© 2000 American Heart Association, Inc.
Cellular Biology |
Transient Induction of Cytokine Production in Human Myocardial Fibroblasts by Coxsackievirus B3
From the Institut für Virologie und Seuchenhygiene (A.H., S.Z., S.W.), Medizinsche Hochschule, Hannover, Germany; Abteilung für Thorax-, Herz-, und Gefässchirurgie (W.R.), Georg-August-Universität Göttingen, Germany; and Department of Medicine (I.M.G.), Section of Hematology/Oncology, University of Illinois at Chicago, Ill.
Correspondence to Dr Albert Heim, Institut für Virologie und Seuchenhygiene, Medizinische Hochschule Hannover, Carl-Neuberg-Strasse 1, D-30625 Hannover, FRG. E-mail ahei{at}virologie.mh-hannover.de
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Abstract |
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Abstract—Cytokine expression in enterovirus infections of the heart may trigger inflammation and have detrimental effects on myocytes. However, the induction of cytokines in human myocardial cells by cardiotropic enteroviruses, for example, Coxsackievirus B3 (CVB3), was not yet demonstrated. Fibroblasts are the predominant cell type of the myocardial interstitium before inflammatory infiltration develops. Hence, we investigated, by enzyme immunoassays, reverse transcription–quantitative polymerase chain reaction (RT-qPCR), and nucleic acid sequence–based amplification (NASBA), whether CVB3 induces cytokine expression in cultured human myocardial fibroblasts. As early as 3 hours after infection, RT-qPCR demonstrated a 2-fold increase of interleukin (IL)–6 and IL-8 mRNA compared with basal transcription, resulting in a significant increase of IL-6 and IL-8 to a median level of 1500 pg/mL (range, 1246 to 1858) and 529 pg/mL (range, 428 to 601) in culture supernatants, respectively. IL-6 and IL-8 expression returned to basal levels within 3 and 5 days, respectively, despite a persistent (carrier-state) CVB3 infection. For comparison, IL-6 and IL-8 were induced in dermal fibroblasts later than 3 days after CVB3 infection. Although the low-level IL-1
transcription of myocardial fibroblasts was not significantly
increased, IL-1 was released from cells to culture supernatants 5
days after infection. Furthermore, a suppression of interferon-ß
transcription was demonstrated up to 24 hours after CVB3 infection of
myocardial fibroblasts by highly sensitive NASBA. In conclusion, our
results demonstrate a heart-specific pattern of a rapid and transient
induction of proinflammatory cytokines after CVB3 infection,
whereas the expression of protective interferon-ß was suppressed
by CVB3.
Key Words: fibroblasts • interferon • interleukin • myocarditis • gene expression
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Introduction |
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Many enterovirus infections are probably subclinical, but infections of the heart with cardiotropic serotypes may cause serious diseases, for example, myocarditis with life-threatening arrhythmias.1 2 Chronic enterovirus myocarditis may result in congestive heart failure, and the only remedy then may be heart transplantation.2 3 Recently, rapid diagnosis of myocardial enterovirus infections was achieved by endomyocardial biopsy with subsequent detection of enterovirus RNA.1 4 5 Despite the progress in diagnosing enterovirus heart disease, its pathogenesis is not yet understood completely. Besides direct lysis of myocytes and induction of autoimmunity to cardiac antigens,6 7 8 9 10 11 induction of cytokine expression by enterovirus replication may be crucial for the pathogenesis of myocarditis and heart failure.12 13 Cytokine expression in the myocardium may result in inflammatory infiltration, autoimmunity, myocyte hypertrophy, fibrosis, and negative inotropic effects.12 13 14 15 Elevated plasma concentrations of interleukin (IL)–1
, IL-1ß, and tumor necrosis factor (TNF)-
were observed in patients suffering from myocarditis.16
Moreover, IL-1, IL-1ß, IL-4, IL-6, IL-8, and TNF- gene
expression was demonstrated by reverse transcription-polymerase chain
reaction (RT-PCR) in the myocardium of patients suffering
from myocarditis or dilated
cardiomyopathy,17 18 but the
cytokine-expressing cells were not yet identified. Immediately after an enterovirus infects the heart, cytokine and chemokine expression by infected myocardial cells may be a first trigger for inflammatory infiltration and immune response. However, it is unknown whether enteroviruses can induce cytokine expression in human myocardial cells. In vivo, an enterovirus infection of both myocytes and myocardial interstitial cells, obviously fibroblasts, was demonstrated by in situ hybridization.1 19 More than 90% of the interstitial cells are fibroblasts before an inflammatory infiltration develops.20 In vitro, cultures of myocardial fibroblasts can be infected with Coxsackievirus B3 (CVB3) but are not lysed as quickly as myocytes, because a persistent (carrier-state type) virus replication evolves.21 Therefore, we suspected that cytokine expression of human myocardial fibroblasts may be significant in enterovirus heart disease. In this study, we demonstrated the induction of proinflammatory cytokines after CVB3 infection of myocardial fibroblasts and the suppression of interferon (IFN)–ß transcription.
