Human IL-3 (multi-CSF): Identification by expression cloning of a novel hematopoietic growth factor related to murine IL-3

Three distinct domains in TEL-AML1 are required for transcriptional repression of the IL-3 promoter

“…Brie¯y, 5 mg of total RNA was incubated with dNTP mix, random primer, RNasin, and MMLV-reverse transcriptase (Gibco/BRL), and 56RT bu er for 60 min at 378C. Commercially available primers were used in the PCR reaction, to detect IL-3 (Yang et al, 1986), or GM-CSF mRNA (Miyatake et al, 1985). The PCR reactions were performed with 0.5 ml of cDNA, dNTP mix, 50 mM each of primers, 2.5 units of Taq polymerase (Promega), and 106Taq.…”

Preservation of immature hematopoietic progenitor cells responding to interleukin 3 in marrow treated with 4-hydroperoxycyclophosphamide.

“…BM cells were cul-Lured in 35 x 10 mm dishes using a modification of the technique previously described (Yang et al 1986;Sieff et al 1987). 5 x 104 to 2 x 105 non-adherent BM mononuclear cells were prated in 1 ml of Iscove's modified Dulbecco's medium (IMDM, Gibco Laboratories), clntaining 0.9% methylcelulose, 30% FCS, 1% bovine serum albumin (BSA, Sigma Chemical Co., Ltd. St. Louis, Mo, USA), 2 X 10-s M 2-mercaptoethanol (2ME, Bio-Rad Laboratories, Richmonds, CA USA), 100 colony forming units (CFU) /ml of human recombinant interleukin 3 (IL3, Genzyme Corp., Boston, MA, USA), and 2 U/ml of human recombinant erythropoietin (EPO, Chugai Pharmaceutical Co. Ltd., Tokyo).…”

Expression of different combinations of interleukins by human T cell leukemic cell lines that are clonally related.

“…DNA fragments used as probes are as follows : Human IL-la cDNA, 2-kb Bam HI fragment of pCDIL-la (11); human IL-lei cDNA, 1-kb Pst I-Acc I fragment of pA-26 (12) (provided by Dr. Webb of Massachusetts Institute of Technology, Cambridge, MA) ; human IL-2 cDNA, 0.8-kb Pvu II-Pst I fragment of pHIG5-3 (13) (provided by Dr. Onoue of Kumamoto University); human IL-3 cDNA, synthetic 31-mer oligonucleotide 5'-GTTGAATGCCTCCAGGTTTGGCCTTCGAAGG-3' corresponding to antisense strand between positions 222 and 252 of the published sequence (14), which is identical to gibbon IL-3 cDNA sequence ; human IL-4 cDNA, 0.3-kb Nhe I-Eco RI fragment (supplied by Ono Pharmaceutical Co., Osaka, Japan) ; human IL-5 cDNA, 1-kb Barn HI fragment ofph-IL-5-30 (15); human IL-6 cDNA, 1.1 kb Bam HI-Eco RI fragment of pBSF2-38 (16) (provided by Dr. T. Hirano of Osaka University); /3-actin gene, 0.4-kb Hinf I fragment which encodes the 4th exon and its 3' intron (17). These fragments, except for human IL-3 synthetic oligonucleotide, were labeled to obtain the specific activity of 600-1,200 cpm/pg using random primers (18) and a-[ 32P]dCTP (3,000 Ci/mmol, Amersham Corp., Arlington Heights, IL).…”