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Departments of
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Molecular Microbiology and Immunology and
Veterinary Pathobiology, University of Missouri, Columbia, MO 65211
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Abstract |
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, IL-4, IL-6, IL-10, IL-12p70, GM-CSF, IFN-
, TNF-
, macrophage inflammatory protein-2, KC, macrophage inflammatory protein-1, and monocyte chemoattractant protein-1 directly from the tibiotarsal joint in arthritis-resistant C57BL/6 (B6) and -susceptible C3H/He (C3H) mice. Only the chemokines KC and monocyte chemoattractant protein-1 were differentially expressed in joints of B6 and C3H mice and correlated with the development of Lyme arthritis. Infection of CXCR2-/- mice on either genetic background resulted in a significant decrease in the development of pathology, although infection of CCR2-/- mice had little or no effect. Neutrophils in CXCR2-/- mice were marginalized within blood vessels and could not enter the joint tissue. These results suggest that chemokine-mediated recruitment of neutrophils into the infected joint is a key requirement for the development of experimental Lyme arthritis. |
Introduction |
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TopAbstractIntroductionMaterials and MethodsResultsDiscussionReferences |
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Numerous chemokines and cytokines have been correlated with resistance or susceptibility to development of experimental Lyme arthritis (6). However, to date there are no reports of cytokine or chemokine expression in mouse ankles, either at the mRNA or protein level, following infection with B. burgdorferi. Levels of cytokines have been measured in serum or plasma (7, 8, 9, 10, 11), cultured cell lines (12, 13, 14, 15, 16, 17), restimulated spleen or lymph node (18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31), or a tissue chamber (32), but not directly from the site of pathology. The importance of cytokine production in determining disease severity has been investigated in several studies by in vivo depletion of cytokines using specific Ab treatment, or by infection of specific gene knockout mice. Several earlier studies suggested that arthritis severity was linked to the development of Th cell subsets, with Th1 cells promoting severe arthritis and Th2 cells promoting disease resistance (18, 19). Treatment of resistant mice with Ab to IL-4 resulted in increased arthritis severity, while treatment of susceptible mice with Ab to IFN- resulted in decreased arthritis severity (18, 19). However, definitive studies using knockout mice demonstrated that these cytokines were not absolutely required for resistance or susceptibility to arthritis development (31, 33, 34). Infection of B6 IL-6-/- mice resulted in an increased incidence of arthritis as compared with wild-type B6 mice, but severity was not reported (26). However, infection of B6 IL-10-/- mice resulted in increased arthritis severity, but 10-fold fewer B. burgdorferi organisms than in the ankles of wild-type B6 mice (16). This suggested that IL-10-mediated control of arthritis severity occurred at the expense of effective bacterial clearance. Conversely, treatment of C3H mice with Ab to IL-12 resulted in decreased arthritis severity while increasing the numbers of spirochetes in tissues (8). Together these reports demonstrate that manipulation of pro- or anti-inflammatory cytokines can modulate the severity of arthritis development in mouse models, but none alone can alter the genetic phenotype of the mouse. Thus, it is likely that a complex balance of pro- and anti-inflammatory mediators ultimately combine to produce arthritis resistance or susceptibility.
In the current study, we examined the temporal production of a panel of 12 pro- and anti-inflammatory cytokines and chemokines directly from the ankles of arthritis-resistant and -susceptible mice following footpad inoculation with B. burgdorferi. To our surprise, we found few differences between resistant and susceptible animals in the production of the pro- and anti-inflammatory cytokines discussed above. Only production of the chemokines KC and monocyte chemoattractant protein (MCP)3-1 correlated with the development of arthritis in susceptible mice. Infection of mice deficient for the MCP-1 receptor, CCR2-/-, revealed few differences from wild-type mice. However, infection of mice deficient for the KC receptor, CXCR2-/-, revealed reduced arthritis severity in both resistant and susceptible mouse strains. Neutrophils from CXCR2-/- mice were unable to cross the vascular endothelium to enter the joint tissue. These results suggest that development of joint pathology in experimental Lyme disease is regulated by chemokine-mediated recruitment of neutrophils into the joint tissue.
