Humoral immune response to Salmonella abortusovis in sheep: in vitro induction of an antibody synthesis from either sensitized or unprimed lymph node cells
| Autor: | Patricia Berthon, Michel Olivier, I. Gohin, Isabelle Lantier |
|---|---|
| Přispěvatelé: | Unité de Pathologie Infectieuse et Immunologie [Nouzilly] (PII), Institut National de la Recherche Agronomique (INRA), ProdInra, Migration |
| Jazyk: | angličtina |
| Rok vydání: | 1994 |
| Předmět: |
[SDV]Life Sciences [q-bio]
Immunology PBS phosphate-buffered saline Enzyme-Linked Immunosorbent Assay Lymphocyte Activation IMMUNOLOGIE Article NLS normal lamb serum Microbiology 03 medical and health sciences 0302 clinical medicine Immune system Antigen Salmonella medicine Animals Antibody-Producing Cells Lymph node Cells Cultured ComputingMilieux_MISCELLANEOUS 030304 developmental biology 0303 health sciences Salmonella Infections Animal Sheep biology SFC spot-forming cells General Veterinary Ig immunoglobulins Primary and secondary antibodies Antibodies Bacterial 3. Good health [SDV] Life Sciences [q-bio] medicine.anatomical_structure Lymphatic system Immunoglobulin M APC antigen-presenting cell HS horse serum Cell culture Immunoglobulin G biology.protein LPS lipopolysaccharide Bacterial antigen Lymph Nodes FCS fetal calf serum Antibody 030215 immunology |
| Zdroj: | Veterinary Immunology and Immunopathology Veterinary Immunology and Immunopathology, Elsevier, 1994, 41, pp.275-294 |
| ISSN: | 0165-2427 |
| Popis: | In vitro culture conditions were determined to induce an anti-Salmonella abortusovis antibody synthesis from lymph node leucocytes of three immunized sheep and two unprimed lambs maintained in culture in the presence of heat-inactivated bacteria for 2 weeks. Humoral immune responses were assessed by enumerating specific antibody-secreting cells using ELISASPOT and by titrating immunoglobulins secreted into culture supernatants using ELISA techniques. Optimal secondary antibody response was observed from cultures performed with fetal calf serum (compared with horse serum) and with an antigen concentration of one to ten bacteria per cell. This kind of antigenic stimulation allowed induction of numerous antibody-secreting cells without adsorption of the secreted antibodies. Maximal numbers of antibody-secreting cells could reach a rate of 1% of the sheep leucocytes initially put into culture. Kinetic profiles of antibody production from boosted lymph node cells were characterized by an ascending phase from the sixth to the twelfth day of culture and then showed a plateau phase until Day 14. Most of the responses were composed of IgM and IgG1 antibodies, traces of IgG2 being detected at the end of experiments. From the twelfth day of antigenic stimulation, the IgM isotype was preferentially expressed with high antigen concentration (100 bacteria per cell), whereas the highest amounts of IgG1 were detected at lower concentration (one to ten bacteria per cell). While anti-Salmonella IgM appeared to be mainly specific for the lipopolysaccharide (LPS) cell wall fraction, some IgG1 recognized other bacterial antigens. Kinetic profiles and magnitudes of primary antibody responses induced in vitro from lamb lymph node cells did not differ from those defined in cultures of sheep boosted leucocytes. But these immune reactions were mainly made up of anti-LPS IgM. Few anti-Salmonella IgG1 were detected from the tenth day of culture. So these in vitro assays allowed induction of an antibody synthesis from either in vivo sensitized or unprimed sheep lymph node leucocytes. This methodology would permit achievement of more detailed studies on interactions between Salmonella and lymph node leucocytes, leading to a better understanding of the mechanisms controlling bacterial dissemination through the lymphoid tissue. |
| Databáze: | OpenAIRE |
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