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* Department of Internal Medicine, University of Texas Medical Branch, Galveston, TX 77555;
Department of General Surgery, Graduate School of Medicine, Chiba University, Chiba, Japan; and
Shriners Hospital for Children, Galveston, TX 77555
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Abstract |
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) have been described as a major effector cell on the host’s innate immunities. However, CAM are not generated in immunocompromised hosts whose alternatively activated macrophages (AAM) predominate. In this study, the mechanism by which AAM suppress the ability of resident macrophages (RM) to generate CAM was investigated. AAM were isolated from peritoneal exudates of mice 2 days after third-degree thermal injuries affecting 15% total body surface area. CAM were generated from RM (peritoneal M from normal mice) through stimulation with CpG DNA, a typical CAM inducer. RM did not polarize to CAM when they were cultured with AAM in a dual-chamber Transwell even when supplemented with CpG DNA. In addition, RM stimulated with CpG DNA did not convert to CAM when they were cultured with the culture fluids of AAM (AAM Culture-Sup). AAM Culture-Sup contained IL-6, IL-10, CCL17, PGE2, and TGF-
. Among these, CCL17 and IL-10 inhibited CAM generation. The ability of AAM Culture-Sup to inhibit CAM generation was eliminated when the Culture-Sup was treated with a mixture of mAbs directed against CCL17 and IL-10. These results indicate that CCL17 and IL-10 released from AAM inhibit CAM generation from RM stimulated with CpG DNA. |
Introduction |
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TopAbstractIntroductionMaterials and MethodsResultsDiscussionReferences |
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)3 have long been recognized as heterogeneous (1). The heterogeneity of M almost certainly reflects their plasticity and versatility in response to microenvironmental signals. Recent studies have suggested five (1) or three (2) pathways of M activation in innate and acquired immunities. However, in addition to resident M (RM), M have generally been divided into two populations, classically activated macrophages (CAM) and alternatively activated macrophages (AAM), based on their immunobiological properties (3, 4, 5). CAM play an important role in the antimicrobial innate immunities of the host (6, 7, 8). CAM are generated from RM stimulated with a microbe, microbial product (CpG DNA, poly(I:C), and LPS) or various biological response modifiers (muramyl dipeptide and poly(styrene-maleic acid) conjugated with neocarzinostatin) (5, 9, 10, 11, 12). CAM exhibit 1) high oxygen consumption, 2) the ability to kill cells infected with intracellular pathogens, 3) cytotoxicity against tumor cells, 4) the ability to express inducible NO synthase (iNOS), and 5) the ability to secrete NO, proinflammatory cytokines (IL-1, IL-6, and TNF-
) and Th1 response-associated cytokines (IFN-
, IL-12, IL-18, CCL3, CXCL9, and CXCL10) (4, 5). In addition, CAM induce Th1 cells by producing IL-12, CCL3, and IFN-. In contrast, AAM play a role in the negative regulation of both CAM and Th1 cell generation (4, 5, 13, 14, 15, 16). AAM preferentially express receptors for foreign Ags, such as mannose receptor, -glucan receptors, and scavenger receptors (17, 18). AAM have been described as a producer cell for IL-1R antagonist, IL-10, CCL17, CCL22, and arginase (4, 5). They also inhibit IFN- production and developmental Th1 responses (15, 16).
Numerous studies have shown that CAM generation is required for a host’s defense against infections with various intracellular pathogens or the growth of tumors (1, 5). In fact, CAM that appeared in mice infected with Mycobacterium bovis bacillus Calmette-Guérin and Listeria monocytogenes were shown to be powerful killer cells for these pathogens, tumor cells, and cells infected with various bacteria and viruses (1). The ability of CAM to kill these targets was displayed nonspecifically (1). In other words, the host resistance of individuals exposed to foreign invasions may be effectively improved if CAM can be induced. However, CAM were not generated in individuals whose AAM predominate, because AAM inhibited CAM generation (19, 20). Hosts susceptible to foreign invasions are generally recognized as individuals with a predominance of AAM.
In the present study, CCL17 and IL-10 released from AAM were shown to be effector molecules that inhibit CAM generation from RM stimulated with CpG DNA. The regulation of the production of these cytokines may improve the resistance to various opportunistic infections in patients whose AAM predominate.
