science pharmaceutical expo biotech jobs
Originally published online as doi:10.1189/jlb.1107794 on February 19, 2008

Published online before print February 19, 2008
This Article
Right arrow Abstract Freely available
Right arrow Full Text (PDF)
Right arrow All Versions of this Article:
jlb.1107794v1
83/5/1194    most recent
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Services
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Download to citation manager
Right arrow reprints & permissions
Citing Articles
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Wildenberg, M. E.
Right arrow Articles by Versnel, M. A.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Wildenberg, M. E.
Right arrow Articles by Versnel, M. A.
(Journal of Leukocyte Biology. 2008;83:1194-1200.)
© 2008 by Society for Leukocyte Biology

Lack of CCR5 on dendritic cells promotes a proinflammatory environment in submandibular glands of the NOD mouse

Manon E. Wildenberg*,1, Cornelia G. van Helden-Meeuwsen*, Joop P. van de Merwe*, Christophe Moreno{dagger}, Hemmo A. Drexhage* and Marjan A. Versnel*

* Department of Immunology, Erasmus MC, Rotterdam, The Netherlands; and
{dagger} Division of Gastroenterology and Hepatopancreatology, Erasme Hospital, Brussels, Belgium

1Correspondence: Dept. of Immunology, Erasmus MC, Dr. Molewaterplein 50, 3015 GE Rotterdam, The Netherlands. E-mail: m.wildenberg{at}erasmusmc.nl


arrow
ABSTRACT
 
Sjögren’s syndrome is an autoimmune disease characterized by lymphocytic infiltration of the salivary glands. In the NOD mouse, a model for this disease, the development of lymphocytic infiltrates in the salivary glands is preceded by an accumulation of dendritic cells (DC). Given the key importance of DC in regulating the immune response, we characterized the DC isolated from NOD salivary glands. These DC lacked membrane expression of CCR5, whereas DC from control salivary glands did express this molecule. The lack of expression was present already prior to the onset of lymphocytic infiltration, indicating that this was not the result of ongoing inflammation. DC from other sources in the NOD mouse also showed a decrease in CCR5 expression. The lack of CCR5 expression in the NOD salivary gland was accompanied by an increase in inflammatory chemokines. Furthermore, DC from CCR5–/– animals or DC treated with a CCR5 antagonist showed increased secretion of IL-12. Interestingly, in Sjögren’s syndrome patients, CCR5 expression on circulating monocytes was decreased and correlated to increased levels of IL-12. These data indicate that CCR5 has regulatory properties and that the lack of CCR5 in NOD DC contributes to the proinflammatory environment in the salivary glands.

Key Words: chemokine receptor • Sjögren’s syndrome • autoimmunity • regulation


arrow
INTRODUCTION
 
The NOD mouse is frequently used as a model to study the development of primary Sjögren’s syndrome (pSjS) [1 , 2 ]. Similar to human patients, these mice develop lymphocytic infiltrates in the salivary glands and finally show a decreased salivary flow. The lymphocytic infiltrates start to develop at approximately 7 weeks of age and show an increasing structural organization [2 ]. Earlier research in our laboratory has shown that the influx of lymphocytes is preceded by an accumulation of dendritic cells (DC) [2 ].

DC are potent APC capable of stimulating naïve and memory T cells and boosting B cell responses, thus playing a key role in the initiation of the immune response [3 , 4 ]. Furthermore, DC are crucial to the development of tolerance, preventing harmful immune responses against harmless antigens [5 , 6 ]. To fully exert their sentinel function, DC migrate from the blood into tissues and further to the draining lymph nodes. This migration is a complex, multi-step process, tightly regulated by numerous molecules including chemokines and their receptors [7 , 8 ].

The chemokine family contains a large number of proteins, characterized by their small size and conserved cysteine residues [9 ]. Most chemokines bind and signal through several receptors, and one receptor usually has multiple ligands [9 , 10 ], creating a highly complex network of interactions. Chemokine receptors are G-protein-coupled 7-transmembrane receptors, which have differential expression patterns on various leukocyte subsets [11 ]. On DC, the expression of chemokine receptors is linked to their state of maturation. For example, immature DC express CXCR4 and CCR1, -2, and -5, most of which are down-regulated after exposure to an activating stimulus [12 , 13 ]. Besides cell trafficking, chemokine receptors have been described to regulate several other processes in DC, including endocytosis, dendrite formation, cell survival, and maturation of DC [14 15 16 17 18 ].

Given the importance of DC in the immune response, the expression of chemokine receptors in the DC accumulating in the NOD salivary glands was determined. Most strikingly, a complete lack of CCR5 expression was observed in these cells. Earlier, it has been shown that mice deficient in CCR5 display excessive immune responses, for example, in an influenza model [19 ]. Therefore, we hypothesized that the lack of CCR5 results in a proinflammatory environment. The results indeed show an immunoregulatory role for CCR5 in DC, both by scavenging of inflammatory ligands and by regulating the secretion of IL-12.


arrow
MATERIALS AND METHODS
 
Animals
Female C57BL/6 and CCR5–/– mice were obtained from Harlan (Horst, The Netherlands) and Jackson ImmunoResearch Laboratories (Bar Harbor, ME, USA), respectively. NOD/LTj mice were bred in our own facility under specified pathogen-free conditions. Animals were used at the age of 5–20 weeks. All mice were supplied with water and standard chow ad libitum. Experimental procedures were approved by the Erasmus University Animal Ethics Committee (Rotterdam, The Netherlands).

