The role of the beta chemokines in experimental obliterative
bronchiolitis
Alexander S. Farivar, Baiya Krishnadasan, Babu V. Naidu, Steven M. Woolley,
and Michael S. Mulligan*
Division of Cardiothoracic Surgery, Department of Surgery, University of Washington Medical Center, Seattle, WA 98195, USA
Received 14 May 2003
Abstract
Beta chemokines have been implicated in cardiac and renal allograft rejection. This study determined if antibody antagonization of beta
chemokines conferred protection against the development of experimental obliterative bronchiolitis (OB) in a heterotopic rat tracheal
allograft model. Rat tracheas were transplanted from Brown-Norway or Lewis donors into Lewis recipients. Rats received 200
g/day of
either anti-RANTES or anti-MCP-1 antibody for 14 days. Luminal obstruction and epithelial loss were calculated. Northern blots for MCP-1
and RANTES mRNA expression were performed, and immunohistochemistry for chemokine protein localization. There was a significant
increase in airway obstruction in allografts compared to isografts (P
0.001). Antibody-treated allografts demonstrated an amelioration of
airway obstruction from 58% (vehicle allografts) to 26% (anti-RANTES) and 12% (anti-MCP-1), both of which were significant (P

0.001). Epithelial preservation was increased in both antibody-treated groups (P
0.001), and increased expression of MCP-1 and RANTES
mRNA was present in tracheal allografts by Day 2 and maximal by Day 6. Beta chemokines are expressed during the development of
experimental OB, as MCP-1 and RANTES mRNA expression increased with time from transplantation. Both MCP-1 and RANTES are
functional in the formation of the fibroproliferative response that characterizes OB in this model, and their antagonization conferred
protection against airway obstruction and epithelial loss.
© 2003 Elsevier Inc. All rights reserved.
Keywords: Beta chemokines; MCP-1; RANTES; Obliterative bronchiolitis; Rat tracheal allografts; Lung transplantation; Heterotopic tracheal transplantation
Introduction
Lung transplantation was introduced into clinical prac-
tice over 20 years ago. Significant improvements in survival
and decreases in morbidity are evident in recent reports
from large centers, with a 3-year survival in the transplanted
population of 70% (Meyer et al., 1999). Despite numerous
improvements in surgical technique, organ preservation so-
lutions, and immunosuppression, the major impediment to
long-term survival in lung and heart-lung transplant recip-
ients is chronic rejection or obliterative bronchiolitis (OB).
OB has been reported to affect 43% of lung transplant
recipients (Sundaresan et al., 1995), and is characterized
clinically by progressive dyspnea, a nonproductive cough,
and reductions in the FEV-1 and midexpiratory flow vol-
umes (Reichenspurner, 1995). Treatment typically consists
of intensification of immunosuppressive therapy or substi-
tution of medications in a standard posttransplant triple
medication protocol. Such therapy is, at best, capable of
only slowing the rate of progression, as OB is typically
progressive and ultimately fatal.
While recent investigations have attempted to define the
mediators involved in the development of OB, experimental
protocols have been limited by the difficulty inherent in
developing a practical and reproducible lung transplant
model. Whole organ transplants are desirable, but studies
are confounded by technical complications and the project
costs can be prohibitive. A technically simpler model for
airway transplantation has gained acceptance over the past 8
years (Hertz et al., 1993). This technique, originally devel-
* Corresponding author. Box 356310, University of Washington Medi-
cal Center, 1959 NE Pacific St., Seattle, WA 98195. Fax: 1-206-543-0325,
E-mail address: msmmd@u.washington.edu (M.S. Mulligan).
R
Available online at www.sciencedirect.com
Experimental and Molecular Pathology 75 (2003) 210–216 www.elsevier.com/locate/yexmp
0014-4800/$ – see front matter © 2003 Elsevier Inc. All rights reserved.
doi:10.1016/S0014-4800(03)00093-5

oped in mice and since adapted to rats (Tazelaar et al.,
1987), produces a lesion that is histologically indistinguish-
able from human OB. Typically, complete airway obstruc-
tion develops by Day 28 following heterotopic implantation
of rat airways into allogeneic recipients.
