tuberculosis induced CD4+ T-cell-dependent responses The key fin

tuberculosis induced CD4+ T-cell-dependent responses. The key findings of the present study are that eight of a total of 157 peptides selected for HLA class I binding induce T-cell responses in PBMC from one or several PPD+ donors (Table 2). In contrast, only four of 10 donors, with low PPD reactivity in ELISPOT

assay, reacted with one or more of the eight antigenic peptides indicating the M. tuberculosis specificity of the responses observed. However, instead of being HLA class I restricted, these responses are apparently restricted buy PD98059 by HLA class II molecules because the responses are all blocked by anti-HLA-DR antibody added to the ELISPOT assay culture. In addition, according to results from cell depletion and FACS analyses anti-M. tuberculosis peptide responses are mediated by CD4+ T cells. The

eight epitopes discovered are derived from five different M. tuberculosis proteins. Three of these [Rv1979, Rv3144c (PPE52) and Rv3532 (PPE61)] each express two positive CTL epitopes whereas the remaining two proteins [Rv0284 and Rv3507 (PE_PGRS)] only harbour a single epitope. Interestingly, six of the eight positive epitopes are derived from the PE/PPE gene family of conserved mycobacterial proteins (Table 2). The PE/PPE gene family is interesting from an immunological point of view because they comprise approximately Compound Library 10% of the M. tuberculosis genome and may be a source of antigenic variation, which the bacterium uses to evade the host immune response.34 These proteins are surface-associated cell wall proteins and may also be accessible to antibodies.40 The B-cell and T-cell

immune responses have been reported against both PE and PPE proteins, but their immunological Adenosine triphosphate significance remains largely unknown.41–44 Only a few T-cell epitopes have been identified for the PE/PPE gene family. Two have been found in PE-PGRS proteins (Rv1818c and Rv3812) and one in PPE protein (Rv3018c). In the present study we report five new epitopes for the PE/PPE gene family: a single epitope for the Rv3507 (PE_PGRS) and four new epitopes for the PPE proteins [Rv3144c (PPE52) and Rv3532 (PPE61)]. Regarding the phenotype of M. tuberculosis peptide-responding T cells, our data from T-cell depletion of PBMC before ELISPOT and FACS analyses showed that the responding T cells are indeed CD4+ T cells. In our previous studies 26–28 in which we probed for specific T-cell immunity in PBMC against pox and flu virus-derived HLA-I binding peptides, respectively, HLA-I and HLA-II antibody blocking experiments and CD4+ and CD8+ T-cell depletion experiments showed that peptide reactivity was initiated by either CD4+ or CD8+ T cells but never by both subsets in the same ELISPOT culture. It is generally accepted that HLA class I binding peptides are composed of 8–11 amino acids, whereas HLA class II binding peptides consist of 15–20 amino acids being recognized by CD8+ and CD4+ T cells, respectively.

A natural hypothesis given these findings

A natural hypothesis given these findings Apoptosis inhibitor is that the diminished exhaustion seen in LTNPs could be dependent on lower expression of Blimp-1. This is the possibility addressed in the paper published in this issue of the European Journal of Immunology, in which Seddiki et al. [18] present experiments measuring Blimp-1 levels in the CD4+ T cells from HIV+ LTNPs, individuals with CHI and healthy controls. These experiments showed that, at both the protein and mRNA levels, Blimp-1 expression is higher in individuals with CHI than in LTNPs. Supporting this was the finding

that the downstream effects of elevated Blimp-1 expression in chronic infection, namely elevated PD-1 and diminished IL-2 expression, were also more pronounced in individuals with CHI relative to levels in LTNPs. This prompted Seddiki et al. [18] AUY-922 to consider the mechanism by which Blimp-1 expression is regulated in T lymphocytes.