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Materials and Methods |
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Eight- to 11-wk-old, pathogen-free, male BALB/c mice purchased from The Jackson Laboratory (Bar Harbor, ME) were used in this study. The Institutional Animal Care and Use Committee of the University of Texas Medical Branch approved all procedures performed in these animal experiments (Institutional Animal Care and Use Committee approval no. 02-04-024).
Reagents and medium
rIL-1, IL-2, IL-4, IL-6, IL-10, IL-13, IFN-, TNF-, and CCL3 were purchased from PeproTech (Rocky Hill, NJ). TGF- and mAbs for IL-1, IL-4, IL-6, IL-10, IFN-, TGF-, TNF-, CD3, and CD28 were purchased from BD PharMingen (San Diego, CA). PGE2 ELISA kit and CCL17 as well as mAbs for CCL3 and CCL17 were purchased from R&D Systems (Minneapolis, MN). CpG DNA (5'-TCCATGACGTTCCTGACGTT-3') was purchased from Sigma-Genosys (Woodlands, TX). CpG DNA was used to induce CAM generation, as previously described (21). M were cultured in RPMI 1640 medium supplemented with 10% heat-inactivated FBS, 2 mM L-glutamine, and antibiotics (complete medium).
Burn injury
Thermally injured mice, BALB/c mice exposed to flame-burn injuries (22), were prepared as follows. Mice were anesthetized with pentobarbital (40 mg/kg i.p.) and electric clippers were used to shave the hair on the back of each mouse from groin to axilla. The mice were then exposed to a gas flame for 9 s by pressing the window of the custom-made insulated mold (with a 2.5 x 3.5-cm window) firmly against the shaved back. A Bunsen burner equipped with a flame-dispersing cap was used as a source of the gas flame. This procedure consistently produced a third-degree burn on 
15% of total body surface area (TBSA) for a 26-g mouse (22). Immediately after thermal injury, physiologic saline (3 ml/mouse i.p.) was administered for fluid resuscitation. Animals were then housed until use for experiments. Control mice had their back hair shaved but were not exposed to the gas flame. They also received physiologic saline (3 ml/mouse i.p.).
Preparation of RM, AAM, and CAM
For the M isolation, 4–10 x 106 cells of peritoneal exudates from mice were centrifuged. The cell pellets were suspended in 2 ml of RPMI 1640 medium supplemented with 2% FBS (maintenance medium) and cultured in fibronectin-coated petri dishes (60 x 15 mm) for 15 min at 37°C (23). At the end of cultivation, the dishes were washed twice with maintenance medium warmed to 37°C. Adherent cells (M populations) were recovered from the dishes using a rubber policeman. M purity was measured as 92% or more when tested by FACSVantage (BD Biosciences, Mountain View, CA). RM were M freshly isolated from peritoneal exudates of normal mice. As described previously (24), AAM were isolated from peritoneal exudates of mice 2 days after severe burn injuries (third-degree flame burn, >15% TBSA). As previously described (4), standard AAM were induced in cultures of RM treated with a mixture of IL-4 and IL-13 (10 ng/ml each) for 48 h. CAM were generated in vitro from RM after stimulation with CpG DNA (24 h stimulation, 10 µg/ml) (21).
