Our results indicated that

motoneurons were protected by

Our results indicated that

motoneurons were protected by VPA against cell death induced by brachial plexus root avulsion through c-Jun inhibition and Bcl-2 induction. © 2013 Wiley Periodicals, Inc. Microsurgery 33:551–559, 2013. “
“The free jejunum has become an important method for reconstructing extensive oncologic defects of the upper esophagus and pharynx. The advantages of a single-staged reconstruction with a low incidence of morbidity have generally outweighed criticisms such as the requirement for a laparotomy and poor voice quality. The aim of the study was to present the technique and outcomes of free jejunal reconstruction of the upper esophagus in RAD001 research buy 31 consecutive cases. We reviewed our experience of free jejunal flaps undertaken over a 6-year period. Our surgical approach, complications, and results of swallow and speech restoration are described. A functional swallow was achieved by 27/31 patients. However, satisfactory voice restoration was seen in only a small proportion of patients. Complications at the donor site occurred in just one patient. The current review confirms the jejunal flap as a reliable reconstructive option with minimal donor site

morbidity. © 2012 Wiley Periodicals, Inc. Microsurgery, 2013. “
“The role of vascularized bone marrow in promoting composite allograft survival can be assessed by intrinsically chimeric flaps. In this study, we introduce a significant modification to a previously described rat model of DNA Damage inhibitor combined superficial inferior epigastric the artery (SIEA) myocutaneous/vascularized femur transplantation. We previously noted autocannibalization in orthotopic myocutaneous SIEA allotransplants, which complicated clinical and histologic evaluation of rejection. We therefore designed syngeneic experiments in eight Lewis (RTl1) rat pairs to explore the feasibility of tunneling the SIEA component of chimeric SIEA myocutaneous/vascularized femur flaps to the recipient dorsum. Vascularized SIEA myocutaneous/femur transplants survived in their entirety to POD 63 study endpoint with patent anastomoses

in seven of eight (87.5%) transplants as confirmed clinically, histologically, and via near-infrared fluorescent angiography. Tunneling of the SIEA component of SIEA myocutaneous/vascularized femur flaps to the recipient dorsum can be achieved with high success rate and acceptable operative times, and is a technically easy method to study the role of vascularized bone marrow in composite allografts. This modification facilitates SIEA component monitoring, removes it from constant contact with cage bedding, and places it in a location where autocannibalization is unlikely. © 2011 Wiley Periodicals, Inc. Microsurgery, 2012. “
“The prevalence of obesity is rising in Western society. The aim of this meta-analysis was to evaluate the available evidence regarding the effect of obesity on outcomes of free autologous breast reconstruction.

We previously found that some transitional B cells in rabbit sple

We previously found that some transitional B cells in rabbit spleen localize to the MZ [13]. Human transitional B cells are CD27− [15], and we found that most rabbit transitional type 1 (T1) B cells were also CD27− (Fig. 1C); surprisingly, however, approximately 50% of the transitional type 2 (T2) GSK-3 beta phosphorylation B cells were CD27+ (Fig. 1C). We suggest that the CD27+ T2 B cells may be precursors to CD27+ mature MZ B cells. T2 B cells in mice are similarly thought to contain precursors for MZ B cells as well

as for FO cells [10]. Functionally, 24 h after anti-Ig and CD40L stimulation, we found more CD27+ B cells in cell cycle than CD27− B cells (Fig. 1D), indicating that CD27+ B cells enter cell cycle more readily than CD27− B cells. Upon stimulation with CD40L and IL-4 for 8 days, we found significantly more total Ig in the culture supernatant of sorted CD27+ B cells than CD27− B cells (Fig. 1E), suggesting that ABT-199 datasheet CD27+ B cells secrete more Ig than CD27− B cells. We conclude that rabbit CD27+ and CD27− B cells represent distinct subsets that differ

by virtue of their anatomical location, phenotype, and functional properties. To determine if there was a perturbation in the splenic B-cell compartment after neonatal removal of GALT, we stained frozen spleen tissues with anti-CD23 and anti-CD27 mAbs to identify FO and MZ B cells, respectively. Unlike control rabbits that had well-defined CD23+ and CD23− areas (Fig. 1F, left), nearly all B cells in the follicles of GALTless

