-
_______________________________________________________________
_______________________________________________________________
Report Information from ProQuest14 January 2015
23:46_______________________________________________________________
-
Dokumen 1 dari 1 Targeting genetically modified macrophages to
the glomerulus Link dokumen ProQuest Abstrak: Macrophages are key
players in the development of the majority of renal diseases and
are thereforeideal cellular vectors for site specifically targeting
gene therapy to inflamed glomeruli. Macrophages can begenetically
modified using viral vectors ex vivo then re-introduced into the
body where they can home to thediseased site. This review
summarises current experience in efficiently targeting modified
macrophages to theinflamed glomerulus focussing on the factors
controlling macrophage localisation, macrophage gene
transfermethods, in vivo gene delivery and results of recent
investigations using modified macrophage gene therapy forglomerular
disease. Teks lengkap: Headnote Key Words Macrophages *
Inflammation * Glomerulonephritis . Adenovirus . Gene Abstract
Macrophages are key players in the development of the majority of
renal diseases and are therefore idealcellular vectors for site
specifically targeting gene therapy to inflamed glomeruli.
Macrophages can begenetically modified using viral vectors ex vivo
then re-introduced into the body where they can home to thediseased
site. This review summarises current experience in efficiently
targeting modified macrophages to theinflamed glomerulus focussing
on the factors controlling macrophage localisation, macrophage gene
transfermethods, in vivo gene delivery and results of recent
investigations using modified macrophage gene therapy forglomerular
disease. Introduction Macrophage infiltration of renal tissue is a
feature of many forms of nephrological disease ranging from
classicalinflammatory diseases such as rapidly progressive
glomerulonephritis to more indolent 'non-inflammatory'processes
such as diabetic nephropathy. The classical view has been that
these cells are primarily injurious,and that inhibiting their
function or preventing entry into renal tissue is the main goal of
therapy. Macrophagesare, however, heterogeneous and perform
distinct functions; increasingly their role in the down-regulation
andresolution of inflammation has been identified [1]. This is most
clearly seen in skin wounds where inhibition ofmacrophage entry
prolongs inflammation and delays healing [2]. This raises the
potential to utilise macrophagesas therapeutic cells to
down-regulate renal inflammation and restore normal function.
Critical to achieving this isto establish what determines their
function, what factors control macrophage localisation to inflamed
renaltissue, and how their properties can be modified to ameliorate
inflammation. Functions of Macrophages in Renal Inflammation
Macrophages arc critical adaptors in the immune response, able to
both drive the immune response by antigenpresentation to T cells
and respond to cytokines produced by the specific immunity; they
are both conductor andplayer in the inflammatory orchestra.
Macrophage infiltration into renal tissue is an early feature
ofglomerulonephritis and contributes to the development of
necrosis, crescent formation and subsequent scarring.Macrophage
function in the early stages of inflammation is geared towards
promoting inflammation as within theinflamed environment
macrophages are exposed to a wide range of stimuli including
pro-inflammatory cytokinessuch as IL-1[beta], TNF-[alpha] and
IFN-[gamma], lipid mediators, reactive oxygen and nitrogen
products,chemokines and necrotic and apoptotic cells. The
importance of a number of these factors in experimentalmodels of
glomerular inflammation has been established with inhibition of
cytokines such as IL-1[beta], TNF-[alpha] and macrophage migration
inhibitory factor (MIF) reducing disease severity. This
pro-inflammatoryactivation of macrophages renders them unresponsive
to anti-inflammatory cytokines such as transforming
http://search.proquest.com/docview/274309492?accountid=166961
-
growth factor-[beta] (TGF-[beta]) or IL-4 [3], and
re-orientating this response is a critical approach to
resolveinflammation. The crosstalk between infiltrating macrophages
and their environment is important ininflammation. Suto et al. [4]
have demonstrated that injection of activated macrophages into
glomeruli resultedin the up-regulation of matrix metalloproteinase
9 (MMP-9) in normal glomeruli but expression was down-regulated
when activated macrophages were injected into regenerating
glomeruli from rats with Thy-1 nephritiswhere TGF-[beta] production
was high. Thus, local expression of TGF-[beta] from resident
mesangial cells maysuppress activated macrophages and macrophage
factors may influence the regulation of genes in residentglomerular
cells [5]. Factors Controlling Macrophage Localisation to Inflamed
Renal Tissue The first stage of monocyte/macrophage involvement in
glomerular inflammation is adhesion to activatedendothelium and
transmigration in response to a chemotactic gradient. There is
increasing evidence thatdifferent vascular beds and different forms
of inflammation utilise distinct molecules to enable
macrophagelocalisation and these need to be understood for
glomerular inflammation to allow efficient homing of
geneticallymodified inflammatory cells. Initial work demonstrated
that blockade of classical integrin molecules,
intracellularadhesion molecule-1 (ICAM-1) and vascular cell
adhesion molecule (VCAM) by antibodies decreasedinflammation in
nephrotoxic nephritis (NTN) [6], however, there was no consistent
evidence of a reduction inglomerular macrophage numbers and the
reduction in injury may have reflected alteration of trafficking
ofinflammatory cells in other organs such as spleen or regional
lymph nodes. Recent studies have focused ondirect visualisation of
leukocyte adhesion. De Vrcise et al. [7] showed that inhibition of
selectins did not affectglomerular leukocyte adhesion, while
inhibition of [beta]^sub 2^ integrin CD11b (counter-receptor to
ICAM-1)prevented glomerular leukocyte accumulation. Recently,
chemokines have also been show to have roles in adhesion.
