Extracts of plant pollens and spores of a fungus that are used for immunotherapy of allergies were obtained commercially from Greer and analyzed for protein composition by SDS‐polyacrylamide gel electrophoresis. The extracts analyzed were pecan (Carya illinoensis), mesquite (Prosopis glandulosa), short ragweed (Ambrosia artemisiifolia), white oak (Quercus alba), mountain cedar (Juniperus ashei), and the fungus Aspergillis fumigatus. The extracts were resuspended in deionized water at a concentration of protein = 1.0 mg/ml. Samples containing 1–5 μg of protein were incubated for 2 min. at 100°C with SDS buffer and loaded on 4–20% gradient polyacrylamide gels. Electrophoresis was performed at 75 V for 2 hrs. Broad range markers (Bio‐Rad) were electrophoresed in two lanes on each gel in parallel with the samples. The gels were stained with Coomassie Brilliant Blue. The values of Mr (kDa) for the markers were 7.1, 21, 35, 49, 80, 124, and 209. The Mr (kDa) of the observed proteins for each of the allergens was determined by comparison to the migration of the markers: Carya illinoensis (10, 13, 17, 20, 27, 39, 41, 50, 51, 56, 71, 105, 119); Prosopis glandulosa (20, 27, 44, 50, 57, 63, 78, 115, 139, 146); Ambrosia artemisiifolia (16, 20, 25, 29, 42, 55, 62, 69, 84, 111); Quercus alba (12, 23, 40, 44, 63, 115, 135, 141) Juniperus ashei (14, 25, 37, 51, 66); Aspergillis fumigatus (10, 24, 27, 51, 59, 79, 85, 91, 100, 108, 124, 129, 140). The extracts will be fractionated in order to isolate and identify the proteins responsible for allergic reactions. This work was supported by a grant from the Welch Foundation to the Department of Chemistry at University of the Incarnate Word and by CA74388 to PDF.

Interleukin‐2 (IL‐2) is a lymphokine that binds to a high affinity receptor (CD 25) on the surface of human T lymphocytes. Transduction of this signal activates T cells by means of biochemical pathways that result in increased transcription, DNA replication, and, ultimately, cell division. HuT 78, a CD 25+ human T cell lymphoma line, was obtained from ATCC and used in these studies. It was previously shown that the specific activities of both DNA topoisomerase I (Topo I) and DNA topoisomerase II (Topo II) increased 10‐fold to 20‐fold in nuclear extracts prepared from HuT 78 cells within the first 24 hrs. after treatment with IL‐2 (Foglesong, 2006). Those results suggested that both Topo I and Topo II function in transcription and DNA replication in activated human T lymphocytes. The polypeptide composition of the nuclear extracts was analyzed by SDS‐polyacrylamide gel electrophoresis in order to determine whether the observed increases in the specific activities of these enzymes were due to increased synthesis of protein. Aliquots of the nuclear extracts containing 1–5 μg protein were incubated at 100o C for 2 min. in SDS buffer and electrophoresed at 75 V for 2 hr. on 4 to 20% gradient polyacrylamide gels along with broad range markers (Bio‐Rad). The gels were stained with Coomassie Brilliant Blue. Under these denaturing conditions the relative molecular masses (Mr) for Topo I, Topo IIα, and Topo IIβ are 100, 170, and 180 kDa, respectively. No changes in the abundance of proteins of these sizes were observed in the extracts prepared in the first 24 hrs. following treatment of HuT 78 cells with IL‐2. These results taken together suggest that the activities of Topo I and Topo II are regulated post‐translationally in IL‐2‐activated human T lymphocytes. This work was supported by CA 74388 to PDF.