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Materials and Methods |
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Female C3H/HeJ, C57BL/6J, and DBA/2J mice, 4–6 wk of age, were purchased from The Jackson Laboratory (Bar Harbor, ME). The CXCR2-/- mice (BALB/c-Cmkar2tm1 Mwm) were purchased as breeders, also from The Jackson Laboratory, and were backcrossed onto C3H (seven generations) or DBA (six generations) genetic backgrounds. Male B6/129 CCR2-/- mice were a kind gift from I. Charo (Gladstone Institute, San Francisco, CA) and were backcrossed onto C3H (six generations) or B6 (seven generations) genetic backgrounds. Heterozygous mice from each of the knockout strains were then intercrossed to produce knockout and wild-type littermates for use in experiments. The mice were housed in a specific pathogen-free facility and given sterile food and water ad libitum.
Bacteria and infections
A virulent low-passage, clonal isolate of the B. burgdorferi N40 strain (a kind gift from S. Barthold, University of California, Davis, CA) was used for all infections. Frozen stocks were placed in 7.5 ml of Barbour, Stoenner, Kelly (BSK) II medium (Sigma-Aldrich, St. Louis, MO) and grown to log phase by incubating for 5 days at 32°C. Spirochetes were enumerated using dark field microscopy and a Petroff-Hausser counting chamber (Hausser Scientific, Horsham, PA). Spirochetes were diluted using sterile BSK II medium and mice were inoculated in both hind footpads with 2.5 x 105 B. burgdorferi organisms contained in 50 µl of medium. Sham-infected mice received injections of medium alone.
Assessment of arthritis
Ankle swelling was determined by measuring the tibiotarsal joints through the thickest anteroposterior diameter of the ankle using a metric caliper (Ralmike’s Tool-A-Rama, South Plainfield, NJ). Baseline ankle diameters were determined immediately before infection and then weekly thereafter. Increases in ankle diameter were determined by subtracting the baseline measurement from the weekly measurement. To determine arthritis severity scores, one ankle from each mouse was chosen at random at sacrifice and excised by removing the skin and cutting just above and below the tibiotarsal joint. The excised joint was placed in 10% buffered zinc-formalin, decalcified, embedded in paraffin, and sections were stained with H&E. The sections were scored on a scale from 0 to 3 (35) for arthritis severity by two independent observers in a blinded manner. Grade 0 represents no inflammation, grades 1 and 2 represent mild to moderate inflammation, and grade 3 represents severe inflammation. Arthritis in histological samples was characterized by neutrophil and monocyte infiltration into the joints, tendons, and ligament sheaths; hyperplasia and hypertrophy of the synovium; and fibrin exudates. The extent of the observed changes provided the basis for the arthritis severity scores. Independent observer severity scores were averaged for each sample.
Direct measurement of cytokines in joint extracts
Cytokines were measured directly from joint extracts by a slight modification of the method described by Kasama et al. (36). Briefly, ankle joints were excised as described above and immediately frozen in liquid N2. Each frozen joint was then wrapped in aluminum foil and pulverized with a hammer. The still frozen, pulverized tissue was then placed immediately into 1 ml of ice cold homogenization buffer consisting of HBSS containing a 0.2% protease inhibitor mixture (Sigma-Aldrich) and 0.4% Triton X-100. The samples were homogenized on ice using a tissue homogenizer (IKA Works, Wilmington, NC). The samples were then centrifuged at 2000 x g for 20 min at 4°C. The supernatant was then filtered through a 0.45-µm filter and the resultant supernatant was diluted to 1.5 ml using homogenization buffer. Levels of cytokines and chemokines in supernatants were quantified by ELISA. ELISA kits for murine IL-1, IL-4, IL-6, IL-10, IL-12p70, GM-CSF, IFN-, TNF-, MCP-1 (OptEIA kits; BD PharMingen, San Diego, CA), macrophage inflammatory protein (MIP)-1, MIP-2, and KC (DuoSet; R&D Systems, Minneapolis, MN) were all performed according to the manufacturer’s instructions. Total protein concentration was measured using the BCA kit (Pierce, Rockford, IL) according to the manufacturer’s instructions. The concentration of cytokines and chemokines was expressed in picograms per milligram of protein.
