Analysis of selected variables in the under-agarose assay for chemotactic responses of canine neutrophils

Hajime Nagahata From the Departments of Veterinary Pathobiology (Nagahata, Kociba, Reiter) and Veterinary Clinical Sciences (Couto), College of Veterinary Medicine, The Ohio State University, 1925 Coffey Rd, Columbus, OH 42310.

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Gary J. Kociba From the Departments of Veterinary Pathobiology (Nagahata, Kociba, Reiter) and Veterinary Clinical Sciences (Couto), College of Veterinary Medicine, The Ohio State University, 1925 Coffey Rd, Columbus, OH 42310.

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Jolene A. Reiter From the Departments of Veterinary Pathobiology (Nagahata, Kociba, Reiter) and Veterinary Clinical Sciences (Couto), College of Veterinary Medicine, The Ohio State University, 1925 Coffey Rd, Columbus, OH 42310.

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C. Guillermo Couto From the Departments of Veterinary Pathobiology (Nagahata, Kociba, Reiter) and Veterinary Clinical Sciences (Couto), College of Veterinary Medicine, The Ohio State University, 1925 Coffey Rd, Columbus, OH 42310.

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SUMMARY

Chemotaxis under agarose was evaluated to establish an assay system and to characterize chemotactic responses of canine neutrophils. A method for the measurement of canine neutrophil chemotaxis was established, with optimal responses obtained with agarose containing 10% pooled canine serum, a concentration of 5 × 105 cells/well, zymosan-activated serum (zas), or autologous serum or plasma as the chemoattractants, and a 120-minute incubation period. Canine neutrophils responded well to zas, heat-inactivated zas, autologous serum and plasma, and heat-inactivated pooled serum. Chemotactic activity was proportional to the concentration of serum used as the chemoattractant. Mean (± sd) random migration, chemotaxis, chemotactic index, and chemotactic differential of neutrophils from 9 healthy Greyhounds were 1.09 (± 0.23), 1.95 (± 0.38), 1.82 (± 0.31), and 0.86 (± 0.32) mm, respectively.

SUMMARY

Chemotaxis under agarose was evaluated to establish an assay system and to characterize chemotactic responses of canine neutrophils. A method for the measurement of canine neutrophil chemotaxis was established, with optimal responses obtained with agarose containing 10% pooled canine serum, a concentration of 5 × 105 cells/well, zymosan-activated serum (zas), or autologous serum or plasma as the chemoattractants, and a 120-minute incubation period. Canine neutrophils responded well to zas, heat-inactivated zas, autologous serum and plasma, and heat-inactivated pooled serum. Chemotactic activity was proportional to the concentration of serum used as the chemoattractant. Mean (± sd) random migration, chemotaxis, chemotactic index, and chemotactic differential of neutrophils from 9 healthy Greyhounds were 1.09 (± 0.23), 1.95 (± 0.38), 1.82 (± 0.31), and 0.86 (± 0.32) mm, respectively.

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