One manner in which gene expression is regulated in cells is via microRNA (miR). These are small noncoding sequences of RNA that bind to untranslated regions of target mRNA and either suppress their translation or accelerate their degradation. The authors assessed the ability of different miRs to suppress Blimp-1 expression. In results consistent with those of other researchers [19], Seddiki et al. [18] found that transfection of miR-9 decreased Blimp-1, while increasing IL-2, expression in CD4+ T cells. The authors also demonstrated that, in CD4+ T cells, TCR stimulation leads to expression of miR-9. Finally, to support the hypothesis that diminished Blimp-1 levels in LTNPs is due to miR-9 expression, the authors measured CD4+ T cell miR-9 levels and found them to be elevated in LTNPs relative to levels

in individuals Sucrase with CHI. This study [18] provides strong evidence that differences in the CD4+ T-cell expression of Blimp-1 can explain the improved anti-viral profile of the CD4+ T cells from LTNPs versus those from individuals with CHI. It supports data gained from the murine system and proposes, together with another recent publication [19], a novel mechanism for Blimp-1 regulation. The therapeutic possibility raised by this work is that a reduction of Blimp-1 levels in individuals with CHI could improve viral control and provide the proof that the improved viral control seen in LTNPs is Blimp-1 dependent. That this is an important consideration in T-cell directed therapies for HIV is underlined by the failure of IL-2 therapy in HIV to produce a clinical benefit despite improving the CD4+ T-cell count [20]. Blimp-1 was initially described as being dependent on IL-2 signalling and the failure of IL-2 therapy may therefore be attributable to the IL-2-induced Blimp-1 expression and the exhausted phenotype that it promotes.

We believe that the accompanying artery of the sciatic nerve may

We believe that the accompanying artery of the sciatic nerve may be a recipient vessel for free-flap transfer in selected patients. © 2014 Wiley Periodicals, Inc. Microsurgery, 2014. “
“Purpose: In this report, we present our experience

on the repair of brachial plexus root avulsion injuries with the use of contralateral C7 nerve root transfers with nerve grafting through a modified prespinal route. Methods: The outcomes of the contralateral C7 nerve root transfer to neurotize the upper trunk and C5/C6 nerve roots of the total or near total brachial plexus nerve root avulsion injury in a series of 41 patients were evaluated. Opaganib research buy The contralateral C7 nerve root that was dissected to the distal end of the divisions, along with the sural nerve graft, were placed underneath the anterior scalene and longus colli muscles, and then passed through the retro-esophageal space to neurotize the recipient nerve. The mean length of the dissected contralateral C7 nerve root was 6.5 ± 0.7 cm, and the mean length of sural nerve graft was 6.8 ± 1.9 cm. The suprascapular

nerve was neurotized additionally by the phrenic nerve or the terminal motor branch of accessory nerve in some patients. Results: The mean length of the follow-up was 47.2 ± 14.5 months. The muscle strength was graded M4 or M3 for the biceps muscle in 85.4% of patients, for the deltoid muscle in 82.9% of patients, and for the upper parts of pectoral major in 92.7% of patients. The functional recovery of shoulder STK38 abduction in the patients with the additional suprascapular nerve neurotization was remarkably improved. Conclusions: CP-868596 datasheet The modified prespinal route could significantly reduced the length of nerve graft in the contralateral C7 nerve root transfer

to the injured upper trunk in brachial plexus root avulsion injury, and it may improve the functional outcomes, which deserves further investigations. © 2011 Wiley Periodicals, Inc. Microsurgery, 2012. “
“Vascular thrombosis is one of the major postoperative complications of free flap microvascular breast reconstruction operations. It is associated with higher morbidity, higher cost, increased length of hospital stay, and potentially flap loss. Our purpose is to evaluate the rate of this complication and whether patient characteristics play a role. Using the Nationwide Inpatient Sample (NIS) database, we examined the clinical data of patients who underwent free flap breast reconstruction between 2009 and 2010 in the United States. Multivariate and univariate regression analyses were performed to identify independent risk factors of flap thrombosis. A total of 15,211 patients underwent free flap breast reconstruction surgery (immediate reconstruction: 43%). The most common flap was the free deep inferior epigastric perforator (DIEP) flap (53.6%), followed by free transverse rectus abdominis myocutaneous (TRAM) flap (43.