rabbits were CD23+ (Fig. 1F, right). Consistent with this observation, we found almost no CD27+ MZ B cells in the GALTless rabbits (Fig. 1G), indicating that GALT is required Oxymatrine for development of MZ B cells. The intestinal microbiota is required for development of GALT [16] and in the absence of intestinal microbiota, follicles of proliferating B cells are not found in GALT, and the number of peripheral B cells is markedly reduced [9]. In GALTless rabbits, only organized GALT, appendix, sacculus rotundus, and Peyer’s patches are removed; isolated lymphoid follicles [17] and cryptopatches would remain in the GALTless rabbits and be exposed to intestinal microbiota. The apparent absence of MZ B cells in GALTless rabbits indicates that isolated lymphoid follicles and cryptopatch B cells either do not mature into MZ B cells, or that they give rise to only small numbers of MZ B cells. Notch 2 is important for both murine and human MZ B-cell development [18-21], and its ligand delta-like-1 (DL1) is expressed by intestinal epithelial cells [22]. We suggest that transitional B cells enter the follicle-associated epithelium and domes of the appendix [13], interact with DL1+ epithelial cells, and become committed to a MZ fate; these cells would then migrate to the spleen and possibly other tissues. The CD27+ T2 B cells in spleen may represent putative MZ precursors derived from T1 B cells in GALT.

Low IgM levels with B cell lymphopenia have been reported

Low IgM levels with B cell lymphopenia have been reported

in X-linked dyskeratosis congenita (X-linked DC), with severe combined immunodeficiency (T + B − NK − SCID) reported in the most severe variant of dyskeratosis congenita (Hoyeraal–Hreidarsson syndrome) [36]. Premature ageing is also a feature of this disease [32–34] and TINF2 gene mutation (a component of the telomere protection complex) [35] leading to short telomeres has been described in X-linked DC. It is not clear whether the immune abnormalities are due to the defective tRNA pseudouridylation or the short telomere length. Turner’s syndrome (45,X0) is postulated to have a ribosomal defect due to haploinsufficiency of ribosomal protein RPS4X [48,49]. Variable degrees of antibody deficiency (panhypogammaglobulinaemia [48], GSK1120212 low IgM [50,52]) Selinexor clinical trial including decreased T and B cell numbers [50,54] and coeliac disease with IgA deficiency have been recognized in this syndrome [53,55]. Some of these patients with Turner’s syndrome have clinical syndromes of recurrent sinopulmonary infections and other features overlapping with CVID [50,51]. We have looked at the evidence for ribosomal defects being associated with

and possibly causative of immune abnormalities with features of CVID. We describe two such patients with different ribosomal defects who subsequently developed a presentation consistent with CVID. A review of the Protein kinase N1 literature indicates that patients with ribosomal defects may share abnormalities of T or B cell development with many features of CVID, and which may not be recognized

as such by non-immunologists. Given that the four established genetic defects account for fewer than a fifth of cases of CVID, this hypothesis could be tested in the future by more detailed studies of ribosome genetics and/or function in CVID. This work was supported by the Centre for Immunoglobulin Therapy and Department of Immunology, Hull Royal Infirmary. WACS is Director of Centre for Immunoglobulin Therapy, which has received unrestricted educational grants from Octapharma, Baxter, Grifols, CSL-Behring. The rest of the authors have no financial interests to disclose. “
“Human cytomegalovirus (HCMV) has been reported to reshape the NK-cell receptor (NKR) distribution, promoting an expansion of CD94/NKG2C+ NK and T cells. The role of NK cells in congenital HCMV infection is ill-defined. Here we studied the expression of NKR (i.e., NKG2C, NKG2A, LILRB1, CD161) and the frequency of the NKG2C gene deletion in children with past congenital infection, both symptomatic (n = 15) and asymptomatic (n = 11), including as controls children with postnatal infection (n = 11) and noninfected (n = 20).

