FRINK Linked Data Fragments server

Matches in SemOpenAlex for { <https://semopenalex.org/work/W73990037> ?p ?o ?g. }

• W73990037 endingPage "150" @default.
• W73990037 startingPage "139" @default.
• W73990037 abstract "Chemokines are low molecular weight cytokines which act as chemoattractants for infiltrating cells bearing appropriate receptors (CCR) to sites of inflammation. It has been proposed that CCR2 on monocytes is responsible for their recruitment into the central nervous system (CNS) in experimental autoimmune encephalomyelitis (EAE), a model for multiple sclerosis, and two previous reports have described resistance of CCR2−/− mice to EAE. The present study examined three different mouse strains with CCR2 deletions for susceptibility to EAE. Animals were studied up to 4 months post-sensitization and were examined by neuropathology, RNase protection assay, in situ hybridization, and in vitro assays. All three strains were found to be susceptible to EAE: C57BL/6 × J129 and Balb c strains, 100%; and C57BL/6, 67%. Unusual in CNS lesions of CCR2−/− mice was an overabundance of neutrophils versus monocytes in wild-type animals. An attempt of the immune system to develop compensatory mechanisms for the lack of CCR2 was evidenced by a corresponding increase in mRNA for other chemokines and CCR. Inasmuch as neutrophils replaced monocytes and led to demyelination, our findings support the concept that promiscuity of chemokines and CCR was able to surmount the deletion of CCR2, still resulting in full expression of this autoimmune disease. Chemokines are low molecular weight cytokines which act as chemoattractants for infiltrating cells bearing appropriate receptors (CCR) to sites of inflammation. It has been proposed that CCR2 on monocytes is responsible for their recruitment into the central nervous system (CNS) in experimental autoimmune encephalomyelitis (EAE), a model for multiple sclerosis, and two previous reports have described resistance of CCR2−/− mice to EAE. The present study examined three different mouse strains with CCR2 deletions for susceptibility to EAE. Animals were studied up to 4 months post-sensitization and were examined by neuropathology, RNase protection assay, in situ hybridization, and in vitro assays. All three strains were found to be susceptible to EAE: C57BL/6 × J129 and Balb c strains, 100%; and C57BL/6, 67%. Unusual in CNS lesions of CCR2−/− mice was an overabundance of neutrophils versus monocytes in wild-type animals. An attempt of the immune system to develop compensatory mechanisms for the lack of CCR2 was evidenced by a corresponding increase in mRNA for other chemokines and CCR. Inasmuch as neutrophils replaced monocytes and led to demyelination, our findings support the concept that promiscuity of chemokines and CCR was able to surmount the deletion of CCR2, still resulting in full expression of this autoimmune disease. Experimental autoimmune encephalomyelitis (EAE) is a T cell-mediated inflammatory disease of the central nervous system (CNS) that serves as an animal model for multiple sclerosis (MS).1Raine CS Biology of disease. The analysis of autoimmune demyelination: its impact upon multiple sclerosis.Lab Invest. 1984; 50: 608-635PubMed Google Scholar, 2Steinman L Assessment to the utility of animal models for MS and demyelinating disease in the design of rational therapy.Neuron. 1999; 24: 511-514Abstract Full Text Full Text PDF PubMed Scopus (216) Google Scholar The disease can be induced by immunization with whole myelin or a variety of myelin antigens, their peptides, or T cells responsive to these antigens.3Raine CS The lesion in multiple sclerosis and chronic relapsing experimental allergic encephalomyelitis: a structural comparison.in: Raine CS McFarland HF Tourtellotte WW Multiple Sclerosis: Clinical and Pathogenetic Basis. Chapman and Hall, London1997: 243-286Google Scholar During induction of EAE, T cells sensitized to myelin antigens migrate across the blood-brain barrier into surrounding white matter,4Cross AH Cannella B Brosnan CF Raine CS Homing to central nervous system vasculature by antigen-specific lymphocytes: I. localization of 14C-labeled cells during acute, chronic, and relapsing experimental allergic encephalomyelitis.Lab Invest. 