The beta chemokines are a family of polypeptide mole-
cules that possess proinflammatory activities. Structurally
these proteins are approximately 7–10 kDa and are charac-
terized by four highly conserved cysteine motifs. The first
two conserved cysteine residues in the beta chemokine
family do not have an intervening amino acid and are
therefore designated C–C chemokines. Alpha chemokines
have an intervening amino acid between the first two con-
served cysteine residues and are designated C–X–C chemo-
kines. The C–C chemokines include MCP-1 (monocyte
chemoattractant protein), MCP-2, MCP-3, MIP-1 (macro-
phage inflammatory protein), MIP-1, RANTES (regulated
upon activation, normal T-cell expressed and secreted), and
1-309. These proteins have been characterized over the past
15 years and have murine equivalents known by the same
names (Miller and Krangel, 1992).
Recent interest has focused on the activity of MCP-1 and
RANTES in the pathogenesis of chronic allograft rejection.
Research in both in vivo and in vitro models has implicated
these chemokines in lymphocyte and monocyte chemotaxis.
RANTES was the first C–C chemokine to be characterized
and was found to induce specific and dose-dependent che-
motaxis of CD4 T cells (Schall et al., 1990). Initial inves-
tigations with MCP-1 revealed that it was a potent mono-
cyte chemoattractant (Leonard et al., 1991), and it was later
found to be a powerful chemotaxin for both CD4 and
CD8 cells (Loetscher et al., 1994). In fact, MCP-1 and
RANTES are the most powerful lymphocyte chemotaxins
among the C–C, or beta, chemokines. The activity of the
C–C chemokines on lymphocytes involves adherence to
glycosaminoglycans on the subendothelial cell matrix and
activation of G-proteins (Gilat et al., 1994).
The involvement of the beta chemokines in chronic re-
jection of lung allografts has not yet been studied. Since
both MCP-1 and RANTES are proinflammatory and appear
to participate in the development of chronic allograft rejec-
tion in other organ systems, we performed the present stud-
ies to examine the role of these chemokines in a heterotopic
rat tracheal model of experimental OB.
Materials and methods
Experimental animals
Inbred male Lewis rats and Brown-Norway rats weigh-
ing 250–275 g were obtained from Charles River Labora-
tories Inc. (Portage, MI). These two strains were chosen
since they represented a complete MHC mismatch. Lewis
rats served as donors in the isograft experiments and as
recipients in the allograft and isograft groups. All animals
were cared for in compliance with the Principles of Labo-
ratory Animal Care formulated by the Institute of Labora-
tory Animal Resources and the Guide for the Care and Use
of Laboratory Animals prepared by the National Academy
of Sciences and published by the National Institutes of
Health (NIH Publication No. 86-23, revised 1985).
Experimental surgery
After induction of deep ketamine anesthesia (200–250
mg/kg) and systemic heparinization (50 units), donor ani-
mals were shaved and prepped. A median sternotomy was
performed with extension of the incision into the neck. The
trachea and mainstem bronchi were dissected to include the
first-order branches to the upper and lower lobes. The ex-
planted graft was then placed into cold physiologic saline.
Under sterile conditions, a dorsolateral incision was
made on anesthetized recipients, a subcutaneous pocket was
fashioned, and the underlying fascia was fenestrated over-
lying the flank musculature. Meticulous hemostasis was
maintained throughout the dissection. The graft was secured
in the pocket with two interrupted 6-O sutures to prevent
migration, and the wound was closed with 4-O absorbable
sutures in the deep tissues and nylon on the skin.
The animals were allowed to recover from surgery and
returned to their cages. The wounds were monitored daily
for infection and dehiscence. There were three allograft
groups that received a daily intraperitoneal injection of
either anti-RANTES antibody (200
g), anti-MCP-1 anti-
body (200
g), or an equivalent volume of vehicle (saline).