The number of cells capable of secreting Ag85b-specific IFN-γ was

The number of cells capable of secreting Ag85b-specific IFN-γ was significantly higher in the Ag+Al+CpG group (154±106) than in Ag, CpG and NS groups (P<0.05) (Fig. 2d). An identical trend was found for the number of cells that secreted HspX-specific IFN-γ and C/E-specific IFN-γ (Fig. 2e and f). The number of antigen-specific IFN-γ-secreting cells in the Ag+Al+CpG group (30±26 and 44±38) was considerably higher than that in Ag, CpG and NS groups (P<0.05). The level of IL-12 was significantly higher in the Ag+Al+CpG group (42.24±26.45 pg mL−1) than in the other groups (Fig. 3a). The relatively high concentration of IL-12 in the Decitabine mouse NS group (10.53±1.58 pg mL−1) and similar levels in

the Ag (13.18±1.88 pg mL−1), Ag+Al (14.92±5.09 pg mL−1), Ag+CpG (19.45±12.32 pg mL−1) and CpG (14.03±3.14 pg mL−1) groups resulted in no significant differences when conducting multiple comparisons among these groups. Similar buy Rapamycin results were observed with IL-12 secretion

in response to HspX and C/E (Fig. 3b and c). The only group that showed an apparently higher concentration of IL-12 was the Ag+Al+CpG group (33.62±18.95 and 23.20±9.09 pg mL−1). No statistical difference in the level of IL-12 was observed among the other groups. Guinea pigs were evaluated for total lesion scores of the liver, spleen and lung and for bacterial load in the spleen [mean log10 bacilli (CFU)±SD] (Fig. 4). Total lesion scores of the tested organs in the Ag+Al+CpG group (42.50±16.72) were lower than those in the other groups, but no significant difference was found (Fig. 4a). Antigen alone in the Ag group (45.45±28.59) resulted in lower (but not statistically significant) scores than in the Ag+Al (46.67±24.96) and Ag+CpG (53.75±25.68) groups. Only the combination of the two adjuvants was capable of modestly controlling disease progression. A similar trend was also observed 3-mercaptopyruvate sulfurtransferase for the bacterial load in the spleen. The Ag+Al+CpG group (4.75±1.65) had the lowest bacterial load of all

of the groups, but no significant difference was found when compared with other groups. The Ag+Al (5.24±1.35) and Ag+CpG (5.13±0.52) groups had a similar level of bacterial load, and the Ag and NS groups were almost the same (Fig. 4b). Due to the weak immunogenicity of recombinant proteins, subunit vaccine formulations require adjuvants to enhance their immunogenicity. Recently, many of these adjuvanted subunit vaccines have entered clinical evaluations (Weinrich Olsen et al., 2001; Skeiky et al., 2004; Dietrich et al., 2005, 2006; Agger et al., 2006; Dietrich et al., 2006). In this study, we combined CpG and aluminum and observed enhanced immunogenicity of Ag85b, HspX and C/E. The combination of adjuvants effectively induced a strong humoral and cellular immune response in mice, and antigen-specific IgG was significantly higher than injection of either CpG or aluminum alone.

31; −0 31) Infants in the no feature condition were 37 5% less l

31; −0.31). Infants in the no feature condition were 37.5% less likely to search for the familiar toy than infants in the identifying feature condition (B2 = −1.15, χ2(1) = 4.97, p < 0.05, 95% CI [−2.16; −0.139]).