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Materials and Methods |
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Cells and Virus
Human myocardial and dermal (FS4) fibroblasts were propagated as described previously.21 CVB3 was generated by transfection from the plasmid CVB3-M1.22 Infectious virus titers (plaque-forming units [PFU]/mL) in culture supernatants of human myocardial fibroblasts were determined by plaque assays.21
Enzyme Immunoassays and Immunocytochemistry
Cytokine concentrations were determined using enzyme
immunoassays for IL-1ß, IL-6, IL-8, soluble IL-6 receptor (sIL-6R),
IFN-ß (Biosource), IL-1, and TNF- (R&D Systems).
Immunostaining for IL-1, IL-6, and IL-8 was
performed as described previously with an alkaline
phosphatase/anti–alkaline phosphatase technique.21
Quantitative PCR (qPCR)
RNA was purified from myocardial fibroblasts, and cDNA was
synthesized. In each cDNA sample, the concentration of a
"housekeeping" gene transcript (GAPDH) was quantified by qPCR
(Cytoxpress kit, Biosource). Before performing IL-1, IL-6, and IL-8
qPCR, all samples were diluted to equal GAPDH cDNA concentrations. IL-6
qPCR was performed using a Cytoxpress kit (Biosource). IL-1 and IL-8
qPCRs were performed with serial 2-fold dilutions of competitive
internal standard DNA ("MIMIC", Clontech) and
analyzed by ethidium bromide–stained agarose gel
electrophoresis. Template concentrations were calculated under the
assumption of equal template-band and MIMIC-band densities for equal
template and MIMIC concentrations.
Nucleic Acid Sequence–Based Amplification (NASBA)
NASBA was performed for sensitive detection of IFN-ß mRNA
using the NASBA amplification kit (Organon Teknika), as described
recently.23
Superinduction of IFN-ß Gene Expression
IFN-ß expression was induced with 10 µg/mL poly-IC
(double-stranded RNA [dsRNA], 100 µg/mL cycloheximide, and 2
µg/mL actinomycin D).24
General Experimental Outline and Statistical Analysis
Briefly, experiments were carried out with 2 different
experimental designs. For the "short-term" design, 16 cultures were
infected with CVB3 with a high (2.0 PFU/cell) multiplicity of infection
(moi), and 16 cultures were mock-infected. At 3, 6, 9, and 24 hours
after infection, supernatants of 4 CVB3-infected and 4 mock-infected
cultures were collected, and cells were trypsinized for RNA extraction.
To determine the influence of CVB3 infection, cytokine
concentrations of CVB3-infected cultures were compared with
time-matched controls. Mann-Whitney tests were performed because of the
heterogeneity of group variances.
For the "long-term" experimental design, 4 cultures were infected
with a low moi (0.1 PFU/cell), and 4 cultures were mock-infected. Media
were changed daily. On day 7, cultures were trypsinized, and RNA was
extracted. Cytokine concentrations were measured in culture
supernatants, and the area under curve (AUC) was calculated. AUC values
of infected cultures were compared with AUC values of controls either
by performing t tests, if F tests did not indicate
heterogeneity of group variances (GraphPad Prism and
Statmate software, version 2.01), or by performing Mann-Whitney tests.