1990; 63: 162-170PubMed Google Scholar re-encounter antigen and become activated to produce soluble mediators, including chemokines, for which there is compelling evidence for roles in lesion pathogenesis.3Raine CS The lesion in multiple sclerosis and chronic relapsing experimental allergic encephalomyelitis: a structural comparison.in: Raine CS McFarland HF Tourtellotte WW Multiple Sclerosis: Clinical and Pathogenetic Basis. Chapman and Hall, London1997: 243-286Google Scholar Chemokines are chemoattractants produced under pathological conditions by tissue elements and infiltrating leukocytes.5Baggiolini M Dahinden CA CC chemokines in allergic inflammation.Immunol Today. 1994; 15: 127-133Abstract Full Text PDF PubMed Scopus (516) Google Scholar, 6Furie MB Randolph GJ Chemokines and tissue injury.Am J Pathol. 1995; 146: 1287-1301PubMed Google Scholar Two main groups of chemokines are recognized, α-chemokines, having two adjacent cysteines (C-C chemokines), and β-chemokines, with two cysteines separated by one amino acid, C-X-C chemokines.7Mantovani A Chemokines: introduction and overview.Chem Immunol. 1999; 72: 1-6Crossref PubMed Scopus (38) Google Scholar Chemokines are involved in leukocyte maturation, lymphocyte trafficking, and renewal of circulating leukocytes.8Baggiolini M Chemokines and leukocyte traffic.Nature. 1998; 392: 565-568Crossref PubMed Scopus (2388) Google Scholar The molecular regulation of leukocyte trafficking is complex and involves not only interactions between adhesion molecules and their receptors, but also between members of the entire superfamily of chemokines.8Baggiolini M Chemokines and leukocyte traffic.Nature. 1998; 392: 565-568Crossref PubMed Scopus (2388) Google Scholar During EAE, involvement and up-regulation of several C-C chemokines, including macrophage inhibitory protein-1α (MIP-1α) and macrophage chemoattractant protein-1 (MCP-1), is well established.9Glabinski AR Ransohoff RM Chemokines and chemokine receptors in CNS pathology.J Neurovirol. 1999; 5: 3-12Crossref PubMed Scopus (153) Google Scholar MCP-1 is associated with monocyte/macrophage recruitment,10Rollins BJ Monocyte chemoattractant protein 1: a potential regulator of monocyte recruitment in inflammatory disease.Mol Med Today. 1996; 2: 198-204Abstract Full Text PDF PubMed Scopus (354) Google Scholar, 11Fuentes ME Durham SK Swerdel MR Lewin AC Barton DS Megill JR Bravo R Lira SA Controlled recruitment of monocytes and macrophages to specific organs through transgenic expression of monocyte chemoattractant protein-1.J Immunol. 1995; 155: 5769-5776PubMed Google Scholar and its target is the chemokine receptor, CCR2, expressed by these cells.12Dzenko KA Andjelkovic AV Kuziel WA Pachter JS The chemokine receptor CCR2 mediates the binding and internalization of monocyte chemoattractant protein-1 along brain microvessels.J Neurosci. 2001; 21: 9214-9223Crossref PubMed Google Scholar Previous studies have reported that disruption of chemokine receptors (CCR) may lead to impaired monocyte function, including chemokine-directed chemotaxis,13Boring L Gosling J Chensue SW Kunkel SL Farese RVJ Broxmeyer HE Charo IF Impaired monocyte migration and reduced type 1 (Th1) cytokine responses in C-C chemokine receptor 2 knockout mice.J Clin Invest. 1997; 100: 2552-2561Crossref PubMed Scopus (872) Google Scholar, 14Kurihara TT Warr G Loy J Bravo R Defects in macrophage recruitment and host defense in mice lacking the CCR2 chemokine receptor.J Exp Med. 1997; 186: 1757-1762Crossref PubMed Scopus (554) Google Scholar and that manipulation of CCR might result in resistance to inflammatory disease. In the case of CCR2, recent communications15Fife BT Huffnagle GB Kuziel WA Karpus WJ CC chemokine receptor 2 is critical for induction of experimental autoimmune encephalomyelitis.J Exp Med. 2000; 192: 899-905Crossref PubMed Scopus (447) Google Scholar, 16Izikson L Klein RS Charo IF Weiner HL Luster AD Resistance to experimental autoimmune encephalomyelitis in mice lacking the CC chemokine receptor (CCR)2.J Exp Med. 2000; 192: 1075-1080Crossref PubMed Scopus (513) Google Scholar have reported that CCR2 knockout (ko) mice are resistant to EAE and that CCR2 is a key susceptibility factor in this disease.15Fife BT Huffnagle GB Kuziel WA Karpus WJ CC chemokine receptor 2 is critical for induction of experimental autoimmune encephalomyelitis.J