This comparison demonstrates that infants’ enhanced performance with the object that had identifying feature on it cannot be explained by infants’ generally stronger and richer representation of the object. It also suggests that the effect of prior location of the target object cannot be ameliorated by providing infants with nonidentifying information MK-2206 about the object or simply by drawing their attention to it. All together, the analyses revealed no differences in infants’ performance with the new toy across the three Small molecule library concentration groups and better performance with the familiar toy in the identifying feature condition than in the two control conditions. These results show that infants have difficulty tracking object identity when an object is moved from room to room. The findings are consistent with the proposal that infants’ confusion about the object identity resulting from such location changes disrupts infants’ ability to reveal understanding of absent reference. In the current study, we investigated the possibility that infants’ difficulty locating a hidden object encountered in a different context before the study is related to their

difficulty establishing the object’s identity across multiple contexts. To facilitate infants’ Isotretinoin ability to track objects across large-scale spatial displacements in this research, we highlighted the same, characteristic feature of the object in both locations where infants encountered the object. This manipulation facilitated infants’ subsequent ability to find the object

in response to a verbal request. When two different features were highlighted or pointed at, infants were less likely to locate the object based on a verbal request for it. When an object was not introduced before the experimental phase, infants had no difficulty locating the object when it was hidden. Together these findings suggest that large-scale spatial displacements may disrupt infants’ ability to locate verbal referents, but that they can be released from this difficulty if attempts are made to clarify that the referent is the same object as the one that they had recently seen in a different context. A limitation of the current study is that toy type was confounded with toy familiarity: The dog was always new to infants, and the pig was always familiar. However, the condition differences found for the familiar toy suggest that the current results cannot be explained by infants’ preference to one toy over the other. If a toy preference were the only factor guiding infants’ responses, they should not have searched for the (familiar) pig in any of the conditions.

The authors thank Inovio Pharmaceuticals for supplying the electr

The authors thank Inovio Pharmaceuticals for supplying the electroporation equipment. This work was supported

by Cancer Research UK no. C1238/A3849 (Vittes, G. E. and Stevenson, F. K.) and Leukaemia and Lymphoma Research UK no. 08025 (Harden, E. L. and Rice, J.). Conflict of interest: The authors declare no financial or commercial conflict of interest. “
“Citation Wu M-H, Huang M-F, Chang F-M, Tsai S-J. Leptin on peritoneal macrophages of patients with endometriosis. Am J Reprod Immunol 2010; 63: 214–221 Problem  The expression of cyclooxygenase (COX)-2 is considered as a marker of macrophage activation and has been implicated in the development of endometriosis. Leptin is an immunomodulator, which may also affect the development

this website of endometriosis. However, how leptin contributes to these pathological processes has not been completely understood. The aim of this study was to investigate the effects of leptin on peritoneal macrophages and its relationship with endometriosis. Methods of study  Peritoneal fluid from 60 women of reproductive age was obtained while they underwent laparoscopy. Forty patients had endometriosis and 20 patients did not have endometriosis. The concentration of leptin in the peritoneal fluid and prostaglandin F2α levels was measured by ELISA, and the other protein expression using Western blot when peritoneal macrophages were stimulated with leptin. Results  Concentration selleck kinase inhibitor of leptin in peritoneal fluid was increased in patients with endometriosis compared with disease-free

normal control. Functional leptin receptor was ADAMTS5 present in peritoneal macrophages. Treatment of peritoneal macrophages with leptin induced COX-2 expression. Production of prostaglandin F2α by peritoneal macrophages was increased after leptin stimulation in women with endometriosis. Conclusion  Elevated concentration of leptin in peritoneal fluid may contribute to the pathological process of endometriosis through activation of peritoneal macrophages. “
“An emerging theme among vacuole-adapted bacterial pathogens is the ability to hijack ubiquitin machinery to modulate host cellular processes and secure pathogen survival. Mono- and polyubiquitination differentially dictate the subcellular localization, activity, and fate of protein substrates. Monoubiquitination directs membrane traffic from the plasma membrane to the endosome and has been shown to promote autophagy. Anaplasma phagocytophilum is an obligate intracellular bacterium that replicates within a host cell-derived vacuole that co-opts membrane traffic and numerous other host cell processes. Here, we show that monoubiquitinated proteins decorate the A. phagocytophilum-occupied vacuolar membrane (AVM) during infection of promyelocytic HL-60 cell, endothelial RF/6A cells, and to a lesser extent, embryonic tick ISE6 cells.

