Cloning, sequencing and characterization of lentiviral-mediated expression of rhesus macaque (Macaca mulatta) interleukin-2 receptor alpha cDNA.
The rhesus macaque CD25 (RhCD25) cDNA isolated from rhesus PBMCs was found to share 95.5 and 91.9% homology with the human orthologue at the nucleotide and amino acid levels, respectively. Comparative sequence analyses suggest that both human CD25 (HuCD25) and RhCD25 share identity for most of the critical amino acids previously identified to be essential for viable folding and IL-2 ligand binding. The human leukemic cell line, HH, deficient for IL-2Ralpha was transduced with a lentiviral vector (LV) engineered to express RhCD25 (HH-RhCD25). RhCD25 was characterized for expression by flow cytometric analyses, ELISA, Western blotting, functional signalling, and biological assays in comparison to HuCD25. In summary, vectors expressing the RhCD25 cDNA can be used as a tool to aid in the characterization of soluble CD25 in non-human primate studies, and to provide a tempting alternative as an autologous cell surface marker in rhesus macaque gene therapy and bone marrow transplantation studies.
Silvertown
Josh D
JD
Division of Experimental Therapeutics, Ontario Cancer Institute, University Health Network, University of Toronto, Toronto, Ont., Canada M5G-2M1.
Walia
Jagdeep S
JS
Medin
Jeffrey A
JA
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D002460:Cell Line; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000235:genetics; D015870:Gene Expression; D005822:Genetic Vectors; D006801:Humans; D053645:Interleukin-2 Receptor alpha Subunit; D016086:Lentivirus / Q000235:genetics; D008253:Macaca mulatta / Q000235:genetics* / Q000276:immunology*; D008969:Molecular Sequence Data; D018123:Receptors, Interleukin / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism; D015375:Receptors, Interleukin-2 / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D011994:Recombinant Proteins / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D017386:Sequence Homology, Amino Acid; D012689:Sequence Homology, Nucleic Acid; D015398:Signal Transduction; D012995:Solubility; D013045:Species Specificity; D014161:Transduction, Genetic
D018076:DNA, Complementary; C508017:IL2RA protein, human; D053645:Interleukin-2 Receptor alpha Subunit; D018123:Receptors, Interleukin; D015375:Receptors, Interleukin-2; D011994:Recombinant Proteins