In this report, we investigated the cell infiltration that expres

In this report, we investigated the cell infiltration that expresses FOXP3 or IL-17 in allograft tissue with biopsy-proven ATCMR, and we intended to appraise whether the ratio between them is associated with allograft outcome after ATCMR. The study population consisted of 71 clinically indicated renal allograft biopsies performed on 56 renal transplant recipients in our transplant centre from August 1999 to August 2008. Of the 71 biopsy samples, 56 biopsies were a first-time ATCMR and the other 15 specimens

were repeat ATCMR biopsy samples (13 specimens were the second ATCMR and two specimens VX-770 clinical trial were third ATCMR). The indication for the allograft biopsy was graft dysfunction defined as a serum creatinine increment of greater than or equal to 10% from the baseline value. These cases were selected only for the diagnosis of ATCMR type I or II according to Banff’s working classification and the availability of sufficient paraffin-embedded tissue.23,24 BK virus or cytomegalovirus nephropathy, click here lymphoproliferative disorder, interstitial fibrosis/tubular atrophy (IF/TA) grade III was not present in these

patients or biopsies. Out of 56 patients, 33 patients (59%) were a living related donor, 13 cases (23%) were a living unrelated donor, and 10 cases (17·9%) were deceased donor transplantation. The HLA mismatch number was 3·7 ± 1·3 and four cases (7%) were a second transplantation. The flow-cross-match test before transplantation was negative and the Panel reactive antibody was less than 20% in all patients. Our centre’s protocol for immune suppression is described

in a previous publication.25 Briefly, the main immunosuppressive agents used were cyclosporine (n = 31, 55%) or tacrolimus (n = 25, 45%). Mycophenolate mofetil was added as a primary immunosuppressant in 42 patients (75%). Basiliximab was used as an additional induction therapy in 22 patients (39%). Patients were followed from the date of transplantation to the date of nephrectomy, permanent dialysis, re-transplantation, or Vorinostat cost death. During the study period, ATCMR was treated with three to five daily boluses of intravenous methylprednisolone (500 mg/day), followed by a 5–7-day oral steroid taper. When the serum creatinine level failed to decrease within 5 days, muromonab-CD3 (OKT3) or anti-thymocyte globulin (ATG) was applied. The Institutional Review Board of Seoul St Mary’s Hospital approved the study. All biopsies were examined for FOXP3+ cell and IL-17+ cell infiltration. Paraffin sections were immersed in three changes of xylene and hydrated using a graded series of alcohols. Antigen retrieval was performed routinely by immersing the sections in sodium citrate buffer (pH 6·0) in a microwave for 15 min.

Endothelin-1, a potent vasoconstrictor peptide, was measured by N

Endothelin-1, a potent vasoconstrictor peptide, was measured by Nakamura et al. [57] in control individuals, along with individuals with Raynauds and also vibration-induced white finger. https://www.selleckchem.com/products/DAPT-GSI-IX.html The authors reported that endothelin-1 levels were elevated rapidly upon

finger cold immersion in both control and Raynauds individuals. In Raynauds, this rise was much higher, and it remained elevated even after immersion. However, there was no correlation between endothelin-1 levels and incidences of CIVD, suggesting that, while endothelin-1 is highly related to sympathetic hyperactivity, it does not directly contribute to the opening of peripheral blood vessels eliciting CIVD [57]. Geurts et al. [35] observed selleck screening library no changes in either endothelin-1 or NO levels in response to repeated hand immersions, but the caveat of no thermal acclimation precluded any conclusions. Overall, while broad improvements in thermal responses in individuals who live or work in cold environments are possible, microcirculatory adaptations and changes in the CIVD response in the fingers and toes appear to be neither guaranteed nor predictable. Much of the evidence for adaptation has involved cross-sectional

studies, but significant gaps remain in understanding the contribution of genetic or morphological differences across different ethnic populations in cold response, along with the role of self-selection when considering comparisons across different occupations. The primary systematic improvement with prolonged acclimation is in a decreased perceptual discomfort or pain. However, with notable exceptions [1,63], longitudinal and laboratory studies have found minimal improvement in actual CIVD measures, with some finding that thermal responses actually became impaired over the acclimation period. Selleck CHIR99021 Given the emphasis on developing strategies for protecting from cold injuries in occupational and recreational settings,

people should not rely on physiological adaptation through repeated local cold exposure. Rather, given the importance of overall body thermal status on CIVD responses, individuals should try to keep their body core warm and wear well-insulated and well-fitted gloves and boots to prevent the occurrence of local cold injuries [9]. One avenue for further research appears to be in understanding the interactions between exercise and hypoxia on local blood flow and CIVD trainability. However, such research should be performed with standardized definitions for CIVD and its measurement rather than with the historic and current wide variability in methodology. An enhanced circulation to the extremities is presumed to occur with repeated exposure to cold, serving as a protective mechanism against peripheral cold injury.