Both experiments (short and long term) were repeated twice with
different preparations of myocardial fibroblasts. For an overall
analysis of the data of long-term experiments, the AUC values
were transformed as follows:
AUCtransformed=AUC/mean
AUCcontrols. Transformed AUCs of CVB3-infected
cultures were compared with transformed AUCs of controls by performing
a t test with Welch correction. Two-sided P
0.05
was regarded as statistically significant.
An expanded Materials and Methods section is available online at http://www.circresaha.org.
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Cytokine Basal Expression
All cultured myocardial fibroblasts stained uniformly positive for IL-6, IL-8, and IL-1
(Figure 1
).
IL-1 staining was associated with the nucleus, whereas IL-6 and IL-8
staining was cytoplasmic (Figure 1). IL-6 and IL-8 were detected
in culture supernatants of all 6 lines of myocardial fibroblasts tested
(both >500 pg/mL after 24 hours), but concentrations varied with each
cell line (compare controls in Figures 2A and 2B with Figure 3). Cytokine concentrations
(Figures 2A and 2B, controls) increased with cell density, as
fibroblasts proliferated from 2x105 cells/well
(
30% confluency, day 1) to 100% confluency on day 7. In contrast
to IL-6 and IL-8, IL-1 concentrations in culture supernatants were
low (0.5 pg/mL), either slightly above or below the sensitivity
threshold of the ELISA, depending on the line of myocardial fibroblasts
and cell density. No basal expression of other cytokines
(IL-1ß, TNF-, sIL-6R, and IFN-ß) was detected in myocardial
fibroblast cultures by enzyme immunoassays.
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Transient Induction of IL-6 and IL-8 by CVB3 Infection
IL-6 and IL-8 concentrations increased rapidly after CVB3
infection of human myocardial fibroblasts (Figures 2A
and 2B).
To determine the influence of CVB3 infection on cytokine
concentrations early after infection, the AUCday
1–4 values of infected cultures were compared with
AUCday 1–4 values of controls, indicating a
significant increase of cytokine concentrations (IL-6, Figure 2A, P=0.026, Mann-Whitney test; IL-8, Figure 2B, P<0.001, t test). Moreover, a
combined analysis of AUCday 1–4 data
(depicted in Figures 2A and 2B) with AUCday
1–4 data from 2 other long-term experiments using
different lines of myocardial fibroblasts (raw data not shown)
confirmed an early increase of IL-6 concentrations
(P=0.0035, t test, n=24) and IL8 concentrations
(P<0.001, t test, n=24) after CVB3 infection.
Despite a carrier-state infection of the cell culture (virus titer in
culture supernatants on day 7, 1.6x105 PFU/mL),
IL-6 and IL-8 concentrations of CVB3-infected cultures returned to
basal expression levels on days 4 and 6, respectively. Subsequently,
IL-6 and IL-8 concentrations of CVB3-infected cultures were even lower
than in mock-infected controls (Figures 2A and 2B). This
indicates a roughly equal cytokine expression per cell in
CVB3-infected and control cultures on day 7, as controls had
proliferated to 100% confluency, whereas cell density remained
unchanged or was slightly reduced in CVB3-infected cultures.
For comparison, human dermal fibroblasts (FS4) were infected with CVB3.
In contrast to human myocardial fibroblasts, a late but persistent
increase of IL-6 (P<0.001, AUC, t test) and IL-8
(P<0.001, AUC, t test) was demonstrated (Figures 2C and 2D).