Criteria of CAM and AAM
M were identified as CAM when the following typical properties were demonstrated: production of CCL3 and IL-12 (25, 26), expression of iNOS mRNA (27), induction of Th1 responses (25), and exhibition of killing activity against bacteria (6). M were identified as AAM when the following criteria were demonstrated: production of CCL17 and IL-10 (3, 28), expression of mannose receptor mRNA (29), and induction of Th2 responses (30). For the production of cytokines, various M populations (1 x 106 cells/ml) were cultured for 24 h without any stimulation. Then, the culture fluids harvested were assayed for the cytokines using ELISA. The detection limits of CCL3, IL-12, IL-10, and CCL17 were 16, 14, 8, and 16 pg/ml, respectively. In addition, M populations were tested for mRNA expression by RT-PCR using primers as follows: for mannose receptor, 5'-CCATCGAGACTGCTGCTGAG-3' (F) and 5'-AGCCCTTGGGTTGAGGATCC-3' (R) and for iNOS, 5'-CCCTCCAGTGTCTGGGAGCA-3' (F) and 5'-TGCTTGTCACCACCAGCAGT-3' (R). Methicillin-resistant S. aureus (MRSA), biotype 21777, was isolated from a clinical specimen from a burn patient in Shriners Hospital for Children and used as target cells to test the antibacterial killing activity of M. MRSA used in this study was defined as a vancomycin-sensitive strain. Using Lissner’s methods (31), MRSA-killing activities of various M populations were examined by culturing 1 x 106 cells/ml M with 1 x 107 CFU/ml MRSA for 20 min at 37°C (multiplicity of infection = 10). After unphagocytized MRSA was washed out, the cells were cultured for an additional 3 h at 37°C. The number of MRSA in the whole culture was determined using a standard colony-counting method (31). The ability of various M populations to induce Th1 or Th2 responses was determined by the methods described in previous reports (23). Naive T cells (1 x 106 cells/ml) were cocultured with M populations (1 x 106 cells/ml) for 6 days in the presence of 50 U/ml IL-2. To induce cytokines, the nonadherent cells were harvested and recultured for 12 h with a mixture of anti-CD3 and anti-CD28 mAbs (2 µg/ml each). The culture fluids were harvested and assayed for IFN- (a Th1 response) or IL-4 (a Th2 response) using ELISA.
Inhibition of CAM generation from RM stimulated with CpG DNA
The inhibitory effect of AAM on CAM generation was analyzed using the following procedures: 1) RM stimulated with CpG DNA were cocultured with AAM, 2) RM were cultured with AAM0 in a dual-chamber Transwell supplemented with CpG DNA, and 3) RM stimulated with CpG DNA were cultured with the culture fluids of AAM (AAM Culture-Sup). After cultivation by these different methods, the RM were harvested and cultured for an additional 24 h to produce CCL3. CCL3 was chosen as a parameter of CAM, because it has been described as such in previous studies (4, 5). Cocultivation experiments (above, procedure 1) were performed as follows. In the presence of 10 µg/ml CpG DNA, RM were cocultured with AAM at a ratio of 1:100 to 64:100 in 96-well microtiter plates. Twenty-four hours after cultivation, the cells were washed three times with complete medium and cultured for an additional 24 h with complete medium. The culture fluids harvested were assayed for CCL3. A Transwell culture (above, procedure 2) was performed as described below. RM were cultured with AAM in a dual-chamber Transwell supplemented with CpG DNA. Six hundred microliters of RM suspension (1 x 106 cells/ml) was placed into the lower chamber of the Transwell (0.4-µm pore size; Costar, Corning, NY). One hundred microliters of AAM suspension (5 x 106 cells/ml) was placed into the upper chamber of the Transwell. Twenty-four hours after cultivation, M harvested from the lower chamber were recultured for 24 h. The resulting culture fluids were assayed for CCL3. RM were also cultured with the AAM Culture-Sup (above, procedure 3). AAM Culture-Sup was the culture fluids of AAM (1 x 106 cells/ml) 24 h after cultivation. RM stimulated with CpG DNA were cultured with complete medium supplemented with AAM Culture-Sup (15%, v/v) for 24 h. The cells harvested were washed three times with complete medium and cultured for an additional 24 h. The resulting culture fluids were assayed for CCL3.
Determination of soluble factors from AAM
To determine the active components in AAM Culture-Sup, AAM were cultured without any stimulation for 24–48 h, and the culture fluids harvested were assayed for IL-1, IL-6, IL-10, TNF-, TGF-, CCL17, and PGE2 using ELISA. These soluble factors have been well described as the products of M or M-related cells (4, 5, 16). The detection limits of IL-1, IL-6, IL-10, TNF-, TGF-, PGE2, and CCL17 were 20, 12, 8, 20, 22, 18, and 16 pg/ml, respectively. Then, the recombinant cytokines, specifically detected in the culture fluids of AAM, were assayed for their ability to inhibit CAM generation. Thus, in the presence of CpG DNA, RM (1 x 106 cells/ml) were cultured with various doses of recombinant cytokines. Twenty-four hours after the cultivation, the cells were washed three times and then cultured for an additional 24 h. The culture fluids harvested were assayed for CCL3. After certain cytokines were determined to inhibit CAM generation, the AAM Culture-Sup was treated with mAbs directed against the appropriate cytokines and applied to the assay of CAM generated from RM stimulated with CpG DNA. Amounts of mAbs (clone 110904; clone JES5-2A5) used in the experiments were determined according to the manufacturer’s protocol.