(A) Sensitised acceptor emission FRET analysis of positive and ne

(A) Sensitised acceptor emission FRET analysis of positive and negative control cell lines. The positive PD0325901 cell line control consisted of cells expressing CFP coupled to YFP. The negative control consisted of cells expressing individual CFP and YFP proteins encoded by different plasmids. While the positive control cells demonstrated high FRET efficiency (47.4%±1.6), the negative control showed 0% FRET efficiency. (B) Equal amount of WT YFP-ζ and MUT YFP-ζ proteins were expressed in COS-7 cells upon transfection as detected by anti-YFP Western blot analysis. (C) Acceptor photobleaching FRET analysis was performed using images collected in three independent experiments,

as described in the supplementary methods section. *P<0.0001. Figure S5. Mutations in the ζ RRR motifs do not affect its conformation but impair its association with actin. (A) Mutations in the RRR motifs restore TCR cell surface expression. ζ-deficient T-cell clones stably expressing the WT (17 and 14) or MUT (8 and 15) ζ were tested for cell surface TCR expression using anti-CD3e antibodies and FACS analysis. WT and MUT ζ expressing T-cell clones were lysed and immunoprecipitated with four different antibodies directed against various epitopes (“a”-“d”) localized within the ζ intracytoplasmic domain (B). Samples were separated on reduced SDS-PAGE

and immunoblotted for ζ (C). (D) T-cells expressing the MUT ζ failed to undergone percipitataion with actin. WT and MUT transfected T-cell clones or splenocytes from WT and transgenic (ζD66–150), mice were lysed, immunoprecipitated with anti-actin antibodies. Samples were immunoblotted with

see more antibodies directed against the indicated proteins. Ab = antibody with no lysate. Figure S6. WT and MUT T-cell clones exhibit a similar immediate TCR-mediated activation pattern. (A) WT and MUT T-cell clones exhibit a similar pattern of ζ isoforms induced upon short activation. Cells were activated with anti-CDe and anti-CD28 antibodies, lysed, and the non-cska fraction was subjected to immunoblotting with anti-ζ antibodies. (B) A similar ZAP-70 phosphorylation pattern was observed in both WT and MUT T-cell clones upon brief activation. The cells were activated as described in (A), lysed, immunoprecipitated with anti-ZAP-70 antibodies, separated on reduced SDS-PAGE and immunoblotted with anti-ZAP-70 or anti-phosphotyrosine antibodies. (C) A similar LAT phosphorylation pattern was observed in both WT and MUT T-cell clones upon brief activation. The cells were activated as described in (A), lysed, separated on reduced SDS-PAGE and immunoblotted with anti-LAT or anti-pLAT antibodies. Figure S7. Cska and non-cska expression during T-cell activation. (A) Total cska and non-cska ζ expression during T-cell activation. Mouse splenocytes were activated with anti-CD antibodies for various intervals, lysed, the cska and non-cska fractions were separated and subjected to immunoblotting with anti-ζ antibodies.

23 Rainer et al [14] developed a selective SceSel+ medium contai

23 Rainer et al. [14] developed a selective SceSel+ medium containing dichloran and benomyl as active compounds, which inhibits a large diversity of filamentous

fungi. The SceSel+ medium prevented growth of Aspergillus in sputum samples; Scedosporium strains are overgrown or outcompeted on full medium by Aspergillus strains, due to faster growth rates of A. fumigatus strains. Blyth et al. [13] shows that benomyl-media are significantly more efficient for selective isolation of Scedosporium species than for routine media such as SGA, with up to 100% see more recovery of Scedosporium from spiked samples when SceSel+ was used. When sputum samples are processed with benomyl-based media, recovery rates