Exp Med. 2000; 192: 899-905Crossref PubMed Scopus (447) Google Scholar, 16Izikson L Klein RS Charo IF Weiner HL Luster AD Resistance to experimental autoimmune encephalomyelitis in mice lacking the CC chemokine receptor (CCR)2.J Exp Med. 2000; 192: 1075-1080Crossref PubMed Scopus (513) Google Scholar, 17Huang DR Wang J Kivisakk P Rollins BJ Ransohoff RM Absence of monocyte chemoattractant protein 1 in mice leads to decreased local macrophage recruitment and antigen-specific T helper cell type 1 immune response in experimental autoimmune encephalomyelitis.J Exp Med. 2001; 193: 713-726Crossref PubMed Scopus (508) Google Scholar Since we had access to three separate mouse strains with CCR2 deletions, we decided to extend a preliminary study on one strain,18Pitt D Kuziel WA Charles PA Cannella B Raine CS Experimental autoimmune encephalomyelitis in chemokine receptor deficient mice. (Abstract).J Neuroimmunol. 1998; 90: 60Abstract Full Text PDF Google Scholar to test further the susceptibility of CCR2−/− mice to EAE. Study of these three different strains with CCR2 disruption has revealed all to be highly susceptible. Interestingly, each strain displayed subtle differences, particularly in regard to the cellular make-up of the CNS infiltrate while the process of demyelination was not disrupted. As an approach toward mechanism, we have investigated cytokine, chemokine, and CCR profiles and found them to be in line with an increase in polymorphonuclear leukocytes, the cell type predominating in CCR2−/− EAE lesions. Therefore, CCR2 is not an all-important determining factor in EAE and it appears that multiple compensatory mechanisms exist in the expression of autoimmune demyelination. Six- to eight-week-old wild-type (wt) and CCR2−/− mice on a C57BL/6 × J129 (C57/J129) background, a C57BL/6 × J129 background back-crossed × 8 onto C57BL/6 (C57BL/6), or a Balb c background (six back-crosses) were used. With the exception of wt Balb c mice, all mice were provided by W. Kuziel. Control wt female Balb c mice were purchased from The Jackson Laboratory, Bar Harbor, ME. CCR2−/− mice were generated by homologous recombination and all were bred under pathogen-free conditions. In addition, 20 mice (10 CCR2−/−, 10 wt) 12- to 24-weeks of age, were tested for the susceptibility of older animals to develop EAE. These animals were studied for clinical susceptibility only. Animal care and use was performed in accordance with National Institutes of Health guidelines and mice were housed in an American Association for Accreditation of Laboratory Animal Care (AAALAC) accredited facility. A total of 214 animals was used; 60 C57/J129, 140 C57BL/6, and 14 Balb c. Myelin was prepared from guinea pig spinal cord (GPSC) according to established procedures.19Norton WT Poduslo SE Myelination in rat brain: method of myelin isolation.J Neurochem. 1973; 21: 749-757Crossref PubMed Scopus (1266) Google Scholar An encephalitogenic peptide corresponding to myelin oligodendrocyte glycoprotein (MOG) residue aa35–55 (MOG35–55) was synthesized by an in-house facility. Amino acid composition was verified by mass spectroscopy: purity was > 98%. Monoclonal antibodies (mAB) were used to CD4 T cells (L3T4) and CD8a T cells (Lyt-2; both from BD PharMingen, La Jolla, CA); to neutrophils (7/4), macrophages (F4/80), and B cells (CD19; Serotec, Kidlington, Oxford, United Kingdom); and to IL-4 and IL-10 (BD PharMingen). Polyclonal antibodies were used to IFN-γ and TNF-α (R&D Systems, Minneapolis, MN). On day 0 (d 0) and day 7 post-immunization (p.i.) C57/J129 and C57BL/6 mice were sensitized for active EAE by subcutaneous (s.c.) injection (two sites, dorsal flank) with a total of 600 μg of encephalitogenic MOG35–55 emulsified in incomplete Freund's adjuvant (IFA; Difco, Detroit, MI), containing 70 μg Mycobacterium tuberculosis, H37Ra (Difco). On day 0 and day 2 p.i., each mouse also received 500 ng pertussis toxin (PTX; List Biological Laboratories, Campbell, CA) intravenously (i.v.), via a tail vein. Balb c mice were sensitized for active EAE with 700 μg GPSC homogenate emulsified in IFA containing 35 μg M. tuberculosis. 