Human materials
Heparinized blood and serum samples were obtained from female pSjS patients (mean age 52.7±16.4; mean disease duration 6.6±7.8 years) at Erasmus MC (Rotterdam, The Netherlands). Inclusion criteria consisted of a positive diagnosis, according to the criteria of the American-European consensus group [20 ], and the absence of other rheumatic or autoimmune disorders. Control blood samples were obtained from female volunteers (mean age 45.8±6.1) not suffering from autoimmune diseases or taking immunosuppressive medication. The Medical Ethical Committee of Erasmus MC approved this study, and all subjects gave informed consent.

PBMC were isolated from peripheral blood using a Ficoll gradient (Amersham, Little Chalfont, UK).

Isolation of CD11c+ cells from tissue
Organs were removed and cleared of adipose tissue and lymph nodes. Tissue was digested by Liberase (Roche, Woerden, The Netherlands) treatment at 37°C and washed twice in PBS containing 10% FCS. Subsequently, cells were cleared from debris using a Ficoll (Amersham Biosciences, Uppsala, Sweden) gradient. Cells appearing at the interface were incubated with mouse CD11c microbeads (Miltenyi, Bergisch Gladbach, Germany), according to the manufacturer’s protocol and isolated using the AutoMACS (Miltenyi) system.

Flow cytometry
Cells were resuspended in PBS containing 1% BSA and incubated with primary antibodies, which were biotinylated MHC class II (C57BL/6 clone ER-TR3; NOD clone 10.2.16), biotinylated CD11c, CD8{alpha}-FITC, CD80-PE, CD86-FITC, CD40-PE, CXCR4-PE, and CCR5-PE (all BD Biosciences, San Jose, CA, USA). For human material, CCR5-PE-Cy5 (BD Biosciences) was used. Afterwards, cells were washed twice and when appropriate, incubated with streptavidin-allophycocyanin (BD Biosciences). For intracellular CCR5 staining, surface CCR5 was blocked by incubation with a tenfold excess of unlabeled anti-CCR5 antibody (BD Biosciences). Next, cells were fixed using 2% paraformaldehyde and permeabilized using 0.5% saponin. Cells were then incubated with labeled CCR5 antibody diluted in 0.5% saponin, washed, and resuspended in 1% BSA. Cell suspensions were analyzed using a FACSCalibur (Becton Dickenson, San Diego, CA, USA) and WinMDI software (Microsoft Co., Redmont, WA, USA). Expression was calculated as mean fluorescence intensity (MFI) specific staining – MFI isotype control. For calculation of relative expression levels, expression of C57Bl/6 was set to 1, and NOD levels were calculated accordingly.

Cytokine determination
Levels of IL-12p40 in supernatant were determined using an IL-12p40 Duoset kit (R&D Systems Inc., Minneapolis, MN, USA), and serum levels of IL-12 were determined using a human cytokine 25-plex AB bead kit (Biosource, Camarillo, CA, USA), according to the manufacturer’s suggestions. Induction of IL-12 was calculated as secretion after LPS stimulation divided by secretion without stimulus.

Bone marrow-derived DC (BMDC) culture
BM was isolated from femur and cultured in RPMI-1640 medium (BioWhittaker, Verviers, Belgium), supplemented with 10% FCS (BioWhittaker), 50 µM β-ME (Sigma Aldrich, Deisenhofen, Germany), and 20 ng/mL recombinant mouse GM-CSF (Biosource) for 9 days. To enrich immature DC, cells were incubated with anti-CD86 antibody (clone GL-1), washed, labeled with goat anti-rat microbeads (Miltenyi), and separated using the AutoMACS (Miltenyi) system. For stimulation experiments, BMDC were incubated with Met-RANTES (10 µg/mL, R&D Systems Inc.), where appropriate, and stimulated using LPS (5 ng/mL, Sigma Aldrich) for 16 h, after which supernatants were collected.

RNA isolation and RT-PCR
RNA was isolated from submandibular glands (SMG) using the RNeasy Mini kit (Qiagen, Valencia, CA, USA), according to the manufacturer’s suggestions. cDNA was generated using Superscript II (Invitrogen, Carlsbad, CA, USA) and random hexamer primers (Amersham). RT-PCR analysis was carried out using predesigned primer/probe sets (Applied Biosystems, Foster City, CA, USA), according to the manufacturer’s suggestions. For calculation of relative expression, all samples were normalized against expression of the household gene Abl.

Statistics
Data were analyzed using Student’s t-test, except for relative data (C57BL/6 set to 1), where the Mann Whitney U-test was used. Error bars represent SD. Correlations were calculated using Spearman’s Rank test. All statistical analyses were performed using SPSS software (SPSS Inc., Chicago, IL, USA) and were considered significant if P < 0.05.


arrow
RESULTS
 
Lack of CCR5 expression on CD11c+ cells in NOD salivary gland
CD11c+ cells were isolated from SMG of NOD and control mice, and the chemokine receptor expression pattern was analyzed by flow cytometry. CD11c+ cells isolated from control mice showed expression of CXCR4 and CCR5 (Fig. 1A ), NOD CD11c+ showed a somewhat decreased expression whereas of CXCR4 and completely lacked expression of CCR5 (Fig. 1A) . As CCR5 is down-regulated during the activation of DC, the lack of CCR5 could be a result of an increased activation of CD11c+ cells in NOD SMG. However, expression of the activation markers CD40, CD80, and CD86 did not show a significant pattern of up-regulation (Fig. 1A and 1B) .