Quantitative real-time PCR
Quantitative real-time PCR was performed using the ABI Prism 7700 Sequence Detection System (Applied Biosystems, Foster City, CA) to analyze levels of cytokine or chemokine mRNA induced by B. burgdorferi infection. Following mouse sacrifice, ankles were collected as described above, flash-frozen in liquid N2, and stored at -80°C. The frozen ankle joints were pulverized to a powder as described above, homogenized using the QIAShredder (Qiagen, Valencia, CA), and total RNA was extracted using the RNEasy Tissue kit (Qiagen). Synthesis of cDNA was performed using 1 µg of total RNA, MuLV reverse transcriptase (Applied Biosystems), and random hexamers (Applied Biosystems). The mouse gene Nidogen is a single copy gene (37) and was used as an endogenous control with a probe concentration of 200 nM and a concentration of 300 nM for each primer in each reaction. Nidogen primer and probe sequences were designed using the Primer Express (Applied Biosystems) primer design software. The primer and probe sequences were as follows: probe, (5'-VIC-AGCTCAGCCTGGCAGACGCAAAGT-TAMRA); forward primer (5'-AGGGCAGAATGCCTGAAC); and reverse primer (5'-AGGATACTGGAGCCCTTCGAG). For the quantification of KC and MCP-1 (38), or for IL-6, IFN-, MIP-1, and MIP-2 (39), primer and probe sequences were as described previously. The cDNA samples (1.5 µl) were mixed with primers, probe, and the TaqMan Universal PCR Master Mix in a total volume of 25 µl as described in the manufacturer’s protocol (Applied Biosystems). The PCR conditions were: 50°C for 2 min, 95°C for 10 min, and 45 cycles of 95°C for 15 s and 60°C for 1 min. Quantitative real-time PCR was performed in duplicate for KC, MCP-1, IL-6, IFN-, MIP-1, and MIP-2 and normalized to the copies of Nidogen mRNA within the same sample (multiplexed PCR). The levels of mRNA were expressed as a fold change in expression level as compared with untreated normal mice.
Quantitative assessment of B. burgdorferi numbers in tissues
Quantitative multiplex real-time PCR was performed using the ABI Prism 7700 Sequence Detection System (Applied Biosystems) to analyze levels of B. burgdorferi DNA present in the tissues of infected mice. Following mouse sacrifice, ankles were excised as described above, snap-frozen in liquid N2, and stored at -80°C. The frozen tissue was pulverized as described above and the DNA was extracted using the DNEasy Tissue kit (Qiagen). The DNA was eluted in 100 µl of elution buffer and diluted to 50 µg/ml using Tris-EDTA buffer. One microliter of the diluted sample was then used in PCRs and has been estimated to contain 1000 copies of the mouse Nidogen gene (37). The mouse Nidogen gene was used as an endogenous control as described above. Quantification of B. burgdorferi DNA in samples was done by detection of the Flagellin gene using primers and probes as described (40). TaqMan PCR conditions were the same as described above. Quantitative multiplex real-time PCR for each sample was performed in duplicate or triplicate for Flagellin and normalized to copies of Nidogen DNA from the same tube. B. burgdorferi DNA within each sample was quantified by comparing to a standard curve consisting of known numbers of B. burgdorferi. Similarly, normalization of mouse DNA within each sample was completed by comparing to a standard curve of dilutions of mouse DNA from the same tissue (ear or ankle).
Immunohistochemistry
Paraffin sections were fixed in xylene and rehydrated through ethanol and 1 M PBS (pH 6.0). Endogenous peroxidases were quenched with 3% H2O2 in methanol, before blocking nonspecific binding with 1.5% normal rabbit serum (ABC Staining System; Santa Cruz Biotechnology, Santa Cruz, CA) diluted in PBS. Sections were incubated overnight at 4°C with a 1/500 dilution of the anti-neutrophil mAb RB6-8C5 (41). Normal rat serum was used as a negative control. Diaminobenzidine tetrahydrochloride (Sigma-Aldrich) was used as the peroxidase substrate and slides were counterstained with Mayer’s hematoxylin solution (Fisher Scientific, Fair Lawn, NJ). Stained sections were then dehydrated and mounted.
