IDENTIFICATION OF CANINE T LYMPHOCYTES WITH ANTIDOG LYMPHOCYTE SERUM AND ANTIRAT LYMPHOCYTE SERUM BY INDIRECT IMMUNOFLUORESCENT STAINING

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Abstract
Reliable methods for the identification of T lymphocytes in the dog have not been available. We determined the proportion of T cells in canine lymphoid tissues by an indirect immuno-fluorescent assay utilizing rabbit anti-rat thymocyte (RART) serum. We found that RART, gave values similar to those of rabbit anti-dog lymphocyte (RADL) serum. RADL, however, required absorption before it was T cell-specific while absorption was unnecessary with RART. Both heterologous antisera stained greater than 90% of the dog thymocytes, 60% of the lymph node cells, 40% of the spleen and peripheral blood cells, and only 5% to 10% of the bone marrow cells. The specificity of RART was further demonstrated by its high reactivity with enriched canine T cell subpopulations obtained from nylon-wool columns, preparative EAC rosette formation, and preparative cell electrophoresis. In contrast, lymphocyte populations enriched for B cells in the same experiments exhibited low reactivity with RART. Also, RART gave low reactivity with B cells obtained from dogs with lymphoproliferative diseases (<90% positive for surface IgG). A double labeling immunofluorescent technique demonstrated no overlap (<1%) between RART-positive cells and cells with surface IgG. These studies indicate that RART is a convenient and reliable reagent for the enumeration of canine T lymphocytes. This methodology should be useful for assessment of immune function in the dog.