tend to increase significantly: Horréet al. [24] noted 14.3% positive samples and Blyth et al. [13] 14.7%. Although our isolation procedure included the use of DRBC-benomyl agar, our isolation rate (8.5% positive) was somewhat in the lower range. When experimental non-culture methods are used to detect Scedosporium in CF sputum samples, significantly higher recovery rates are obtained. Cimon et al. [15], using counterimmuno-electrophoresis were able to raise the detection rate to 21.1% positive samples, compared to 8.6% with classical JAK inhibitor culture. In the present study, we found 62.7% of the samples positive by PCR-RLB. The phenomenon that PCR-based diagnostic Pazopanib mouse assays detect substantially more positives is an often-encountered problem.

A disadvantage of PCR is the risk of false-positive results, caused by either pre-PCR contamination of samples, non-specific amplification or by amplification of DNA from dead cells. Careful precautions against cross-contamination were taken during sample collection and preparation by using separated rooms and filtered tips. No cross reaction or false positive in tester strains was found. All clinical samples were processed twice and identical results were obtained. Therefore, the chance of false-positivity due to contamination or non-specific amplification is negligible. In 10 samples two or three species were detected. These results suggest the regular inhalation of fungal spores belonging to different species of the P. apiosperma/P. boydii complex and the subsequent colonisation of the respiratory tract or at least their persistence in the airways.10 The prevalence of Scedosporium DNA in the environment and the air presently is unknown, which hampers to ascertain whether or not real colonisation has taken place in patients with positive samples. To clarify this, further study is necessary. In one case, PCR-RLB was negative although the sample was proven to be positive by culture. The single deviating Scedosporium culture-positive sample was repeatedly negative using PCR-RLB and remained so with a 1 : 5 dilution of the DNA extract.

Supernatants from stimulated DCs were collected and stored at

Supernatants from stimulated DCs were collected and stored at

−80° until cytokine assays were performed. PrestoBlue Cell Viability Reagent (Invitrogen), diluted 1 : 10 with medium, was added to generated DCs (2 × 105 cells/100 μl diluted solution) in a 96-well plate. Samples were then incubated for 30 min at 37°. PrestoBlue is reduced from blue resazurin to red resorufin in the presence of viable cells. We then read the fluorescence (excitation 570 nm, emission 600 nm) with a Benchmark plus (Bio-Rad Laboratories Inc., Hercules, CA). The supernatants of DC cultures were measured for cytokine content by cytometric bead array (CBA) assays. A human inflammation CBA kit (BD Pharmingen, selleck chemicals San Jose, CA) was used to quantify IL-12p70 and tumour necrosis factor-α (TNF-α) levels. Samples were analysed using a FACS Caliber flow cytometer (BD Pharmingen). Cell

surface marker fluorescence intensity was assessed using a FACS Caliber analyser and analysed using CellQuest (BD Pharmingen) or FlowJo (TreeStar Inc., Ashland, OR) software. Dead cells were excluded with propidium iodide staining. Monoclonal antibodies against CD14, CD80, CD83, CD86, CD40, CD1a, CD209 and CD205 were purchased from BD Pharmingen. Anti-TGR5 monoclonal antibody was purchased from R&D Systems. Total MLN0128 price RNA was extracted from cells using an RNeasy Micro kit (Qiagen, Hilden, Germany), and cDNA was synthesized using a Quantitect RT kit (Qiagen) according to the manufacturer’s instructions. Quantitative real-time PCR (qPCR) was performed using TaqMan Universal PCR Master Mix (Applied Biosystems, Foster City, CA) and on-demand gene-specific primers, designed using the DNA Engine Opticon 2 System (Bio-Rad Laboratories, Inc.) and analysed with Opticon monitor software (MJ Research, Waltham, MA). The primers were as follows: BSEP (Hs00184824_m1), NTCP (Hs00161820_m1), Farnesyltransferase OATP (Hs00366488_m1), ASBT (Hs01001557_m1),