Statistical analysis
Statistical analysis of the temporal expression of cytokines was completed using the SAS System (SAS Institute, Cary, NC) general linear models procedure, least squares means. All other analysis was completed using SigmaStat software (SPSS, Chicago, IL) for Student’s t test. Critical values for statistical significance were set at = 0.05.
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Results |
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The production of several cytokines and chemokines has been correlated with the development of experimental Lyme arthritis (6). However, none of these inflammatory modulators have been measured directly from the site of pathology. To determine the temporal expression of a panel of pro- and anti-inflammatory cytokines and chemokines directly from the tibiotarsal joint, we infected arthritis-resistant B6 and -susceptible C3H mice in the hind footpads with B. burgdorferi. Because we were harvesting ankle joints at early time points, and because the inoculation itself might cause cytokine or chemokine production, some mice from both strains were sham-infected by inoculating sterile BSK medium alone into their hind footpads. At time points of 0.3 (8 h), 1, 3, 5, 7, 10, 14, and 21 days postinfection, two sham-infected and five B. burgdorferi-infected mice from each mouse strain were sacrificed, and both ankle joints were removed and processed separately for cytokine analysis. To follow the development of arthritis during the experimental time course, at days 0, 7, 14, and 21 postinfection five sham-infected and B. burgdorferi-infected mice from each mouse strain were chosen at random and ankle diameter measurements were taken. Fig. 1 shows the increase in ankle diameters for C3H (Fig. 1A) and B6 (Fig. 1B) mice over the 21-day experiment. As is typical for this model system, the susceptible C3H mice developed ankle swelling during the second week of infection that began to resolve by day 21 postinfection. Ankle diameters for the sham-inoculated C3H mice did not change during the experiment. Arthritis-resistant B6 mice developed only a mild increase in ankle diameter during the infection, while there was no change in the sham-infected B6 mice. Three extra sham- and B. burgdorferi-infected mice per mouse strain were inoculated on day 0 for use in determining arthritis severity scores (Fig. 1C). The sham-infected mice had none or very low levels of inflammatory cells in their tibiotarsal joints at day 21 postinfection, while the B. burgdorferi-infected B6 and C3H mice developed mild and severe arthritis, respectively.
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, IL-4, IL-6, IL-10, IL-12, IFN-, TNF-, GM-CSF, MIP-2, KC, MIP-1, and MCP-1 in ankle joints over the 21-day experimental period following inoculation of B. burgdorferi into the footpads of arthritis-resistant B6 and -susceptible C3H mice. The data are expressed on a per milligram of protein basis with the average protein concentration per ankle being 
1 mg/ml. Curves for sham-infected mice were completed but are not shown for clarity. Surprisingly, we found few significant differences in cytokine or chemokine expression between B6 and C3H mice over the course of infection. To be considered significant, the expression levels from B. burgdorferi-infected animals had to be significantly different from the sham-infected animals, and also significantly different from the B. burgdorferi-infected animals from the other mouse strain. Using these criteria, the expression of only KC and MCP-1 in joints from C3H mice were significantly different from the B6 mice. The increased expression of KC and MCP-1 in C3H mice also correlated with the development of arthritis (see Fig. 1). This experiment was completed three times using B6 mice, and twice using DBA mice as the arthritis-resistant strain, with similar results. These data suggest that the expression of KC and MCP-1 may be required for the development of experimental Lyme arthritis.
