Genetically Modified Bone Marrow-Derived Vehicle Cells Site Specifically Deliver an Anti-Inflammatory Cytokine to Inflamed Interstitium of Obstructive Nephropathy This information is current as of September 27, 2021. Hiroko Yamagishi, Takashi Yokoo, Toshiyuki Imasawa, Tetsuya Mitarai, Tetsuya Kawamura and Yasunori Utsunomiya J Immunol 2001; 166:609-616; ; doi: 10.4049/jimmunol.166.1.609 Downloaded from http://www.jimmunol.org/content/166/1/609 References This article cites 35 articles, 7 of which you can access for free at: http://www.jimmunol.org/ http://www.jimmunol.org/content/166/1/609.full#ref-list-1 Why The JI? Submit online. • Rapid Reviews! 30 days* from submission to initial decision • No Triage! Every submission reviewed by practicing scientists by guest on September 27, 2021 • Fast Publication! 4 weeks from acceptance to publication *average Subscription Information about subscribing to The Journal of Immunology is online at: http://jimmunol.org/subscription Permissions Submit copyright permission requests at: http://www.aai.org/About/Publications/JI/copyright.html Email Alerts Receive free email-alerts when new articles cite this article. Sign up at: http://jimmunol.org/alerts The Journal of Immunology is published twice each month by The American Association of Immunologists, Inc., 1451 Rockville Pike, Suite 650, Rockville, MD 20852 Copyright © 2001 by The American Association of Immunologists All rights reserved. Print ISSN: 0022-1767 Online ISSN: 1550-6606. Genetically Modified Bone Marrow-Derived Vehicle Cells Site Specifically Deliver an Anti-Inflammatory Cytokine to Inflamed Interstitium of Obstructive Nephropathy1 Hiroko Yamagishi,* Takashi Yokoo,*† Toshiyuki Imasawa,* Tetsuya Mitarai,‡ Tetsuya Kawamura,* and Yasunori Utsunomiya2* In this study, we used genetically modified bone marrow-derived CD11b؉CD18؉ vehicle cells to deliver IL-1 receptor antagonist (IL-1ra) for treatment of inflamed renal interstitium in an animal model of unilateral ureteral obstruction (UUO). Vehicle cells that expressed the ICAM-1 ligands, CD11b and CD18, were obtained from bone marrow cells of DBA/2j mice and adenovirally transduced with the IL-1ra gene or glucocerebrosidase (GC) gene ex vivo. In kidneys treated to develop UUO, levels of ICAM-1, IL-1␤, and IL-1R expression increased within 3 days compared with contralateral untreated kidneys in the same mice. Similarly, Downloaded from the macrophage infiltration in the cortical interstitium increased after 3 days in UUO kidneys, but not untreated kidneys. After UUO developed, DBA/2j mice were injected i.v. with either IL-1ra؉ vehicle cells (IL-1ra-treated mice) or GC؉ vehicle cells GC-treated mice) at 24 h after UUO. Six days after the injection of these vehicle cells, marked increase of CD11b؉ IL-1ra؉ vehicle) cells was observed in the ICAM-1-positive interstitium of UUO kidneys from IL-1ra-treated mice. In contrast, no CD11b؉ IL-1ra؉ cells appeared in ICAM-1-negative contralateral kidneys from these mice. Furthermore, the infiltration of macrophages (p < /in the interstitium of UUO kidneys http://www.jimmunol.org (0.005 ؍ expression of ICAM-1 (p < 0.005), and presence of ␣-smooth muscle actin (p ,(0.001 were significantly decreased in IL-1ra-treated mice compared with GC-treated mice. These findings suggest that IL-1 may con- tribute to the development of renal interstitial injury and that our method can deliver a functioning gene encoding an antiin- flammatory cytokine gene specifically at that site by interacting with local adhesion molecules. The Journal of Immunology, 2001, 166: 609–616. ubulointerstitial injury is the final common pathway for As a modulator of IL-1 activity, the IL-1 receptor antagonist progressive renal disease of several types. Although cy- (IL-1ra)3 can suppress IL-1 activity, as evident by the ability of tokines, infiltrating cells, and adhesion molecules may all this receptor to block experimental glomerulonephritis (10, 12, T by guest on September 27, 2021 be involved in the pathogenesis of this interstitial injury (1–4), the 13). Based on this concept, we previously established a novel sys- underlying mechanisms are not fully understood. Moreover, no tem for using bone marrow-derived cells as vehicles for site-spe- effective therapy currently exists for the related disease, interstitial cific delivery of an IL-1ra gene into inflamed glomeruli. This pro- fibrosis. cedure, which suppressed local IL-1 action (14, 15), successfully IL-1 is an important proinflammatory cytokine with a wide prevented the progression of glomerular injury evoked by Ab to range of effects, including activation of endothelial cells, stimula- the glomerular basement membrane (GBM). tion of tissue infiltration by neutrophils and macrophages, and in- With that background, we initiated this two-part study. For the duction of other mediators of inflammation such as TNF-␣, IL-8, first part, we examined the time course of macrophage infiltration ICAM-1, and NO (5, 6). As an example of