphocytes Molecular Sequence Data Immunology Biology Lymphocyte Activation Monoclonal antibody Recombinant virus Peripheral blood mononuclear cell law.invention Adjuvants Immunologic Western blot law Virology Complementary DNA medicine Animals Humans Amino Acid Sequence Cloning Molecular Interleukin 4 B-Lymphocytes Sheep medicine.diagnostic_test Lymphoblast Cell Biology Molecular biology Recombinant DNA Cattle Female Interleukin-4 Cell Division |
| Zdroj: | Journal of Interferon & Cytokine Research. 20:419-425 |
| ISSN: | 1557-7465 1079-9907 |
| DOI: | 10.1089/107999000312360 |
| Popis: | Using the reverse-transcriptase polymerase chain reaction (RT-PCR), cDNA encoding ovine (Ov) interleukin-4 (OvIL-4) was generated from mitogen-stimulated peripheral blood mononuclear cells (PBMC). Two identical clones generated from separate RT-PCR reactions differed from a published OvIL-4 sequence, although they had a high degree of identity with the bovine and human homologs. We show by sequence analysis that the OvIL-4 cDNA retained the four alpha-helix structure and disulfide bonds identified in human IL-4 (HuIL-4). Moreover, the cDNA encoding OvIL-4 was expressed in insect cells using the baculovirus Autographa californica nuclear polyhedrosis virus (AcMNPV) as a vector. Supernatants from insect cells infected with the recombinant virus secreted an additional protein with a relative molecular mass of 17,000. This protein was recognized by an anticervine IL-4 monoclonal antibody (mAb) in a Western blot and did not react with any proteins in supernatants from uninfected insect cells or cells infected with the wild-type AcMNPV. Supernatants from insect cells infected with the recombinant virus induced the proliferation of activated B cells in a dose-dependent manner and typically demonstrated 5 x 105 dilution U/ml of activity. However, OvIL-4 had no effect on the proliferation of resting T cells isolated from efferent lymph and actually inhibited the ability of a mitogen to stimulate these resting lymphocytes. In contrast, OvIL-4 induced the proliferation of mitogen-activated lymphoblast, demonstrating the complex role(s) OvIL-4 plays in the regulation of B and T cells. |
| Databáze: | OpenAIRE |
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