| HOME | HELP | FEEDBACK | SUBSCRIPTIONS | ARCHIVE | SEARCH | TABLE OF CONTENTS |
|
|
|
|||||||
|
|||||||||
Diabetes Research Institute, Heinrich-Heine-University, Düsseldorf, Germany
 |
Abstract |
|---|
 Top Abstract IntroductionMaterials and MethodsResultsDiscussionReferences |
|---|
; mouse macrophages in addition produce nitric oxide.
The proinflammatory macrophage response is hsp60 dose dependent and
similar in kinetics and extent to LPS stimulation. Human hsp60 was
found to synergize with IFN-
in its proinflammatory activity.
Finally, human hsp60 induces gene expression of the Th1-promoting
cytokines IL-12 and IL-15. These findings identify autologous hsp60 as
a danger signal for the innate immune system, with important
implications for a role of local hsp60 expression/release in chronic
Th1-dependent tissue inflammation. |
Introduction |
|---|
|
TopAbstractIntroductionMaterials and MethodsResultsDiscussionReferences |
|---|
In recent years, evidence has been accumulating for an important role of the innate immune system in initiating and guiding responses of the adaptive immune system, as carried by T and B cells 15, 16, 17, 18, 19 ; therefore, we wondered whether the innate immune system may recognize endogenous hsp60 and respond to it.
In the present study, we analyzed the outcome of interactions between mammalian hsp60 and mouse or human macrophages. To our surprise, we observed that human hsp60 has potent immunostimulatory properties, when added to macrophage cultures. Hsp60 thus may serve as danger signal to the innate immune system when expressed on or released from stressed autologous cells.
|
Materials and Methods |
|---|
|
TopAbstractIntroductionMaterials and MethodsResultsDiscussionReferences |
|---|
The mouse macrophage cell line J774 A.1 was purchased from German Collection of Microorganism and Cell Culture (Braunschweig, Germany). J774 cells were cultured in RPMI 1640 medium (Sigma, Deisenhofen, Germany) supplemented with 10% (v/v) FCS (Life Technologies, Eggeustein, Germany), ampicillin (25 mg/L), penicillin (120 mg/L), streptomycin (270 mg/L), 1 mM sodium pyruvate, 2 mM L-glutamine, nonessential amino acids (10 ml/L, 100x), 24 mM NaHCO3, and 10 mM HEPES.
Mouse bone marrow-derived macrophages
Mice of both sexes of C57BL/6JBom mice purchased from Breeding & Research Center A/S (Bomholtgård, Ry, Denmark) were used. Bone marrow cells were obtained by flushing femurs and tibias with ice-cold PBS under sterile conditions and washed by centrifugation (500 x g, 10 min). A total of 2.5 x 106 bone marrow cells was incubated in tissue culture dishes with 10 ml of Pluznik medium (5% heat-inactivated horse serum, 15% FCS, 15% L929 cell-conditioned medium 20 , and 65% RPMI 1640 supplemented as described above) at 37°C in humidified 5% CO2 in air. After 7 or 8 days, adherent bone marrow-derived macrophages were detached by incubation with ice-cold Ca2+-Mg2+-free HBSS for 10 min, followed by twice washing with HBSS (500 x g, 5 min).
Mono Mac 6 cell
Mono Mac 6, a human monocyte line 21 , was kindly provided by Dr. H. W. Ziegler-Heitbrock (Institute for Immunology, University of Munich, Munich, Germany). Mono Mac 6 cells were cultured in RPMI 1640 medium containing the OPI (oxaloacetate, pyruvate, insulin) supplement (Sigma), 2 mM L-glutamine, antibiotics (120 mg/L penicillin and 200 mg/L streptomycin), and 10% FCS.
Stimulation of macrophage
For analysis of TNF- or nitrite production, mouse macrophages
were adjusted to a density of 1 x 106/ml and placed
on a flat-bottom 96-well plate (200 µl/well). Mono Mac 6 cells were
cultured at a concentration of 2 x 106/ml in 24-well
plates (total volume 500 µl). For analysis of cytokine gene
expression, 1.5 x 106 of macrophages per 2 ml were
seeded in 12-well plates. After incubation at 37°C and 5%
CO2 for 18 h, macrophages were incubated in the
presence or absence of recombinant human hsp60 (StressGen
Biotechnologies, Victoria, Canada), Escherichia coli LPS
(Sigma), or recombinant mouse IFN- (Sigma). To test for endotoxin
contaminations, 0.1 µg/ml polymyxin B sulfate (PmB; Sigma) was added
to cell cultures. At the end of experiments, culture supernatants were
collected and stored at -20°C until further analyses.
TNF- measurements
TNF- released into culture supernatants was determined by
sandwich ELISA, as described previously 22 . A 96-well
NUNC-Immuno-plate (Nunc, Wiesbaden, Germany) was coated overnight at
4°C with 50 µl rat anti-mouse TNF- mAb (5 µg/ml) or mouse
anti-human TNF- mAb (2 µg/ml) (PharMingen, San Diego, CA)
diluted with coating buffer (0.1 mM NaHCO3, pH 8.2). After
discarding the coating solution and two washes with washing buffer
(PBS/0.05% Tween-20, pH 7.4), the wells were blocked with 250 µl of
blocking buffer (PBS/2% (w/v) milk powder) for 1 h at 37°C. The
plates were washed twice with washing buffer thereafter. Binding of
TNF- was performed by incubation of 50 µl of culture supernatants
diluted 1/5- and 1/10-fold with PBS/2% (w/v) milk powder or serial
standard dilutions of rTNF- (Genzyme, Kent, U.K.) for 2 h at
37°C. Subsequently, the wells were washed four times. A total of 50
µl of matching biotinylated rat mAb against mouse TNF- (2 µg/ml)
or biotinylated mouse mAb against human TNF- (1 µg/ml) was placed
in each well. The plates were incubated for 1 h at 37°C. After
extensive washing, the samples were incubated for 45 min at 37°C with
50 µl of 1 µg/ml peroxidase-conjugated avidin (Dianova, Hamburg,
Germany) in blocking buffer and subsequently washed six times. A total
of 100 µl of 1 mM ABTS
(2,2'-azino-bis(3-ethylbenzo-thiazoline-6-sulfonic
acid)diammonium salt)/0.1 M citrate buffer, pH 4.35, containing 0.02%
H2O2 was added to each well for determining the
residual peroxidase activity as a measure of Abs bound. After 15–30
min of incubation at 37°C, the OD was measured at 405 nm against a
reference wavelength of 492 nm on a microplate reader. The TNF-
content was calculated by using a standard curve of rTNF- with RPMI
1640 medium alone as a blank.
