Two days later, 30 IU/ml of human rIL-2 was added to the medium. After 5 days, the cultured cells were collected and used as CTL effector cells. To detect B16 melanoma-specific CTL activity, we used TRP-2-peptide-pulsed EL-4 target cells or EL-4 cells pulsed with lymphocytic choriomeningitis virus (LCMV) glycoprotein (GP)34–41 peptide (H-2Kb-restricted peptide AVYNFATCGI; produced by Genenet) as a third-party control. To detect CT26-specific CTL activity, we used CT26 target cells or J558L target cells as a third-party control. The target cells were labelled with 100 μCi Na251CrO4 for 1.5 h, and the 51Cr release assay was performed as previously

described [15]. The percentage of specific 51Cr release was calculated as follows:

% cytotoxicity = [(Cr release of experimental medium − culture medium background)/(maximum Cr release − culture medium background)] × 100. Each data point was obtained from triplicate wells. Statistical analysis.  Bortezomib Tumour growth was analysed using two-way anova, and the significance was calculated using Bonferroni’s post hoc test. The number of tumour-specific IFN-γ-producing CD8+ T cells was analysed BMS-354825 molecular weight by one-way anova, and the significance was calculated using Bonferroni’s multiple comparison post hoc tests. Survival rates were analysed using a log-rank comparison test. A probability value of P < 0.05 was considered significant. All data were analysed using Graphpad Prism®4 software (version 4; GraphPad Software, Inc., San Diego, CA, USA). Our group and others previously reported that i.t. injection of syngeneic DC without pulsation with tumour lysates could induce efficient antitumour responses to various cancers with TAA-specific CTL responses in murine s.c. tumour models [14, 15]. In this study, we referred to this DC-based cancer immunotherapy as ITADT. We investigated whether allogeneic DC could be used for cancer immunotherapy Rebamipide in the setting of ITADT. First, we used a B16 melanoma model. C57BL/6 mice were subcutaneously injected

with B16.F1v cells, and an i.t. injection of DC was given 3 days later followed by two additional injections at 1- week intervals. Consistent with previous reports [14, 15], ITADT using syngeneic female C57BL/6 DC (BL6 F DC; H-2b) induced an efficient antitumour effect, resulting in significant suppression of tumour growth, with 2/10 tumours being totally eradicated. The BL6 F DC-treated mice also showed significantly improved survival rates compared with PBS-treated controls (Fig. 1A,B and supplementary Fig. S1A, P < 0.01). We then tested semi-allogeneic DC (C57BL/6 × DBA/2 F1: BDF1 DC; H-2b/d) or minor disparate allogeneic DC (male C57BL/6: BL6 M DC; H-2b) and found that ITADT using these DC could induce antitumour effects similar to ITADT using syngeneic DC (Fig. 1A,B). In 2/11 mice treated with BL6 M DC and 1/11 mice treated with BDF1 DC, the B16.F1v tumours were eradicated (supplementary Fig. S1A).

01). Conclusion: Our studies confirmed the role of Gd-IgA1-IgG IC in the pathogenesis of IgAN and induction of proteinuria and hematuria.

Furthermore, the Gd-IgA1-IgG IC may bind to glomerular endothelial cells and induce release of pathogenic cytokines and chemokines. SUZUKI HITOSHI1, SUZUKI YUSUKE1, MAKITA YUKO1, YANAGAWA HIROYUKI1, JULIAN BRUCE A2,3, NOVAK JAN3, TOMINO YASUHIKO1 1Division of Nephrology, Department of Internal Medicine, Juntendo University Faculty of Medicine; 2Departments of Medicine, PLX3397 price University of Alabama at Birmingham; 3Departments of Microbiology, University of Alabama at Birmingham Introduction: IgA1 in circulating immune complexes and mesangial deposits of patients with IgA nephropathy (IgAN) is aberrantly glycosylated, galactose-deficient in O-glycans (Gd-IgA1), and is bound to anti-glycan IgG/IgA autoantibodies. However, the origin of cells producing Gd-IgA1 and the autoantibodies is not certain. Upper respiratory tract infections and tonsillitis are frequently associated with clinical presentation and exacerbation of IgAN, suggesting a link with disease pathogenesis. In some patients, tonsillectomy and glucocorticoids (TSP) may slow disease progression PD0325901 in early clinical stages. Therefore, we assessed whether