100 ng PTX was also given i.v. on day 0, day 2, and day 7 p.i. Of the 12- to 24-week-old group, 10 (5 CCR2−/−, 5 wt) were sensitized as above and 10 (5 CCR2−/− and 5 wt) were sensitized according to the technique of Izikson et al,16Izikson L Klein RS Charo IF Weiner HL Luster AD Resistance to experimental autoimmune encephalomyelitis in mice lacking the CC chemokine receptor (CCR)2.J Exp Med. 2000; 192: 1075-1080Crossref PubMed Scopus (513) Google Scholar with 100 μg MOG in complete Freund's adjuvant (CFA) and intraperitoneal administration of PTX, for comparison of methods of induction. Animals were assessed daily for clinical signs and evaluated according to the following scale: grade 0, no abnormalities; grade 1, weak tail; grade 2, limp tail and weakness in hind-limbs; grade 3, hind-limb paraparesis; grade 4, tetraplegia; grade 5, moribund or death. Light microscopy studies were performed on glutaraldehyde/osmium-fixed tissue from optic nerve, cerebrum, cerebellum, and spinal cord. The tissue was dehydrated and embedded in epoxy resin from which 1-μm sections were cut and stained with toluidine blue. Inflammation, demyelination, Wallerian degeneration (WD), and remyelination were scored on a scale of 0 to 5, as described previously.20Cannella B Hoban CJ Gao YL Garcia-Arenas R Lawson D Marchionni M Gwynne D Raine CS The neuregulin, glial growth factor 2, diminishes autoimmune demyelination and enhances remyelination in a chronic relapsing model for multiple sclerosis.Proc Natl Acad Sci USA. 1998; 95: 10100-10105Crossref PubMed Scopus (148) Google Scholar Ultrastructural studies were conducted on thin sections contrasted with lead and uranium salts, carbon-coated, and scanned by electron microscopy (EM) in a Hitachi HS600 (Tokyo, Japan). For immunohistochemistry, blocks of phosphate-buffered saline (PBS)-perfused lumbar spinal cord were snap-frozen in liquid nitrogen. Cryostat sections (10-μm) were fixed with cold acetone and methanol, and primary antibodies were applied overnight at 4°C. For evaluation of bound antibody, the Vectastain Elite kit (Vector, Burlingame, CA), was used. 3′, 3′ diaminobenzidine (KPL, Gaithersburg, MD), served as the substrate for horseradish peroxidase. Immunoreactive cells were counted in spinal cord sections from six to eight different areas at × 10 and × 63 magnification. Spinal cords were collected from mice in each group and snap-frozen at different time-points after sensitization. Total RNA was prepared using standard procedures. RNA probes were used to detect chemokines and chemokine receptors, and cytokine mRNAs were generated by in vitro transcription. RNase protection assays (RPAs) were performed using RiboQuant template sets, mCK1, mCK3b, and mCR5 (BD Pharmingen), in combination with RPA II kit from Ambion (Austin, TX). Protected fragments were visualized by autoradiography and quantified by phosphoimaging the gels with a Storm 860 scanner and Image QuaNT V3.01 software (Molecular Dynamics, Sunnyvale, CA). For each sample, a ratio of the intensity of the cytokine, chemokine, or chemokine receptor band was obtained using the value of the band for the housekeeping genes, mL32 and GAPDH. In situ hybridization (ISH) was performed on 10-μm frozen sections using 300- to 500-bp PCR products of different cytokines, chemokines, and chemokine receptors, labeled with digoxigenin by nick-translation (Roche, Indianapolis, IN), according to the manufacturer's instructions. PCR products representing the following mRNA sequences were applied: CCR1, bp 309–650; CCR2, bp434–710; CCR5, bp706–1047; IL-8R, bp252–661; RANTES, bp199–491; MCP-1, bp163–501; MCP-5, bp78–378; MIP-1β, bp239–578; IL-8, bp118–651; TNF-α, bp1083–1434; IFN-γ, bp232–539; TGF-β, bp1327–1579; and IL-4, bp128–407. Negative controls for ISH consisted of hybridization solution without labeled probe. C57BL/6 mice were sensitized s.c. with a total of 600 μg MOG35–55/CFA on day 0 and day 7 p.i. On day 15 p.i., mice were anesthetized with ether and sampled, and popliteal and axillary lymph nodes and spleen excised. Single-cell suspensions from draining lymph nodes and spleen were made and activated by culturing at 37°C in 7% CO2 in RPMI medium supplemented with 1% mouse serum (Rockland, Gilbertsville, PA), 1 mmol/L glutamate, 1 mmol/L pyruvate, 100 units/ml penicillin, 100 μg/ml streptomycin (all from Invitrogen; Grand Island, NY), 0.02 mmol/L β-mercaptoethanol (Sigma, St. Louis, MI), to which were added different antigens, such as 30 μg/ml of MOG35–55, 30 μg/ml MBP, 2.5 μg/ml concanavalin A (ConA), and 500 ng/ml LPS (all from Sigma). To determine proliferation after 2 days, cells were treated with 25 μCi 3H-thymidine (NEN Life Science Prod, Pittsburgh, PA), for an additional 16 hours, before being harvested and assessed for radioactivity.21Sung S Rose CE Fu SM Intratracheal priming with ovalbumin- and ovalbumin 323–339 peptide-pulsed dendritic cells induces airway hyper-responsiveness, lung eosinophilia, goblet cell hyperplasia, and inflammation.J Immunol. 