Fractalkine is a CX3C chemokine thatexists as a soluble chemotactic
agent but it also has a transmembrane domain and is expressed on
activatedendothelium. Inhibition of fractalkine receptor (CX3CR1)
in NTN reduced glomerular macrophage infiltration andattenuated
histological and biochemical measures of injury [8]. The chemokine
GRO-[alpha] can also beimmobilised to cell surface proteoglycans to
support adhesion of monocytes to mesangial cells in vitro.
Inaddition, blockade of GRO-[alpha] or fractalkine receptors on
macrophages injected directly into the renal arteryof rats with NTN
suppressed their localisation to inflamed glomeruli [9]. Thus,
adhesion to glomerularendothelium in experimental models appears to
involve chemoadhesin interaction followed by
integrin-mediatedadhesion. Once adherent to endothelium,
macrophages transmigrate across the glomerular basement membrane
inresponse to chemokines. A wide range of approaches have been used
to study which chemokines are pre-eminent in the control of renal
inflammation [reviewed in 10] and as with adhesion, it appears to
depend on boththe nature of injury and at what point during the
immune response they are assessed. Two chemokines forwhich the
clearest role is defined are monocyte chemotactic protein-1 (MCP-1
- binds to CCR2) and regulatedon activation normal T cell expressed
and secreted, RANTES (binds to CCR1, 3 and 5). Early studies
showedthat inhibition of MCP-1 by anti-bodies reduced macrophage
glomerular infiltration and proteinuria in NTN andexpression of
MCP-1 and CCR2 are early features of immune complex
glomerulonephritis, NTN and murinelupus nephritis. We have also
demonstrated that direct inhibition of CCR2 receptor on macrophages
reducedtheir glomerular localisation in rats with NTN [9].
Similarly, RANTES is found in many models of glomerularinflammation
and inhibition by the antagonists met-RANTES or
aminoxypentane-RANTES reduces macrophageinfiltration in NTN and
anti-Thy-1 nephritis. Expression of chemokines has also been
identified in human renal biopsies and a recent study by Segerer et
al.[11] showed that in crescentic glomerulonephritis, CCR2 was
mainly expressed on glomerular macrophageswhile CCR5 was found
mainly on interstitial T cells. This suggests that specific
chemokine expression directsmacrophages and T cells to separate
compartments with distinct functional consequences. Thus, at
present,
-
MCP-1 and its counter-receptor CCR2 are prominent in the early
stages of glomerular inflammation and providea target to direct
macrophages to inflamed glomeruli. Further work is required to
determine the orchestration ofchemokines during glomerulonephritis,
particularly those expressed during the resolution of inflammation.
Macrophage Manipulation Macrophages are ideally suited to alter
inflammatory disease due to their preferential localisation to
inflamedtissue, the properties they can develop and their
interaction with other cells of the immune response. Thus far,work
has focused on altering macrophage function ex vivo before
re-administration into animals with renalinflammation. A number
approaches exist to transduce macrophages to express specific
transgenes includingviral vectors such as adenoviruses,
retroviruses and lentiviruses as well as non-viral methods
including cationicDNA carrier molecules, electroporation and
receptor-mediated DNA uptake [reviewed in 12]. Non-viralapproaches
give a low efficiency of transfection compared to viral methods,
thus the latter have been favouredfor in vivo manipulation.