tonsillar cells produce Gd-IgA1 or anti-glycan autoantibodies. Methods: Tonsillar

cells obtained from 29 patients with IgAN were cultured 72 hours. Gd-IgA1 and anti-glycan IgG secreted by these cells were measured by ELISA. Proteinuria and hematuria, and serum levels of Gd-IgA1, Gd-IgA1-specific IgG and IgA, and IgG-IgA immune complexes (IC) were measured before and Olopatadine after TSP. Results: Proteinuria and hematuria improved after TSP (P < 0.05). Eighteen of 29 patients had proteinuria less 0.3 g/g and 5 red blood cells/HPF after TSP (Remission group). Eleven patients did not clinically improve (non-Remission group). Serum levels of Gd-IgA1, Gd-IgA1-specific autoantibodies, and IgG-IgA IC decreased during glucocorticoid therapy after tonsillectomy (P < 0.01). The rates of decrease in the levels of Gd-IgA1, Gd-IgA1-specific antibodies and IgG-IgA IC were greater in the Remission group (P < 0.01). Tonsillar cells from Remission group produced more Gd-IgA1 and anti-glycan IgG than those from non-Remission group (P < 0.01). Conclusion: Tonsillar cells may contribute to the circulating Gd-IgA1 and anti-glycan IgG in patients with IgAN. These biomarkers may be useful for guiding therapy of IgAN. YAMADA KOSHI1,2, HUANG ZHI-QIANG1, RASKA MILAN1,3, REILY COLIN R.1, SUZUKI HITOSHI1,2, MOLDOVEANU ZINA1, KIRYLUK KRZYSZTOF4, SUZUKI YUSUKE2, TOMINO YASUHIKO2, GHARAVI ALI G.4, WILLEY CHRISTOPHER D.1, JULIAN BRUCE A.

2a). Interestingly, no production or secretion of FhaB was detected buy Luminespib under the iron-starved conditions (Fig. 2b). On the other hand, production and secretion of CyaA, Prn, and DNT were not significantly affected by the iron concentration (Fig. 2b). These results clearly indicate that BvgAS-regulated gene expression is not always enhanced by iron-starved conditions. To further investigate BvgAS-regulated gene expression

under iron-starved conditions, total RNA was prepared from B. bronchiseptica cultured under iron-replete or -depleted conditions. The cDNA samples reverse-transcribed from the total RNA samples were subjected to quantitative RT-PCR analysis to quantify the relative amounts of bsp22 and fhaB mRNA as a hallmark of the BvgAS-regulated

gene that is positively or negatively regulated by iron-starved conditions (Fig. 3). The Bsp22 gene was transcriptionally activated by iron starvation. In contrast, the fhaB gene was repressed in response to iron starvation, demonstrating that the relative amounts of mRNAs are correlated with protein production, as shown in Fig. 2b. It has been reported that B. bronchiseptica induces necrotic cell death of various mammalian cultured cells in a T3SS-dependent manner (6, 8). To examine whether this phenotype is affected by iron-depleted conditions, L2 rat lung epithelial cells infected with B. bronchiseptica precultured under iron-replete or -depleted conditions selleck products were fixed and stained with Giemsa solution to analyze cell morphology (Fig. 4a). Approximately 60–70% of cells infected with B. bronchiseptica under iron-replete conditions were detached from the substrata and the remainder of adherent cells