There was one isograft group that received saline daily. At
the time of graft harvest on Day 14, the animals were
anesthetized with lethal doses of pentobarbital (50 mg/kg)
and explanted grafts were placed into an appropriate pre-
servative solution. Each group contained at least eight ani-
mals.
Reagents
Antibodies against MCP-1 and RANTES were obtained
by immunizing rabbits with recombinant cytokines that had
been expressed in Escherichia coli and purified to homoge-
neity and high specific activity with ion-exchange and gel-
filtration chromatography. Polyclonal rabbit antibodies
were then titered for reactivity to MCP-1 and RANTES by
indirect ELISA. This reactivity was noted to be highly
specific with no cross-reactivity to other tested chemokines
(Bless et al., 2000).
Histological evaluation
Allograft and isograft airways were explanted at post-
transplant Day 14 and fixed in 10% phosphate-buffered
formalin for subsequent sectioning, hematoxylin and eosin
211A.S. Farivar et al. / Experimental and Molecular Pathology 75 (2003) 210–216

(H&E) staining, and examination by light microscopy. The
degree of intraluminal and peritracheal inflammation was
noted.
Computerized morphometry: Airway obstruction
Images of the H&E-stained tracheal sections were taken
with a high-resolution video camera attached to a micro-
scope. These images were imported to a computer and
analyzed using NIH Image software. The percentage of
luminal obstruction in transplanted airways was calculated
in two steps. First the outline of the inner surface of the
cartilage was traced. The line representing the membranous
trachea was drawn straight by connecting the two ends of
the cartilage. In the second step the cursor was used to trace
the inner surface of the actual residual lumen. The cross-
sectional area of the actual residual lumen was subtracted
from the area contained within the inner circumference of
the cartilage. This value was then divided by the area within
the cartilage to determine the degree of luminal obstruction.
The formula for calculating the percentage airway obstruc-
tion is expressed as follows:
percentage obstruction 
area within cartilage  area of residual lumen
area within the cartilage  100.
In normal unmanipulated airways the respiratory epithelium
and submucosa lie within the cartilaginous rings. Therefore,
using this calculation, normal airways will demonstrate a
“baseline airway obstruction” of about 3%. Multiple sec-
tions (n  10) were taken from the middle 1 cm of each
graft, and the mean calculated percentage obstruction was
determined.
Computerized morphometry: Epithelial preservation
Using the same computerized imaging program, H&E-
stained sections of the transplanted airways were examined.
The inner circumference of the original airway was deter-
mined and the cursor was used to trace areas of intact
normal respiratory epithelium, variant epithelium, or absent
epithelium. The percentage of the original circumference
was then determined for each of these three possibilities.
Once again, numerous sections (n  10) were taken from
the central portion of each graft and mean values were
determined.
Northern blot analysis
Additional allografts and isografts were harvested at
Days 2 and 6 following transplantation. Airways were dis-
sected, explanted, and snap-frozen in liquid nitrogen. RNA
was extracted using a phenol-chloroform extraction method.
Cytoplasmic RNA was fractionated electrophoretically in a
1% formaldehyde gel and transferred to a nylon blot mem-
brane. Blots were then probed with a high-affinity primer
specific for RANTES and MCP-1.
Immunohistochemistry (IHC)
Additional sections of allograft airways were harvested
at Day 6 following transplantation and embedded in OCT
compound in order to localize chemokine protein. Frozen
sections were obtained and mounted on poly-L-lysine-
coated slides. These slides were fixed in methanol contain-
ing 0.3% hydrogen peroxide to quench endogenous perox-
idase activity. Slides were then washed with PBS and
incubated with purified anti-RANTES or anti-MCP-1 IgG.
Following a 45-min incubation period, the slides were
washed with PBS and stained for antibody using the biotin-
avidin-peroxidase system from rat IgG (Vectastain; Vector
Laboratories, Inc., Burlingame, CA). After H&E counter-
staining, sections were coated with Permount (Lemmer
Laboratories, Pittsburgh, PA.) and examined by light mi-
croscopy for RANTES or MCP-1 protein. This was dem-
onstrated by the presence of reddish brown reaction prod-
ucts on hematoxylin-counterstained sections.