Measurement of nitrite production
The amount of nitrite (NO2-) released by macrophages was detected in cell-free supernatants by the colorimetric Griess reaction, as described previously 23 . Briefly, 50 µl of the supernatant and serial dilutions of NaNO3 standard solution was placed in 96-well plastic plate, and then mixed with equal volume of Griess reagent containing 1% sulfanilamide, 0.1% naphthylethylene-diamide-dihydrochloride, and 2.5% H3PO4. After incubation for 10 min at room temperature, the OD of reaction products reflecting the concentration of NO2- was assessed at 550 nm on a Microplate reader. The results were expressed in micromoles of NO2- per ml.
Cytokine mRNA analysis
RNA was isolated immediately after termination of the experiments by adding 1 ml Trizol-Reagent (Life Technologies) to the culture plates, and following the procedure suggested by the manufacturer. Determination and quantification of specific mRNA were performed by RT-PCR, as described elsewhere 24 . In brief, first-strand cDNA synthesis was performed using the Life Technologies RT-PCR kit. Specific primers for ß-actin, IL-12 p40, and IL-12 p35 were as described previously 25 . The specific primers for IL-15 were as follows: 5' primer, 5'-CCATCTCGTGCTACTTGTGT-3'; 3' primer, 5'-CTGTTTGCAAGGTAGAGCAC-3'. A total of 4-µl aliquots of cDNA was added to 23.4-µl reaction mixture containing MgCl2 (1.25 mM final concentration, for IL-15, only 0.8 mM), all four dNTPs (4 mM), and specific primers for IL-12p40, IL-12p35, IL-15, and ß-actin. After hot start at 78°C, 1 U of Taq polymerase was added to the reaction pool. The first cycle of each PCR was started after 3 min at 95°C of denaturation time. In the next cycles, denaturation was performed for 1 min. Primer annealing was done for 1 min at 60°C (ß-actin), 58°C (IL-12p35), 55°C (IL-12p40), and 45°C (IL-15). The elongation steps were done at 72°C for 80 s (ß-actin), 40 s (IL-12p35), 30 s (IL-12p40), and 45 s (IL-15). The PCR products were separated on a 2% agarose gel in 0.5x TBE buffer (Tris, boric acid, EDTA). PCR products were blotted onto nylon membrane, followed by hybridization with specific 32P-labeled probes binding at sites between the primer sequences. Signals were quantified by measuring 32P-stimulated luminescence (PSL) with a phosphor imager. Relative PSL of cytokine mRNA was calibrated to the strength of ß-actin signal, which was set equal to 1. Controls verified a linear relationship between mRNA quantity analyzed and signals obtained.
Statistical analysis
Data were expressed as mean ± SEM. Statistical analysis was performed using the Student’s t test, two-sided. Differences were considered statistically significant with p < 0.05.
|
Results |
|---|
|
TopAbstractIntroductionMaterials and MethodsResultsDiscussionReferences |
|---|
production by human hsp60 in mouse macrophages
When mouse macrophages J774 were exposed to 10 ng/ml of LPS for
7 h, substantial amounts of TNF- were detected in culture
supernatants. Culture of cells with 10 µg/ml of human hsp60 elicited
a TNF- response of similar magnitude (Fig. 1
A). TNF- secretion in
response to human hsp60 was found dose dependent, with 1 µg/ml being
the lowest effective concentration. Neutralization of LPS by 0.1
µg/ml PmB completely suppressed the TNF- response to LPS, but not
to hsp60 (Fig. 1A). The suppression of the response to LPS
by PmB was observed down to a concentration of 0.01 µg PmB/ml. There
was no suppression of the response to human hsp60 by 0.01–0.1 µg/ml
PmB. Only the highest concentration is depicted in Fig. 1A.
This finding ruled out a contribution of endotoxin to the
macrophage-stimulatory property of human hsp60. Heat treatment
destroyed hsp60 activity (not shown).
|
production induced by human
hsp60, we analyzed the amounts of TNF- produced by J774 cells after
2, 7, 24, and 48 h of incubation. Fig. 1
B illustrates
the kinetics of TNF- production in response to human hsp60. TNF-
was demonstrable in culture supernatants as early as 2 h after
addition of human hsp60 (Fig. 1B). After stimulation with
the highest hsp60 concentration, TNF- production peaked at 24
h, with TNF- still demonstrable at 48 h. With the lower dose of
hsp60, peak levels were seen earlier, at 7 h, and TNF-
concentrations were close to control levels already by 24 h of
culture. The kinetics of the TNF- response to LPS was similar to the
one seen with 10 µg/ml human hsp60 (Fig. 1B). Induction of NO2- production by human hsp60 in mouse macrophages
To determine whether human hsp60 affected inducible NO formation
in mouse macrophages, experiments were conducted examining the effect
of human hsp60 on NO2- production, including dose
dependency and kinetics. As shown in Fig. 2A, J774 cells treated with 1
and 10 µg of human hsp60/ml released significant amounts of
NO2- after incubation for 24 h (8.4 ± 0.6
and 13.5 ± 0.5 µM, respectively; p < 0.001
compared with the control group). At a concentration of 0.1 µg/ml
hsp60/ml, NO2- formation was not induced. J774 cells
treated with 10 ng/ml LPS also responded with nitrite production at
levels comparable with those seen for human hsp60 (Fig. 2A).