19 vs 2 30 mm) (P = 0 002) The range of stiffness is between 4,

19 vs. 2.30 mm) (P = 0.002). The range of stiffness is between 4,339 and 4,697 N mm−1. Stiffness tends to decrease significantly (P < 0.001) with increasing flap size. Harvest of flap sizes greater or equal than 9 cm results in significantly lower stiffness compared to the 3-cm flap. In this composite femur model, when stressed with supraphysiologic forces, the femur retains its axial stability even after harvest of large corticocancellous

flaps from its medial aspect. Statistical significance detected in deformation and stiffness may not be clinically relevant if the femur does not fracture after flap harvest. Such was the case in this experiment. The possibility exists of safely harvesting large flaps from this donor site. Corticocancellous flaps FK228 purchase from the medial aspect of the femur may serve as an alternative to standard flaps used in medium and large osseous reconstructions.

The size of flap that can be safely raised without compromising the stability of the femur has not yet been delineated. © 2012 Wiley Periodicals, Inc. Microsurgery, 2012. “
“Background: The fasciocutaneous internal mammary artery perforator (IMAP) island flap allows for superior esthetical and functional skin cover in the head and neck region in combination with limited donor site morbidity. Its modification as a free flap allows reconstruction of more cranial defects. Patients and methods: Three IMAP free flaps varying from 7 × 4 cm2 to 10 × 6 cm2 were transplanted in three patients with a mean age of 59 years (range, 54–69 years). Enhancement of the flap’s vascular pedicle DMXAA manufacturer at least doubles the diameter of the internal mammary vessels to be anastomosed. Results: (-)-p-Bromotetramisole Oxalate Coverage with excellent texture and color match was uneventfully obtained and the flaps’ donor sites were primarily closed in all three cases. Conclusions: Our experience proves the consistent feasibility of successful transplantation of the IMAP free flap. Because of its characteristics, we suggest

contemplating the use of this flap in the upper head and neck region. © 2010 Wiley-Liss, Inc. Microsurgery, 2010. “
“In case blood perfusion compromises, vascular enhancement with arterial supercharge or venous superdrainage can increase viability of the flap. In this study, vascular pressure monitorization was used in a rat extended abdominal perforator flap model to reveal intraoperative vascular compromise and the need for vascular augmentation. A rat abdominal perforator flap was designed, which was based on the right second cranial perforator of epigastric artery. Vascular pressures of the flap were monitored continuously for 60 min, by catheters placed in the right superficial inferior epigastric artery and vein. Forty rats were divided into four experimental groups, as follows: group 1 (n = 10, no vascular augmentation), group II (n = 10, arterial supercharge), group III (n = 10, venous superdrainage), and group IV (n = 10, arterial and venous augmentation).

HIV sexual transmission is very inefficient, and a number of biol

HIV sexual transmission is very inefficient, and a number of biological factors are critical in determining whether an unprotected sexual exposure to HIV results in productive infection. This review will focus on ways in which biology, rather than behaviour,

may contribute to regional and racial differences in HIV epidemic spread. Specific areas of focus are viral factors, host genetics, and the impact of co-infections and host https://www.selleckchem.com/products/Aloxistatin.html immunology. Considering biological causes for these racial disparities may help to destigmatize the issue and lead to new and more effective strategies for prevention. It was famously said by Kofi Annan that ‘in Africa, AIDS has a woman’s face’,1 but gender is by no means the most marked imbalance when it comes to the effects of HIV. While women now bear over half of the global HIV burden,2 it is only in the continent of Africa that women constitute the majority of infected persons. In contrast, there is a tremendous disparity in the effects of HIV along racial and ethnic lines that is apparent throughout the world. This imbalance is most marked at a continental level, given that approximately two-thirds of all HIV-infected persons are in Africa, but is also apparent within most regional subepidemics. The reasons underlying the racial and geographical imbalances