Statistical analysis
Standard deviation and standard error of the mean were
calculated for each group. Comparisons between groups
were accomplished using Student’s t test. Differences were
considered statistically significant at P
0.05.
Results
Luminal obstruction
In isografts harvested at 14 days the cross-sectional area
of the tracheal lumen was narrowed by approximately 3%.
As noted previously, the analytical technique rather than a
specific posttransplant response accounted for this narrow-
ing. In the saline-treated allografts harvested at 14 days, the
degree of luminal obstruction was 58  5% (Fig. 1). The
difference between the occlusion noted in the isografts and
allografts was statistically significant (P
0.001). In con-
trast, the luminal obstruction in allografts treated with 200

g/day of anti-RANTES (Fig. 2) or anti-MCP-1 (Fig. 3)
antibody was 26 2.7 and 12 1.6%, respectively (Fig. 4).
The decrease in obstruction noted in both antibody-treated
groups was significantly less (P
0.001) than that seen in
the saline-treated allografts.
Epithelial loss
Isografts examined at 14 days demonstrated near com-
plete preservation of the respiratory epithelium. Only 0.6 
212 A.S. Farivar et al. / Experimental and Molecular Pathology 75 (2003) 210–216

0.1% was lost, with 96  0.7% of the epithelium appearing
normal and 3  0.7% variant. In contrast, the saline allo-
grafts harvested at 14 days revealed a 98  0.6% loss of
epithelium. Furthermore, of the 2% of epithelium that re-
mained, all was morphologically variant.
In the allografts treated with anti-MCP-1 antibody, epi-
thelial loss was 54  3.7%. Twenty-one  3.6% of the
airway was lined with variant epithelium, while normal
epithelium was present in 24  4% of the specimen. Sim-
ilarly, in animals treated with anti-RANTES antibody, epi-
thelial loss was 54  2.5%, with 23  2.9% of the epithelial
lining variant and 22.6  1.9% normal (Fig. 5). Treatment
with anti-MCP-1 and anti-RANTES antibody was highly
protective in preserving the airway epithelium, and in both
cases statistically significant (P
0.001).
Immunohistochemistry
IHC was performed to determine the cellular location of
chemokine proteins. In analyzing the rat tracheal allografts
at Day 6 posttransplant for both MCP-1 and RANTES,
chemokine protein clearly localized to the subepithelial
layer of the allografts. A representative picture for MCP-1 is
shown (Fig. 6).
Northern blot analysis
MCP-1 mRNA increased progressively after Day 2 in
harvested allografts. RANTES mRNA was also upregu-
lated, but less markedly than seen with MCP-1. The expres-
sion for both was detectable at Day 2 on Northern blot and
Fig. 1. Photograph of an H&E-stained saline-treated tracheal allograft at
Posttransplant Day 14. Note the fibroproliferative response within the
lumen leading to partial occlusion, which on average was 58% in this
experimental group.
Fig. 2. Photograph of an H&E-stained anti-RANTES-treated tracheal al-
lograft at Day 14. There was a statistically significant decrease in airway
obstruction, as well as a preservation of respiratory epithelium when
compared to saline-treated allografts.
Fig. 3. Photograph of an H&E-stained anti-MCP-1-treated tracheal allo-
graft at Day 14. Treatment with anti-MCP-1 antibody afforded a greater
degree of protection from luminal obstruction, but similar preservation of
epithelium as compared to the anti-RANTES-treated allografts.
Fig. 4. There was a statistically significant decrease (P
0.001) in airway
obstruction in both the anti-RANTES and anti-MCP-1-treated allografts
when compared to untreated allografts.
213A.S. Farivar et al. / Experimental and Molecular Pathology 75 (2003) 210–216

maximal by Day 6. The isografts did not demonstrate in-
creased mRNA expression for either MCP-1 or RANTES
regardless of the time point examined.