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In the mouse, MCP-1 appears to recruit monocytes/macrophages solely through the CCR2 (45). To further characterize the role of MCP-1 in the development of Lyme arthritis, we infected CCR2-/- mice on both B6 and C3H genetic backgrounds and followed the development of arthritis for 21 days. Ankle swelling curves and ankle severity scores are shown for B6 and C3H CCR2-/- and CCR2+/+ mice in Fig. 3. In three of five experiments, the B6 CCR2-/- mice had a slight but significant increase (p < 0.001) in ankle swelling compared with the B6 CCR2+/+ mice (Fig. 3A). However, no significant differences were seen in ankle swelling between the C3H CCR2-/- and CCR2+/+ mice (Fig. 3B). Deletion of CCR2 had no significant effect on the development of Lyme arthritis in disease-resistant or -susceptible strains. Arthritis severity scores were similar between the C3H CCR2-/- and CCR2+/+ mice and also between the B6 CCR2-/- and CCR2+/+ mice (Fig. 3C). Histologic examination of joints from C3H CCR2-/- mice revealed the presence of numerous cells with macrophage morphology within the inflammatory infiltrate, similar to the C3H CCR2+/+ mice (data not shown), indicating the continued recruitment of macrophages into the joint despite the absence of CCR2. Joint cytokine expression for IL-4, IL-6, IL-10, IL-12, MCP-1, KC, and MIP-2 at day 21 postinfection was also similar between the CCR2-/- and CCR2+/+ mice (data not shown). Levels of B. burgdorferi DNA in ankle joints 21 days postinfection tended to be higher in the CCR2-/- as compared with the CCR2+/+ mice (Table II). In the B6 mice, the difference in B. burgdorferi DNA levels between the CCR2-/- and CCR2+/+ mice approached significance (p = 0.065) and may signal decreased host control of spirochetes when monocyte recruitment is impaired. These results suggest that MCP-1 may be playing a role in the development of Lyme arthritis, but other mechanisms exist to recruit macrophages in the absence of CCR2.
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The CXCR2 appears to be the sole receptor for KC in the mouse (46). To further characterize the role of KC in the development of Lyme arthritis, we infected CXCR2-/- mice on both DBA and C3H genetic backgrounds, and followed the development of arthritis for 21 days. Ankle swelling curves and arthritis severity scores for the CXCR2-/- mice are shown in Fig. 4. Ankle swelling curves for the DBA CXCR2-/- mice were no different from the DBA CXCR2+/+ mice as neither of the groups developed significant swelling. However, the C3H CXCR2-/- mice developed significantly less ankle swelling (p < 0.001) than the C3H CXCR2+/+ mice (Fig. 4B). Arthritis severity scores were significantly less (p < 0.03) for both the DBA CXCR2-/- mice and the C3H CXCR2-/- mice when compared with their CXCR2+/+ controls (Fig. 4C).
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Discussion |
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and IL-4, in resistance and susceptibility to development of Lyme arthritis (18, 19). Restimulation of lymph node or spleen cells from susceptible mouse strains resulted in the production of IFN-, while those from resistant mouse strains resulted in higher levels of IL-4. The linkage of arthritis resistance with IL-4, and susceptibility with IFN- production, was weakened through studies demonstrating arthritis resistance in mice deficient in IL-4 or IL-4R (33, 34) and arthritis susceptibility in mice deficient for IFN- or IFNR (31, 33). Although there was no absolute requirement for IL-4 or IFN- for arthritis resistance or susceptibility, respectively, treatment of mice with recombinant cytokines (20, 27) or anti-cytokine Abs (8, 9, 18, 19) was able to modulate disease development. This suggests that the balance of pro- and anti-inflammatory mediators produced during the host immune response regulates the severity of arthritis. In the current study, we characterized the temporal expression of a number of pro- and anti-inflammatory cytokines and chemokines directly from the site of pathology. Surprisingly, we found few differences in the production of IL-1, IL-4, IL-6, IL-10, IL-12, GM-CSF, TNF-, IFN-, MIP-2, and MIP-1 in joints between resistant and susceptible mouse strains. Only the chemokines KC and MCP-1 were differentially expressed at significantly higher levels in joints from susceptible, as compared with resistant, mouse strains and correlated with the onset of arthritis.