MLN0128 cost in HIV prevalence are complex and have led to myths, stereotypes, stigma and discrimination that may impede the development of better HIV prevention tools and programs. As is the case for all sexually transmitted infections (STIs), socio-economic and cultural factors have been hypothesized to be critical contributors to HIV transmission Farnesyltransferase and increased HIV prevalence in Africa.3,4 Many of these sociocultural factors are potentially stigmatizing and include higher per-capita rates of commercial sex,5 increased partner exchange/concurrency,6,7 intimate partner violence,8–10 and traditions such as wife inheritance.11 There are data supporting the causal association of HIV with at least some of these factors, but

it is unfortunate that a focus on the cultural and behavioural aspects of HIV transmission tends to implicitly lay blame for infection on affected communities or individuals.12 While a discussion of the sociocultural associations of HIV is beyond the scope of this review, our goal is to emphasize that there may be other causes for the geographical and racial imbalances in HIV prevalence that are equally important. Specifically, our goal is to explore possible biological cofactors that may enhance vulnerability and contribute to the substantial global racial disparities in HIV prevalence. Our hope is that a better understanding of such cofactors may allow the development of new HIV prevention tools while reducing stigma. There are major racial and geographical disparities in HIV prevalence.

Recent reports have shown that RP105-deficient B cells are defect

Recent reports have shown that RP105-deficient B cells are defective in their response to TLR2 and TLR4 ligands, whereas it is likely that RP105/MD-1 positively regulate TLR2/TLR4 responses in B cells.39 In contrast, Divanovic et al.40 reported that RP105 negatively regulates LPS-induced responses in macrophages and dendritic cells.

In the present learn more study, we examined RP105 to ascertain the expression of innate immune-related molecules in B cells. The major population of peritoneal B cells has been well reported to be B-1a cells and the immune function of this subset is essentially different from that of the conventional B-cell subset (B-2 cells) that exists in other organs. The present results obtained by flow cytometry suggest that the major population of intestine-related B cells (MLNs, PPs, colon lamina propria) has a B-2 lineage. Next, we examined the production of IL-10 and TGF-β1 in TLR-mediated B KPT-330 ic50 cells. Mononuclear cells were isolated from several

parts of BALB/c mice and magnetically purified using microbeads. Next, purified B cells (B220+ PDCA-1−) were cultured with or without TLR ligands, then cytokine concentrations in the culture supernatants were measured by EIA. The B-cell fractions used in the experiments were confirmed to be > 95% pure by flow cytometry (Fig. 2a). Although IL-10 production was induced in TLR ligand-mediated B cells, the level of production in CpG-DNA-stimulated cells was significantly higher than that in LPS-stimulated cells (Fig. 2b). In addition, IL-10 production by TLR-mediated PerC B cells was remarkably higher than that by B cells isolated from other parts

of the mice. These results may have been dependent on the unique characteristics of PerC B cells derived from a B-1 lineage. However, when compared with the results of IL-10, lower production levels of TGF-β1 in response to TLR ligands were observed in all DNA ligase of the tested samples (Fig. 2b). In the body systems, TGF-β1 occurs in two physiological forms: latent and active. Although TGF-β1 is important in regulating crucial cellular activities, in most cases an activated TGF-β1 ligand will initiate the TGF-β1 signalling cascade. In our present system, the majority of TGF-β1 as assessed was solely inactive or latent. We also measured the active form of TGF-β1 but the amount was too low to demonstrate any effects of TLR ligands on their secretion (data not shown). Following our experimental results, we investigated the presence of a regulatory B-cell subset producing IL-10 and TGF-β1 in the intestines of BALB/c mice. Furthermore, we conducted additional experiments to elucidate the role of this intestinal regulatory B-cell subset in the pathogenesis of CD using SAMP1/Yit mice. Development of ileitis in the SAMP1/Yit mice was confirmed by histological examinations.

Recent reports have also suggested a role for B cells in the path

Recent reports have also suggested a role for B cells in the pathogenesis of the disease [26, 27, 46, 47], and autoantibodies have been used to define the autoimmune manifestations. Finally, transferring bulk lymphocytes allowed us to define the behaviour of Treg cells during the proliferation. Indeed, we noticed clear signs of immune dysregulation in the recipients

that received cells from Aire−/− donors, and some of the findings were similar to those found in Aire−/− mice themselves. One such perturbation was Hydroxychloroquine supplier the hyperproliferation of T cells, particularly the CD8+ population, which was observed both systemically and in the gut-associated lymphoid tissues. A Th1 dominance was also observed within the colon tissue of the Aire group recipients; Palbociclib mw previous studies have implicated Th1 cells in the immunopathology of Aire−/− mice [39] and also in colitis [38]. A higher incidence of autoantibodies in the Aire group was evident, as well. These data support the view that T cells that have developed in the absence of Aire are more autoreactive, and readily induce some manifestations