To evaluate the kinetics of IL-6 and IL-8 induction, human myocardial
fibroblasts were infected in the short-term experimental design with a
high moi (2 PFU/cell) to assure that every cell had contact with the
virus during the incubation period (1 hour). A significant increase of
IL-6 and IL-8 concentrations was noted as early as 3 hours after
infection compared with control cultures (Figure 3). This result
was confirmed with 2 different lines of human myocardial fibroblasts
(data not shown). Furthermore, IL-6 and IL-8 mRNA concentrations were
significantly increased 3 hours (and 6 hours) after infection, as
determined by qPCR, but returned to basal concentrations as
early as 9 hours after infection (Figure 4, Table). As qPCRs
were performed from identical cell numbers and after predilution of
cDNA samples to equal concentrations of a housekeeping gene mRNA, they
could be used to estimate the cytokine expression per cell. The
approximately equal IL-6 and IL-8 mRNA concentrations of infected
cultures and controls on day 7 (Table) confirmed that the higher
IL-6 and IL-8 concentrations of controls on day 7 (Figures 2A and 2B) were a result of a higher cell density, but not of an increased
cytokine production per cell.
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IL-1, IL-1ß, sIL-6R, and TNF-
IL-1 concentrations in culture supernatants were starting to
increase 4 days after CVB3 infection of myocardial fibroblasts compared
with control cultures (Figure 5).
However, IL-1 mRNA concentrations were unchanged in CVB3-infected
cultures (data not shown), indicating an unchanged IL-1 gene
transcription and a release of intracellular IL-1 from lysed cells.
Furthermore, an induction of IL-1ß, sIL-6R, and TNF- expression
was not observed after CVB3 infection of myocardial fibroblasts.
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IFN-ß Expression
As replicative CVB3 dsRNA should induce IFN-ß in fibroblasts, we
searched for IFN-ß expression in myocardial fibroblasts. However,
IFN-ß was not detected in culture supernatants after CVB3 infection
or mock infection (sensitivity 1 IU/mL). To exclude biologically active
IFN-ß concentrations <1 IU/mL in CVB3-infected cultures, an IFN-ß
neutralizing antibody was added to CVB3-infected cultures.
Neither virus titers nor cytokine concentrations were changed,
indicating the absence of biologically active IFN-ß in CVB3-infected
cultures. Moreover, we searched for IFN-ß gene transcription by
highly sensitive NASBA. Thus, IFN-ß gene transcription was
demonstrated in mock-infected controls, but not in CVB3-infected
cultures (moi 2 PFU/cell) up to 24 hours after infection (48 hours
after trypsinization) (Figures 6A and 6B). Forty-eight hours after infection, IFN-ß transcription was
switched off in control cultures, but IFN-ß mRNA was detected in
CVB3-infected myocardial fibroblasts (Figures 6A and 6C). In
contrast to CVB3 infection, superinduction of myocardial fibroblasts by
dsRNA (poly-IC) demonstrated the capability of myocardial fibroblasts
to express IFN-ß (20 IU/mL culture supernatant).
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Discussion |
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TopAbstractIntroductionMaterials and MethodsResultsDiscussionReferences |
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In this study, we demonstrated for the first time that a cardiotropic enterovirus, CVB3, transiently induced proinflammatory cytokines, IL-6 and IL-8, in human myocardial fibroblasts, a simplified but species-specific model to detect both cytokine expression and actions of cytokines.21 25 Moreover, the kinetics of CVB3-induced cytokine expression in myocardial fibroblasts was clearly divergent from dermal fibroblasts (Figure 2
), indicating an organ-specific pattern of
induction.
Previously, the expression of IL-1ß, IL-4, IL-6, IL-8, and TNF-
had been demonstrated by RT-PCR in the myocardium of
patients suffering from enterovirus myocarditis, cryptogenic
myocarditis, and dilated
cardiomyopathy,17 18 but neither the
cytokine-expressing cells (myocardial or infiltrating) nor the
stimulus (virus or secondary to infiltration) had been unequivocally
identified. Recently, Seko et al26 compared the
cytokine expression in murine CVB3 myocarditis with that of
CVB3-infected murine heart cells in vitro. It was concluded that
IL-1, IL-5, IL-6, and IL-7 were predominantly expressed by heart
cells, whereas IL-1ß, IL-2, IL-3, IL-4, IL-10, TNF-, TNF-ß, and
IFN-
were predominantly expressed by inflammatory
cells.26 However, the cytokine-expressing heart
cells were not identified, as heart cell cultures consisted of
myocytes, fibroblasts, and endothelial
cells.26 By contrast, the propagation method used by us
yielded human myocardial fibroblasts cultures that were free of other
heart cells,21 and therefore fibroblasts were identified
as cytokine-producing cells.