Figure 1
View larger version (19K):
[in this window]
[in a new window]

  		Figure 1. Chemokine receptor and activation markers on SMG CD11c+ cells, which were isolated from SMG by Ficoll gradient followed by magnetic bead isolation. Cells were then stained for surface markers and analyzed by flow cytometry. (A) Gray lines represent NOD mice; black lines represent C57BL/6 mice; one representative sample of each strain is shown. (B) Average expression in C57BL/6 mice was set to 1, and expression levels of NOD mice were calculated accordingly (n=4–5); error bars represent SD; *, P < 0.05. (C) CD11c+ cells were isolated from SMG of mice at 5 or 15 weeks of age. Expression of CCR5 was calculated as MFI specific staining – MFI control.

As expression of CCR5 can be influenced by inflammatory environmental stimuli, expression levels may be influenced by the ongoing inflammatory process in the salivary glands of NOD mice. Therefore, CD11c+ cells isolated from SMG prior to the development of lymphocytic infiltrates (5 weeks) were compared with CD11c+ from SMG with full-blown inflammatory infiltrations (15 weeks). At both ages, CD11c+ from control mice showed clear expression of CCR5 (Fig. 1C) , whereas expression in NOD was already below detection level at 5 weeks of age. The absence of activation marker up-regulation in combination with the fact that CCR5 expression is undetectable already prior to the onset of lymphocytic infiltration indicates that the lack of expression is not caused by the ongoing inflammation in the SMG.

Decreased CCR5 expression in NOD DC derived from other sources
To determine if the lack of CCR5 expression is tissue-specific or a general phenomenon in NOD mice, CD11c+ cells from other in vitro and in vivo sources were studied. CD11c+/autofluorescencelow DC derived from the lungs of NOD mice showed a clearly decreased expression of CCR5 when compared with control mice (Fig. 2A ). Similarly, DC cultured from BM (BMDC) in the presence of GM-CSF showed a significantly decreased expression of CCR5 (Fig. 2B) .


Figure 2
View larger version (25K):
[in this window]
[in a new window]

  		Figure 2. CCR5 expression on lung and BM CD11c+ cells, which were isolated from lungs of C57BL/6 and NOD mice (A) or generated in vitro from BM (B) and analyzed by flow cytometry. Results of gated autofluorescentlow cells (A) or gated CD11c+ cells (B) are shown; black lines represent specific staining; gray lines represent isotype control. One representative image for each strain is shown. Bars represent average expression; error bars represent SD. (C, D) CD11c+ cells were isolated from salivary glands (C) or cultured from BM (D) and analyzed for intracellular expression of CCR5 by flow cytometry. Average expression in C57BL/6 mice was set to 1, and expression levels of NOD mice were calculated accordingly (n=3); error bars represent SD; *, P < 0.05.

Lack of surface CCR5 is accompanied by increased intracellular CCR5 in NOD SMG
CCR5 is a cycling receptor, undergoing rounds of ligand-binding, internalization, uncoupling of the receptor, and relocalization to the cell surface [21 ]. Therefore, the lack of CCR5 on CD11c+ cells in NOD SMG could be a result of an altered localization of this molecule. Indeed, when intracellular expression of CCR5 was studied, an increased expression was found in CD11c+ cells isolated from NOD SMG when compared with CD11c+ from control mice (Fig. 2C) . However, in BMDC, no increase in intracellular CCR5 was found (Fig. 2D) , indicating that the altered CCR5 localization in SMG may be dependent on the microenvironment.

Lack of CCR5 receptor expression may lead to increased free ligand
The lack of CCR5 expression on the surface of CD11c+ cells in the SMG of NOD mice may influence biologically available levels of CCR5 ligands CCL3, CCL4, and CCL5. When measured in total salivary gland, mRNA levels of these chemokines tended to be somewhat higher in NOD than in control mice (Fig. 3A ). Furthermore, in control mice, these chemokines can be scavenged from the environment by surface CCR5 on tissue CD11c+ cells, thus decreasing the level of free, active ligand. This results in a balanced ratio between CCR5 ligands and CCR5 itself. As a result of the lack of surface CCR5 expression, this mechanism is not available in NOD mice, which would be indicated by an increased CCR5 ligand:CCR5 ratio. Indeed, ratios of CCL3, CCL4, and CCL5 relative to the level of CCR5 were increased significantly in the NOD salivary gland (Fig. 3B) .


Figure 3
View larger version (31K):
[in this window]
[in a new window]

  		Figure 3. Ratio of CCR5 ligands:CCR5 is increased in NOD SMG. Total mRNA was isolated from SMG, and expression levels of CCL3, CCL4, CCL5, and CCR5 were determined by RT-PCR (n=5). Error bars represent SD; *, P < 0.05.

Lack of CCR5 signaling leads to increased IL-12 secretion
CCR5 has been linked to the regulation of IL-12 secretion in an oral-tolerance model [22 ]. To test whether CCR5 also regulates IL-12 specifically in DC, BMDC were cultured from C57BL/6 and CCR5–/– mice and stimulated with LPS. Secretion of IL-12, as well as relative induction, indeed was higher in CCR5–/– mice (Fig. 4A ). To exclude a role for the knockout procedure, wild-type BMDC treated with the CCR5 antagonist Met-RANTES [23 ] were compared with untreated cells. Again, secretion and induction of IL-12 were higher in BMDC treated with Met-RANTES (Fig. 4B) . This supports the idea that CCR5 is involved in the regulation of IL-12 secretion.


Figure 4
View larger version (10K):
[in this window]
[in a new window]

  		Figure 4. IL-12 secretion in the absence of CCR5 signaling. (A) BMDC were generated in the presence of GM-CSF from wild-type (WT) and CCR5–/– animals. Cells were then stimulated using LPS (5 ng/mL), and IL-12p40 was measured in the supernatant by ELISA (n=6). Error bars represent SD. (B) BMDC were generated in the presence of GM-CSF from wild-type animals, treated with met-RANTES (met), and stimulated using LPS (5 ng/mL). IL-12p40 levels in the supernatants were measured by ELISA (n=3). Bars represent mean values; error bars represent SD.