The stimulatory effect of human hsp60 could not be inhibited by the
addition of PmB, while the effect of 10 ng/ml of LPS was almost
neutralized by the same amount of PmB.
|
A,
demonstrating the biological variability of the nitrite response (Fig. 2B).
Human hsp60 also stimulates primary mouse macrophages and
synergizes with IFN-
Next, we analyzed whether the results obtained with a transformed
mouse macrophage line could be reproduced in primary cultures of mouse
macrophages. Bone marrow-derived macrophages from C57BL/6 mice were
stimulated with human hsp60 or LPS, as above. As shown in Fig. 3A, treatment with 1 or 10
µg/ml of human hsp60 elicited dose-dependent formation of nitrite.
Again, the response was similar as induced by 10 ng/ml LPS. Incubation
of macrophages with IFN- induced less nitrite than in response to
hsp60. There was a strong synergistic effect between human hsp60 and
IFN- on nitrite production. Culture of macrophages with 1 µg of
hsp60/ml plus IFN- also resulted in significantly enhanced
production of nitrite (32.8 ± 1.1 µM) compared with that
produced by 1 µg of hsp60/ml (3.3 ± 0.5 µM) or by IFN-
alone (3.6 ± 0.1 µM, Fig. 3B; p <
0.001). The synergistic effect was also observed in the presence of
PmB, excluding a contribution of endotoxin (Fig. 3B).
|
To determine whether the stimulatory properties of human hsp60 are
confined to a xenogeneic system, we also analyzed the response of a
human monocyte line, Mono Mac 6. Fig. 4A shows that human hsp60 dose
dependently induced the production of TNF- in Mono Mac 6 cells. The
two higher concentrations of human hsp60 (3 and 10 µg/ml) induced
significant TNF- production in Mono Mac 6 (p
< 0.001 and p < 0.0001, respectively, compared with
the control group). Slightly elevated levels of TNF- were also found
in cultures with 1 µg/ml hsp60 (p < 0.05).
The amount of TNF- produced by macrophages incubated with 0.1 µg
of hsp60 did not differ from that produced by the controls
(p > 0.05).
|
within 2
h of exposure to human hsp60 (Fig. 4B). Peak levels were
reached at 6 h, and levels declined rapidly thereafter. These
kinetics were similar to the ones obtained for stimulation with LPS
(Fig. 4B). Induction of IL-12 gene expression by human hsp60
In a further series of experiments, we determined whether human
hsp60 might not only induce mediators characteristic of nonspecific
immune defense, such as TNF- or NO, but also mediators linking the
primitive immune system to T cell responses, such as IL-12. Bone
marrow-derived macrophages from C57BL/6 were cultured for 24 h in
the presence of varying concentration of human hsp60. Subsequently, RNA
was isolated and the amount of message for IL-12p35 and IL-12p40 was
determined by semiquantitative RT-PCR, which included quantitation of
PCR products by phosphor imaging and calibration to the amount of
ß-actin mRNA observed. As shown in Fig. 5, A and B, the
mRNAs encoding both IL-12 subunits were up-regulated in a
dose-dependent manner following exposure to 10 µg/ml of human hsp60.
In macrophages cultured only in the presence of medium, minimal IL-12
mRNA was detectable. Mean increases in IL-12p35 and IL-12p40 mRNA
expression in cultures treated with 10 µg/ml human hsp60 were 13- and
9.8-fold, respectively, compared with levels in medium controls. In the
presence of 1 µg/ml human hsp60, the increase of IL-12p35 mRNA was
5.8-fold and that of IL-12p40 mRNA was 5-fold. The induction of IL-12
gene expression by human hsp60 could not be ascribed to contamination
with endotoxin, since the effect induced by 10 µg/ml of human hsp60
could not be inhibited by the addition of PmB. As a positive control,
10 ng/ml of LPS induced IL-12p35 and IL-12p40 mRNA production to a
similar extent as seen for hsp60 (Fig. 5, A and
B). To illustrate the mean data of several experiments
presented in Fig. 5, A and B, bands of a single
experiment obtained after gel electrophoresis and blotting are
demonstrated in Fig. 5C.
|
, A and B,
message for both IL-12 subunits increased within 7 h. While mRNA
levels for IL-12p40 remained high at 24 and 72 h (Fig. 6A), there was a more transient production of IL-12p35 mRNA,
which was strongly decreased already at 24 h (Fig. 6B).
|
To identify whether human hsp60 would induce IL-15 gene
expression, we repeated the PCR reaction with IL-15-specific primers
and quantified the amount of mRNA as described for IL-12. IL-15 mRNA
was up-regulated about fourfold following exposure to 10 µg/ml of
human hsp60 with or without PmB (Fig. 7A). In response to 1 µg/ml
of hsp60, there was a 2.5-fold increase of mRNA detectable. IL-15 mRNA
expression was also induced by 10 ng/ml of LPS.
|
B presents the kinetics of hsp60-induced IL-15 mRNA
expression. After incubation for 7 h, IL-15 mRNA levels had
increased about threefold. Maximal message expression was observed at
24 h, and increased mRNA expression was still noted at 72 h.