ELISPOT assay
To determine the percentage of AAM in the CAM preparation or the percentage of CAM in the AAM preparation, an ELISPOT assay was performed as previously described (32). A 96-well, Millititer HA plate (Millipore, Bedford, MA) was coated with 2 µg/ml anti-mouse CCL3 mAb or anti-mouse CCL17 mAb in 0.1 M carbonate buffer (pH 9.6) overnight at 4°C. The plates were then blocked with complete medium for 1 h. M suspensions diluted serially with complete medium were incubated in the plates for 24 h at 37°C. Next, the plates were washed with PBS containing 0.05% Tween 20 and overlaid with biotinylated anti-mouse CCL3 mAb or anti-mouse CCL17 mAb overnight at 4°C. Subsequently, the plates were washed and treated with a 1/2000 dilution of avidin-conjugated alkaline phosphatase (Sigma-Aldrich, St. Louis, MO). CCL3 or CCL17, secreted by single cells, was visualized by adding a solution of 5-bromo-4-chloro-3-indolyl phosphate/nitroblue tetrazolium (Sigma-Aldrich). The colorimetric reaction was halted after 30 min by washing the plates with water, and the spots were enumerated under x40 magnification. The dilution of the cells producing spots was used to calculate the total numbers of CCL3- or CCL17-producing M per sample. The results were displayed by the ratio between CCL3-producing M and CCL17-producing M.
Statistical analysis
Data are presented as mean ± SEM. Comparisons of the results between the experimental and control groups were made by ANOVA. Analysis was accomplished using Statview 4.5 (Abacus Concepts, Berkeley, CA). If a p value was <0.05, the result was considered to be significant.
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Results |
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generation by AAM or AAM products
The inhibitory effect of AAM on CAM generation was examined. Three kinds of M populations were used in these experiments: RM, freshly isolated peritoneal M from normal mice; CAM, RM stimulated with CpG DNA; AAM, peritoneal M from thermally injured mice. As shown in Table I, the three M populations used in this study exhibited typical properties for RM, CAM, and AAM, respectively. In the following studies, M with the ability to produce CCL3 are considered CAM, because CCL3 has been described as a typical chemokine specifically produced by CAM (4, 5). As shown in Table II, CAM were not found in cultures of RM stimulated with CpG DNA when they were cocultured with AAM at percentages of 39% or more. In addition, no CAM were generated when RM (lower chamber) were cultured with AAM (upper chamber) in a dual-chamber Transwell supplemented with CpG DNA (Fig. 1A). In the presence of CpG DNA, conversion of RM to CAM drastically decreased when the cultivation was performed with AAM Culture-Sup (culture supernatants of 1 x 106 cells/ml AAM 24 h after cultivation, 15%, v/v; Fig. 1B). Similar results were obtained when standard AAM induced by a mixture of IL-4 and IL-13 or culture fluids of these M were subjected to the same test (Fig. 1). The results shown in Table II and Fig. 1 indicate that the ability of AAM to inhibit CAM generation from RM stimulated with CpG DNA is evidenced by the soluble factors released from AAM.
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To determine which components inhibited CAM generation, AAM were cultured at the cell density of 1 x 106 cells/ml for 24 or 48 h. The culture fluids were assayed for various cytokines and PGE2, which have been described as typical products of M or M-related cells (4, 5, 16). As a control, the culture fluids of RM were used for the experiments. The results are shown in Fig. 2. PGE2, CCL17, IL-6, IL-10, and TGF- were all found in AAM culture fluids. In addition, almost the same amount of PGE2 and TGF- was detected in the culture fluids of RM. These results suggest that CCL17, IL-6, and IL-10 all may inhibit the generation of CAM from RM stimulated with CpG DNA.
