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Mitogenic responses of the head-associated lymphoid tissues of the chicken

D M Maslak1, D L Reynolds

  • 1Department of Microbiology, Immunology, and Preventive Medicine, Iowa State University, Ames, Iowa 50011, USA.

Avian Diseases
|January 1, 1995
PubMed

Insights

This study adapted a microassay to measure chicken lymphocyte responses in head-associated lymphoid tissues. The harderian gland showed significant responses to T- and B-cell mitogens, indicating potential for immune function assessment.

Area of Science:

  • Immunology
  • Avian Biology
  • Cellular Immunology

Background:

  • Head-associated lymphoid tissues (HALT) play a crucial role in avian immunity.
  • Assessing immune responses in specific HALT, like the harderian gland, is vital for understanding local immunity.
  • Existing methods for measuring lymphocyte blastogenesis in avian HALT can be challenging.

Purpose of the Study:

  • To adapt and validate a 3-(4,5-dimethylthiazol-2-yl)-2,5 diphenyl tetrazolium bromide (MTT) blastogenesis microassay for chicken head-associated lymphoid tissues.
  • To evaluate the mitogenic responses of lymphocytes from the harderian gland and conjunctiva-associated lymphoid tissue (CALT).
  • To assess the utility of the MTT assay for measuring cell-mediated immunity in these tissues.

Main Methods:

  • Adaptation of an MTT-based blastogenesis microassay.
  • Isolation of lymphocytes from chicken peripheral blood, spleen, harderian gland, and CALT.
  • Stimulation of lymphocytes with T-cell (concanavalin A) and B-cell (Salmonella typhimurium lipopolysaccharide) mitogens.
  • Quantification of lymphocyte proliferation using MTT assay.

Main Results:

  • Lymphocytes from peripheral blood, spleen, and harderian gland exhibited significant blastogenic responses (P < 0.01) to both T- and B-cell mitogens compared to controls.
  • Harderian gland lymphocytes showed significant dose-dependent responses to concanavalin A and Salmonella typhimurium lipopolysaccharide.
  • The MTT assay was not suitable for CALT lymphocytes due to a low lymphocyte yield, limiting assay parameters.

Conclusions:

  • The MTT blastogenesis microassay is effective for measuring immune responses in the chicken harderian gland.
  • The harderian gland possesses functional lymphocytes responsive to T- and B-cell mitogens.
  • The assay's limitations in CALT highlight challenges in studying immune responses in tissues with low lymphocyte density.
  • This adapted MTT assay shows promise for evaluating cell-mediated immunity in avian head-associated lymphoid tissues.

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