its potential for damage, as well as ICAM-1, IL-1 mRNA, and IL-1R expression in mice a pathological role has been identified for IL-1 in experimental and treated to develop a unilateral ureteral obstruction (UUO) in the human glomerulonephritis (7–10). Glomerular as well as tubular cortical interstitium. This model is a well-established archetype of epithelial cells may synthesize and release IL-1 (8, 10, 11), yet no renal interstitial injury (2–4). For the second study, we use genet- proof exists to implicate IL-1 as a cause of tubulointerstitial injury. ically modified bone marrow-derived CD11bϩ CD18ϩ vehicle cells to deliver IL-1ra to inflamed interstitium of UUO kidneys as a new therapeutic approach for controlling tubulointerstitial injury. *Department of Internal Medicine, Division of Nephrology and Hypertension, and †Department of Gene Therapy, Institute of DNA Medicine, Jikei University School of Materials and Methods Medicine, Tokyo, Japan; and ‡Department of Internal Medicine, Saitama Medical Animals School, Kawagoe, Saitama, Japan Thirty DBA/2j female mice were purchased from Nippon Crea (Tokyo, Received for publication June 9, 2000. Accepted for publication October 6, 2000. Japan). All animals used in this study were maintained in our animal fa- The costs of publication of this article were defrayed in part by the payment of page cility on standard laboratory chow. charges. This article must therefore be hereby marked advertisement in accordance with 18 U.S.C. Section 1734 solely to indicate this fact. 1 This work was supported by a grant from the Study Group on IgA Nephropathy (to H.Y.) and a grant from the Ministry of Education (Japan) (to Y.U.). 2 Address correspondence and reprint requests to Dr. Yasunori Utsunomiya, Depart- 3 Abbreviations used in this paper: IL-1ra, IL-1 receptor antagonist; GBM, glomerular ment of Internal Medicine, Division of Nephrology and Hypertension, Jikei Univer- basement membrane; GC, glucocerebrosidase; hpf, high power field; IL-1Rt1, IL-1R sity School of Medicine, 3-25-8 Nishi-shinbashi, Minato-ku, Tokyo, 105-8461 Japan. type I; ␣-SMA, ␣-smooth muscle actin; UUO, unilateral ureteral obstruction. Copyright © 2001 by The American Association of Immunologists 0022-1767/01/$02.00 610 SITE-SPECIFIC GENE DELIVERY INTO INFLAMED INTERSTITIUM FIGURE 1. Expression of ICAM-1 in renal cortexes 3 days after treatment to cause to UUO. Original magnification, ϫ400. ICAM-1 was clearly observed in cortical tubular epithelial cells, interstitia, and vessels in UUO kidneys (A), whereas contralateral untreated kidneys had no obvious ICAM-1 staining (B). Downloaded from http://www.jimmunol.org/ Unilateral ureteral obstruction ␮g/ml amphotericin B. Cells were seeded onto unprocessed 10-cm dishes at a concentration of 1 ϫ 107 cells/dish and cultured in a humidified atmosphere of At 8 wk of age, 20 mice were anesthetized by the i.p. injection of pento- 5% CO2 for 1 wk. These vehicle cells were verified as expressing CD11b and barbital, and their right ureters were ligated and cut down as described (16) CD18, both of which are ligand of ICAM-1 by FACS (14). to cause UUO. Five of these mice with UUO kidneys were sacrificed for histological examination and RT-PCR analyses at posttreatment days 3, 5, 7, and 14. Recombinant adenovirus preparation and in vivo injection of IL-1ra Establishment of vehicle cells Replication-defective recombinant adenoviruses carrying IL-1ra by guest on September 27, 2021 Bone marrow-derived CD11bϩ and CD18ϩ vehicle cells were established (AxCAmIL-1RA) were purchased from Riken DNA Bank (Ibaraki, Japan), as previously described (14). Briefly, bone marrow cells were harvested and adenoviruses carrying glucocerebrosidase (GC) cDNA (AxACGC) from the femur and tibia of the 7- to 8-wk-old DBA/2j mice and suspended (17) were kindly provided by Dr. T. Ohashi (Department of Gene Therapy, in DMEM (Life Technologies, Grand Island, NY) supplemented with 10% Jikei University School of Medicine, Tokyo, Japan). Both were under the heat-inactivated FBS, 20% heat-inactivated horse serum, 20% L-929-con- control of a CMV enhancer chicken ␤-actin hybrid promoter (18). Recom- ditioned medium, 100 U/ml penicillin G, 100 ␮g/ml streptomycin, and 0.25 binant viruses were propagated and isolated from 293 host cells. Bone FIGURE 2. Immunohistochemical detection of F4/80-positive cells in the interstitia of UUO kidneys at day 3. Original magnification, ϫ400. F4/80- positive cells were recruited into the in- terstitia of a UUO kidney (A). In con- trast, few F4/80-positive cells were found in the periglomerular space or interstitia of an unobstructed kidney from the same mouse (B). The Journal of Immunology 611 FIGURE 3. Time course of interstitial ICAM-1 expression and F4/80-positive cell infiltration in UUO-treated mice. In UUO kidneys (left), intersti- tial ICAM-1 expression peaked at day 5. The num- ber of F4/80-positive macrophages in the intersti- tium of UUO kidneys increased by day 3 and peaked at day 7.