exhibited shrunken cytoplasm and condensed nuclei (Fig. 4a). The L2 cells exposed to the T3SS mutant strain showed normal morphology that was identical to that of uninfected cells. In contrast, more than 90% of cells infected with B. bronchiseptica under iron-depleted conditions were detached, and their morphological changes were more pronounced than those of bacteria cultured under iron-replete conditions. Furthermore, HeLa cells were infected with B. bronchiseptica and the relative amounts of LDH released into the extracellular medium measured (Fig. 4b). The cytotoxicity evident in host O-methylated flavonoid cells infected with B. bronchiseptica under iron-depleted conditions was statistically greater than that of those infected with B. bronchiseptica under iron-replete conditions. T3SS-dependent hemolytic activity was also evaluated using RBCs (Fig. 4c). Again, hemolytic activity of B. bronchiseptica grown under iron-depleted conditions was statistically greater than that of B. bronchiseptica grown under iron-replete conditions. Collectively, these results suggest that B. bronchiseptica is able to recognize iron-starved conditions and exert the T3SS function in response to them.

cruzi and L. major (15). The majority of species-specific genes – of which T. cruzi (32%) and T. brucei (26%) have a much greater proportion than L. major (12%) – occur at non-syntenic chromosome-internal

and subtelomeric regions and consist of members of large surface antigen families. These gene family expansions, along with structural RNAs and retroelements, are often associated with breaks in synteny. Gene divergence, acquisition and loss, and rearrangements within and between syntenic regions have shaped the genomes of the trypanosomatids (15). A remarkable Fulvestrant cell line feature of the T. brucei and T. cruzi genomes is the extensive expansion of species-specific genes, the large majority encoding surface proteins, such as Variant Surface Glycoproteins (VSGs) in T. brucei, trans-sialidase superfamily, mucin-associated surface proteins and mucins (TcMUC) among others in T. cruzi, all of them likely involved in important host-parasite interactions (15). These surface protein-encoding genes are often clustered into large arrays that can be as large as 600 kb and are/were subjected to intense rearrangements during the parasites’ evolution (15,20). It is likely therefore

that much of the striking polymorphism among the T. cruzi and T. brucei isolates that are reflected in several epidemiological and pathological aspects of Chagas disease and African sleeping sickness may be in part because of variability within these regions. Whole genome comparisons Compound Library of distinct trypanosomatid lineages through would allow further investigation of this. A wide range of pathologies is found within trypanosomatid parasite lineages. Thus, there remains a considerable evolutionary and pathological

space yet to be explored through additional comparative sequencing (we define pathogenomics as the genome analysis of pathogens). With the advent of massively parallel sequencing technologies, sequencing of additional trypanosomatid strains can now be performed at a fraction of the cost of the sequencing of the reference genomes. The Wellcome Trust Sanger Institute (WTSI) has initiated such efforts. The recent sequencing of the genomes of several Leishmania species, causative agents of cutaneous, mucocutaneous and visceral leishmaniasis, is beginning to unravel many features of potential relevance to parasite virulence and pathogenesis in the host. When compared to L. major, the genomes of Leishmania braziliensis and L. infantum displayed a highly conserved gene content and order. However, two hundred genes with a differential distribution between the three species were identified (21,22). Perhaps most unexpected was the discovery that L. braziliensis genome retained the components (Argonaute and Dicer) of a putative RNA interference pathway, which are absent in L. major and L. infantum. A subsequent functional study demonstrated the presence of a strong RNAi activity in L. braziliensis (23).

Because of the timing of serum EMA and NFR antibodies, circulating ANA were evaluated at three time-points: during EMA-positive results, under EMA disappearance/NFR-positive results and after NFR disappearance. At all time-points, serum ANA were positive in two of 20 CD see more patients in group 1. In both cases, an ANA-S antibody pattern (subpattern: fine speckled) was visible. None of the 15 subjects in group