2001; 166: 1261-1271PubMed Google Scholar Clinical and histological scores in all age groups were compared with the Mann-Whitney rank sum test, using Instat (Graph Pad). Results were expressed as mean ± SD. In both C57 strains (C57/J129 and C57BL/6), CCR2−/− mice sensitized at 8 weeks of age developed clinical signs of MOG peptide-induced EAE (Figures 1, A and B), which were characterized by a delay in onset of 3 to 5 days and reduced clinical severity in comparison to wt mice. In general, CCR2−/− mice developed a milder EAE, between grade 2 (C57/J129) and grade 1.5 (C57BL/6), versus 3.0 to 3.5 in wt mice. However, Balb c CCR2−/− mice showed a more typical response (grade 3) (Figure 1C). As in both C57 strains, Balb c CCR2−/− mice displayed a delayed onset. Also noteworthy were differences in the course of disease; C57/J129 developed a relapsing EAE while C57BL/6 and Balb c mice showed a monophasic pattern. Significantly, 100% of 8-week-old sensitized C57/J129 and Balb c CCR2−/− mice displayed clinical signs, whereas in C57BL/6 CCR2−/− mice, the average incidence was 67%. Of the 12- to 24-week-old sensitized groups, four of five CCR2−/− mice from the group sensitized by the present technique developed signs of EAE (grades 1 to 3) versus two of five wt (grades 1 to 2), while of these sensitized according to Izikson et al,16Izikson L Klein RS Charo IF Weiner HL Luster AD Resistance to experimental autoimmune encephalomyelitis in mice lacking the CC chemokine receptor (CCR)2.J Exp Med. 2000; 192: 1075-1080Crossref PubMed Scopus (513) Google Scholar one of five CCR2−/− (grade 1) and two of five wt (grade 2) developed disease. In C57/J129 mice (CCR2−/− and wt) sensitized at 8 weeks of age, spinal cord lesions typical of acute EAE were seen (Figure 2, Figure 3). However, wt C57/J129 mice showed more severe pathology with widespread WD and a monocyte-rich infiltrate (Figure 2D). In contrast, CNS lesions in C57/J129 CCR2−/− mice revealed a striking overabundance of infiltrating neutrophils and a paucity of monocytes (Figure 2C). The lesion in CCR2−/− mice was less destructive and fewer axons showed damage. In C57BL/6 mice, comparable high numbers of neutrophils in CNS infiltrates and a similar degree of demyelination in both ko and wt (Figure 2, G–J), were encountered. Similar to C57/J129 ko mice, CCR2−/− C57BL/6 mice displayed less WD in comparison to wt mice (Figure 2, I–J).Figure 3Histopathological differences between CCR2−/− and wt mice with disease progression. A–C: Acute EAE. Balb c CCR2−/− mice revealed decreased pathology compared to wt. D–F: Remission. CCR2−/− mice of both C57 strains displayed less pathology (D and E), which was less prominent in Balb c ko animals (F). Interestingly, CCR2−/− mice with a C57/J129 background showed decreased axonal damage (D), in contrast to C57BL/6 (E) and Balb c CCR2 ko mice (F). G–I. Chronic EAE. Balb c mice showed less pathology, but increased demyelination and WD in CCR2−/− mice (I), in contrast to both CCR2−/− from C57 strains (G and H). Data represent average histological scores of three animals (C57 strains) or a single animal (Balb c), evaluated from semi-thin cross sections of lower spinal cord (four levels, eight slides). *, ± P < 0.001; **, P < 0.01; ***, P < 0.05View Large Image Figure ViewerDownload Hi-res image Download (PPT) Balb c wt mice with acute EAE displayed widespread lesion activity with extensive cellular infiltration and demyelination (Figure 2L). CCR2−/− Balb c mice, on the other hand, displayed small CNS lesions with only a few infiltrating cells, comprised mainly of neutrophils (Figure 2K). Occasionally, in CCR2−/− animals, the CNS portion of the PNS in the anterior root entry zone was also affected. Also of interest in Balb c and C57/J129 CCR2−/− mice was the appearance of lymph node-like high endothelial venules in the CNS (Figure 2, C and K), indicative of active trafficking and homing of lymphocytes, described previously in EAE.4Cross AH Cannella B Brosnan CF Raine CS Homing to central nervous system vasculature by antigen-specific lymphocytes: I. localization of 14C-labeled cells during acute, chronic, and relapsing experimental allergic encephalomyelitis.Lab Invest. 