A technique for inducing B-cell ablation in chickens ...

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Title A technique for inducing B-cell ablation in chickens by in ovo injection of cyclophosphamide
Author(s) D. L. Reynolds, A. D. Maraqa
Journal Avian Diseases
Date 1999
Volume 43
Issue 3
Start page 367
End page 375
Abstract Cyclophosphamide (CY) was injected in ovo on the 16th, 17th and 18th days of incubation. Blood samples were collected periodically from CY-treated and non-treated birds after hatch and were used to measure blood lymphocyte responses to the T-cell and B-cell mitogens, concanavalin A and lipopolysaccharide (LPS), respectively. Additionally, flow cytometric analysis was used to determine the presence of B and T cells in peripheral blood, and birds were vaccinated with Newcastle disease virus (NDV) antigen at 3 weeks of age and booster vaccinated at 5 weeks of age. CY treatment reduced hatchability by 35-40%, increased mortality by 3-5% within the first 2 weeks of life, and induced a significant retardation in body weight gains. At 2 weeks of age, approximately 50% of CY-treated birds were devoid of B-cell mitogenic responsiveness while demonstrating significant T-cell mitogenic responsiveness. However, B-cell responses were observed at 4 and 6 weeks from a small percentage of birds that were originally T-cell responsive and B-cell nonresponsive at 2 weeks of age. Flow cytometric analysis of peripheral blood lymphocytes revealed that CY-treated birds had significantly less B cells (or were devoid of B cells) than the corresponding non-treated control birds. However, no significant difference in the T-cell percentage was observed between CY-treated and nontreated birds. CY-treated birds did not produce detectable antibodies specific for NDV during the first and second weeks postvaccination, as demonstrated by haemagglutination inhibition assay. However, antibodies were detected in some CY-treated birds 10 days postbooster. Those antibody-positive birds were the same birds that had subsequently responded to the LPS mitogen on the blastogenesis microassay. It is concluded that it is important to monitor the B- and T-cell responses in CY-treated birds to identify those birds in which B-cell regeneration may have occurred.

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