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from RM stimulated with CpG DNA. In these experiments, various doses of murine rCCL17, rIL-10, and rIL-6 were individually added to cultures of RM stimulated with CpG DNA. rIL-10 at a dose of 100 pg/ml or more inhibited CAM generation (Fig. 3). CAM generation was also inhibited by rCCL17 at doses ranging from 20 to 100 pg/ml. The maximum inhibition was demonstrated when 20 pg/ml rCCL17 was added to cultures of RM stimulated with CpG DNA. However, rIL-6 at doses ranging from 20 to 1000 pg/ml did not inhibit CAM generation. Since 500–600 pg/ml IL-10 and CCL17 were detected in AAM-Culture Sup (culture fluids of AAM 24 h after cultivation, Fig. 2) and 15% (v/v) of AAM Culture-Sup was added to RM cultures (Fig. 1B), the individual amounts (75–90 pg/ml) of IL-10 or CCL17 contained in this assay system were not enough to completely inhibit CAM generation. When a mixture of rIL-10 and rCCL17 (100 pg/ml each) was added to cultures of RM stimulated with CpG DNA, CAM generation was completely inhibited (Fig. 3). These results suggest that both IL-10 and CCL17 contained in AAM Culture-Sup play a cooperative role in inhibiting CAM generation. These results were reproduced when the experiment was performed with AAM Culture-Sup and mAbs directed against CCL17 and IL-10. As shown in Fig. 4, AAM Culture-Sup inhibited CAM generation from RM (89% inhibition). This activity of AAM Culture-Sup was not completely abrogated when it was treated with anti-IL-10 mAb (clone 110904) and anti-CCL17 mAb (clone JES5-2A5) individually. Anti-IL-10 mAb treatment eliminated the activity of AAM Culture-Sup by 15%. Anti-CCL17 mAb treatment eliminated the activity by 50%. AAM Culture-Sup treated with isotype control Ab inhibited CAM generation from RM (90% inhibition). When AAM Culture-Sup was treated with a mixture of mAbs for IL-10 and CCL17, the ability of AAM Culture-Sup to inhibit CAM generation was eliminated by 78%. These results indicate that IL-10 and CCL17 may cooperatively inhibit CAM generation. Data shown in Figs. 3 and 4 suggest that small amounts of CCL17 released from AAM early after cultivation may have an influence on IL-10 production from AAM or induce expanded expression of IL-10R on RM.
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or AAM in various RM cultures
In the presence or absence of CpG DNA, RM (lower chamber) were cultured with AAM (upper chamber) in a dual-chamber Transwell supplemented with or without a mixture of mAbs for IL-10 (50 ng/ml) and CCL17 (500 ng/ml). M harvested from the lower chamber 24 h after cultivation were assayed using ELISPOT to determine the number of CCL3-producing M or CCL17-producing M. As shown in Fig. 5, CCL3-producing M were not found in the lower chamber of the Transwell (RM stimulated with CpG DNA) 24 h after cultivation with AAM (upper chamber). A majority of cells in the lower chamber was shown to be CCL17-producing M. However, a majority of cells in the lower chamber (RM stimulated with CpG DNA) converted to CCL3-producing M when Transwell cultures were performed with AAM (upper chamber) in the presence of a mixture of mAbs directed against CCL17 and IL-10. These results indicate that CCL3-producing M and CCL17-producing M are not mixed with each other in the same cell populations.
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Discussion |
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have been described as effector cells for the host’s innate immunities (6, 7, 8). It has been suggested that individuals whose AAM predominate, such as severely burned patients, are susceptible to various opportunistic pathogens due to impaired CAM generation (5, 16, 19). Therefore, the mechanism by which AAM inhibit CAM generation was investigated using RM (peritoneal M from normal mice), AAM (peritoneal M from burn mice), and CAM (RM stimulated with CpG DNA). As shown in Table I, these M preparations were typical cellular populations for RM, CAM, and AAM. When RM stimulated with CpG DNA were cocultured with AAM, CAM generation was clearly suppressed depending on the percentage of AAM added to the cocultures. In addition, RM stimulated with CpG DNA did not convert to CAM when they (lower chamber) were cultured with AAM (upper chamber) in a dual-chamber Transwell. A recent report has noted that AAM suppress mitogen-stimulated lymphocytic proliferations through cell-cell contact, implicating a receptor-mediated mechanism (33). However, the results in this study indicated that cell-cell contact between AAM and RM was not necessary for AAM to inhibit the generation of CAM. Furthermore, in the presence of CpG DNA, RM did not convert to CAM when they were cultured with culture fluids of AAM (AAM Culture-Sup). Since IL-6, IL-10, and CCL17 were specifically detected in AAM Culture-Sup, recombinant murine preparations of these cytokines were tested for their ability to inhibit CAM generation. The results showed that only IL-10 and/or CCL17 were shown to inhibit CAM generation from RM stimulated with CpG DNA, while IL-6 did not. TGF- was found in the culture fluids of RM and AAM. The various immunosuppressive activities of TGF- have already been documented (34, 35). NO production and arginase induction by M were inhibited when >1 ng/ml TGF- was added to the cultures (36). In these results, however, the amount of TGF- detected in the culture fluids of AAM was <100 pg/ml. This amount of TGF- may not represent inhibitory activity on CAM generation. Together, all of these results indicate that AAM inhibit the generation of CAM from RM by producing IL-10 and CCL17.