1990; 63: 162-170PubMed Google Scholar In the remission phase, during which the most statistically significant (Mann-Whitney) differences were observed, C57/J129 CCR2−/− mice displayed fewer demyelinated fibers and less WD than wt (Figure 3). Both C57 strains displayed less infiltration and WD in the CCR2−/− groups. In Balb c CCR2−/−, there were similar degrees of demyelination and remyelination and WD was less common than in wt, but WD was more extensive. During the chronic phase, except for some differences in the amount of WD, both ko and wt mice from all strains was comparable. As in MS,3Raine CS The lesion in multiple sclerosis and chronic relapsing experimental allergic encephalomyelitis: a structural comparison.in: Raine CS McFarland HF Tourtellotte WW Multiple Sclerosis: Clinical and Pathogenetic Basis. Chapman and Hall, London1997: 243-286Google Scholar with time, the plaque area in EAE animals was largely taken over by fibrous astrocytes and reactive microglia. This phenomenon was seen in ko and wt animals from C57BL/6 and Balb c (not shown). The same plaques were devoid of CNPase immunoreactivity due to depletion of oligodendrocytes and myelin and gliosis appeared more pronounced in C57BL/6 than Balb c mice, perhaps a reflection of the smaller CNS lesions in Balb c. Ultrastructural study confirmed the predominance of neutrophils in EAE lesions in CCR2−/− C57/J129 mice. Neutrophils and some eosinophils were found in both the leptomeningeal space and CNS compartment and were seen to traverse into the CNS between subpial astrocytic processes through gaps in the glia limitans (Figure 4A). Occasionally, neutrophils were seen to flank nerve fibers with attenuated degenerating myelin sheaths (Figure 4B). Significantly, EM study showed the process of demyelination to be intact in CCR2−/− mice. As is well documented in mice and other species,3Raine CS The lesion in multiple sclerosis and chronic relapsing experimental allergic encephalomyelitis: a structural comparison.in: Raine CS McFarland HF Tourtellotte WW Multiple Sclerosis: Clinical and Pathogenetic Basis. Chapman and Hall, London1997: 243-286Google Scholar disrupted myelin formed an extracellular vesicular network around affected axons, resulting in the formation of lamellar myelin droplets (Figure 4C). These were then internalized by macrophages following their attachment to clathrin-coated pits on the macrophage surface (Figure 4D), described in detail in EAE22Epstein LG Prineas JW Raine CS Attachment of myelin to coated pits on macrophages in experimental allergic encephalomyelitis.J Neurol Sci. 1983; 61: 341-348Abstract Full Text PDF PubMed Scopus (76) Google Scholar and MS.23Prineas JW Connell F The fine structure of chronically active multiple sclerosis plaques.Neurology. 1978; 28: 68-75Crossref PubMed Google Scholar This process has been shown to involve immunoglobulin whereby opsonized myelin droplets bind to Fc receptors expressed in coated pits on the surface of macrophages (Figure 4B).24Moore GR Raine CS Immunogold localization and analysis of IgG during immune-mediated demyelination.Lab Invest. 1988; 59: 641-648PubMed Google Scholar This phenomenon, known as receptor-mediated phagocytosis of myelin, is peculiar to autoimmune demyelinating models and MS.3Raine CS The lesion in multiple sclerosis and chronic relapsing experimental allergic encephalomyelitis: a structural comparison.in: Raine CS McFarland HF Tourtellotte WW Multiple Sclerosis: Clinical and Pathogenetic Basis. Chapman and Hall, London1997: 243-286Google Scholar Presumably, immunoglobulin mediation in the process of demyelination was not disrupted by CCR2 deletion. Immunohistochemical study of EAE lesions in C57BL/6 mice also confirmed the predominance of neutrophils compared to macrophages and T cells in both groups (data not shown). Among T cell populations, CD4+ lymphocytes were common in both CCR2−/− and wt