Biochemical and immunological properties of cytokines conjugated to dendritic polymers.
Here we describe a post-translational modification of SC-63032, a variant of the species restricted, multi-lineage hematopoeitic factor human interleukin-3 (hIL-3). We have made two new dendritic polymer (polyamidoamine or PAMAM dendrimers, generation 5)-SC-63032 bioconjugates. Using two distinct chemistries (one of which is novel to this work), we achieved site-specific conjugation with respect to the amino acid in the proteins ligated to the dendrimers. In both bioconjugates, conjugated cytokine maintains its ability to bind the hIL-3 alpha receptor subunit, but is significantly (about 10-fold) less potent in inducing hIL-3 dependent in vitro cell proliferation than is the free cytokine. In vivo data indicates that conjugation decreases the immunogenicity of the conjugated cytokine modestly. In the absence of pharmacokinetic or biodistribution effects associated with the bioconjugates that increase their potency in vivo (which can only be tested in a higher primate, due to the species restriction of hIL-3 and its derivatives), these immune mitigation effects may be too small to be therapeutically significant. Though unmodified PAMAM dendrimers fail to elicit an antibody response in mice, protein conjugation to dendrimers haptenizes them, and a dendrimer-specific antibody response is produced. In toto, the principal limitation of the dendrimer-cytokine bioconjugates herein is in their reduced receptor affinity and potency in vitro. Were the in vivo potency of the bioconjugates to parallel the in vitro potency of the conjugates reported here, it is likely that particular dendrimer bioconjugates could not justify their higher costs of goods relative to the parent SC-63032 molecule, though retention of SC-63032 biological activities in conjugates suggests that other cytokine-dendrimer bioconjugates may be bioactive. This is good news to the nanotechnology community, in as much as PAMAM dendrimers are among the monodisperse polymeric nanomaterials available, and these results show that they can be used successfully in conjugates to bioactive proteins.
Lee
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SC
Department of Cellular and Molecular Biochemistry, and the Dorothy M. Davis Heart and Lung Research Institute, 473 W. 12th Avenue, Ohio State University, Columbus, OH 43210, USA. Lee.1996@osu.edu
Parthasarathy
R
R
Botwin
K
K
Kunneman
D
D
Rowold
E
E
Lange
G
G
Klover
J
J
Abegg
A
A
Zobel
J
J
Beck
T
T
Miller
T
T
Hood
W
W
Monahan
J
J
McKearn
J P
JP
Jansson
R
R
Voliva
C F
CF
D000818:Animals; D001672:Biocompatible Materials / Q000737:chemistry; D002460:Cell Line; D006224:Cricetinae; D016207:Cytokines / Q000737:chemistry / Q000276:immunology / Q000378:metabolism; D005260:Female; D007668:Kidney / Q000378:metabolism*; D051379:Mice; D008807:Mice, Inbred BALB C; D011073:Polyamines / Q000737:chemistry*; D011108:Polymers / Q000737:chemistry; D011485:Protein Binding; D015202:Protein Engineering / Q000379:methods*; D011506:Proteins / Q000737:chemistry / Q000276:immunology* / Q000378:metabolism*; D016185:Receptors, Interleukin-3 / Q000378:metabolism*; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000276:immunology / Q000378:metabolism
D001672:Biocompatible Materials; D016207:Cytokines; D011073:Polyamines; D011108:Polymers; D011506:Proteins; D016185:Receptors, Interleukin-3; D011993:Recombinant Fusion Proteins; C500269:SC-63032