Recently, suppressor of cytokine signaling (SOCS) 1 and SOCS3 were described as key inhibitors of M activation (37, 38). Especially, IL-10 has been described as an activator of SOCS3, which inhibits the M activation stimulated by LPS (39). In addition, IL-6 has been identified as an activator of SOCS3 (37, 38). In this study, however, IL-6 did not inhibit CAM generation. These findings appear to stem from the different sensitivities of IL-10- and IL-6-induced signal transduction toward inhibitory mechanisms. SOCS3 inhibits IL-6-induced signal transduction; however, IL-10-induced signal transduction is not inhibited by SOCS3 (38, 40). The role played by CCL17 in the induction of SOCS3 is not known. The effect of this chemokine on the various functions of M is also unknown. However, the results of this study show that CCL17 may inhibit the immunobiological functions expressed by CAM. CCL17 has been suggested as an effector molecule on Th2 responses associated with AAM (41, 42). CCL17 migrates Th2 cells into inflammatory sites through the CCR4 displayed by Th2 cells (43, 44). Th2 responses enhanced by AAM stimulate further generation of AAM by producing IL-4, IL-10, and IL-13. These reactions severely impair the generation of CAM and Th1 cells. Actually, in an OVA-induced murine asthma model, anti-CCL17 mAb treatment reduced the production of Th2 cytokines (IL-4 and IL-13) in bronchoalveolar lavage fluids (45). Further studies are required to explore the inhibitory mechanism of CCL17 on CAM generation.
CpG DNA has been shown to activate dendritic cells, B cells, and NK cells as well as M (46, 47). In this study, we used CpG DNA as an inducer of CAM, because RM treated with CpG DNA produced CCL3, CCL5, and IL-12, expressed iNOS mRNA, showed strong killing activity against S. aureus, and induced Th1 responses (24). We have also reported that all of these parameters shown by CAM were not significantly demonstrated when RM were cultured with AAM (peritoneal M from burn mice, RM treated in vitro with a mixture of IL-4 and IL-13) (24). Among these properties shown by CAM, in this article we presented CCL3 production as a representative maker of CAM. In addition to CpG DNA, LPS, dsRNA, and IFN- have been reported to induce CAM generation (5, 11, 48). In our preliminary studies, when RM were cultured with AAM in the presence of LPS (10 ng/ml), dsRNA (2 µg/ml), or IFN- (1 ng/ml), CAM were not generated. In contrast, AAM, induced by 1 acid glycoprotein (15), a mixture of IL-4 and IL-13 (24) or glucocorticoids (49), equally inhibited CAM generation from RM stimulated with CpG DNA. These results indicate that the AAM, which appeared in various patients, might randomly inhibit the generation of CAM from RM. Thus, individuals with AAM predominating are nonspecifically susceptible to wide-ranging foreign invasions.
In this article, TNF-, IL-1, IL-6, PGE2, IL-10, CCL17, and TGF- were measured as possible soluble factors released from AAM. TNF-, IL-1, and IL-6 have been described as the products of M or M-related cells (4, 5, 16). PGE2, IL-10, and TGF- have also been described as M products with immunosuppressive activities (16). Therefore, we tested these soluble factors as possible effector molecules of AAM. Also, we tested CCL17 as a possible effector molecule of AAM. CCL17 and CCL22 have been described as chemokines equally associated with the induction of Th2 responses (1). This means there is a possibility that these two chemokines may be equally produced by AAM and equally cooperate with IL-10 on inhibiting CAM generation. However, due to availability, in this study we only measured CCL17. Further studies for CCL22 will be required.
