INVITRO EFFECTS OF ADENOSINE ON LYMPHOCYTES AND ERYTHROCYTES FROM HORSES WITH COMBINED IMMUNODEFICIENCY

The effect of adenosine on the mitogenic response of peripheral blood lymphocytes (PBL) and on the nucleotide pools of erythrocytes from normal horses, horses heterozygous for the combined immunodeficiency (CID) trait (carriers), and foals with CID was studied. When PBL from normal, carrier, and CID horses were stimulated by phytohemagglutinin (PHA), concanavalin A, or pokeweed mitogen, [3H]thymidine uptake was inhibited by adenosine (0.1 μM to 1.0 mM) in a dose-dependent manner. Adenosine (100 μM) mediated inhibition of [3H]thymidine uptake was prevented in both normal and carrier horse PBL by incubation with uridine. Uridine had no sparing effect on PBL from horses wit CID. Differences were detected between human and horse PBL in response to adenosine and erythro-9-(2-hydroxy-3-nonyl) adenine (EHNA), a competitive inhibitor of adenosine deaminase. In the first assay, mitogen-stimulated PBL from horses were more sensitive to adenosine. In the second assay, adenosine was added to PBL cultures at various times after PHA addition. Adenosine inhibited mitogenesis in horse PBL if added within the first 24 h. In human PBL cultures, adenosine inhibited mitogenesis only if added within the first 4 h. The third assay measured capacity of PHA-stimulated human and horse lymphocytes to escape inhibition by adenosine or EHNA. At the end of a 72-h culture period, horse PBL were still inhibited by adenosine and EHNA, but human PBL were not. EHNA and adenosine together caused synergistic inhibition of mitogenesis in both human and horse PBL. With prolonged incubation (72 h), synergistic inhibition was detected only in horse PBL. With high-pressure liquid chromatography, nucleotide levels in erythrocytes of normal, carrier, and CID horses were found to be similar. Incubation with adenosine produced a 1.5- to 2-fold increase in total adenine nucleotide pools in erythrocytes form all horses. However, these increases were accompanied by alterations in the relative amount of the nucleotide components. This was seen as a significant decrease in the ATP: (AMP plus ADP plus ATP) ratio and energy charge in erythrocytes from normal horses. In contrast, the ATP: (AMP plus ADP plus ATP) ratio decreases only slightly in erythrocytes from CID horses, whereas no change in the energy charge was detected. The data from these studies indicate a difference in adenosine metabolism exists between human and horse lymphocytes, and an abnormality may exist in purine metabolism or in an interconnecting pathway in horses with CID.