Decreased levels of CCR5 in pSjS patients
The relevance of decreased CCR5 expression in human pSjS was explored by measuring CCR5 expression on monocytes, which are generally considered the most important blood precursors for DC. Similar to the mouse model, a significant decrease in surface expression of CCR5 was found in monocytes of pSjS patients when compared with monocytes of controls (Fig. 5A ). Interestingly, the level of CCR5 expression was negatively correlated to the serum concentration of IL-12 (Fig. 5B ; Spearman’s Rho –0.483; P=0.004).


Figure 5
View larger version (16K):
[in this window]
[in a new window]

  		Figure 5. Decreased CCR5 expression in pSjS patients. (A) PBMC were isolated from pSjS patients and healthy controls and analyzed by flow cytometry. CCR5 expression was determined as MFI-specific staining – MFI isotype control. Each dot represents one individual patient; *, P < 0.01 (B) Serum levels of IL-12p40 were determined by the AB bead kit and correlated to the monocyte expression of CCR5, as determined by flow cytometry. Each dot represents one individual patient; *, P < 0.05.


arrow
DISCUSSION
 
No expression of CCR5 was found on CD11c+ cells in the salivary glands of NOD mice, whereas control mice did show expression. As the lack of CCR5 was present prior to the development of lymphocytic infiltrates, and expression of activation markers was not increased, this was not the result of an increased state of DC activation as a result of the ongoing inflammation. In CD11c+ cells isolated from NOD mice, an increase in intracellular CCR5 was detected, indicating that the lack of surface expression may at least partly be explained by an altered subcellular localization of the molecule. However, BMDC and lung DC also showed decreased CCR5 expression, which was not accompanied by increased intracellular levels, suggesting a different cause such as a genetic deficiency. Therefore, the complete lack of CCR5 on SMG CD11c+ cells might be partially explained by an altered subcellular localization and partially by another, possibly genetic factor. The lack of surface CCR5 expression appears to have functional consequences, as NOD mice show defective recruitment in response to a CCR5 ligand in an in vivo migration model [24 ].

CCR5 was initially thought to be involved in directing immature DC to tissue sites [13 ]. Surprisingly, in the steady-state, CCR5–/– mice develop normally and show cell numbers and lymphoid architecture similar to wild-type animals [25 ]. Furthermore, several reports show normal macrophage recruitment in CCR5–/– mice during inflammation [19 , 25 26 27 ]. These findings may be explained in light of the high redundancy in the chemokine system. In CCR5-deficient cells, migration is probably regulated by other chemokine molecules, such as CCR1 and CCR2. RNA levels of these receptors were comparable in NOD and control SMG (data not shown), which may explain the presence of DC in NOD SMG, despite the lack of CCR5.

In accordance with previous studies in CCR5–/– mice [19 , 26 , 28 ], mRNA of CCR5 ligands tended to be increased in NOD SMG. In addition, the lack of expression of CCR5 in DC isolated from NOD SMG leads to a lack of scavenging by this receptor, thus contributing to increased amounts of free, biologically available ligands. Such scavenging by CCR5 has been reported to play a role in the control as well as the resolution of immune responses [29 , 30 ]. The increased availability of free CCL3, -4, and -5 may lead to an enhanced influx of other leukocytes, such as T cell and macrophages, as is seen in the salivary glands of NOD mice. One could argue that the increased levels of CCR5 ligand are causative rather than the result of decreased expression of CCR5 as a result of receptor internalization. However, as DC cultured from BM of NOD mice in vitro also show strong defects in expression of CCR5, this appears to be unlikely. Furthermore, extended exposure of wild-type DC to CCR5 ligands does not lead to decreased expression of the receptor, with the exception of CCL3 at very high concentrations (data not shown). However, even is this case, expression of CCR5 remained at clearly detectable levels, unlike NOD DC in vivo.

In a mouse model for experimental autoimmune encephalitis, oral tolerance could be induced in wild-type, but not in CCR5–/– mice [22 ]. Furthermore, in the CCR5–/– mice, increased levels of IL-12 were observed in the GALT. The current study shows that DC derived from CCR5–/– or DC treated with a CCR5 antagonist show increased secretion of IL-12p40 compared with wild-type BMDC. Although levels were low, results showed a similar trend for IL-12p70 (data not shown). Earlier, increased secretion of IL-12 in response to various stimuli was reported in NOD DC and NOD macrophages, which also show decreased expression of CCR5, and this was indeed the result of autocrine signaling [31 , 32 ]. Interestingly, in pSjS patients, decreased levels of CCR5 were observed in peripheral monocytes, and this correlated negatively with the level of IL-12 in the serum, supporting the hypothesis that CCR5 plays a role in the regulation of IL-12 in DC.

IL-12 secreted by DC is a well-known inducer of Th1 cells and plays an important role in the development of various immune responses [33 ]. In the NOD mouse model, addition of IL-12 leads to acceleration of the development of diabetes, whereas inhibition of IL-12 using an antagonist has a protective effect [34 , 35 ]. Unfortunately, the salivary glands were not studied in these experiments, but increased expression of IL-12 is found in the salivary glands of pSjS patients [36 ]. Furthermore, transgenic mice overexpressing IL-12 develop infiltrations in their salivary glands similar to those seen in NOD mice, indicating an important role for this cytokine in the pathogenesis of pSjS [37 ].

