Species specificity of human interleukin-3 demonstrated by cloning and expression of the homologous rhesus monkey (Macaca mulatta) gene

Abstract

To enable preclinical studies on homologous interleukin-3 (IL-3) in primate species, we isolated the gene encoding Rhesus monkey IL-3 (RhIL- 3). The nucleotide sequence of the RhIL-3 gene displayed 92.9% homology with that of the human IL-3 (hIL-3) gene. The isolated RhIL-3 gene encodes a 143-amino acid (aa) precursor polypeptide, nine C-terminal residues shorter than the human protein. Protein homology was found to be 89.5% for the signal peptide (19 aa) and 80.5% for the mature protein (124 aa). Comparison of the human and RhIL-3 coding sequences showed that the majority of substitutions had occurred at amino acid replacement sites indicating a rapid evolution of the IL-3 protein. After expression of a genomic fragment in COS cells, RhIL-3 cDNA was constructed, which enabled large-scale production of the RhIL-3 polypeptide, RhIL-3 produced by Bacillus licheniformis and purified to homogeneity appeared to be approximately 100-fold more effective in stimulating Rhesus monkey hematopoietic progenitors than hIL-3, whereas RhIL-3 and hIL-3 showed comparable stimulatory activity on normal as well as malignant human hematopoietic cells. Thus, the rapid evolution of hIL-3 has resulted in a unidirectional species specificity, which most likely restricts the in vivo effects of hIL-3 in Macaca species.

Interleukin‐3 Promotes Proliferation and Differentiation of Human Hematopoietic Stem Cells but Reduces Their Repopulation Potential in NOD/SCID Mice

“…To optimize engraftment rates, supplementation with human cytokines seems to be beneficial [10, 11]. While several hematopoietic active cytokines are cross‐reactive between mice and humans and can provide human hematopoietic cells with growth factors, interleukin‐3 (IL‐3), formerly known as multi‐colony‐stimulating factor, is highly species specific [12]. The support of early human hematopoiesis in the chimeric bone marrow with IL‐3 is probably crucial for human cell engraftment.…”

Large Animal Models for Stem and Progenitor Cell Analysis

“…They can be bred in captivity, and they are not endangered, although the surge in vaccine research for HIV and bioterrorism agents has produced a worldwide shortage of captive pathogen-free animals. Macaque and baboon hematopoietic cells (HSCs) respond to all human hematopoietic cytokines, with similar concentrations required for maximal in vitro stimulation, with the exception of human interleukin 3 (IL-3), which must be added to macaque cells at a 10-fold higher concentration to yield the same degree of colony formation as in baboons (Burger et al, 1990).…”

Upregulation of microRNA‑16 alters the response to inhaled β‑agonists in patients with asthma though modulating expression of ADRB2

“…A comparison of 16 asthmatics to 12 healthy controls identified that miRNA (miR)-221 was discovered to be increased (17). The study of Burger et al (18) revealed that decreased airway inflammation may be triggered by blocking miR-221 in a mouse asthma model induced by ovalbumin, demonstrating the function of miR-221 in asthma. T cells have been the focus in the majority of investigations on miRNAs in the blood for asthma risk.…”