Adenoviral vectors are at present the most popular method of
genetically modifyingmacrophages; large genes (up to ~ 10 kb) can
be transferred in standard vectors and a high level of expressionof
the transgene achieved. Adenoviruses are relatively inefficient at
transducing monocytes due to the lowexpression of adenoviral
receptor integrins [alpha]^sub v^[beta]^sub 3^ and [alpha]^sub
v^[beta]^sub 5^, butinfection rates can be increased to greater
than 90% if primary cultures of monocytes are incubated
withmacrophage colony-stimulating factor which up-regulates
integrin expression. Adenoviral transduced primarycultures of
bone-derived macrophages (BMDM) or macrophage cell lines such as
RAW 267.4 and NR8383 cellsexpress high levels of transgene within
24 h of transduction and this remains stable for 7-10 days [13]
allowingefficient short-term expression in vivo. Retroviruses
transduce rapidly dividing cells by stably incorporating DNA into
the host cell genome, however,the transduction efficiency is low
due to the low rate of proliferation. To overcome the problem,
CD34^sup +^bone marrow stem cells, precursors of macrophages, have
been used that are capable of cell division andretroviral
infection. This allows reconstitution with genetically modified
bone marrow cells that permits long-lasting expression of the
relevant transgene. Recombinant lentiviruses have a natural tropism
for macrophagesand can stably transduce these cells. Despite their
high level of gene transfer and expression, concernsregarding
safety issues such as generation of replicon-competent virus during
production of vectors have so farlimited their use to in vitro
work. In vivo Localisation of Genetically Modified Macrophages Site
specifically targeting genetically modified macrophages to the
glomerulus and limiting expression in non-target tissues is a
fundamental requirement for their use as vehicles for gene therapy.
Macrophages have beeninjected intravenously or directly into the
renal artery where they have a concentrated exposure to
theinflammatory site. The main concern with intravenous injection
is that only a small proportion of the modifiedmacrophages localise
to inflamed glomeruli due to their hold up in the lung and
trafficking to the spleen. Directinjection into the renal artery by
contrast, allows highly efficient first-pass localisation and we
have shown thatadenoviral transduced macrophages localise
preferentially in the presence of glomerular inflammation in
ratsand when macrophages are activated with lipopolysaccharide
[13]. Yokoo et al. [14] have shown that CD11b^sup +^CD18^sup +^
(ligand for ICAM-1) bone marrow-derived cellstransduced with
recombinant adenovirus then re-injected systemically into mice
localise to glomeruli when theanimals were treated with
lipopolysaccharide, which up-regulates ICAM-1 expression, and on
induction ofglomerulonephritis [15]. Thus the state of activation
of macrophages and the endothelium, as well as themethod of
delivery, affects the efficiency of glomerular macrophage
localisation. The transgene expressed bymacrophages can also affect
the properties of cells as we found that transduction with
adenoviruses expressingIL-4 markedly enhanced the ability of
macrophages to localise to inflamed glomeruli compared to
expression ofinert transgene [16]. The most likely mechanism for
this is up-regulation of CXCR1 and 2 expression bymacrophages,
which adhere to GRO-[alpha], a recognised chemoadhesin on
glomerular endothelium. Thus,
-
glomerular-specific genetically modified macrophage localisation
can be achieved effectively.
Effect of Genetically Modified Macrophages on Renal Inflammation
The transduction of macrophages to alter the development of
inflammatory disease has thus far focused onexpression of cytokines
with principally anti-inflammatory and/or regulatory effects.
