Related Experiment Videos
Isolation of the turkey heterophil and measurement of its migratory functions under agarose
Abstract:
A procedure for isolation of the turkey heterophil was developed. The procedure involves preliminary lysis of erythrocytes, using ammonium chloride and centrifugation of the leukocytes over a discontinuous gradient of 18% and 24% Ficoll. A minimum of 30% of the heterophils was consistently recovered from whole blood by application of this procedure. Spontaneous and chemotactic migratory responses of the isolated heterophils were assessed, using the migration under agarose technique. Supplementation of the agarose with 10% heat-inactivated turkey plasma and extension of the incubation period to 3 hours at 41 C in a 5% CO2-humidified atmosphere were required for optimal chemotaxis in response to zymosan-activated chicken serum. The heterophils did not respond chemotactically to the oligopeptide attractant, N-formyl-methionyl-leucyl-phenylalanine, and did not specifically bind a radiolabeled form of this ligand.
Insights
A new method effectively isolates turkey heterophils, crucial white blood cells. These isolated cells show optimal chemotaxis with specific conditions, aiding avian immunology research.
Area of Science:
- Avian immunology
- Cellular immunology
- Hematology
Background:
- Turkey heterophils are vital immune cells, but their isolation and functional assessment present challenges.
- Understanding turkey heterophil function is crucial for avian disease research and vaccine development.
Purpose of the Study:
- To develop a reliable procedure for isolating functional turkey heterophils from whole blood.
- To characterize the migratory responses of isolated turkey heterophils.
Main Methods:
- Isolation involved erythrocyte lysis with ammonium chloride and density gradient centrifugation over Ficoll.
- Cell migration was assessed using the migration under agarose technique.
- Chemotaxis was optimized by supplementing agarose with turkey plasma and adjusting incubation conditions.
Main Results:
- The developed procedure consistently recovered at least 30% of heterophils from whole blood.
- Optimal chemotaxis required 10% heat-inactivated turkey plasma and a 3-hour incubation at 41°C with 5% CO2.
- Isolated turkey heterophils did not exhibit chemotaxis or specific binding to N-formyl-methionyl-leucyl-phenylalanine.
Conclusions:
- A robust method for turkey heterophil isolation was established, enabling further functional studies.
- Turkey heterophils demonstrate chemotaxis towards zymosan-activated chicken serum under specific laboratory conditions.
- The lack of response to N-formyl-methionyl-leucyl-phenylalanine suggests species-specific or cell-specific migratory cues.