KC is a CXC chemokine that is the murine homologue of human growth-related oncogene- (47) and is a potent neutrophil chemoattractant (48). MCP-1 is a CC chemokine and is a potent chemoattractant and activator of monocytes (48). The inflammatory infiltrate in the joints of susceptible mice consists mainly of neutrophils and monocytes (2). The high levels of KC and MCP-1 found in the joint tissue from C3H mice correlate well with the recruitment of these two cell types into the site of infection. Because the joints from C3H mice were highly inflamed, it is surprising that other proinflammatory cytokines were not expressed at levels higher than the mildly inflamed joints from DBA or B6 mice. Several of the cytokines tested were reported to be differentially expressed in tissues from resistant and susceptible mouse strains, although they were not measured directly from infected ankle tissue (6, 42). Quantitative measurement of mRNA levels from mouse joints confirmed the results obtained measuring cytokine protein. Others have also found good correlation between protein and mRNA data measuring cytokines directly from arthritic joints using virtually identical methods (49).
In the mouse, KC binds only to the CXCR2 (50), while MCP-1 binds only to the CCR2 (45). We infected CXCR2-/- and CCR2-/- mice on both arthritis-resistant and -susceptible genetic backgrounds to determine the requirement for signaling through these receptors for the development of Lyme arthritis. Infection of CCR2-/- mice on either a B6 or C3H background had little effect on the development of arthritis or on spirochete clearance. B6 CCR2-/- mice did have significantly greater ankle swelling during the peak of arthritis than did the CCR2+/+ mice. The fact that monocytes were still present within the inflammatory infiltrates in the joints of CCR2-/- mice indicates that redundant recruitment mechanisms must act in these mice. One possibility might be MCP-3, which is a monocyte chemoattractant known to signal through other receptors besides CCR2 (48). We have noted the consistent up-regulation of mRNA levels of MCP-3 during microarray analysis of joints from susceptible mouse strains (C. R. Brown, unpublished observations). However, other researchers have reported success in blocking monocyte/macrophage-mediated responses using the CCR2-/- mice, although not in arthritis models (51, 52, 53, 54, 55, 56, 57, 58).
In contrast to our results using the CCR2-/- mice, infection of CXCR2-/- mice on both arthritis-resistant and -susceptible genetic backgrounds resulted in a significant decrease in arthritis severity, but had little effect on spirochete loads in joint tissue. Immunohistochemical staining revealed that neutrophils present in joint sections were marginalized in blood vessels and could not traverse the vascular endothelium to enter the joint tissue. These results are similar to those using CXCR2-/- mice in a model of acute experimental polynephritis. Neutrophils were unable to traverse the mucosal epithelium and accumulated in the tissues leading to neutrophil abscesses, increased bacterial loads, and renal scarring (59, 60). In the Lyme arthritis model, it appears that killing spirochetes is secondary to the role of immune regulation by neutrophils. CXCR2-/- and CXCR2+/+ mice contained similar numbers of spirochete DNA within their joints. However, without CXCR2 neutrophils could not enter the joint tissue, and the inflammatory response was poorly developed. Similar results were reported in a model of hepatitis B virus infection, whereby depletion of neutrophils had no effect on viral clearance by CTL but rather profoundly inhibited the recruitment of other Ag-nonspecific cells into the liver resulting in diminished tissue damage (61). Thus, in a similar way neutrophil entry into the joint tissue may result in the subsequent recruitment of other inflammatory cells and ultimately in the development of pathology.
Joints from CXCR2-/- mice contained high levels of KC at 21 days postinfection, a time when KC levels in CXCR2+/+ mice were returning to near baseline levels. This may indicate the presence of a negative feedback loop during inflammation to limit the recruitment of neutrophils to the site of infection. As neutrophils accumulate in the tissue, the production of KC may be down-regulated, limiting the numbers of new neutrophils entering the site, thereby minimizing tissue damage and promoting inflammation resolution. In the joints from the CXCR2-/- animals, neutrophils could not cross the vascular endothelium to enter the joint tissue and KC levels remained elevated. Because joints from CXCR2-/- and CXCR2+/+ mice contained similar levels of spirochetes 21 days postinfection, the mechanism of KC down-regulation is not likely mediated by bacterial clearance, but rather by an interaction between the recruited neutrophils and the KC-producing cells within the joint. An alternative explanation is that neutrophils bind and metabolize or sequester KC as they enter the tissues.