Discussion
Since the original description of RANTES in 1988
(Schall et al., 1988) and the subsequent characterization of
MCP-1 (Matsushima et al., 1989), research and interest in
chemokines have blossomed. In vitro experiments have
attempted to characterize both the molecular mechanisms
and the target cells involved in RANTES and MCP-1 acti-
vation. Although considerable progress has been made in
elucidating the mechanism mediating the activity of these
chemokines, there still persist fundamental questions about
the nature of these proteins. For example, it is believed that
the target cells for RANTES are T cells of the memory
phenotype (Bacon et al., 1995). However, recent experi-
ments suggest that CD8 cells may be the primary cells
activated by RANTES (Koga et al., 1999). The mechanism
by which the beta chemokines activate cells is also an area
of intense research. It is clear that G proteins are involved in
chemokine signaling and that a subsequent increase in in-
tracellular calcium is produced through a signal transduc-
tion pathway. However, the nature of the precise molecular
events mediating these changes awaits further research.
Chronic inflammatory diseases including rheumatoid ar-
thritis (Koch et al., 1992) and idiopathic pulmonary fibrosis
(Antoniades et al., 1992) have been linked to the production
of chemokines. Recent animal studies have also implicated
chemokines in allograft rejection. Chronic rejection of solid
organ transplants is a problem for the majority of allograft
recipients. The cells that mediate chronic rejection, namely
T cells and monocytes, are also the target cells of the
chemokines.
In the allografts that received RANTES antibody, the
percentage of luminal obstruction was nearly half that of
saline-treated allografts, reflecting a significant attenuation
of posttransplant airway occlusion. There was also en-
hanced preservation of airway epithelium, half of which
appeared normal. In contrast, saline-treated allografts ex-
hibited loss of the entire respiratory epithelium. Many
groups have investigated the expression of RANTES
mRNA and the localization of RANTES protein using im-
munohistochemistry. In skin allografts, the expression of
RANTES appears to occur late in the rejection process
(Kondo et al., 1997) and to be associated with CD8 T
cells. (Koga et al., 1999) A similar pattern of late RANTES
expression has also been observed in cardiac allografts
(Fairchild et al., 1997) and renal allografts (Pattison et al.,
1994). Despite a relative wealth of descriptive studies, few
interventional studies have reported the effects of chemo-
kine blockade in vivo. A recent study, utilizing a murine
kidney transplant model, antagonized endogenous RAN-
TES with a methionine-labeled RANTES and observed a
decrease in vascular and tubular injury in the met-RAN-
TES-treated allografts (Grone et al., 1999).
RANTES expression during this experiment occurred
early and was persistent throughout the rejection process, as
revealed by Northern blot. Possible reasons for the discrep-
ancy between RANTES production in this model and the
others referenced may relate to organ-specific expression of
Fig. 5. While untreated allografts retained only 2% of respiratory epithelium at 14 days, all of which was variant in morphology, both antibody-treated groups
exhibited significant increases in preserved epithelium, half of which were normal appearing.
Fig. 6. In the development of the OB lesions, MCP-1 protein was strongly
evident at Posttransplant Day 6 at the interface between the inner surface
of the cartilage and the advancing front of fibrovascular connective tissue.
Arrows depict areas of most intense staining.
214 A.S. Farivar et al. / Experimental and Molecular Pathology 75 (2003) 210–216

chemokines. Additionally, revascularization of the tracheal
allografts likely occurs at a different pace than that seen in
skin and heart allografts. The fact that RANTES mRNA was
detectable as early as Day 2 suggests that the source is likely
the infiltrating recipient leukocytes. As revascularization
proceeds, a graft-derived source becomes more likely.
In a murine renal transplant model of chronic rejection,
Nadeau and colleagues found that RANTES was the first
inducible marker of rejection and peaked 2 weeks following
rejection (Nadeau et al., 1995). Such a delayed pattern of
expression could be consistent with a regulatory participa-
tion in chronic allograft rejection responses.