In vivo, both cytokine expression and action are supposed to be a complex network of interactions between myocardial cells and infiltrating cells. For example, sIL-6R may be released from infiltrating monocytes27 28 and interact with IL-6 produced by myocardial fibroblasts to induce myocyte hypertrophy.29 Although several cytokines, most importantly IL-1 and TNF, have several detrimental effects on the heart,12 13 14 15 30 IL-6 (if applied early) was protective in murine virus myocarditis by stimulating the immune response.31 Therefore, both the immediate expression of IL-6 and the rapid switch off, which may limit detrimental effects on the myocardium, seem to be an appropriate response of myocardial fibroblasts to a CVB3 infection and may be protective in vivo.
Nevertheless, chronic inflammatory infiltration and tissue destruction
can follow the infection of the heart in vivo, and in vitro a
persistent carrier-state infection of myocardial fibroblasts evolves.
For example, IL-8 produced by fibroblasts may have detrimental effects,
because IL-8 enhances cytopathic effects of enterovirus replication and
suppresses the antiviral activity of IFN-,32 33 which
is active against CVB3 replication in myocardial
fibroblasts.21 Moreover, IL-1 was released from
infected fibroblasts (Figure 5). Although IL-1 is
predominantly an intracellular cytokine, it has the same
activities as IL-1ß if released from cells.34 In vivo,
IL-1 may trigger a postinfectious autoimmune
myocarditis.13
In addition to CVB3-induced expression, we observed a basal expression
of IL-6, IL-8, and IL-1 in myocardial fibroblasts. For comparison,
IL-8 expression was described in explanted healthy donor hearts, but an
IL-1 expression was not detected, and hearts were not tested for
IL-6.18 However, a basal expression of both IL-1 and
IL-6 was detected in heart tissue samples of healthy
mice.26
Interferons have a powerful antiviral activity compared with all other
cytokines. Virus infection and dsRNA are well-established
inducers of interferon production. For example, CVB3 infection
induced interferon effectively (200 IU/mL culture supernatant) in human
monocytes,35 and it was suspected that CVB3 infection
should induce fibroblast IFN, IFN-ß, in human myocardial fibroblasts.
IFN-ß is highly active against CVB3 replication in myocardial
fibroblasts,25 probably because of a Janus kinase (Jak)-1
tyrosine kinase activation, which is not observed after exposure of
myocardial fibroblasts to IFN-.36 However, IFN-ß was
not detected in culture supernatants of CVB3-infected myocardial
fibroblasts either early after infection or during carrier-state CVB3
persistence. As "superinduction" with dsRNA (poly-IC) demonstrated
that myocardial fibroblasts are capable of expressing IFN-ß, our
results suggest that CVB3 has evolved strategies to avoid IFN-ß
induction in myocardial fibroblasts. To confirm this hypothesis, we
searched for low-level IFN-ß gene transcription. In case of
intronless genes, as, for example, the IFN-ß gene, intron-spanning
primer pairs cannot be used to recognize false-positive RT-PCR results
caused by amplification of genomic DNA. NASBA, which is as sensitive as
RT-PCR, permits selective amplification of single-stranded mRNA in a
background of genomic dsDNA.23 IFN-ß mRNA was detected
by NASBA in human myocardial fibroblasts as long as 24 hours after mock
infection (48 hours after trypsinization), which may be nonspecific
"stress" procedures inducing low levels of IFN-ß transcription.
On the other hand, IFN-ß gene transcription was not observed
(sensitivity, 10 copies of mRNA/100 ng cellular RNA) during the first
24 hours after CVB3 infection (Figures 6A and 6B), supporting
the hypothesis that CVB3 has evolved strategies to avoid IFN-ß
induction in myocardial fibroblasts.
