Eotaxin triggers eosinophil-selective chemotaxis and calcium flux via a distinct receptor and induces pulmonary eosinophilia in the presence of interleukin 5 in mice.
BACKGROUND
Understanding the processes that control selective eosinophilia is of fundamental importance in a variety of human diseases (e.g., allergies, parasitic infections, malignancy). Interleukin 5, an eosinophil-specific growth and activating factor, and eotaxin appear to collaborate in this process. Eotaxin is a recently described chemotactic factor that belongs to the C-C (or beta) chemokine family and has been implicated in animal and human eosinophilic inflammatory states. We have recently reported the molecular characterization of murine eotaxin and now report the biological properties of purified recombinant murine eotaxin in vitro and in vivo in the presence or absence of interleukin 5 (IL-5) in mice.


METHODS
Murine eotaxin was expressed in bacteria and purified by affinity chromatography and HPLC. Activity was tested in vitro by examining chemotactic and calcium flux responses of purified murine leukocytes. Additionally, desensitization of calcium flux responses to other chemokines, eosinophil survival assays, and basophil histamine release were examined. Finally, eotaxin was delivered to wild-type or IL-5 transgenic mice and the host response was examined.


RESULTS
Eotaxin had activity only when the recombinant molecule had the native mature amino terminus and contained the first 25 amino acids of the mature protein. It was active in vitro at an effective concentration between 10 and 100 ng/ml in both chemotaxis and calcium flux assays toward eosinophils, but not macrophages or neutrophils. Furthermore, intranasal or subcutaneous application of eotaxin selectively recruited large numbers of eosinophils into the mouse lung and skin, respectively, only in the presence of interleukin 5. Macrophage inflammatory protein-1 alpha, a related C-C chemokine active on eosinophils, and eotaxin were not able to cross-desensitize. Eotaxin had no affect on the in vitro survival of eosinophils and did not induce basophil histamine release.


CONCLUSIONS
Mouse eotaxin is an eosinophil specific chemoattractant that has a markedly enhanced effect in vivo in the presence of another eosinophil selective cytokine IL-5, and utilizes a signal transduction receptor pathway that is distinct from that utilized by macrophage inflammatory protein-1 alpha. This data suggests that the development of tissue eosinophilia in vivo involves a two-step mechanism elicited by interleukin 5 and eotaxin.
Rothenberg
M E
ME
Department of Genetics, Howard Hughes Medical Institute, Harvard Medical School, Boston, Massachusetts, USA.
Ownbey
R
R
Mehlhop
P D
PD
Loiselle
P M
PM
van de Rijn
M
M
Bonventre
J V
JV
Oettgen
H C
HC
Leder
P
P
Luster
A D
AD
D000818:Animals; D001491:Basophils / Q000187:drug effects / Q000276:immunology*; D002118:Calcium / Q000097:blood*; D002470:Cell Survival / Q000187:drug effects; D002478:Cells, Cultured; D054413:Chemokine CCL11; D019742:Chemokines, CC; D002631:Chemotactic Factors, Eosinophil / Q000494:pharmacology*; D002634:Chemotaxis, Leukocyte / Q000187:drug effects*; D003001:Cloning, Molecular; D016207:Cytokines / Q000302:isolation & purification / Q000494:pharmacology; D004802:Eosinophilia / Q000097:blood / Q000139:chemically induced / Q000276:immunology*; D004804:Eosinophils / Q000166:cytology / Q000187:drug effects / Q000502:physiology*; D004926:Escherichia coli; D015951:Factor Xa / Q000378:metabolism; D006636:Histamine Release / Q000187:drug effects; D006801:Humans; D015848:Interleukin-5 / Q000096:biosynthesis / Q000235:genetics / Q000502:physiology*; D008171:Lung Diseases / Q000097:blood / Q000139:chemically induced / Q000276:immunology*; D008264:Macrophages / Q000187:drug effects / Q000502:physiology; D051379:Mice; D008808:Mice, Inbred CBA; D008815:Mice, Inbred Strains; D008822:Mice, Transgenic; D054397:Receptors, CCR3; D019707:Receptors, Chemokine; D018121:Receptors, Cytokine / Q000502:physiology*; D011994:Recombinant Proteins / Q000302:isolation & purification / Q000494:pharmacology
C515456:CCL11 protein, human; C515350:CCR3 protein, human; C515457:Ccl11 protein, mouse; C515351:Ccr3 protein, mouse; D054413:Chemokine CCL11; D019742:Chemokines, CC; D002631:Chemotactic Factors, Eosinophil; D016207:Cytokines; D015848:Interleukin-5; D054397:Receptors, CCR3; D019707:Receptors, Chemokine; D018121:Receptors, Cytokine; D011994:Recombinant Proteins; D015951:Factor Xa; D002118:Calcium