Fig. 7C depicts bands obtained for PCR of a single
experiment. |
Discussion |
|---|
|
TopAbstractIntroductionMaterials and MethodsResultsDiscussionReferences |
|---|
The initial studies were performed with a mouse macrophage line, allowing for the possibility that the inflammatory response toward homologous mammalian hsp60 may be an abnormal property acquired during cell transformation and adaption to long-term in vitro propagation. However, the same response to human hsp60 was observed with primary macrophages. Finally, human hsp60 was also added to a human monocyte line, and induction of proinflammatory cytokine secretion was seen also in this nonxenogeneic system.
The stimulatory action of human hsp60 in mammalian macrophages may be related to a similar property of bacterial hsp60 26, 27, 28, 29, 30, 31, 32 . The latter studies concur that the minimal effective dose for eliciting a cytokine response is in the microgram range, as found in this study for human hsp60. In addition, these studies excluded a role of endotoxin contamination in the effects observed. We conclude that mammalian hsp60 shares with the microbial homologues the ability to activate macrophages, and probably other cells of the innate immune system, as has been reported for microbial hsp60 31, 32 .
The mechanisms involved in the stimulatory action of microbial hsp60 have not yet been identified so that we could not test for an analogous mode of action for human hsp60. Two opposing mechanisms may be considered. Hsp60 may act as chaperone by binding to immature or partially denatured proteins 33 on the surface of macrophages or in the cell interior, after endocytosis. Hence, hsp60 would exert its physiologic function 1, 2, 3, 4 , only that the site of action and target molecules is different for exogenous hsp60 in comparison with its endogenously synthesized counterpart. The other conceivable mechanism would be that macrophages express a receptor on the cell surface, which recognizes a defined epitope of hsp60. This epitope would have to be common between bacterial and mammalian hsp60, and therefore associated with highly conserved parts of the molecule. Indeed, human hsp60 is more than 50% identical to mycobacterial hsp65 5 . Because of the similarity in action to LPS, the LPS receptor may be involved. However, it has been reported that the stimulatory action of mycobacterial hsp65 could not be inhibited by an Ab to the LPS receptor CD14 26 .
The physiologic or pathophysiologic relevance of the proinflammatory macrophage response to either microbial or autologous hsp60 presently has not been determined. In eukaryotic cells, most of the hsp60 is localized to mitochondria; cell surface expression was found increased in inflammatory conditions or during wound healing 9, 34, 35, 36, 37, 38, 39, 40, 41, 42 . Both hsp60 on cell surfaces or in mitochondria becoming accessible during cell necrosis might provide high local concentrations sufficient for stimulation of an innate immune response. Further insight will come from analyzing the stimulatory effect of membrane-bound versus free hsp60. Hsp60 may also be released from cells in soluble form, and the increased expression of hsp60 in peritoneal fluids from women with endometriosis was found associated with proinflammatory cytokines and activated macrophages 43 .
In nonstimulated monocytes or macrophages, the induction of cytokine or NO production requires prior transcriptional activity 44, 45, 46 so that it is safe to assume that human hsp60 exerts at least some of its stimulatory effects at the level of gene transcription. Direct evidence comes from the observation of significantly elevated IL-12 and IL-15 mRNA levels in macrophages exposed to hsp60. Analysis of the time course revealed peak or close to peak mRNA levels after 7 h of incubation with human hsp60, which fits with the time course of classical receptor-mediated signal transduction pathway as reported for the LPS-induced transcription of the IL-12 47 and IL-15 genes 48 .
Taken together, the available evidence supports the concept that macrophages respond to autologous hsp60 when it becomes accessible to their cell surface. Such may happen when hsp60 is set free during necrosis of tissue cells during inflammation or when hsp60 is partially translocated to the plasma membrane 8, 9 in response to diverse types of stress. Increased local expression of autologous hsp60 has been noted in many inflammatory conditions, such as rheumatoid arthritis, insulitis, Crohn’s disease, or atherosclerosis 49, 50, 51, 52, 53, 54 . Furthermore, the local overexpression of autologous hsp60 is found simultaneously with the formation of cellular infiltrates in tissues 50, 53, 54, 55 .
Autologous hsp60 when translocated to the cell surface or released
during cell necrosis hence may qualify as danger Ag, as proposed by P.
Matzinger 56 , which alerts the innate immune system to sites of cell
stress or nonprogrammed cell death, i.e., necrosis. Local exposure of
hsp60 would account for an early-onset response, which eventually would
be followed by activation of the adaptive T cell-dependent immune
system. In this context, it is noteworthy that human hsp60 did not only
induce nonspecific proinflammatory mediators such as NO or TNF-, but
also the expression of IL-12 and IL-15. The latter two cytokines play a
key role in the induction of cellular Th1-type immune responses and
link the innate immune system to T cell immunity 57, 58 .
Interestingly, human hsp60 was found to synergize with IFN- in its
stimulatory action of macrophages. This finding is in accord with
previous studies of mycobacterial hsp65 28 and underscores a role of
hsp60 in regulatory circuits between innate and adaptive immunity.
It may therefore not come as a surprise that hsp60 is a key Ag also of adaptive immune responses. This holds true for both the immune defense against infectious pathogens 10 , as well as the chronic tissue inflammation of assumed autoimmune nature 11, 12, 13, 14 . Experimentally induced sterile inflammation readily induces self hsp60-specific T cells 59 .