In a recent report, CCR5–/– mice showed an increased antibody production in the setting of a cardiac transplant [38 ]. These antibodies were highly functional, as CCR5–/– mice showed acute humoral rejection, which could be transferred by serum [39 ]. Similar increases in levels of antibody were observed earlier in an infection model in CCR5–/– animals [25 ]. In both studies, numbers of IL-4-producing T cells were increased, suggesting a skewing of the immune system toward antibody production in the absence of CCR5. Increased levels of autoantibodies are a well-known phenomenon in human pSjS patients as well as NOD mice [40 41 42 ]. The lack of CCR5 on DC may play a role in this by increasing the numbers of IL-4-producing T cells, thus stimulating the generation of autoantibodies. Indeed, in NOD mice, IL-4 is of crucial importance to the development of pSjS-like symptoms, as NOD.IL-4–/– mice do not develop salivary dysfunction [43 ]. Interestingly, in the patient cohort described in this study, CCR5 expression was lower in patients who were positive for the well-known autoantibody SS-B, although this did not reach statistical significance (P=0.067; data not shown).

The hypothesis that the lack of CCR5 plays a role in the increased expression of IL-12 as well as IL-4 appears somewhat at odds with the original Th1/Th2 paradigm. However, it has now been clearly shown that the two responses are not as mutually exclusive as previously thought, and in pSjS patients, increased numbers of IFN-{gamma}- and IL-4-producing T cells are found [44 45 46 ]. Furthermore, the NOD mouse, which is regarded as a strongly Th1-biased animal, displays a more aggressive form of the stereotypical Th2 condition asthma [47 ].

In humans, the role of CCR5 polymorphisms [48 , 49 ] has been studied in various autoimmune diseases. In diabetes and systemic lupus erythematosus, no association was found between the frequency of CCR5 polymorphisms and disease incidence [50 51 52 ], but more complications and interestingly, higher levels of autoantibodies were found in patients with polymorphic CCR5 alleles [52 53 54 ]. Only one earlier study focused on CCR5 in pSjS and concluded that a CCR5 mutation may somewhat protect against pSjS, contradicting our results [55 ]. However, in that study, only one polymorphism was included, leaving the possibility that other polymorphisms influenced the results. In the current study, CCR5 was determined at the protein level, thus resulting in actual expression levels rather than genetics.

The function of CCR5 in the migration of various cell types has been studied extensively. This study shows that in addition, CCR5 is a regulatory molecule and that its expression on immature DC is likely to contribute to the tolerogenic phenotype of this subset. In the salivary gland of the NOD mouse model for pSjS, the lack of CCR5 on DC contributes to a more proinflammatory environment.

Received November 27, 2007; revised January 14, 2008; accepted January 25, 2008.