However, IFN-ß transcription was detected in CVB3 carrier-state
cultures as late as day 7 after infection, but not in mock-infected
cultures (Figures 6A and 6C). Considering this result, it should
be noted that IFN-ß gene transcription in CVB3
carrier-state–infected cultures does not prove that IFN-ß is
transcribed by infected cells, as only 10% of the cells in a
carrier-state culture are infected.21 For example, IFN-ß
transcription may be suppressed in CVB3-infected cells similar to the
first 24 hours after infection, and IFN-ß may be transcribed by
noninfected fibroblasts of a CVB3-infected carrier-state culture as a
reaction to cell components and factors released by infected cells.
Hence, IFN-ß transcription in late carrier-state infection may be a
reaction of cells to nonviral stimuli, similar to the stress that
induced IFN-ß "basal" transcription shortly after the mock
infection procedure. Analogously, it may be suspected that IL-6 and
IL-8 are induced in noninfected cells after release of a speculative
factor from the infected cells of a culture. However, the rapid
increase of the IL-6 and IL-8 mRNA concentration as early as 3 hours
after infection of a fibroblast culture (Table) strongly
indicates either an increased transcription or a blocked IL-6 and IL-8
mRNA degradation by CVB3 in infected cells.
In conclusion, our results with species-specific heart cells demonstrated the induction of proinflammatory cytokines, but the suppression of protective IFN-ß by a cardiotropic enterovirus, CVB3.
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Acknowledgments |
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This study was supported in part by Organon Teknika (Turnhout, Belgium).
Received September 13, 1999; accepted January 11, 2000.
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References |
|---|
|
TopAbstractIntroductionMaterials and MethodsResultsDiscussionReferences |
|---|
1. Kandolf R, Hofschneider PH. Viral heart disease. Springer Semin Immunopathol. 1989;11:1–13.
2.
Why HJ, Meany BT, Richardson PJ, Olsen EG, Bowles NE,
Cunningham L, Freeke CA, Archard LC. Clinical and prognostic
significance of detection of enteroviral RNA in the
myocardium of patients with myocarditis or dilated
cardiomyopathy. Circulation. 1994;89:2582–2589.
3. Figulla HR, Stille-Siegener M, Mall G, Heim A, Kreuzer H. Myocardial enterovirus infection with left ventricular dysfunction: a benign disease compared with idiopathic dilated cardiomyopathy. J Am Coll Cardiol. 1995;25:1170–1175.[Abstract]
4. Tracy S, Chapman NM, McManus BM, Pallansch MA, Beck MA, Carstens J. A molecular and serologic evaluation of enteroviral involvement in human myocarditis. J Mol Cell Cardiol. 1990;22:403–414.[Medline] [Order article via Infotrieve]
5.
Jin O, Sole MJ, Butany JW, Chia WK, McLaughlin PR, Liu
P, Liew CC. Detection of enterovirus RNA in myocardial biopsies from
patients with myocarditis and cardiomyopathy using
gene amplification by polymerase chain reaction.
Circulation. 1990;82:8–16.
6.
Klingel K, Hohenadl C, Canu A, Albrecht M, Seemann M,
Mall G, Kandolf R. Ongoing enterovirus-induced myocarditis is
associated with persistent heart muscle infection: quantitative
analysis of virus replication, tissue damage, and inflammation.
Proc Natl Acad Sci U S A. 1992;89:314–318.
7. Gauntt CJ, Arizpe HM, Higdon AL, Wood HJ, Bowers DF, Rozek MM, Crawley R. Molecular mimicry, anti-Coxsackievirus B3 neutralizing monoclonal antibodies, and myocarditis. J Immunol. 1995;154:2983–2995.[Abstract]
8.
Cunningham MW, Antone SM, Gulizia JM, McManus BM,
Fischetti VA, Gauntt CJ. Cytotoxic and viral neutralizing antibodies
crossreact with streptococcal M protein, enteroviruses, and human
cardiac myosin. Proc Natl Acad Sci U S A. 1992;89:1320–1324.