In conclusion, we show in this study that human hsp60 induces a proinflammatory response in mouse macrophages as well as in human monocytes. Both proinflammatory mediators as well as Th1-inducing cytokines were induced by human hsp60, which identifies hsp60 as a danger Ag on autologous cells, eliciting an innate immune response and preparing for an adaptive cellular, Th1-type, immune reaction.
|
Acknowledgments |
|---|
|
Footnotes |
|---|
2 Address correspondence and reprint requests to Dr. Hubert Kolb, Clinical Department, Diabetes Research Institute, Auf'm Hennekamp 65, D-40225 Düsseldorf, Germany. E-mail address:
3 Abbreviations used in this paper: hsp, heat-shock protein; NO, nitric oxide; PmB, polymyxin B; PSL, 32P-stimulated luminescence.
Received for publication August 24, 1998. Accepted for publication December 11, 1998.
|
References |
|---|
|
TopAbstractIntroductionMaterials and MethodsResultsDiscussionReferences |
|---|
, but not interleukin-4. Diabetologia 37:1154.[Medline]
by Mycobacterium tuberculosis components. J. Clin. Invest. 91:2076.
, interleukin 6, reactive nitrogen intermediates, and toxoplasmastatic activity in murine peritoneal macrophages. Infect. Immun. 63:3454.[Abstract]
B/Rel in induction of nitric oxide synthase. J. Biol. Chem. 269:4705.This article has been cited by other articles:
|
|
 |
|
  M. Cohen-Sfady, M. Pevsner-Fischer, R. Margalit, and I. R. Cohen Heat Shock Protein 60, via MyD88 Innate Signaling, Protects B Cells from Apoptosis, Spontaneous and Induced J. Immunol., July 15, 2009; 183(2): 890 - 896. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 M.-F. Tsan and B. Gao Heat shock proteins and immune system J. Leukoc. Biol., June 1, 2009; 85(6): 905 - 910. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 M. Caroprese, A. Marzano, G. Entrican, S. Wattegedera, M. Albenzio, and A. Sevi Immune response of cows fed polyunsaturated fatty acids under high ambient temperatures J Dairy Sci, June 1, 2009; 92(6): 2796 - 2803. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 L. Calderon-Garciduenas, M. Macias-Parra, H. J. Hoffmann, G. Valencia-Salazar, C. Henriquez-Roldan, N. Osnaya, O. C.-d. Monte, G. Barragan-Mejia, R. Villarreal-Calderon, L. Romero, et al. Immunotoxicity and Environment: Immunodysregulation and Systemic Inflammation in Children Toxicol Pathol, February 1, 2009; 37(2): 161 - 169. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
A. Iannello, O. Debbeche, S. Samarani, and A. Ahmad Antiviral NK cell responses in HIV infection: II. viral strategies for evasion and lessons for immunotherapy and vaccination J. Leukoc. Biol., July 1, 2008; 84(1): 27 - 49. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 S. Lehnardt, E. Schott, T. Trimbuch, D. Laubisch, C. Krueger, G. Wulczyn, R. Nitsch, and J. R. Weber A Vicious Cycle Involving Release of Heat Shock Protein 60 from Injured Cells and Activation of Toll-Like Receptor 4 Mediates Neurodegeneration in the CNS J. Neurosci., March 5, 2008; 28(10): 2320 - 2331. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 A. Osterloh, A. Veit, A. Gessner, B. Fleischer, and M. Breloer Hsp60-mediated T cell stimulation is independent of TLR4 and IL-12 Int. Immunol., March 1, 2008; 20(3): 433 - 443. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
S. Lehnardt, J. Wennekamp, D. Freyer, C. Liedtke, C. Krueger, R. Nitsch, I. Bechmann, J. R. Weber, and P. Henneke TLR2 and Caspase-8 Are Essential for Group B Streptococcus-Induced Apoptosis in Microglia J. Immunol., November 1, 2007; 179(9): 6134 - 6143. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 A. Osterloh, U. Kalinke, S. Weiss, B. Fleischer, and M. Breloer Synergistic and Differential Modulation of Immune Responses by Hsp60 and Lipopolysaccharide J. Biol. Chem., February 16, 2007; 282(7): 4669 - 4680. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 A. Y. W. Chang, J. Y. H. Chan, J. L. J. Chou, F. C. H. Li, K.-Y. Dai, and S. H. H. Chan Heat shock protein 60 in rostral ventrolateral medulla reduces cardiovascular fatality during endotoxaemia in the rat J. Physiol., July 15, 2006; 574(2): 547 - 564. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
S. Lehnardt, P. Henneke, E. Lien, D. L. Kasper, J. J. Volpe, I. Bechmann, R. Nitsch, J. R. Weber, D. T. Golenbock, and T. Vartanian A Mechanism for Neurodegeneration Induced by Group B Streptococci through Activation of the TLR2/MyD88 Pathway in Microglia J. Immunol., July 1, 2006; 177(1): 583 - 592. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 Q. Xiao, K. Mandal, G. Schett, M. Mayr, G. Wick, F. Oberhollenzer, J. Willeit, S. Kiechl, and Q. Xu Association of Serum-Soluble Heat Shock Protein 60 With Carotid Atherosclerosis: Clinical Significance Determined in a Follow-Up Study Stroke, December 1, 2005; 36(12): 2571 - 2576. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
M. Cohen-Sfady, G. Nussbaum, M. Pevsner-Fischer, F. Mor, P. Carmi, A. Zanin-Zhorov, O. Lider, and I. R. Cohen Heat Shock Protein 60 Activates B Cells via the TLR4-MyD88 Pathway J. Immunol., September 15, 2005; 175(6): 3594 - 3602. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