arrow
REFERENCES
 
    1
  1. Hu, Y., Nakagawa, Y., Purushotham, K. R., Humphreys-Beher, M. G. (1992) Functional changes in salivary glands of autoimmune disease-prone NOD mice Am. J. Physiol. 263,E607-E614[Medline]
    2. 2
  2. van Blokland, S. C., van Helden-Meeuwsen, C. G., Wierenga-Wolf, A. F., Drexhage, H. A., Hooijkaas, H., van de Merwe, J. P., Versnel, M. A. (2000) Two different types of sialoadenitis in the NOD- and MRL/lpr mouse models for Sjogren’s syndrome: a differential role for dendritic cells in the initiation of sialoadenitis? Lab. Invest. 80,575-585[Medline]
    3. 3
  3. Shortman, K., Liu, Y. J. (2002) Mouse and human dendritic cell subtypes Nat. Rev. Immunol. 2,151-161[CrossRef][Medline]
    4. 4
  4. Dubois, B., Bridon, J. M., Fayette, J., Barthelemy, C., Banchereau, J., Caux, C., Briere, F. (1999) Dendritic cells directly modulate B cell growth and differentiation J. Leukoc. Biol. 66,224-230[Medline]
    5. 5
  5. Ueno, H., Klechevsky, E., Morita, R., Aspord, C., Cao, T., Matsui, T., Di Pucchio, T., Connolly, J., Fay, J. W., Pascual, V., Palucka, A. K., Banchereau, J. (2007) Dendritic cell subsets in health and disease Immunol. Rev. 219,118-142[CrossRef][Medline]
    6. 6
  6. Steinman, R. M., Hawiger, D., Nussenzweig, M. C. (2003) Tolerogenic dendritic cells Annu. Rev. Immunol. 21,685-711[CrossRef][Medline]
    7. 7
  7. Dieu, M. C., Vanbervliet, B., Vicari, A., Bridon, J. M., Oldham, E., Ait-Yahia, S., Briere, F., Zlotnik, A., Lebecque, S., Caux, C. (1998) Selective recruitment of immature and mature dendritic cells by distinct chemokines expressed in different anatomic sites J. Exp. Med. 188,373-386[Abstract/Free Full Text]
    8. 8
  8. Weis, M., Schlichting, C. L., Engleman, E. G., Cooke, J. P. (2002) Endothelial determinants of dendritic cell adhesion and migration: new implications for vascular diseases Arterioscler. Thromb. Vasc. Biol. 22,1817-1823[Abstract/Free Full Text]
    9. 9
  9. Zlotnik, A., Yoshie, O., Nomiyama, H. (2006) The chemokine and chemokine receptor superfamilies and their molecular evolution Genome Biol. 7,243[CrossRef][Medline]
    10. 10
  10. Murdoch, C., Finn, A. (2000) Chemokine receptors and their role in inflammation and infectious diseases Blood 95,3032-3043[Abstract/Free Full Text]
    11. 11
  11. Patel, L., Charlton, S. J., Chambers, J. K., Macphee, C. H. (2001) Expression and functional analysis of chemokine receptors in human peripheral blood leukocyte populations Cytokine 14,27-36[CrossRef][Medline]
    12. 12
  12. Sallusto, F., Schaerli, P., Loetscher, P., Schaniel, C., Lenig, D., Mackay, C. R., Qin, S., Lanzavecchia, A. (1998) Rapid and coordinated switch in chemokine receptor expression during dendritic cell maturation Eur. J. Immunol. 28,2760-2769[CrossRef][Medline]
    13. 13
  13. Vecchi, A., Massimiliano, L., Ramponi, S., Luini, W., Bernasconi, S., Bonecchi, R., Allavena, P., Parmentier, M., Mantovani, A., Sozzani, S. (1999) Differential responsiveness to constitutive vs. inducible chemokines of immature and mature mouse dendritic cells J. Leukoc. Biol. 66,489-494[Abstract]
    14. 14
  14. Yanagawa, Y., Onoe, K. (2003) CCR7 ligands induce rapid endocytosis in mature dendritic cells with concomitant up-regulation of Cdc42 and Rac activities Blood 101,4923-4929[Abstract/Free Full Text]
    15. 15
  15. Niess, J. H., Brand, S., Gu, X., Landsman, L., Jung, S., McCormick, B. A., Vyas, J. M., Boes, M., Ploegh, H. L., Fox, J. G., Littman, D. R., Reinecker, H. C. (2005) CX3CR1-mediated dendritic cell access to the intestinal lumen and bacterial clearance Science 307,254-258[Abstract/Free Full Text]
    16. 16
  16. Flanagan, K., Moroziewicz, D., Kwak, H., Horig, H., Kaufman, H. L. (2004) The lymphoid chemokine CCL21 costimulates naive T cell expansion and Th1 polarization of non-regulatory CD4+ T cells Cell. Immunol. 231,75-84[CrossRef][Medline]
    17. 17
  17. Zou, W., Machelon, V., Coulomb-L’Hermin, A., Borvak, J., Nome, F., Isaeva, T., Wei, S., Krzysiek, R., Durand-Gasselin, I., Gordon, A., Pustilnik, T., Curiel, D. T., Galanaud, P., Capron, F., Emilie, D., Curiel, T. J. (2001) Stromal-derived factor-1 in human tumors recruits and alters the function of plasmacytoid precursor dendritic cells Nat. Med. 7,1339-1346[CrossRef][Medline]
    18. 18
  18. Marsland, B. J., Battig, P., Bauer, M., Ruedl, C., Lassing, U., Beerli, R. R., Dietmeier, K., Ivanova, L., Pfister, T., Vogt, L., Nakano, H., Nembrini, C., Saudan, P., Kopf, M., Bachmann, M. F. (2005) CCL19 and CCL21 induce a potent proinflammatory differentiation program in licensed dendritic cells Immunity 22,493-505[CrossRef][Medline]
    19. 19
  19. Dawson, T. C., Beck, M. A., Kuziel, W. A., Henderson, F., Maeda, N. (2000) Contrasting effects of CCR5 and CCR2 deficiency in the pulmonary inflammatory response to influenza A virus Am. J. Pathol. 156,1951-1959[Abstract/Free Full Text]
    20. 20
  20. Vitali, C., Bombardieri, S., Jonsson, R., Moutsopoulos, H. M., Alexander, E. L., Carsons, S. E., Daniels, T. E., Fox, P. C., Fox, R. I., Kassan, S. S., Pillemer, S. R., Talal, N., Weisman, M. H. (2002) Classification criteria for Sjogren’s syndrome: a revised version of the European criteria proposed by the American-European Consensus Group Ann. Rheum. Dis. 61,554-558[Abstract/Free Full Text]
    21. 21