The animals treated with antibody to MCP-1 also dem-
onstrated a significant decrease in the percentage of luminal
obstruction in comparison to the allograft controls and the
RANTES antibody-treated animals. However, the amount
of normal epithelium preserved, although more than the
saline-treated allografts, was equivalent to the anti-RAN-
TES-treated animals. MCP-1 has been extensively investi-
gated in allograft rejection, particularly in kidney transplan-
tation. In Europe, investigators have assayed the levels of
MCP-1 in the urine of human kidney recipients to monitor
for rejection (Prodjosudjadi et al., 1996). MCP-1 has also
been localized to monocytes associated with allograft rejec-
tion in a murine model of heart transplantation (Russell et
al., 1993). The primary target cell for MCP-1 has consis-
tently been the monocyte, although studies of kidney allo-
graft rejection have also localized MCP-1 to tubular epithe-
lial cells. In the development of the airway lesions in this
model, MCP-1 protein was strongly evident at Posttrans-
plant Day 6 at the interface between the inner surface of the
cartilage and the advancing front of fibrovascular connec-
tive tissue.
The expression of MCP-1 mRNA appears to occur early
in the rejection process. In the aforementioned cardiac al-
lograft model (Fairchild et al., 1997), MCP-1 mRNA was
detected with the first tissue examination at 7 days and
detectable levels persisted throughout the rejection process.
A similar pattern for MCP-1 was observed in murine skin
grafts where the MCP-1 equivalent in the murine model, JE,
was noted to be elevated within 3 days following transplan-
tation (Kondo et al., 1997) All of these models describe an
influx of monocytes following the increase in MCP-1
mRNA. This pattern of early and persistent expression of
MCP-1 was also evident in the Northern blots done on
allografts in this model.
The expression of the beta chemokines during tracheal
allograft rejection suggests that infiltration of the graft by
monocytes is partially mediated through the action of
MCP-1. The blockade of MCP-1 activity with a monoclonal
antibody specific for MCP-1 decreased the percentage of
luminal obstruction and preserved the airway epithelium.
RANTES may also help to mediate this phase of rejection
that primarily involves both monocytes and lymphocytes.
This paper reports a pattern of chemokine expression for
RANTES and MCP-1 that supports previous efforts in re-
nal, cardiac, and skin allograft rejection models. The allo-
geneic response in the airways is pathologically similar to
chronic cardiac allograft atherosclerosis and vanishing bile
duct syndrome in liver allografts. Previous data demonstrat-
ing roles for chemokines in fibrotic and inflammatory lung
injury coupled with the present findings in our heterotopic
tracheal allograft model suggest that MCP-1 and RANTES
are likely phlogistic mediators involved in the development
of posttransplant obliterative bronchiolitis.
References
Antoniades, H.N., Neville-Golden, J., Galanopoulos, T., et al., 1992. Ex-
pression of monocyte chemoattractant protein 1 mRNA in human
idiopathic pulmonary fibrosis. Proc. Natl. Acad. Sci. USA 89, 5371–
5375.
Bacon, K.B., Premack, B.A., Gardner, P., et al., 1995. Activation of dual
T cell signaling pathways by the chemokine RANTES. Science 269,
1727–1730.
Bless, N.M., Huber-Lang, M., Guo, R.F., et al., 2000. Role of C-C che-
mokines in acute lung injury in rats. J. Immunol. 164, 2650–2659.
Fairchild, R.L., Van Buskirk, A.M., Kondo, T., et al., 1997. Expression of
chemokine genes during rejection and long term acceptance of cardiac
allografts. Transplantation 63, 1807–1812.
Gilat, D., Hershovitz, R., Mekori, Y.A., 1994. Regulation of adhesion of
CD4 T lymphocytes to intact or heparinase treated subendothelial
extracellular matrix by diffusible or anchored RANTES and MIP-1.
J. Immunol. 153, 4899–4906.
Grone, H.J., Weber, C., Weber, K.S., et al., 1999. Met- RANTES reduces
vascular and tubular damage during acute renal transplant rejection:
blocking monocyte arrest and recruitment. FASEB J. 13, 1371–1383.
Hertz, M.I., Jessurun, J., King, M., 1993. Reproduction of obliterative
bronchiolitis lesion after heterotopic transplantation of mouse airways.