mice. However, the number of macrophages and CD4+ T cells were increased in CCR2−/− mice, while the number of neutrophils was lower. Lesions in Balb c animals were comparable in ko and wt mice. Although CCR2−/− Balb c mice displayed lower numbers of neutrophils and F4/80+ macrophages, the numbers of CD4+ and CD8+ T cells were comparable in ko and wt. However, CD8+ cells were generally a minor component of the T cell population. CD19+ B cells were rarely seen during the acute phase of EAE, particularly in CCR2−/− C57BL/6 and Balb c mice. Lumbar spinal cord lesions in C57BL/6 CCR2−/− mice showed IFN-γ, TNF-α, and IL-6 protein and mRNA expression to be marginally decreased (from wt values). Infiltrating cells expressing anti-inflammatory cytokines, like IL-4 and IL-10, were found scattered around CNS blood vessels in both ko and wt mice. In Balb c CCR2−/− mice with EAE, proinflammatory cytokines, such as IFN-γ, TNF-α, and IL-6, were slightly elevated at both protein and mRNA levels, whereas IL-10 expression was lower. However, anti-inflammatory IL-4 (protein and mRNA) and TGF-β (mRNA), could not be detected in ko and wt animals (not shown). RNA study using RPA in C57/J129 mice showed that in CNS tissue, message for CCR2 and CCR5 was decreased in CCR2−/− mice, whereas the mRNA level of CCR1 was comparable in ko and wt animals (Figure 5). In addition, message levels of MIF and cytokines like TGF-β, TNF-α, and IFN-γ were also similar. ISH of CNS tissue from C57BL/6 revealed a visible increase in message for MCP-1, a ligand of CCR2, in CCR2−/− mice versus wt (Figure 6, A and B). Unexpectedly, mRNA expression for CCR2 was seen in ko mice to the same extent as wt C57BL/6 (Figure 6, C and D). This can be explained from the nature of the probe (see Discussion). However, expression of the CCR1-ligand, RANTES, was similar in ko and wt, as was that of the CCR5-ligand, MIP-1β (not shown). CCR1 and CCR5 mRNA-expression was slightly lower in CCR2−/− C57BL/6 mice. Interestingly, mRNA expression for IL-8, a chemokine involved in neutrophil recruitment, was elevated in CCR2−/− animals (Figure 6, E and F), perhaps contributing to the unusual neutrophil population in the CNS. In general, lesions in Balb c CCR2−/− mice displayed higher numbers of cells expressing mRNA for chemokine receptors and their ligands, like CCR1/RANTES and CCR5/MIP-1β (Figure 6, G and H). Moreover, loss of expression of CCR2 mRNA was reflected by reduced message for MCP-5, a CCR2 ligand. In contrast, transcripts for IL-8 receptor were increased in CCR2−/− mice. Negative controls showed a lack of detectable signal (not shown). In vitro proliferation assays on primary mixed lymphocyte cultures from ko and wt MOG35–55-primed C57BL/6 mice showed no differences. Lymphocytes and splenocytes from both groups displayed a high potential for MOG35–55-specific proliferation, whereas the response to MBP, an irrelevant myelin antigen, appeared inhibitory compared to controls (no antigen applied to cell culture), and proliferation indices of MOG35–55-specific lymphocyte populations from different lymphoid organs, lymph node and spleen, differed slightly (Table 1). Therefore, at the level of lymphocyte responsiveness, no abnormalities were seen.Table 1Proliferative Response to MOGwtCCR2−/−Lymphocytes n610 Mean3.272.15 Median3.652.03 SD1.41.22 SE0.570.39 Mann-Whitneyp < 0.09Splenocytes n64 Mean1.51.97 Median0.781.38 SD0.320.69 SE1.351.34 Mann-Whitneyp < 0.76 Open table in a new tab Previous reports have concluded that mice lacking CCR2 are resistant to the induction of EAE16Izikson L Klein RS Charo IF Weiner HL Luster AD Resistance to experimental autoimmune encephalomyelitis in mice lacking the CC chemokine receptor (CCR)2.J Exp Med. 2000; 192: 1075-1080Crossref PubMed Scopus (513) Google Scholar or display much reduced disease.15Fife BT Huffnagle GB Kuziel WA Karpus WJ CC chemokine receptor 2 is critical for induction of experimental autoimmune encephalomyelitis.J Exp Med." @default.
• W73990037 created "2016-06-24" @default.
• W73990037 creator A5015749443 @default.
• W73990037 creator A5018416735 @default.
• W73990037 creator A5019823731 @default.
• W73990037 creator A5021896445 @default.
• W73990037 creator A5057467113 @default.
• W73990037 date "2003-01-01" @default.
• W73990037 modified "2023-10-14" @default.