9. McManus BM, Chow LH, Wilson JE, Anderson DR, Gulizia JM, Gauntt CJ, Klingel KE, Beisel KW, Kandolf R. Direct myocardial injury by enterovirus: a central role in the evolution of murine myocarditis. Clin Immunol Immunopathol. 1993;68:159–169.[Medline] [Order article via Infotrieve]
10. Badorff C, Noutsias M, Kuhl U, Schultheiss HP. Cell-mediated cytotoxicity in hearts with dilated cardiomyopathy: correlation with interstitial fibrosis and foci of activated T lymphocytes. J Am Coll Cardiol. 1997;29:429–434.[Abstract]
11.
Wessely R, Henke A, Zell R, Kandolf R, Knowlton KU.
Low-level expression of a mutant coxsackieviral cDNA induces a
myocytopathic effect in culture: an approach to the study of
enteroviral persistence in cardiac myocytes. Circulation. 1998;98:450–457.
12.
Kelly RA, Smith TW. Cytokines and cardiac
contractile function. Circulation. 1997;95:778–781.
Editorial.
13.
Lane JR, Neumann DA, Lafond-Walker A, Herskowitz A,
Rose NR. Interleukin 1 or tumor necrosis factor can promote Coxsackie
B3-induced myocarditis in resistant B10.A mice. J Exp
Med. 1992;175:1123–1129.
14.
Gulick T, Chung MK, Pieper SJ, Lange LG, Schreiner GF.
Interleukin 1 and tumor necrosis factor inhibit cardiac myocyte
beta-adrenergic responsiveness. Proc Natl Acad Sci U S A. 1989;86:6753–6757.
15. Palmer JN, Hartogensis WE, Patten M, Fortuin FD, Long CS. Interleukin-1 beta induces cardiac myocyte growth but inhibits cardiac fibroblast proliferation in culture. J Clin Invest. 1995;95:2555–2564.
16.
Matsumori A, Yamada T, Suzuki H, Matoba Y, Sasayama S.
Increased circulating cytokines in patients with myocarditis
and cardiomyopathy. Br Heart J. 1994;72:561–566.
17. Satoh M, Tamura G, Segawa I, Tashiro A, Hiramori K, Satodate R. Expression of cytokine genes and presence of enteroviral genomic RNA in endomyocardial biopsy tissues of myocarditis and dilated cardiomyopathy. Virchows Arch. 1996;427:503–509.[Medline] [Order article via Infotrieve]
18. Han RO, Ray PE, Baughman KL, Feldman AM. Detection of interleukin and interleukin-receptor mRNA in human heart by polymerase chain reaction. Biochem Biophys Res Commun. 1991;181:520–523.[Medline] [Order article via Infotrieve]
19. Koide H, Kitaura Y, Deguchi H, Ukimura A, Kawamura K, Hirai K. Genomic detection of enteroviruses in the myocardium: studies on animal hearts with Coxsackievirus B3 myocarditis and endomyocardial biopsies from patients with myocarditis and dilated cardiomyopathy. Jpn Circ J. 1992;56:1081–1093.[Medline] [Order article via Infotrieve]
20. Eghbali M, Czaja MJ, Zeydel M, Weiner FR, Zern MA, Seifter S, Blumenfeld OO. Collagen chain mRNAs in isolated heart cells from young and adult rats. J Mol Cell Cardiol. 1988;20:267–276.[Medline] [Order article via Infotrieve]
21. Heim A, Brehm C, Stille-Siegener M, Muller G, Hake S, Kandolf R, Figulla HR. Cultured human myocardial fibroblasts of pediatric origin: natural human interferon-alpha is more effective than recombinant interferon-alpha 2a in carrier-state Coxsackievirus B3 replication. J Mol Cell Cardiol. 1995;27:2199–2208.[Medline] [Order article via Infotrieve]
22.
Kandolf R, Hofschneider PH. Molecular cloning of the
genome of a cardiotropic Coxsackie B3 virus: full-length
reverse-transcribed recombinant cDNA generates infectious virus in
mammalian cells. Proc Natl Acad Sci U S A. 1985;82:4818–4822.
23.