Systemic injection ofmacrophages transduced with recombinant
adenovirus to express IL-1 receptor antagonist (IL-1ra) reduced
theseverity of glomerular inflammation in mice with NTN and reduced
interstitial macrophage infiltration in a modelof unilateral
urcteric obstruction [15, 17]. We have studied the effects of
macrophages transduced to expressanti-inflammatory cytokincs IL-4,
IL-10 and TGF-[beta]. Initial work transduced the rat alveolar
macrophage cellline NR8383 with adenovirus to express IL-4. The
cells localised to glomeruli of rats with NTN, produced thecytokine
in vivo and reduced the level of albuminuria, histological markers
of glomerular inflammation andmacrophage infiltration [16]
demonstrating that macrophage cell lines can be modified
efficiently and havetherapeutic effects in vivo. By contrast,
injection of NR8383 cells transduced to express active TGF-[beta]
hadlimited impact on the development of renal injury in rats with
NTN [unpubl. data]. The properties of macrophagecell lines arc
different from native macrophages and may express transgenes in a
different manner andsubsequent work focused on utilising primary
cultures of BMDM from inbred strains of rats. Macrophagestransduced
to express IL-10 localised preferentially to glomeruli of rats with
NTN after renal artery injection andproduced a profound reduction
in the severity of glomerular inflammation reducing both macrophage
infiltrationand their state of activation as assessed by both MHC
class II and ED3 expression [18]. Interestingly, in all these
experiments, delivery of macrophages transduced with an inert gene
([beta]-galactosidase) to inflamed glomeruli resulted in a
reduction in injury as assessed by albuminuria but only up to48 h
after disease induction. The most likely mechanisms for this are
either transduction of macrophagescauses them to release
antiinflammatory molecules (e.g. IL-10) or that infusion of the
transduced macrophagesprevents infiltration of pathogenic
macrophages by competition for adhesion sites. This further
highlights howgenetically modified macrophages have a complex
interaction with glomerular inflammation. The injection of IL-4 and
IL-10 expressing macrophages resulted in highly effective
localisation to glomeruli of asingle kidney with transduced cells
seen infrequently in the contralateral kidney. Despite this, we
have foundthat injection into a single kidney markedly attenuated
the development of inflammation in the contralateralkidney. This
implies that the expression of cytokine expressing macrophages at
one inflammatory site candecrease injury both at that site and at
distant sites. These changes could not be mimicked by systemic
-
administration of IL-10 or IL-4 expressing macrophages and there
was no evidence of increased systemic levelsof the relevant
cytokines. In light of the specific requirement of glomerular
localisation in producing thisresponse, we hypothesise the effect
is due to high local concentrations of cytokines altering the
properties ofmacrophages, dendritic cells and T cells trafficking
through the glomerulus that consequently modify thedevelopment of
the immune response or that the transduced macrophages themselves
migrate to regionallymph tissue and regulate the systemic
development of the immune response (fig. 1). As many
autoimmunediseases arc multifocal, these results raise the
potential that disease modified by macrophages at one site maybe
able to ameliorate inflammation at distant sites, a potentially
powerful tool in therapy. Regulation of Macrophage Gene Expression
The work so far has demonstrated the feasibility of macrophages to
down-regulate inflammation and has largelyused adenoviral
transduction to achieve this. However, a critical issue in
macrophage-mediated gene therapy ishow to control gene expression,
in particular, how macrophage activation can lead to regulated
transgeneexpression. Yokoo et al. [19] used double transduced
macrophages that only express target gene at the site
ofinflammation. To achieve this, macrophages were transduced with
an adenovirus containing the IL-1[beta]promoter which controls
expression of Cre recombinase that, in turn, excised DNA from a
second targetadenovirus carrying a reporter gene under the control
of strong viral promoter, thus leading to reporter geneexpression.
Following injection, the vehicle cells were detected in various
organs including the kidney but thereporter gene was expressed only
in the kidney following the induction of nephritis. The double
transductionrequirement for this method, however, reduce its
general applicability and an alternative approach usingtetracycline
sensitive promoters in a single vector could control transgene
expression in vivo. An additional problem with use of adenovirus is
that expression of the transgene is short-lived. Thus to continueto
suppress ongoing inflammation would potentially require repeat
injections of modified macrophages.However, retroviral
transformation of bone marrow stem cells leads to stable
incorporation of the gene ofinterest into the chromosomal DNA and
subsequent daughter cells. This approach has been used to
generateCD34^sup +^ stem cells from mice expressing IL-1ra and
following re-infusion in irradiated mice around 10-20%of bone
marrow and spleen cells were derived from donor cells [20]. When
NTN was induced in these micethere was a reduced severity of
disease, although it is unclear to what extent this effect was a
consequence ofIL-1ra expression by glomerular macrophages as
opposed to effects elsewhere in the immune system.However,
combining long-term expression with regulation of transgene
activation would provide the optimumsystem to control genetically
modified macrophages in inflammation. Future The experimental work
to date has shown the potential for macrophages to ameliorate
immune-mediatedglomerular disease (fig. 1). They provide a powerful
tool to study the effects of both macrophages and cytokineson the
evolution of inflammatory disease. Therapies using genetically
modified macrophages are in thepreliminary stages and many problems
need to be addressed before moving to the clinical setting. In
spite ofthis, cell-based therapy to treat immune-mediated diseases
is a clear objective. With the increasing use of stemcells and
other bone marrow-derived cells in the treatment of autoimmune
diseases, it could be feasible toaugment the anti-inflammatory and
reparative properties of macrophages using genetic manipulation.