of immune dysregulation. However, despite the conditions favouring autoimmunity, created by the LIP, no symptomatic autoimmune disease was observed, and all the animals remained clinically healthy. Also, one prevalent feature of Aire-related autoimmune syndrome, lymphocyte infiltration into Cediranib (AZD2171) solid tissues, was almost completely absent. This finding differs from previous reports in which the phenotype of Aire−/− animals, including the infiltration of lymphocytes to target tissues, was fully transferable

to lymphopenic recipients [28]. All these previous studies, however, were carried out using large numbers of mature lymphocytes, so that very little or no homeostatic proliferation took place. It has been clearly demonstrated that the skewing of peripheral T cell repertoire and autoimmunity is more pronounced with the transfer of small cell numbers [48]. For example, in non-obese diabetic mice, the prevalence of LIP-induced autoimmune diabetes is higher if adoptive cell transfers are carried out with small cell numbers [49]. On the other hand, the number of cells we transferred is not so small as to protect from autoimmunity because of insufficient cell numbers. Indeed, cell numbers as low as 3 × 104 have been reported to cause severe autoimmunity [48]. Therefore, our results indicate that the relative importance of defective thymic negative selection might be lower than previously thought in the development of autoimmunity in the Aire−/− animals. In our model, the Treg cell population originating from Aire−/− donors showed distinct hyperproliferation, as compared to the Treg cells transferred from Aire+/+ donors.

e Toxoplasma encephalitis [23, 36] Consistently, blocking NF-κB

e. Toxoplasma encephalitis [23, 36]. Consistently, blocking NF-κB signaling, which is required for astrocyte activation in EAE, by tissue-specific ablation of key signaling molecules including NEMO, IKK2, and

Act1 in the CNS impaired astrocytic production of inflammatory cytokines and chemokines ameliorating EAE as evidenced by decreased leukocyte infiltration and reduced demyelination [5, 37, 38]. Interestingly, in sharp contrast to the proinflammatory function of most astrocyte-derived chemokines, CXCL12, which is upregulated in the CNS of MS patients, particularly produced by astrocytes, suppressed ongoing EAE by redirecting the polarization of effector Th1 cells into IL10-producing Treg cells [39]. Collectively, the present study extends find more the in vivo selleck products function of astrocytes and illustrates that astrocytes also confer protection against EAE by the

FasL-dependent apoptotic elimination of activated CD25+ Foxp3− and GM-CSF-producing CD4+ T cells and the concomitant inhibition of proinflammatory cytokine production. Thus, augmentation of astrocytic FasL may provide a favorable strategy for treatment of clinically active MS. GFAP-Cre+/− FasLfl/fl mice were generated by crossing C57BL/6 GFAP-Cre transgenic mice [40] with C57BL/6 FasLfl/fl mice [41] and the colony was maintained by breeding of GFAP-Cre+/− FasLfl/fl mice with GFAP-Cre−/− FasLfl/fl mice. Genotyping of offsprings was carried out by PCR of tail DNA with primers targeting GFAP-Cre and FasLfl/fl. Deletion of FasL was analyzed by PCR in various organs and cell types with Del-FasL primers (5′-GTACTTCTTCTGATAAGGACC-3′ filipin and 5′-GGAGTTGAACGAGTAGCCTC-3′). C57BL/6 WT mice were obtained from Harlan (Borchen, Germany). Animal care and experimental procedures were performed according to European regulations and approved by state authorities (Landesverwaltungsamt Halle, Germany; IMMB/G/02–994/10). MOG35–55 (MEVGWYRSPFSRVVHLYRNGK) was purchased from JPT (Berlin, Germany). Active EAE was induced in 8- to 12-week-old

mice by s.c. immunization with 200 μg of MOG35–55 emulsified in complete Freund’s adjuvant (Sigma, Taufkirchen, Germany) supplemented with 800 μg of killed Mycobacterium tuberculosis (Sigma). In addition, mice also received two i.p. injections of 200 ng pertussis toxin (Sigma), dissolved in 200 μL PBS, at the time of immunization as well as 48 h thereafter. Clinical signs of EAE were monitored daily and scored according to a scale of severity from 0 to 5 as described previously [23]. Daily clinical scores were calculated as the average of all individual disease scores within each group. Leukocytes were isolated from the spinal cord and stained for CD4+ T cells, CD8+ T cells, and CD45high inflammatory leukocytes as described before [42].