Heim A, Grumbach IM, Zeuke S, Top B. Highly sensitive
detection of gene expression of an intronless gene: amplification of
mRNA, but not genomic DNA by nucleic acid sequence based amplification
(NASBA). Nucleic Acids Res. 1998;26:2250–2251.
24. Havell EA, Vilcek J. Production of high-titered interferon in cultures of human diploid cells. Antimicrob Agents Chemother. 1972:2:476–484.
25. Heim A, Stille-Siegener M, Pring-Akerblom P, Grumbach I, Brehm C, Kreuzer H, Figulla HR. Recombinant interferons beta and gamma have a higher antiviral activity than interferon-alpha in Coxsackievirus B3-infected carrier state cultures of human myocardial fibroblasts. J Interferon Cytokine Res. 1996;16:283–287.[Medline] [Order article via Infotrieve]
26. Seko Y, Takahashi N, Yagita H, Okumura K, Yazaki Y. Expression of cytokine mRNAs in murine hearts with acute myocarditis caused by Coxsackievirus b3. J Pathol. 1997;183:105–108.[Medline] [Order article via Infotrieve]
27. Vollmer P, Walev I, Rose-John S, Bhakdi S. Novel pathogenic mechanism of microbial metalloproteinases: liberation of membrane-anchored molecules in biologically active form exemplified by studies with the human interleukin-6 receptor. Infect Immun. 1996;64:3646–3651.[Abstract]
28. Walev I, Weller U, Strauch S, Foster T, Bhakdi S. Selective killing of human monocytes and cytokine release provoked by sphingomyelinase (beta-toxin) of Staphylococcus aureus. Infect Immun. 1996;64:2974–2979.[Abstract]
29.
Hirota H, Yoshida K, Kishimoto T, Taga T. Continuous
activation of gp130, a signal-transducing receptor component for
interleukin 6-related cytokines, causes myocardial
hypertrophy in mice. Proc Natl Acad Sci U S A. 1995;92:4862–4866.
30. Kishimoto C, Kuroki Y, Kurokawa M, Ohiai H. Lymphokine activated killer cells in murine Coxsackievirus B3 myocarditis. Basic Res Cardiol. 1997;92:402–409.[Medline] [Order article via Infotrieve]
31.
Kanda T, McManus JE, Nagai R, Imai S, Suzuki T, Yang D,
McManus BM, Kobayashi I. Modification of viral myocarditis in mice by
interleukin-6. Circ Res. 1996;78:848–856.
32.
Khabar KS, Al-Zoghaibi F, Al-Ahdal MN, Murayama T,
Dhalla M, Mukaida N, Taha M, Al-Sedairy ST, Siddiqui Y, Kessie G,
Matsushima K. The alpha chemokine, interleukin 8, inhibits the
antiviral action of interferon alpha. J Exp Med. 1997;186:1077–1085.
33. Khabar KS, Al-Zoghaibi F, Murayama T, Matsushima K, Mukaida N, Siddiqui Y, Dhalla M, Al-Ahdal MN. Interleukin-8 selectively enhances cytopathic effect (CPE) induced by positive-strand RNA viruses in the human WISH cell line. Biochem Biophys Res Commun. 1997;235:774–778.[Medline] [Order article via Infotrieve]
34.
Dinarello CA. Biologic basis for interleukin-1 in
disease. Blood. 1996;87:2095–2147.
35. Henke A, Mohr C, Sprenger H, Graebner C, Stelzner A, Nain M, Gemsa D. Coxsackievirus B3-induced production of tumor necrosis factor-alpha, IL-1 beta, and IL-6 in human monocytes. J Immunol. 1992;148:2270–2277.[Abstract]
36. Grumbach IM, Fish EN, Uddin S, Majchrzak B, Colamonici OR, Figulla HR, Heim A, Platanias LC. Activation of the Jak-Stat pathway in cells that exhibit selective sensitivity to the antiviral effects of IFN-beta compared with IFN-alpha. J Interferon Cytokine Res. 1999;19:797–801.[Medline] [Order article via Infotrieve]
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