TGR5 (Hs01937849_s1), TNFα (Hs00174128_m1), IL-12p35 (Hs00168405_m1) and IL-12p40 (Hs00233688_m1). Monocytes (2 × 105 cells) were treated with lithocholic acid, TCDCA, glycoursodeoxycholic acid (GUDCA) and TGR5 agonist (5 μm) for 5 min in the presence of 1 mm 3-isobutyl-1-methylxanthine. The amount of cAMP was determined with a cAMP-Screen System (Applied Biosystems). For intracellular phosphoprotein staining in monocytes we used a PhosFlow assay (BD Biosciences, Franklin Lakes, NJ). Cells in suspension were stimulated by TCDCA or with control medium for the indicated times, fixed with pre-warmed PhosFlow Cytofix solution for 10 min and permeabilized with ice-cold PhosFlow Perm buffer III for 30 min. Phycoerythrin-conjugated mouse anti-cAMP response element-binding protein (CREB) (pS133)/ATF-1 (pS63) or mouse anti-IgG isotype antibody was added to each tube and incubated at room temperature for 30 min in the dark. The cells were washed with 10 volumes of staining buffer and analysed by flow cytometry.

In comparison with HC, significantly higher percentages of circul

In comparison with HC, significantly higher percentages of circulating IgD+CD27−CD19+ naive B, CD86+CD19+ and CD95+CD19+ activated B, CD3+CD4+CXCR5+,

CD3+CD4+CXCR5+ICOS+, CD3+CD4+CXCR5+PD-1+ and CD3+CD4+CXCR5+ICOS+PD-1+ Tfh cells but lower IgD+CD27+CD19+ preswitch memory B cells were detected, accompanied by significantly higher levels of serum IL-21 in the RA patients. Furthermore, the percentages of CD95+ B cells were correlated positively with the frequency of PD-1+ Tfh cells, but negatively with ICOS+ Tfh cells. The percentages of CD86+ B cells and ICOS+ Tfh cells were correlated positively with the values of disease activity score 28 (DAS28). Following the drug therapies for 1 month, the percentages Imatinib datasheet of CD86+ B and PD-1+ Tfh cells were reduced significantly in the drug-responding patients. Our data suggest that activated B and Tfh cells may contribute to the pathogenesis of RA and the frequency of activated B and Tfh cells may be used as biomarkers

for evaluating the therapeutic responses of individual patients with RA. Rheumatoid arthritis (RA) is a severe chronic autoimmune inflammatory disease. RA is characterized by symmetric polyarthritis associated with pain and swelling in multiple joints. Importantly, most RA patients eventually develop cartilage lesions and bone destruction, leading to functional incapacity. In addition, RA patients are affected by an increased frequency of other co-morbidities

and decreased life expectancy [1]. Currently, the pathogenic process of RA is still unclear. The pathogenesis of RA is attributed to the interaction of many types of immunocompetent cells, such as antigen-specific T and B cells, aberrant activation of antigen-presenting cells (APC) and autoantibodies [2]. Although antigen-specific Thiamet G T cells are crucial for the pathogenesis of RA, recent evidence suggests that B cells play an important role in the development and progression of RA [3]. CD27 is expressed on somatically mutated B cells and the distinct subsets of B cells can be defined as naive immunoglobulin (Ig)D+CD27−, preswitch memory IgD+CD27+, post-switch memory IgD−CD27+ and double-negative IgD−CD27− B cells [4, 5]. Activation of B cells up-regulates CD86, CD95 and major histocompatibility complex (MHC) class II expression and some activated B cells differentiate into plasma cells which express CD38 [6], while others become memory B cells which express CD27 [5]. The up-regulated CD95 expression in activated B cells makes them sensitive to ligand-mediated apoptosis [7, 8]. However, little is known about the frequency of these different subsets of activated B cells in patients with new-onset RA. The activation and functional differentiation of B cells are regulated by CD4+ T cells, particularly by T follicular helper (Tfh) cells [9, 10].