3 presented serum EMA-positive results, while two showed an NFR-like pattern on monkey oesophagus sections. The latter two subjects were put on a GFD for 12 months. Serum EMA and NFR antibodies were evaluated each month, showing no changes in the NFR-like pattern. The characterization of this NFR-like pattern showed that it belonged simultaneously to IgA1 and IgA2 subclasses, and that it was localized in the nucleus. The results of the present study demonstrate that serum IgA from CD patients are able to react with two nuclear antigens determining the appearance of a nuclear fluorescence selleck inhibitor reactivity (NFR) antibody pattern on monkey oesophagus sections used routinely for EMA detection. Moreover, as NFR antibodies are detectable

in serum as long as the CD patients consume gluten and disappear after gluten withdrawal from the diet, they are gluten-dependent and related strictly to CD. The autoimmune nature of CD is understood clearly [5–7], and the main autoantigen is well known to be tTG [11]. However, tTG is not the only CD-related autoantigen, as other tissue components have been shown to be a target of coeliac autoimmunity [12–15]. In serum of active CD patients, antibodies against thyroid and pancreas structures, cytoskeleton molecules and central nervous

Dimethyl sulfoxide system-related antigens have been found previously [14]. The present study adds a new antigen type to the list, as we found that serum IgA from untreated CD patients react with two NFR-related nuclear antigens of 65 and 49 kDa. The identity of NFR-related autoantigens is as yet unknown, but based on the different distribution of EMA and NFR reaction sites on monkey oesophagus sections it is reasonable to hypothesize that these reactivities are due to distinct antigenic specificity. Indeed, EMA and NFR antibody patterns are never observable simultaneously during total IgA EMA detection but, using secondary mAbs against IgA subclasses (IgA1 and IgA2) coupled with different fluorochromes (FITC and TRITC), the presence of two different and not overlapping fluorescence signals becomes evident. That the main endomysial antigen, known to be tTG [11], has a different molecular weight with respect to the newly identified autoantigens (85 versus 65 and 49 kDa), further confirms the hypothesis that EMA and NFR are two distinct antibodies.

However, the use of an echinocandin + liposomal amphotericin B formulation is a better option as indicated by both animal and human data.[31-35] All authors declare no conflicts of interest. “
“Immunocompromised patients have Decitabine mw a high risk for invasive fungal diseases (IFDs). These infections are mostly life-threatening and an early diagnosis and initiation of appropriate antifungal therapy are essential for the clinical outcome. Empirical treatment is regarded as the standard of care for granulocytopenic

patients who remain febrile despite broad-spectrum antibiotics. However, this strategy can bear a risk of overtreatment and subsequently induce toxicities and unnecessary treatment costs. Pre-emptive antifungal therapy is now increasingly used to close the time gap between delayed initiation for proven disease and empirical treatment for anticipated infection without further laboratory or radiological evidence of fungal disease. Currently, some new non-invasive microbiological and laboratory methods, like the Aspergillus-galactomannan sandwich-enzyme immunoassay (Aspergillus GM-ELISA), 1,3-β-d-glucan assay or PCR techniques

have been developed for a better diagnosis Nutlin-3 datasheet and determination of target patients. The current diagnostic approaches to fungal infections and the role of the revised definitions for invasive fungal infections, now IFDs, will be discussed in this review as well as old and emerging approaches to empirical, pre-emptive and targeted antifungal therapies in patients with haemato-oncological malignancies. “
“Prosthetic joint infections (PJI) are rarely due to fungal agents and if so they are mainly caused by Candida strains. This case represents a PJI caused by a multi-drug resistant Pseudallescheria apiosperma, with poor in vivo response to itraconazole and voriconazole. This case differs also by the way of infection, since the Sorafenib chemical structure joint infection did

not follow a penetrating trauma. In the majority of cases, Scedosporium extremity infections remain local in immunocompetent individuals. We report a persistent joint infection with multiple therapeutic failures, and subsequent amputation of the left leg. Detailed clinical data, patient history, treatment regime and outcome of a very long-lasting (>4 years) P. apiosperma prosthetic knee infection in an immunocompetent, 61-year-old male patient are presented with this case. The patient was finally cured by the combination of multiple and extensive surgical interventions and prolonged antifungal combination therapy with voriconazole and terbinafine. Prosthetic joint infection (PJI) is mainly caused by bacteria and rarely by human-pathogenic yeast such as Candida strains.1–4Aspergillus fumigatus5 or other filamentous fungi are only exceptionally involved.