In conclusion, we have shown that the recruitment of neutrophils into the infected joint appears to be a prerequisite for the development of Lyme arthritis and that neutrophils play a nonphagocytic immunoregulatory role in the development of pathology.
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Acknowledgments |
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Footnotes |
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2 Address correspondence and reprint requests to Dr. Charles R. Brown, University of Missouri, 315 Connaway Hall, Columbia, MO 65211. E-mail address: BrownChar{at}missouri.edu
3 Abbreviations used in this paper: MCP-1, monocyte chemoattractant protein 1; BSK, Barbour, Stoenner, Kelly; MIP, macrophage inflammatory protein.
Received for publication February 20, 2003. Accepted for publication May 2, 2003.
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References |
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interferon and interleukin-10 production in draining lymph nodes from mice infected with Borrelia burgdorferi. Infect. Immun. 68:7162. Interferon is not required for arthritis resistance in the murine Lyme disease model. Infect. Immun. 69:3737. interferon. Infect. Immun. 67:3329. and MCP-1. Br. J. Pharmacol. 134:1166.[Medline]
and TNF-, and the noninvolvement of IL-6, in the development of monoclonal antibody-induced arthritis. J. Immunol. 169:1459. are overcome to control infection with Leishmania donovani in CC chemokine receptor (CCR) 5-, macrophage inflammatory protein-1-, or CCR2-deficient mice. J. Immunol. 163:5519.This article has been cited by other articles:
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H. Crandall, D. M. Dunn, Y. Ma, R. M. Wooten, J. F. Zachary, J. H. Weis, R. B. Weiss, and J. J. Weis Gene Expression Profiling Reveals Unique Pathways Associated with Differential Severity of Lyme Arthritis J. Immunol., December 1, 2006; 177(11): 7930 - 7942. [Abstract] [Full Text] [PDF]
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J. J. Lazarus, M. J. Meadows, R. E. Lintner, and R. M. Wooten IL-10 Deficiency Promotes Increased Borrelia burgdorferi Clearance Predominantly through Enhanced Innate Immune Responses J. Immunol., November 15, 2006; 177(10): 7076 - 7085. [Abstract] [Full Text] [PDF]
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M. Guerau-de-Arellano, J. Alroy, D. Bullard, and B. T. Huber Aggravated Lyme Carditis in CD11a-/- and CD11c-/- Mice Infect. Immun., November 1, 2005; 73(11): 7637 - 7643. [Abstract] [Full Text] [PDF]
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J. Glasner, H. Blum, V. Wehner, H. U. Stilz, J. D. Humphries, G. P. Curley, A. P. Mould, M. J. Humphries, R. Hallmann, M. Rollinghoff, et al. A Small Molecule {alpha}4{beta}1 Antagonist Prevents Development of Murine Lyme Arthritis without Affecting Protective Immunity J. Immunol., October 1, 2005; 175(7): 4724 - 4734. [Abstract] [Full Text] [PDF]
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T. Iyoda, K. Nagata, M. Akashi, and Y. Kobayashi Neutrophils Accelerate Macrophage-Mediated Digestion of Apoptotic Cells In Vivo as Well as In Vitro J. Immunol., September 15, 2005; 175(6): 3475 - 3483. [Abstract] [Full Text] [PDF]
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K. Chen, Y. Wei, A. Alter, G. C. Sharp, and H. Braley-Mullen Chemokine expression during development of fibrosis versus resolution in a murine model of granulomatous experimental autoimmune thyroiditis J. Leukoc. Biol., September 1, 2005; 78(3): 716 - 724. [Abstract] [Full Text] [PDF]
