A technique for inducing B-cell ablation in chickens by in ovo injection of cyclophosphamide

Donald Reynolds, A. D. Maraqa

Research output: Contribution to journalReview article

11 Citations (Scopus)

Abstract

The effect of cyclophosphamide (CY) treatment in ovo on avian B and T cells was studied. CY was injected in ovo on the 16th, 17th, and 18th days of incubation. Blood samples were collected periodically from CY-treated and nontreated 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 wk of age and booster vaccinated at 5 wk of age. CY treatment reduced hatchability by 35%-40%, increased mortality by 3%-5% within the first 2 wk of life, and induced a significant retardation in body weight gains. At 2 wk 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 wk from a small percentage of birds that were originally T-cell responsive and B-cell nonresponsive at 2 wk 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 nontreated 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 hemagglutination inhibition assay. However, antibodies were detected in some CY-treated birds 10 days postbooster. Those antibody-positive birds were found to be the same birds that had subsequently responded to the LPS mitogen on the blastogenesis microassay. This study indicates the importance of monitoring the B- and T-cell responses in CY-treated birds to identify those birds in which B-cell regeneration may have occurred.

Original languageEnglish (US)
Pages (from-to)367-375
Number of pages9
JournalAvian diseases
Volume43
Issue number3
DOIs
StatePublished - Jan 1 1999

Fingerprint

cyclophosphamide
Cyclophosphamide
B-lymphocytes
Birds
Chickens
B-Lymphocytes
chickens
injection
Injections
birds
T-lymphocytes
T-Lymphocytes
methodology
Newcastle disease virus
blood
lipopolysaccharides
Mitogens
antibodies
Lipopolysaccharides
Antibodies

Keywords

  • Cyclophosphamide
  • Humoral immune response
  • Immunosuppressive
  • Newcastle disease virus

ASJC Scopus subject areas

  • Food Animals
  • Animal Science and Zoology
  • Immunology and Microbiology(all)

Cite this

A technique for inducing B-cell ablation in chickens by in ovo injection of cyclophosphamide. / Reynolds, Donald; Maraqa, A. D.

In: Avian diseases, Vol. 43, No. 3, 01.01.1999, p. 367-375.

Research output: Contribution to journalReview article

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abstract = "The effect of cyclophosphamide (CY) treatment in ovo on avian B and T cells was studied. CY was injected in ovo on the 16th, 17th, and 18th days of incubation. Blood samples were collected periodically from CY-treated and nontreated 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 wk of age and booster vaccinated at 5 wk of age. CY treatment reduced hatchability by 35{\%}-40{\%}, increased mortality by 3{\%}-5{\%} within the first 2 wk of life, and induced a significant retardation in body weight gains. At 2 wk 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 wk from a small percentage of birds that were originally T-cell responsive and B-cell nonresponsive at 2 wk 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 nontreated 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 hemagglutination inhibition assay. However, antibodies were detected in some CY-treated birds 10 days postbooster. Those antibody-positive birds were found to be the same birds that had subsequently responded to the LPS mitogen on the blastogenesis microassay. This study indicates the importance of monitoring 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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