1). Selectins are a family of three cell adhesion molecules known as L-, P- and E-selectin. Their primary role in recruitment involves weak binding Opaganib concentration to their specific ligand on the surface of monocytes and the

endothelium, which reduces their flow rate velocity and mediates rolling along the endothelium (Fig. 1). During this low-affinity rolling phase, monocytes are exposed to a plethora of secreted cytokines and chemoattractants, which subsequently induces the activation of integrins, which are a large family of heterodimeric transmembrane glycoproteins that connect cells to their microenvironment mediating cell-to-cell adhesion. Integrins present on the surface of monocytes include leukocyte SRT1720 purchase functioning associated antigen (LFA)-1, macrophage adhesion ligand (Mac)-1 commonly referred to as CD11b, and very late activation antigen (VLA)-4.

These integrins interact with their endothelial counter-receptors, intercellular adhesion molecule (ICAM)-1 and vascular cell adhesion molecule (VCAM)-1. Binding of LFA-1 and Mac-1 to ICAM-1, and VLA-1 to VCAM-1 mediates firm adhesion of monocytes to the endothelium allowing for diapedesis to occur into surrounding tissue (Fig. 1). Blockade of E- and P-selectins in rodent models of ischaemia–reperfusion (IR) injury reduces renal macrophage recruitment, which subsequently leads to amelioration of the pro-inflammatory response and reduced tubular damage and interstitial fibrosis production.[44-47] Knockout (KO) mice and neutralizing antibodies against ICAM-1 and its binding partners, LFA-1

and CD11b, also prevent monocyte recruitment medroxyprogesterone and consequently induce less severe damage in several renal disease models including glomerulonephritis (GN),[48-51] diabetic nephropathy,[52-54] unilateral ureteral obstruction (UUO)[55] and IR injury.[56] Following selectin-mediated adhesion of monocytes to the endothelium, increased expression of chemokines and chemokine receptors induce a chemotactic gradient that promotes firm integrin-mediated adhesion and transmigration across the vasculature and into tissue (Fig. 1). Most kidney cells including tubular epithelial cells (TECs), podocytes, mesangial and endothelial cells have the potential to produce chemokines and express chemokine receptors, with a rapid expression induced by the following pro-inflammatory cytokines and mediators TNF-α, IL-1β, interferon (IFN)-γ, lipopolysaccharide (LPS) and reactive oxygen species. CCL2 is the most important chemokine in mobilizing monocytes to the kidney following damage. CCL2 binds to its receptor CCR2, which is highly expressed on inflammatory monocytes.[16] Along with CCL2/CCR2 signalling, CX3CL1, CCL5, CCL3, CCL4, CXCL8, and their corresponding receptors CX3CR1, CCR1, CCR5 and CXCR2 have also been implicated in monocyte recruitment during renal inflammation as recently reviewed.

Using OVA peptide variants with different affinity for the OVA-specific OT-I TCR, it was shown that peptides with high affinity induce high amounts of IRF4 [22, 25], whereas peptides with intermediate or low affinity provoke intermediate or low quantities of IRF4, respectively. This dependency of IRF4 expression amounts on the peptide affinity for OT-I TCR was demonstrated in vitro and also in vivo during infection with recombinant Listeria monocytogenes that expressed the respective peptide variants [22]. At the molecular level, IRF4 expression levels seem to depend on the activity of mammalian target of rapamycin (mTOR). Thus, high IRF4 expression following strong TCR stimulation by high-affinity

ligands correlated with elevated activity of mTOR, whereas inhibition of the mTOR pathway caused downregulation of IRF4 [25]. As recently shown, IRF4 expression is also dependent on the activity of IL-2-inducible T-cell kinase (ITK) [26]. Using inhibitors for both selleck ITK and mTOR, it was demonstrated that these two signaling pathways cooperate for IRF4 induction [25]. Earlier studies had already concluded that the transcription factor C-REL, a member of the NF-κB family, is also crucial for the induction of IRF4 in response to TCR