Theadministration of such genetically enhanced macrophages could
provide a powerful complimentary therapy forglomerulonephritis and
other forms of inflammation. Sidebar Copyright (C) 2003 S. Karger
AG, Basel Sidebar KARGER Fax + 41 61 306 12 34 E-Mail
[email protected]
-
www.karger.com (C) 2003 S. Karger AG, Basel
1660-2129/03/0944-0113$19.50/0 Accessible online at:
www.karger.com/nee References References 1 Mosser DM: The many
faces of macrophage activation. J Leuk Biol 2003;73:209-212. 2
Nagaoka T, Kaburagi Y, Hamaguchi Y, Hasegawa M, Takehara K, Steeber
DA, Tedder TF, Sato S: Delayedwound healing in the absence of
intercellular adhesion molecule-1 or L-selectin expression. Am J
Pathol2000;157:237-247. 3 Erwig LP, Stewart K, Rees AJ: Macrophages
from inflamed but not normal glomeruli are unresponsive to
anti-inflammatory cytokines. Am J Pathol 2000;156:295-301. 4 Suto
TS, Fine LG, Shimizu F, Kitamura M: In vivo transfer of engineered
macrophages into the glomerulus:Endogenous TGF-[beta]-mediated
defence against macrophage-induced glomerular cell activation. J
Immunol1997;159:2476-2483. 5 Kitamura M: Identification of an
inhibitor targeting macrophage production of monocyte
chemoattractantprotein-1 asTGF-[beta]^sub 1^. J Immunol
1997;159:1404-1411. 6 Allen AR, McHale J, Smith J, Cook HT, Karkar
A, Haskard DO, Lobb RR, Pusey CD: Endothelial expressionof VCAM-1
in experimental crescentic nephritis and effect of antibodies to
very late antigen-4 or VCAM-1 onglomerular injury. J Immunol
1999;162:5519-5527. 7 De Vriese AS, Endlich K, Elger M, Lameire NH,
Atkins RC, Lan HY, Rupin A, Kriz W, Steinhausen MW: Therole of
selectins in glomerular leukocyte recruitment in rat
anti-glomerular basement membraneglomerulonephritis. J Am Soc
Ncphrol 1999; 10:2510-2517. 8 Feng L, Chen S, Garcia GE, Xia Y,
Siani MA, Botti P, Wilson CB, Harrison JK, Bacon KB: Prevention
ofcrescentic glomerulonephritis by immunoneutralization of the
fractalkine receptor CX3CR1. Kidney Int1999;56:612-620. 9 Zernecke
A, Weber KS, Erwig LP, Kluth DC, Schropel B, Rees AJ, Weber C:
Combinatorial model ofchemokine involvement in glomerular monocyte
infiltration. J Immunol 2001; 166:5755-5762. 10 Segerer S, Nelson
PJ, Schlondorff D: Chemokines, chemokine receptors and renal
disease: From basicscience to pathophysiologic and therapeutic
studies. J Am Soc Nephrol 2000; 11:152-176. 11 Segerer S, Cui Y,
Hudkins KL, Goodpaster T, Eitner F, MacK M, Schlondorff D, Alpers
CE: Expression of thechemokine monocyte chemoattractant protein-1
and its receptor chemokine receptor-2 in human
crescenticglomerulonephritis. J Am Soc Nephrol 2000;11:2231-2242.
12 Burke BS: Macrophages in gene therapy: Cellular delivery
vehicles and in vivo targets. J Leuk Biol2002;72:417-428. 13 Kluth
DC, Erwig LP, Rees AJ: Gene transfer into inflamed glomeruli using
macrophages transfected withadenovirus. Gene Ther 2000; 7:263-270.