M. T. Pritchard, Z. Li, and E. A. Repasky Nitric oxide production is regulated by fever-range thermal stimulation of murine macrophages J. Leukoc. Biol., September 1, 2005; 78(3): 630 - 638. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
F. J. Quintana and I. R. Cohen Heat Shock Proteins as Endogenous Adjuvants in Sterile and Septic Inflammation J. Immunol., September 1, 2005; 175(5): 2777 - 2782. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
C. Habich, K. Kempe, R. van der Zee, R. Rumenapf, H. Akiyama, H. Kolb, and V. Burkart Heat Shock Protein 60: Specific Binding of Lipopolysaccharide J. Immunol., February 1, 2005; 174(3): 1298 - 1305. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 A. Lang, D. Benke, F. Eitner, D. Engel, S. Ehrlich, M. Breloer, E. Hamilton-Williams, S. Specht, A. Hoerauf, J. Floege, et al. Heat Shock Protein 60 Is Released in Immune-Mediated Glomerulonephritis and Aggravates Disease: In Vivo Evidence for an Immunologic Danger Signal J. Am. Soc. Nephrol., February 1, 2005; 16(2): 383 - 391. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
A. Osterloh, F. Meier-Stiegen, A. Veit, B. Fleischer, A. von Bonin, and M. Breloer Lipopolysaccharide-free Heat Shock Protein 60 Activates T Cells J. Biol. Chem., November 12, 2004; 279(46): 47906 - 47911. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
A. Ahmad and F. Alvarez Role of NK and NKT cells in the immunopathogenesis of HCV-induced hepatitis J. Leukoc. Biol., October 1, 2004; 76(4): 743 - 759. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 W. Ren, R. Strube, X. Zhang, S.-Y. Chen, and X. F. Huang Potent Tumor-Specific Immunity Induced by an In vivo Heat Shock Protein-Suicide Gene-Based Tumor Vaccine Cancer Res., September 15, 2004; 64(18): 6645 - 6651. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
M.-F. Tsan and B. Gao Endogenous ligands of Toll-like receptors J. Leukoc. Biol., September 1, 2004; 76(3): 514 - 519. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
A. V. Bocharov, I. N. Baranova, T. G. Vishnyakova, A. T. Remaley, G. Csako, F. Thomas, A. P. Patterson, and T. L. Eggerman Targeting of Scavenger Receptor Class B Type I by Synthetic Amphipathic {alpha}-Helical-containing Peptides Blocks Lipopolysaccharide (LPS) Uptake and LPS-induced Pro-inflammatory Cytokine Responses in THP-1 Monocyte Cells J. Biol. Chem., August 20, 2004; 279(34): 36072 - 36082. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 A. Jalili, M. Makowski, T. Switaj, D. Nowis, G. M. Wilczynski, E. Wilczek, M. Chorazy-Massalska, A. Radzikowska, W. Maslinski, L. Bialy, et al. Effective Photoimmunotherapy of Murine Colon Carcinoma Induced by the Combination of Photodynamic Therapy and Dendritic Cells Clin. Cancer Res., July 1, 2004; 10(13): 4498 - 4508. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 M. Fleshner and M. L. Laudenslager Psychoneuroimmunology: Then and Now Behav Cogn Neurosci Rev, June 1, 2004; 3(2): 114 - 130. [Abstract] [PDF]
|
|
|
|
 |
|
 M.-F. Tsan and B. Gao Cytokine function of heat shock proteins Am J Physiol Cell Physiol, April 1, 2004; 286(4): C739 - C744. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
K. Vandal, P. Rouleau, A. Boivin, C. Ryckman, M. Talbot, and P. A. Tessier Blockade of S100A8 and S100A9 Suppresses Neutrophil Migration in Response to Lipopolysaccharide J. Immunol., September 1, 2003; 171(5): 2602 - 2609. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
R. C. Reed, B. Berwin, J. P. Baker, and C. V. Nicchitta GRP94/gp96 Elicits ERK Activation in Murine Macrophages: A ROLE FOR ENDOTOXIN CONTAMINATION IN NF-{kappa}B ACTIVATION AND NITRIC OXIDE PRODUCTION J. Biol. Chem., August 22, 2003; 278(34): 31853 - 31860. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
H. Amir-Kroll, G. Nussbaum, and I. R. Cohen Proteins and Their Derived Peptides as Carriers in a Conjugate Vaccine for Streptococcus pneumoniae: Self-Heat Shock Protein 60 and Tetanus Toxoid J. Immunol., June 15, 2003; 170(12): 6165 - 6171. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
B. Gao and M.-F. Tsan Recombinant Human Heat Shock Protein 60 Does Not Induce the Release of Tumor Necrosis Factor {alpha} from Murine Macrophages J. Biol. Chem., June 13, 2003; 278(25): 22523 - 22529. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 B. Metzler, R. Abia, M. Ahmad, F. Wernig, O. Pachinger, Y. Hu, and Q. Xu Activation of Heat Shock Transcription Factor 1 in Atherosclerosis Am. J. Pathol., May 1, 2003; 162(5): 1669 - 1676. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
S. B. Flohe, J. Bruggemann, S. Lendemans, M. Nikulina, G. Meierhoff, S. Flohe, and H. Kolb Human Heat Shock Protein 60 Induces Maturation of Dendritic Cells Versus a Th1-Promoting Phenotype J. Immunol., March 1, 2003; 170(5): 2340 - 2348. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