In conclusion, CCL17 and IL-10 released from AAM were shown to inhibit CAM generation from RM stimulated with CpG DNA. The impairment of the host’s innate immunities associated with the function of CAM may be remedied by controlling AAM or AAM products (CCL17 and IL-10).
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Footnotes |
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2 Address correspondence and reprint requests to Dr. Fujio Suzuki, Department of Internal Medicine, University of Texas Medical Branch, 301 University Boulevard, Galveston, TX 77555-0435. E-mail address: fsuzuki{at}utmb.edu
3 Abbreviations used in this paper: M, macrophages; CAM, classically activated M; AAM, alternatively activated M; RM, resident M; iNOS, inducible NO synthase; TBSA, total body surface area; MRSA, methicillin-resistant S. aureus; SOCS, suppressor of cytokine signaling.
Received for publication August 11, 2003. Accepted for publication November 14, 2003.
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References |
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TopAbstractIntroductionMaterials and MethodsResultsDiscussionReferences |
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1-Acid glycoprotein (AGP) stimulates resident macrophages to generate alternatively activated macrophages (AAM). Part I. Biological properties of AGP-induced AAM. FASEB J. 17:C160.
-deprived donor environment. J. Interferon Cytokine Res. 16:957.[Medline]
production is dependent on macrophage secretion of IL-12. Clin. Immunol. Immunopathol. 84:185.[Medline]
receptors. J. Immunol. 168:3697.. J. Exp. Med. 178:605. suppresses interferon--induced toxoplasmastatic activity in murine macrophages by inhibition of tumor necrosis factor- production. Parasite Immunol. 23:169.[Medline]
stimulates arginase activity in macrophages: implications for the regulation of macrophage cytotoxicity. J. Immunol. 155:2077.[Abstract]
-dependent, IFN- secretion by bone marrow-derived macrophages controls an intracellular bacterial infection. J. Immunol. 167:6453.This article has been cited by other articles:
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  M. Ishii, H. Wen, C. A. S. Corsa, T. Liu, A. L. Coelho, R. M. Allen, W. F. Carson IV, K. A. Cavassani, X. Li, N. W. Lukacs, et al. Epigenetic regulation of the alternatively activated macrophage phenotype Blood, October 8, 2009; 114(15): 3244 - 3254. [Abstract] [Full Text] [PDF]
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 J. Day, A. Friedman, and L. S. Schlesinger Modeling the immune rheostat of macrophages in the lung in response to infection PNAS, July 7, 2009; 106(27): 11246 - 11251. [Abstract] [Full Text] [PDF]
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 D. M. Higgins, R. J. Basaraba, A. C. Hohnbaum, E. J. Lee, D. W. Grainger, and M. Gonzalez-Juarrero Localized Immunosuppressive Environment in the Foreign Body Response to Implanted Biomaterials Am. J. Pathol., July 1, 2009; 175(1): 161 - 170. [Abstract] [Full Text] [PDF]
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 C. Kadoch, E. B. Dinca, R. Voicu, L. Chen, D. Nguyen, S. Parikh, J. Karrim, M. A. Shuman, C. A. Lowell, P. A. Treseler, et al. Pathologic Correlates of Primary Central Nervous System Lymphoma Defined in an Orthotopic Xenograft Model Clin. Cancer Res., March 15, 2009; 15(6): 1989 - 1997. [Abstract] [Full Text] [PDF]
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 T. Raymond, M. Schaller, C. M. Hogaboam, N. W. Lukacs, R. Rochford, and S. L. Kunkel Toll-like Receptors, Notch Ligands, and Cytokines Drive the Chronicity of Lung Inflammation Proceedings of the ATS, December 1, 2007; 4(8): 635 - 641. [Abstract] [Full Text] [PDF]
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 R. Hussain, N. Talat, F. Shahid, and G. Dawood Longitudinal Tracking of Cytokines after Acute Exposure to Tuberculosis: Association of Distinct Cytokine Patterns with Protection and Disease Development Clin. Vaccine Immunol., December 1, 2007; 14(12): 1578 - 1586. [Abstract] [Full Text] [PDF]