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 H. Crandall, Y. Ma, D. M. Dunn, R. S. Sundsbak, J. F. Zachary, P. Olofsson, R. Holmdahl, J. H. Weis, R. B. Weiss, C. Teuscher, et al. Bb2Bb3 Regulation of Murine Lyme Arthritis Is Distinct from Ncf1 and Independent of the Phagocyte Nicotinamide Adenine Dinucleotide Phosphate Oxidase Am. J. Pathol., September 1, 2005; 167(3): 775 - 785. [Abstract] [Full Text] [PDF]
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M. Guerau-de-Arellano, J. Alroy, and B. T. Huber {beta}2 Integrins Control the Severity of Murine Lyme Carditis Infect. Immun., June 1, 2005; 73(6): 3242 - 3250. [Abstract] [Full Text] [PDF]
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M. R.-E.-I. Benhnia, D. Wroblewski, M. N. Akhtar, R. A. Patel, W. Lavezzi, S. C. Gangloff, S. M. Goyert, M. J. Caimano, J. D. Radolf, and T. J. Sellati Signaling through CD14 Attenuates the Inflammatory Response to Borrelia burgdorferi, the Agent of Lyme Disease J. Immunol., February 1, 2005; 174(3): 1539 - 1548. [Abstract] [Full Text] [PDF]
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A. Ray, D. Kumar, A. Shakya, C. R. Brown, J. L. Cook, and B. K. Ray Serum Amyloid A-Activating Factor-1 (SAF-1) Transgenic Mice Are Prone to Develop a Severe Form of Inflammation-Induced Arthritis J. Immunol., October 1, 2004; 173(7): 4684 - 4691. [Abstract] [Full Text] [PDF]
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C. R. Brown, V. A. Blaho, and C. M. Loiacono Treatment of Mice with the Neutrophil-Depleting Antibody RB6-8C5 Results in Early Development of Experimental Lyme Arthritis via the Recruitment of Gr-1- Polymorphonuclear Leukocyte-Like Cells Infect. Immun., September 1, 2004; 72(9): 4956 - 4965. [Abstract] [Full Text] [PDF]
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C. L. Galindo, A. A. Fadl, J. Sha, and A. K. Chopra Microarray Analysis of Aeromonas hydrophila Cytotoxic Enterotoxin-Treated Murine Primary Macrophages Infect. Immun., September 1, 2004; 72(9): 5439 - 5445. [Abstract] [Full Text] [PDF]
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D. D. Bolz, R. S. Sundsbak, Y. Ma, S. Akira, C. J. Kirschning, J. F. Zachary, J. H. Weis, and J. J. Weis MyD88 Plays a Unique Role in Host Defense but Not Arthritis Development in Lyme Disease J. Immunol., August 1, 2004; 173(3): 2003 - 2010. [Abstract] [Full Text] [PDF]
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J. L. Lomas-Neira, C.-S. Chung, P. S. Grutkoski, E. J. Miller, and A. Ayala CXCR2 inhibition suppresses hemorrhage-induced priming for acute lung injury in mice J. Leukoc. Biol., July 1, 2004; 76(1): 58 - 64. [Abstract] [Full Text] [PDF]
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N. Liu, R. R. Montgomery, S. W. Barthold, and L. K. Bockenstedt Myeloid Differentiation Antigen 88 Deficiency Impairs Pathogen Clearance but Does Not Alter Inflammation in Borrelia burgdorferi-Infected Mice Infect. Immun., June 1, 2004; 72(6): 3195 - 3203. [Abstract] [Full Text] [PDF]
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A. I. Medeiros, A. Sa-Nunes, E. G. Soares, C. M. Peres, C. L. Silva, and L. H. Faccioli Blockade of Endogenous Leukotrienes Exacerbates Pulmonary Histoplasmosis Infect. Immun., March 1, 2004; 72(3): 1637 - 1644. [Abstract] [Full Text] [PDF]
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, C3H; •, B6 animals. All values are on a per milligram of protein basis. Data are expressed as means ± SD. Five mice were sacrificed at each time point and each ankle was processed individually. Data shown are representative of three separate experiments. For statistical significance, values from the B. burgdorferi-infected mice must be significantly different from the sham-infected mice of the same mouse strain and be significantly different from the infected mice from the opposite mouse strain. Statistical significance: *, p < 0.001.