• W73990037 title "Experimental Autoimmune Encephalomyelitis (EAE) in CCR2−/− Mice" @default.
• W73990037 cites W1496902169 @default.
• W73990037 cites W1501778305 @default.
• W73990037 cites W1516141774 @default.
• W73990037 cites W1544915536 @default.
• W73990037 cites W1560737490 @default.
• W73990037 cites W1567987955 @default.
• W73990037 cites W1583693385 @default.
• W73990037 cites W1602835962 @default.
• W73990037 cites W1906710068 @default.
• W73990037 cites W1925136148 @default.
• W73990037 cites W1964002710 @default.
• W73990037 cites W1975375148 @default.
• W73990037 cites W1977739000 @default.
• W73990037 cites W1984123968 @default.
• W73990037 cites W2005659566 @default.
• W73990037 cites W2013310648 @default.
• W73990037 cites W2017132882 @default.
• W73990037 cites W2019577957 @default.
• W73990037 cites W2020852722 @default.
• W73990037 cites W2027494021 @default.
• W73990037 cites W2036609995 @default.
• W73990037 cites W2066991415 @default.
• W73990037 cites W2072309750 @default.
• W73990037 cites W2078297463 @default.
• W73990037 cites W2080932190 @default.
• W73990037 cites W2081099685 @default.
• W73990037 cites W2090742115 @default.
• W73990037 cites W2099768074 @default.
• W73990037 cites W2102628016 @default.
• W73990037 cites W2105925708 @default.
• W73990037 cites W2106509445 @default.
• W73990037 cites W2111718223 @default.
• W73990037 cites W2116248267 @default.
• W73990037 cites W2116495710 @default.
• W73990037 cites W2117342688 @default.
• W73990037 cites W2119216843 @default.
• W73990037 cites W2133121320 @default.
• W73990037 cites W2133793569 @default.
• W73990037 cites W2140538238 @default.
• W73990037 cites W2141701917 @default.
• W73990037 cites W2142361972 @default.
• W73990037 cites W2146440741 @default.
• W73990037 cites W2160129587 @default.
• W73990037 cites W2165299403 @default.
• W73990037 cites W2168263358 @default.
• W73990037 cites W2168424667 @default.
• W73990037 cites W2320853752 @default.
• W73990037 cites W2399788710 @default.
• W73990037 cites W2410982626 @default.
• W73990037 cites W2416053426 @default.
• W73990037 cites W2423874902 @default.
• W73990037 cites W39405201 @default.
• W73990037 cites W4244761631 @default.
• W73990037 cites W92198176 @default.
• W73990037 cites W990932059 @default.
• W73990037 doi "https://doi.org/10.1016/s0002-9440(10)63805-9" @default.
• W73990037 hasPubMedCentralId "https://www.ncbi.nlm.nih.gov/pmc/articles/1851120" @default.
• W73990037 hasPubMedId "https://pubmed.ncbi.nlm.nih.gov/12507897" @default.
• W73990037 hasPublicationYear "2003" @default.
• W73990037 type Work @default.
• W73990037 sameAs 73990037 @default.
• W73990037 citedByCount "124" @default.
• W73990037 countsByYear W739900372012 @default.
• W73990037 countsByYear W739900372013 @default.
• W73990037 countsByYear W739900372014 @default.
• W73990037 countsByYear W739900372015 @default.
• W73990037 countsByYear W739900372016 @default.
• W73990037 countsByYear W739900372017 @default.
• W73990037 countsByYear W739900372018 @default.
• W73990037 countsByYear W739900372019 @default.
• W73990037 countsByYear W739900372020 @default.
• W73990037 countsByYear W739900372021 @default.
• W73990037 countsByYear W739900372022 @default.
• W73990037 countsByYear W739900372023 @default.
• W73990037 crossrefType "journal-article" @default.
• W73990037 hasAuthorship W73990037A5015749443 @default.
• W73990037 hasAuthorship W73990037A5018416735 @default.
• W73990037 hasAuthorship W73990037A5019823731 @default.
• W73990037 hasAuthorship W73990037A5021896445 @default.
• W73990037 hasAuthorship W73990037A5057467113 @default.
• W73990037 hasBestOaLocation W739900371 @default.
• W73990037 hasConcept C142724271 @default.
• W73990037 hasConcept C203014093 @default.
• W73990037 hasConcept C2777899865 @default.
• W73990037 hasConcept C2778486448 @default.
• W73990037 hasConcept C2780640218 @default.
• W73990037 hasConcept C71924100 @default.
• W73990037 hasConcept C86803240 @default.

• next