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M. Weng, D. Huntley, I-F. Huang, O. Foye-Jackson, L. Wang, A. Sarkissian, Q. Zhou, W. A. Walker, B. J. Cherayil, and H. N. Shi Alternatively Activated Macrophages in Intestinal Helminth Infection: Effects on Concurrent Bacterial Colitis J. Immunol., October 1, 2007; 179(7): 4721 - 4731. [Abstract] [Full Text] [PDF]
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D.-M. Kuang, Y. Wu, N. Chen, J. Cheng, S.-M. Zhuang, and L. Zheng Tumor-derived hyaluronan induces formation of immunosuppressive macrophages through transient early activation of monocytes Blood, July 15, 2007; 110(2): 587 - 595. [Abstract] [Full Text] [PDF]
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G.-H. Chen, M. A. Olszewski, R. A. McDonald, J. C. Wells, R. Paine III, G. B. Huffnagle, and G. B. Toews Role of Granulocyte Macrophage Colony-Stimulating Factor in Host Defense Against Pulmonary Cryptococcus neoformans Infection during Murine Allergic Bronchopulmonary Mycosis Am. J. Pathol., March 1, 2007; 170(3): 1028 - 1040. [Abstract] [Full Text] [PDF]
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 D. Jankovic, M. C. Kullberg, C. G. Feng, R. S. Goldszmid, C. M. Collazo, M. Wilson, T. A. Wynn, M. Kamanaka, R. A. Flavell, and A. Sher Conventional T-bet+Foxp3- Th1 cells are the major source of host-protective regulatory IL-10 during intracellular protozoan infection J. Exp. Med., February 19, 2007; 204(2): 273 - 283. [Abstract] [Full Text] [PDF]
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T. L. Ness, J. L. Ewing, C. M. Hogaboam, and S. L. Kunkel CCR4 Is a Key Modulator of Innate Immune Responses J. Immunol., December 1, 2006; 177(11): 7531 - 7539. [Abstract] [Full Text] [PDF]
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H. Takahashi, Y. Tsuda, M. Kobayashi, D. N. Herndon, and F. Suzuki CCL2 as a trigger of manifestations of compensatory anti-inflammatory response syndrome in mice with severe systemic inflammatory response syndrome J. Leukoc. Biol., April 1, 2006; 79(4): 789 - 796. [Abstract] [Full Text] [PDF]
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K. J. Carpenter, K. F. Buckland, Z. Xing, and C. M. Hogaboam Intrapulmonary, Adenovirus-Mediated Overexpression of KARAP/DAP12 Enhances Fungal Clearance during Invasive Aspergillosis Infect. Immun., December 1, 2005; 73(12): 8402 - 8406. [Abstract] [Full Text] [PDF]
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K. J. Carpenter and C. M. Hogaboam Immunosuppressive Effects of CCL17 on Pulmonary Antifungal Responses during Pulmonary Invasive Aspergillosis Infect. Immun., November 1, 2005; 73(11): 7198 - 7207. [Abstract] [Full Text] [PDF]
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J. S. Satoguina, E. Weyand, J. Larbi, and A. Hoerauf T Regulatory-1 Cells Induce IgG4 Production by B Cells: Role of IL-10 J. Immunol., April 15, 2005; 174(8): 4718 - 4726. [Abstract] [Full Text] [PDF]
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C. Jakubzick, H. Wen, A. Matsukawa, M. Keller, S. L. Kunkel, and C. M. Hogaboam Role of CCR4 Ligands, CCL17 and CCL22, During Schistosoma mansoni Egg-Induced Pulmonary Granuloma Formation in Mice Am. J. Pathol., October 1, 2004; 165(4): 1211 - 1221. [Abstract] [Full Text] [PDF]
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Y. Tsuda, H. Takahashi, M. Kobayashi, T. Hanafusa, D. N. Herndon, and F. Suzuki CCL2, a product of mice early after systemic inflammatory response syndrome (SIRS), induces alternatively activated macrophages capable of impairing antibacterial resistance of SIRS mice J. Leukoc. Biol., August 1, 2004; 76(2): 368 - 373. [Abstract] [Full Text] [PDF]
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) or RM
) were cultured without any stimulation. The culture fluids harvested 24–48 h after cultivation were assayed for the soluble factors (listed below), that were known to be products of M