H. Cwiklinska, M. P. Mycko, O. Luvsannorov, B. Walkowiak, C. F. Brosnan, C. S. Raine, and K. W. Selmaj Heat shock protein 70 associations with myelin basic protein and proteolipid protein in multiple sclerosis brains Int. Immunol., February 1, 2003; 15(2): 241 - 249. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 J. Campisi and M. Fleshner Role of extracellular HSP72 in acute stress-induced potentiation of innate immunity in active rats J Appl Physiol, January 1, 2003; 94(1): 43 - 52. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 H. Feng, Y. Zeng, M. W. Graner, A. Likhacheva, and E. Katsanis Exogenous stress proteins enhance the immunogenicity of apoptotic tumor cells and stimulate antitumor immunity Blood, January 1, 2003; 101(1): 245 - 252. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 J. Michaelsson, C. Teixeira de Matos, A. Achour, L. L. Lanier, K. Karre, and K. Soderstrom A Signal Peptide Derived from hsp60 Binds HLA-E and Interferes with CD94/NKG2A Recognition J. Exp. Med., December 2, 2002; 196(11): 1403 - 1414. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
K. Bethke, F. Staib, M. Distler, U. Schmitt, H. Jonuleit, A. H. Enk, P. R. Galle, and M. Heike Different Efficiency of Heat Shock Proteins (HSP) to Activate Human Monocytes and Dendritic Cells: Superiority of HSP60 J. Immunol., December 1, 2002; 169(11): 6141 - 6148. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
E. Noessner, R. Gastpar, V. Milani, A. Brandl, P. J. S. Hutzler, M. C. Kuppner, M. Roos, E. Kremmer, A. Asea, S. K. Calderwood, et al. Tumor-Derived Heat Shock Protein 70 Peptide Complexes Are Cross-Presented by Human Dendritic Cells J. Immunol., November 15, 2002; 169(10): 5424 - 5432. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 W van Eden, R van der Zee, P van Kooten, S E Berlo, P M Cobelens, A Kavelaars, C J Heijnen, B Prakken, S Roord, and S Albani Balancing the immune system: Th1 and Th2 Ann Rheum Dis, November 1, 2002; 61(90002): ii25 - 28. [Full Text] [PDF]
|
|
|
|
 |
|
 Q. Xu Role of Heat Shock Proteins in Atherosclerosis Arterioscler. Thromb. Vasc. Biol., October 1, 2002; 22(10): 1547 - 1559. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
B. Billack, D. E. Heck, T. M. Mariano, C. R. Gardner, R. Sur, D. L. Laskin, and J. D. Laskin Induction of cyclooxygenase-2 by heat shock protein 60 in macrophages and endothelial cells Am J Physiol Cell Physiol, October 1, 2002; 283(4): C1267 - C1277. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
R. Ulmansky, C. J. Cohen, F. Szafer, E. Moallem, Z. G. Fridlender, Y. Kashi, and Y. Naparstek Resistance to Adjuvant Arthritis Is Due to Protective Antibodies Against Heat Shock Protein Surface Epitopes and the Induction of IL-10 Secretion J. Immunol., June 15, 2002; 168(12): 6463 - 6469. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
R. M. Vabulas, S. Braedel, N. Hilf, H. Singh-Jasuja, S. Herter, P. Ahmad-Nejad, C. J. Kirschning, C. da Costa, H.-G. Rammensee, H. Wagner, et al. The Endoplasmic Reticulum-resident Heat Shock Protein Gp96 Activates Dendritic Cells via the Toll-like Receptor 2/4 Pathway J. Biol. Chem., May 31, 2002; 277(23): 20847 - 20853. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
G. B. Johnson, G. J. Brunn, Y. Kodaira, and J. L. Platt Receptor-Mediated Monitoring of Tissue Well-Being Via Detection of Soluble Heparan Sulfate by Toll-Like Receptor 4 J. Immunol., May 15, 2002; 168(10): 5233 - 5239. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
R. M. Vabulas, P. Ahmad-Nejad, S. Ghose, C. J. Kirschning, R. D. Issels, and H. Wagner HSP70 as Endogenous Stimulus of the Toll/Interleukin-1 Receptor Signal Pathway J. Biol. Chem., April 19, 2002; 277(17): 15107 - 15112. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
N. N. Panjwani, L. Popova, and P. K. Srivastava Heat Shock Proteins gp96 and hsp70 Activate the Release of Nitric Oxide by APCs J. Immunol., March 15, 2002; 168(6): 2997 - 3003. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
V. Burkart, Y.-E. Kim, B. Hartmann, I. Ghiea, U. Syldath, M. Kauer, W. Fingberg, P. Hanifi-Moghaddam, S. Muller, and H. Kolb Cholera Toxin B Pretreatment of Macrophages and Monocytes Diminishes Their Proinflammatory Responsiveness to Lipopolysaccharide J. Immunol., February 15, 2002; 168(4): 1730 - 1737. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
C. Habich, K. Baumgart, H. Kolb, and V. Burkart The Receptor for Heat Shock Protein 60 on Macrophages Is Saturable, Specific, and Distinct from Receptors for Other Heat Shock Proteins J. Immunol., January 15, 2002; 168(2): 569 - 576. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
R. K. Kanwar, J. R. Kanwar, D. Wang, D. J. Ormrod, and G. W. Krissansen Temporal Expression of Heat Shock Proteins 60 and 70 at Lesion-Prone Sites During Atherogenesis in ApoE-Deficient Mice Arterioscler. Thromb. Vasc. Biol., December 1, 2001; 21(12): 1991 - 1997. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 B.B. Moore, T.A. Moore, and G.B. Toews Role of T- and B-;lymphocytes in pulmonary host defences Eur. Respir. J., November 1, 2001; 18(5): 846 - 856. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 A. Fortin, J. Lagace, and H.-M. Therien Trafficking of Surface-linked and Encapsulated Liposomal Antigens in Macrophages: an Immunocytochemical Study J. Histochem. Cytochem., November 1, 2001; 49(11): 1407 - 1420. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 A. Lamikanra, Z.-K. Pan, S. N. Isaacs, T.-C. Wu, and Y. Paterson Regression of Established Human Papillomavirus Type 16 (HPV-16) Immortalized Tumors In Vivo by Vaccinia Viruses Expressing Different Forms of HPV-16 E7 Correlates with Enhanced CD8+ T-Cell Responses That Home to the Tumor Site J. Virol., October 15, 2001; 75(20): 9654 - 9664. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