14 Yokoo T, Utsunomiya Y, Ohashi T, Imasawa T, Kogure T, Futagawa
Y, Kawamura T, Eto Y, Hosoya T:Inflamed site-specific gene delivery
using bone marrow-derived CD11b^sup +^ CD18^sup +^ vehicle cells
inmice. Hum Gene Ther 1998;9: 1731-1738. 15 Yokoo T, Ohashi T,
Utsunomiya Y, Kojima H, Imasawa T, Kogure T, Hisada Y, Okabe M, Eto
Y, KawamuraT, Hosoya T: Prophylaxis of antibody-induced acute
glomerulonephritis with genetically modified bone marrow-derived
vehicle colls. Hum Gene Ther 1999;10: 2673-2678. 16 Kluth DC,
Ainslie CV, Pearce WP, Clarke D, Anegon I, Rees AJ: Macrophages
transfected with adenovirusto express IL-4 reduce inflammation in
experimental glomerulonephritis. J Immunol 2001;166:4728-4736.
-
17 Yamagishi H, Yokoo T, Imasawa T, Mitarai T, Kawamura T,
Utsunomiya Y: Genetically modified bonemarrow-derived vehicle cells
site specifically deliver an anti-inflammatory cytokine to inflamed
interstitium ofobstructive nephropathy. J Immunol 2001;166:609-616.
18 Wilson HM, Stewart K, Brown PAJ, Anegon I, Chettibi S, Rees AJ,
Kluth DC: Bone marrow-derivedmacrophages genetically modified to
produce IL-10 reduce injury in experimental glomerulonephritis. Mol
Ther2002;6:710-717. 19 Yokoo T, Ohashi T, Utsunomiya Y, Shiba H,
Shen JS, Hisada Y, Eto Y, Kawamura T, Hosoya T:
Inflamedglomeruli-specific gene activation that uses recombinant
adenovirus with the Cre/loxP system. J Am SocNephrol 2001;12:
2330-2337. 20 Yokoo T, Ohashi T, Utsunomiya Y, Shen JS, Hisada Y,
Eto Y, Kawamura T, Hosoya T: Genetically modifiedbone marrow
continuously supplies anti-inflammatory cells and suppresses renal
injury in mouse Goodpasturesyndrome. Blood 2001;98:57-64.
AuthorAffiliation H.M. Wilson D.C. Kluth Department of Medicine and
Therapeutics, Institute of Medical Sciences, Foresterhill,
Aberdeen, Scotland AuthorAffiliation H.M. Wilson Department of
Medicine and Therapeutics Institute of Medical Sciences
Foresterhill, Aberdeen AB25 2ZD (UK) Tel. +44 1224 55835, Fax +44
1224 555766, E-Mail [email protected] MeSH: Adenoviridae --
genetics, Animals, Cell Movement -- physiology, Gene Transfer
Techniques, GeneticEngineering -- methods, Glomerulonephritis --
pathology, Glomerulonephritis -- therapy, Humans,Immunotherapy,
Adoptive -- methods, Macrophages -- virology, Cell Movement --
genetics (utama), KidneyGlomerulus -- pathology (utama), Macrophage
Activation -- genetics (utama), Macrophages -- physiology(utama),
Macrophages -- transplantation (utama) Judul: Targeting genetically
modified macrophages to the glomerulus Pengarang: Wilson, H M;
Kluth, D C Judul publikasi: Nephron Volume: 94 Edisi: 4 Halaman:
e113-8 Tahun publikasi: 2003 Tanggal publikasi: Aug 2003 Tahun:
2003 Bagian: Minireview Penerbit: S. Karger AG Tempat publikasi:
Basel Negara publikasi: Switzerland Subjek publikasi: Medical
Sciences
-
ISSN: 00282766 CODEN: NPRNAY Jenis sumber: Scholarly Journals
Bahasa publikasi: English Jenis dokumen: Journal Article Nomor
aksesi: 12972709 ID dokumen ProQuest: 274309492 URL Dokumen:
http://search.proquest.com/docview/274309492?accountid=166961 Hak
cipta: Copyright S. Karger AG Aug 2003 Terakhir diperbarui:
2014-04-18 Basis data: ProQuest Medical Library
http://search.proquest.com/docview/274309492?accountid=166961
-
Daftar PustakaCitation style: APA 6th - American Psychological
Association, 6th Edition
Wilson, H. M., & Kluth, D. C. (2003). Targeting genetically
modified macrophages to the glomerulus. Nephron,94(4), e113-8.
Retrieved from
http://search.proquest.com/docview/274309492?accountid=166961
_______________________________________________________________
Hubungi ProQuest Hak cipta 2015 ProQuest LLC. Semua hak cipta
dilindungi. - Syarat dan Ketentuan
http://www.proquest.com/go/contactsupporthttp://search.proquest.com/info/termsAndConditions
Targeting genetically modified macrophages to the
glomerulusDaftar Pustaka