  21. Mueller, A., Strange, P. G. (2004) Mechanisms of internalization and recycling of the chemokine receptor, CCR5 Eur. J. Biochem. 271,243-252[Medline]
    22. 22
  22. DePaolo, R. W., Lathan, R., Karpus, W. J. (2004) CCR5 regulates high dose oral tolerance by modulating CC chemokine ligand 2 levels in the GALT J. Immunol. 173,314-320[Abstract/Free Full Text]
    23. 23
  23. Proudfoot, A. E., Power, C. A., Hoogewerf, A. J., Montjovent, M. O., Borlat, F., Offord, R. E., Wells, T. N. (1996) Extension of recombinant human RANTES by the retention of the initiating methionine produces a potent antagonist J. Biol. Chem. 271,2599-2603[Abstract/Free Full Text]
    24. 24
  24. Bouma, G., Nikolic, T., Coppens, J. M., van Helden-Meeuwsen, C. G., Leenen, P. J., Drexhage, H. A., Sozzani, S., Versnel, M. A. (2005) NOD mice have a severely impaired ability to recruit leukocytes into sites of inflammation Eur. J. Immunol. 35,225-235[CrossRef][Medline]
    25. 25
  25. Zhou, Y., Kurihara, T., Ryseck, R. P., Yang, Y., Ryan, C., Loy, J., Warr, G., Bravo, R. (1998) Impaired macrophage function and enhanced T cell-dependent immune response in mice lacking CCR5, the mouse homologue of the major HIV-1 coreceptor J. Immunol. 160,4018-4025[Abstract/Free Full Text]
    26. 26
  26. Algood, H. M., Flynn, J. L. (2004) CCR5-deficient mice control Mycobacterium tuberculosis infection despite increased pulmonary lymphocytic infiltration J. Immunol. 173,3287-3296[Abstract/Free Full Text]
    27. 27
  27. Andres, P. G., Beck, P. L., Mizoguchi, E., Mizoguchi, A., Bhan, A. K., Dawson, T., Kuziel, W. A., Maeda, N., MacDermott, R. P., Podolsky, D. K., Reinecker, H. C. (2000) Mice with a selective deletion of the CC chemokine receptors 5 or 2 are protected from dextran sodium sulfate-mediated colitis: lack of CC chemokine receptor 5 expression results in a NK1.1+ lymphocyte-associated Th2-type immune response in the intestine J. Immunol. 164,6303-6312[Abstract/Free Full Text]
    28. 28
  28. Moreno, C., Nicaise, C., Gustot, T., Quertinmont, E., Nagy, N., Parmentier, M., Louis, H., Deviere, J. (2006) Chemokine receptor CCR5 deficiency exacerbates caerulein-induced acute pancreatitis in mice Am. J. Physiol. Gastrointest. Liver Physiol. 291,G1089-G1099[Abstract/Free Full Text]
    29. 29
  29. Ariel, A., Fredman, G., Sun, Y. P., Kantarci, A., Van Dyke, T. E., Luster, A. D., Serhan, C. N. (2006) Apoptotic neutrophils and T cells sequester chemokines during immune response resolution through modulation of CCR5 expression Nat. Immunol. 7,1209-1216[CrossRef][Medline]
    30. 30
  30. D’Amico, G., Frascaroli, G., Bianchi, G., Transidico, P., Doni, A., Vecchi, A., Sozzani, S., Allavena, P., Mantovani, A. (2000) Uncoupling of inflammatory chemokine receptors by IL-10: generation of functional decoys Nat. Immunol. 1,387-391[CrossRef][Medline]
    31. 31
  31. Poligone, B., Weaver, D. J., Jr, Sen, P., Baldwin, A. S., Jr, Tisch, R. (2002) Elevated NF-{kappa}B activation in nonobese diabetic mouse dendritic cells results in enhanced APC function J. Immunol. 168,188-196[Abstract/Free Full Text]
    32. 32
  32. Alleva, D. G., Pavlovich, R. P., Grant, C., Kaser, S. B., Beller, D. I. (2000) Aberrant macrophage cytokine production is a conserved feature among autoimmune-prone mouse strains: elevated interleukin (IL)-12 and an imbalance in tumor necrosis factor-{alpha} and IL-10 define a unique cytokine profile in macrophages from young nonobese diabetic mice Diabetes 49,1106-1115[Abstract]
    33. 33
  33. Langrish, C. L., McKenzie, B. S., Wilson, N. J., de Waal Malefyt, R., Kastelein, R. A., Cua, D. J. (2004) IL-12 and IL-23: master regulators of innate and adaptive immunity Immunol. Rev. 202,96-105[CrossRef][Medline]
    34. 34
  34. Trembleau, S., Penna, G., Bosi, E., Mortara, A., Gately, M. K., Adorini, L. (1995) Interleukin 12 administration induces T helper type 1 cells and accelerates autoimmune diabetes in NOD mice J. Exp. Med. 181,817-821[Abstract/Free Full Text]
    35. 35
  35. Trembleau, S., Penna, G., Gregori, S., Gately, M. K., Adorini, L. (1997) Deviation of pancreas-infiltrating cells to Th2 by interleukin-12 antagonist administration inhibits autoimmune diabetes Eur. J. Immunol. 27,2330-2339[Medline]
    36. 36
  36. Kolkowski, E. C., Reth, P., Pelusa, F., Bosch, J., Pujol-Borrell, R., Coll, J., Jaraquemada, D. (1999) Th1 predominance and perforin expression in minor salivary glands from patients with primary Sjogren’s syndrome J. Autoimmun. 13,155-162[CrossRef][Medline]
    37. 37
  37. McGrath-Morrow, S., Laube, B., Tzou, S. C., Cho, C., Cleary, J., Kimura, H., Rose, N. R., Caturegli, P. (2006) IL-12 overexpression in mice as a model for Sjogren lung disease Am. J. Physiol. Lung Cell. Mol. Physiol. 291,L837-L846[Abstract/Free Full Text]
    38. 38
  38. Amano, H., Bickerstaff, A., Orosz, C. G., Novick, A. C., Toma, H., Fairchild, R. L. (2005) Absence of recipient CCR5 promotes early and increased allospecific antibody responses to cardiac allografts J. Immunol. 174,6499-6508[Abstract/Free Full Text]
    39. 39
  39. Nozaki, T., Amano, H., Bickerstaff, A., Orosz, C. G., Novick, A. C., Tanabe, K., Fairchild, R. L. (2007) Antibody-mediated rejection of cardiac allografts in CCR5-deficient recipients J. Immunol. 179,5238-5245[Abstract/Free Full Text]