Am. J. Pathol. 142, 1945–1951.
Koch, A.E., Kunkel, S.L., Harlow, L.A., et al., 1992. Enhanced production
of monocyte chemoattractant protein 1 in Rheumatoid Arthritis. J. Clin.
Invest. 90, 772–779.
Koga, S., Novick, A.C., Toma, H., et al., 1999. CD8 cells produce
RANTES during acute rejection of murine allogeneic skin grafts.
Transplantation 67, 854–864.
Kondo, T., Watarai, Y., Novick, A.C., et al., 1997. T cell dependent
acceleration of chemoattractant cytokine gene expression during sec-
ondary rejection of allogeneic skin grafts. Transplantation 63, 732–734.
Leanord, E.J., Skeel, A., Yoshimura, T., 1991. Biological aspects of mono-
cyte chemoattractant protein 1. Adv. Exp. Med. Biol. 305, 67–74.
Loetscher, P., Seitz, M., Clark-Lewis, I., et al., 1994. Monocyte chemo-
tactic proteins: MCP-1, MCP-2, and MCP-3 are major chemoattrac-
tants for CD4 and CD8 cells. FASEB J. 8, 1055–1060.
Matsushima, K.C., Larsen, C.G., Dubois, G.C., et al., 1989. Purification
and characterization of a novel monocyte chemotactic and activating
factor produced by a human myelomonocytic cell line. J. Exp. Med.
169, 1485–1456.
Meyer, B.F., Lynch, J., Trulcock, E.P., et al., 1999. Lung transplantation:
a decade of experience. Ann. Surg. 230, 362–367.
Miller, M.D., Krangel, M.S., 1992. Biology and biochemistry and the
chemokines: a family of chemotactic and inflammatory chemokines.
Crit. Rev. Immunol. 12, 17–46.
215A.S. Farivar et al. / Experimental and Molecular Pathology 75 (2003) 210–216

Nadeau, K.C., Azuma, H., Tilney, N.L., et al., 1995. Sequential cytokine
dynamics in chronic rejection of rat renal allografts: role of cytokines
RANTES and MCP-1. Proc. Natl. Acad. Sci. USA 92, 8729–8733.
Pattison, J., Nelson, P.J., Huie, P., et al., 1994. RANTES chemokine
expression in cell mediated transplant rejection of the kidney. Lancet
343, 209–211.
Prodjosudjadi, W., Daha, M.R., Gerritsma, J.S., et al., 1996. Increased
urinary excretion of monocyte chemoattractant protein-1 during acute
renal allograft rejection. Nephrol. Dial. Transplant. 11, 1096–1103.
Reichenspurner, H., Adams, B., Soni, V., et al., 1995. Obliterative bron-
chiolitis after lung and heart-lung transplantation. Ann. Thorac. Surg.
60, 1845–1853.
Russell, M.E., Adams, D.H., Wyner, L.R., et al., 1993. Early and persistent
induction of monocyte chemoattractant protein-1 in rat cardiac allo-
grafts. Proc. Natl. Acad. Sci. USA 90, 6086–6090.
Schall, T.J., Bacon, K., Toy, K.J., et al., 1990. Selective attraction of
monocytes and T lymphocytes of the memory phenotype by cytokine
RANTES. Nature 347, 669–671.
Schall, T.J., Jongstra, J., Dyer, J.D., et al., 1988. A human T cell-specific
molecule is a member of a new gene family. J. Immunol. 141, 1018–
1025.
Sundaresan, S., Trulock, E.P., Mohanakumar, T., et al., 1995. Prevalence
and outcome of bronchiolitis obliterans after lung transplantation. Ann.
Thorac. Surg. 60, 1341–1347.
Tazelaar, H.D., Prop, P., Nieuwenhuis, P., et al., 1987. Obliterative bron-
chiolitis in the transplanted rat lung. Transplant. Proc. 19, 1052–1061.
216 A.S. Farivar et al. / Experimental and Molecular Pathology 75 (2003) 210–216