M. J. Gough, A. A. Melcher, A. Ahmed, M. R. Crittenden, D. S. Riddle, E. Linardakis, A. N. Ruchatz, L. M. Emiliusen, and R. G. Vile Macrophages Orchestrate the Immune Response to Tumor Cell Death Cancer Res., October 1, 2001; 61(19): 7240 - 7247. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
S. H. More, M. Breloer, and A. von Bonin Eukaryotic heat shock proteins as molecular links in innate and adaptive immune responses: Hsp60-mediated activation of cytotoxic T cells Int. Immunol., September 1, 2001; 13(9): 1121 - 1127. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
S. T. Smiley, J. A. King, and W. W. Hancock Fibrinogen Stimulates Macrophage Chemokine Secretion Through Toll-Like Receptor 4 J. Immunol., September 1, 2001; 167(5): 2887 - 2894. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
H. Feng, Y. Zeng, L. Whitesell, and E. Katsanis Stressed apoptotic tumor cells express heat shock proteins and elicit tumor-specific immunity Blood, June 1, 2001; 97(11): 3505 - 3512. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
A. K. De, K. M. Kodys, B. S. Yeh, and C. Miller-Graziano Exaggerated Human Monocyte IL-10 Concomitant to Minimal TNF-{alpha} Induction by Heat-Shock Protein 27 (Hsp27) Suggests Hsp27 Is Primarily an Antiinflammatory Stimulus J. Immunol., October 1, 2000; 165(7): 3951 - 3958. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
R. E. Vance Cutting Edge Commentary: A Copernican Revolution? Doubts About the Danger J. Immunol., August 15, 2000; 165(4): 1725 - 1728. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
D. M. Hall, L. Xu, V. J. Drake, L. W. Oberley, T. D. Oberley, P. L. Moseley, and K. C. Kregel Aging reduces adaptive capacity and stress protein expression in the liver after heat stress J Appl Physiol, August 1, 2000; 89(2): 749 - 759. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 Q. Xu, G. Schett, H. Perschinka, M. Mayr, G. Egger, F. Oberhollenzer, J. Willeit, S. Kiechl, and G. Wick Serum Soluble Heat Shock Protein 60 Is Elevated in Subjects With Atherosclerosis in a General Population Circulation, July 4, 2000; 102(1): 14 - 20. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 J S H Gaston Immunological basis of chlamydia induced reactive arthritis Sex Transm Inf, June 1, 2000; 76(3): 156 - 161. [Full Text] [PDF]
|
|
|
|
|
|
S. El mir and F. Triebel A Soluble Lymphocyte Activation Gene-3 Molecule Used as a Vaccine Adjuvant Elicits Greater Humoral and Cellular Immune Responses to Both Particulate and Soluble Antigens J. Immunol., June 1, 2000; 164(11): 5583 - 5589. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
C.-H. Chen, T.-L. Wang, C.-F. Hung, Y. Yang, R. A. Young, D. M. Pardoll, and T-C. Wu Enhancement of DNA Vaccine Potency by Linkage of Antigen Gene to an HSP70 Gene Cancer Res., February 1, 2000; 60(4): 1035 - 1042. [Abstract] [Full Text]
|
|
|
|
|
|
Q. Huang, J. F.L. Richmond, K. Suzue, H. N. Eisen, and R. A. Young In Vivo Cytotoxic T Lymphocyte Elicitation by Mycobacterial Heat Shock Protein 70 Fusion Proteins Maps to a Discrete Domain and Is CD4+ T Cell Independent J. Exp. Med., January 17, 2000; 191(2): 403 - 408. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
K. Ohashi, V. Burkart, S. Flohe, and H. Kolb Cutting Edge: Heat Shock Protein 60 Is a Putative Endogenous Ligand of the Toll-Like Receptor-4 Complex J. Immunol., January 15, 2000; 164(2): 558 - 561. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
A. Kol, A. H. Lichtman, R. W. Finberg, P. Libby, and E. A. Kurt-Jones Cutting Edge: Heat Shock Protein (HSP) 60 Activates the Innate Immune Response: CD14 Is an Essential Receptor for HSP60 Activation of Mononuclear Cells J. Immunol., January 1, 2000; 164(1): 13 - 17. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
R. M. Vabulas, P. Ahmad-Nejad, C. da Costa, T. Miethke, C. J. Kirschning, H. Hacker, and H. Wagner Endocytosed HSP60s Use Toll-like Receptor 2 (TLR2) and TLR4 to Activate the Toll/Interleukin-1 Receptor Signaling Pathway in Innate Immune Cells J. Biol. Chem., August 10, 2001; 276(33): 31332 - 31339. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
B. Dybdahl, A. Wahba, E. Lien, T. H. Flo, A. Waage, N. Qureshi, O. F.M. Sellevold, T. Espevik, and A. Sundan Inflammatory Response After Open Heart Surgery: Release of Heat-Shock Protein 70 and Signaling Through Toll-Like Receptor-4 Circulation, February 12, 2002; 105(6): 685 - 690. [Abstract] [Full Text] [PDF]
|
|
| |||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
| HOME | HELP | FEEDBACK | SUBSCRIPTIONS | ARCHIVE | SEARCH | TABLE OF CONTENTS |