    40. 40
  40. Harley, J. B., Alexander, E. L., Bias, W. B., Fox, O. F., Provost, T. T., Reichlin, M., Yamagata, H., Arnett, F. C. (1986) Anti-Ro (SS-A) and anti-La (SS-B) in patients with Sjogren’s syndrome Arthritis Rheum. 29,196-206[Medline]
    41. 41
  41. Watanabe, T., Tsuchida, T., Kanda, N., Mori, K., Hayashi, Y., Tamaki, K. (1999) Anti-{alpha}-fodrin antibodies in Sjogren syndrome and lupus erythematosus Arch. Dermatol. 135,535-539[Abstract/Free Full Text]
    42. 42
  42. Yanagi, K., Ishimaru, N., Haneji, N., Saegusa, K., Saito, I., Hayashi, Y. (1998) Anti-120-kDa {alpha}-fodrin immune response with Th1-cytokine profile in the NOD mouse model of Sjogren’s syndrome Eur. J. Immunol. 28,3336-3345[CrossRef][Medline]
    43. 43
  43. Gao, J., Killedar, S., Cornelius, J. G., Nguyen, C., Cha, S., Peck, A. B. (2006) Sjogren’s syndrome in the NOD mouse model is an interleukin-4 time-dependent, antibody isotype-specific autoimmune disease J. Autoimmun. 26,90-103[CrossRef][Medline]
    44. 44
  44. Kelso, A. (1995) Th1 and Th2 subsets: paradigms lost? Immunol. Today 16,374-379[CrossRef][Medline]
    45. 45
  45. Fitzpatrick, D. R., Kelso, A. (1998) Independent regulation of cytokine genes in T cells: the paradox in the paradigm Transplantation 65,1-5[Medline]
    46. 46
  46. Mitsias, D. I., Tzioufas, A. G., Veiopoulou, C., Zintzaras, E., Tassios, I. K., Kogopoulou, O., Moutsopoulos, H. M., Thyphronitis, G. (2002) The Th1/Th2 cytokine balance changes with the progress of the immunopathological lesion of Sjogren’s syndrome Clin. Exp. Immunol. 128,562-568[CrossRef][Medline]
    47. 47
  47. Araujo, L. M., Lefort, J., Nahori, M. A., Diem, S., Zhu, R., Dy, M., Leite-de-Moraes, M. C., Bach, J. F., Vargaftig, B. B., Herbelin, A. (2004) Exacerbated Th2-mediated airway inflammation and hyperresponsiveness in autoimmune diabetes-prone NOD mice: a critical role for CD1d-dependent NKT cells Eur. J. Immunol. 34,327-335[CrossRef][Medline]
    48. 48
  48. Samson, M., Libert, F., Doranz, B. J., Rucker, J., Liesnard, C., Farber, C. M., Saragosti, S., Lapoumeroulie, C., Cognaux, J., Forceille, C., Muyldermans, G., Verhofstede, C., Burtonboy, G., Georges, M., Imai, T., Rana, S., Yi, Y., Smyth, R. J., Collman, R. G., Doms, R. W., Vassart, G., Parmentier, M. (1996) Resistance to HIV-1 infection in caucasian individuals bearing mutant alleles of the CCR-5 chemokine receptor gene Nature 382,722-725[CrossRef][Medline]
    49. 49
  49. Mummidi, S., Bamshad, M., Ahuja, S. S., Gonzalez, E., Feuillet, P. M., Begum, K., Galvis, M. C., Kostecki, V., Valente, A. J., Murthy, K. K., Haro, L., Dolan, M. J., Allan, J. S., Ahuja, S. K. (2000) Evolution of human and non-human primate CC chemokine receptor 5 gene and mRNA. Potential roles for haplotype and mRNA diversity, differential haplotype-specific transcriptional activity, and altered transcription factor binding to polymorphic nucleotides in the pathogenesis of HIV-1 and simian immunodeficiency virus J. Biol. Chem. 275,18946-18961[Abstract/Free Full Text]
    50. 50
  50. Gambelunghe, G., Ghaderi, M., Brozzetti, A., Del Sindaco, P., Gharizadeh, B., Nyren, P., Hjelmstrom, P., Nikitina-Zake, L., Sanjeevi, C. B., Falorni, A. (2003) Lack of association of CCR2–64I and CCR5-{Delta} 32 with type 1 diabetes and latent autoimmune diabetes in adults Hum. Immunol. 64,629-632[Medline]
    51. 51
  51. Dubois-Laforgue, D., Hendel, H., Caillat-Zucman, S., Zagury, J. F., Winkler, C., Boitard, C., Timsit, J. (2001) A common stromal cell-derived factor-1 chemokine gene variant is associated with the early onset of type 1 diabetes Diabetes 50,1211-1213[Abstract/Free Full Text]
    52. 52
  52. Aguilar, F., Nunez-Roldan, A., Torres, B., Wichmann, I., Sanchez-Roman, J., Gonzalez-Escribano, M. F. (2003) Chemokine receptor CCR2/CCR5 polymorphism in Spanish patients with systemic lupus erythematosus J. Rheumatol. 30,1770-1774[Abstract/Free Full Text]
    53. 53
  53. Mlynarski, W. M., Placha, G. P., Wolkow, P. P., Bochenski, J. P., Warram, J. H., Krolewski, A. S. (2005) Risk of diabetic nephropathy in type 1 diabetes is associated with functional polymorphisms in RANTES receptor gene (CCR5): a sex-specific effect Diabetes 54,3331-3335[Abstract/Free Full Text]
    54. 54
  54. Yang, B., Houlberg, K., Millward, A., Demaine, A. (2004) Polymorphisms of chemokine and chemokine receptor genes in Type 1 diabetes mellitus and its complications Cytokine 26,114-121[CrossRef][Medline]
    55. 55
  55. Petrek, M., Cermakova, Z., Hutyrova, B., Micekova, D., Drabek, J., Rovensky, J., Bosak, V. (2002) CC chemokine receptor 5 and interleukin-1 receptor antagonist gene polymorphisms in patients with primary Sjogren’s syndrome Clin. Exp. Rheumatol. 20,701-703[Medline]




This Article
Right arrow Abstract Freely available
Right arrow Full Text (PDF)
Right arrow All Versions of this Article:
jlb.1107794v1
83/5/1194    most recent
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Services
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Download to citation manager
Right arrow reprints & permissions
Citing Articles
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Wildenberg, M. E.
Right arrow Articles by Versnel, M. A.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Wildenberg, M. E.
Right arrow Articles by Versnel, M. A.