stimulation [27]. Moreover, treatment with cyclosporine Buparlisib in vitro A blocked upregulation of IRF4, suggesting that NFAT signaling also contributes to this process [3]. Finally, FOXP3 regulates IRF4 expression in regulatory T (Treg) cells [19], as do STAT3 in T helper 17 (Th17) cells [28] and STAT6 in Th9 cells [29], whereas T-BET directly represses IRF4 expression in Th1 and Th17 cells [30]. In response to signals induced by antigen recognition

and cytokines, naïve CD4+ T cells differentiate into distinct subpopulations that are characterized by specific effector functions and cytokine profiles. This subdivision is based on the expression of lineage-specific transcription factors, which function as “master regulators” for specific Th-subset properties (Fig. 1). IL-12 drives the differentiation of Th1 cells, which produce IFN-γ, express the transcription factor T-BET (encoded by T-box 21), and clear intracellular Gemcitabine concentration pathogens. Th2 cells are induced by IL-4, secrete IL-4, IL-5, and IL-13, and express the master regulator GATA-binding protein 3 (GATA3). IL-4 in combination with transforming growth factor-β (TGF-β) induces the differentiation of Th9 cells, which produce high levels of IL-9 and IL-10. The lineage-specifying transcription factor for Th9 cells was suggested to be PU.1, which however was previously considered by the same group to characterize an IL-4 low producing subset of Th2 cells [31]. Although Th2 and Th9 cell subsets both contribute to immunity against helminths, Th9 cells are additionally involved in antitumor immunity. The cytokines IL-6 or IL-21 can act alone to induce T follicular helper (Tfh) cells, which express the master regulator BCL-6.

However, we showed that anti-M3R antibodies against these linear epitopes exactly influenced Ca influx via M3R in HSG cells. Therefore, we suggest that these linear peptides might consist of the conformational epitopes on the M3R. Several B cell epitopes were identified on the extracellular domains, and some SS patients were reactive to several extracellular domains other than the second extracellular loop. The second extracellular loop of M3R has been the focus of our interest in epitopes and function of anti-M3R antibodies [4,5,9,10]. Recently, Koo et al.[6] reported that the third extracellular loop represents a functional ZVADFMK epitope bound by SS-IgG. Selleckchem GSK3 inhibitor In contrast

to these results, we found in the present study that antibodies to the second extracellular loop of M3R inhibited the increase of (Ca2+)i induced by cevimeline hydrochloride in a functional assay using HSG cells. This inhibitory effect of anti-M3R antibodies might explain the reduction in salivary secretion in some SS patients. Our data also demonstrated that antibodies against the third extracellular loop did not have an effect on the increase in (Ca2+)i, while antibodies against the N-terminal and first extracellular

loop enhanced the increase in (Ca2+)i. These results indicate that the effects of anti-M3R antibodies on the secretion of saliva could be different from these epitopes, although further experiments using antibodies from more patients are necessary. Although the molecular mechanisms on the difference among individual B cell epitopes have not been elucidated, we could propose the following three possibilities. The first is that antibodies against the second extracellular domain GNAT2 of M3R directly inhibit

the intracellular signal pathway, resulting in the decrease of Ca2+ influx and reduction of saliva. In contrast, antibodies against N-terminal region and the first extracellular domain of M3R might enhance the intracellular signalling and increase of Ca2+ influx. The second is that anti-M3R antibodies binding to the second extracellular domain could inhibit the M3R agonist, and then antibodies suppress indirectly the stimulation of Ca2+ influx. The third is that anti-M3R antibodies influence the expression of M3R molecules on HSG. Some antibodies which target the N-terminal region or the first extracellular loop of M3R may be able to up-regulate expression of M3R and enhance Ca2+ influx, whereas the other antibodies against the second extracellular domain might down-regulate the expression of M3R on HSG, resulting in a reduction of Ca2+ influx. It has been reported that the expression of M3R in salivary glands could be affected by anti-M3R antibodies in patients with SS [1].

Thus, this study reveals that pneumolysin induces the proinflammatory cytokine expression in a time-dependent manner. Inflammation triggered by infections is one of the counteractions that occur

in the host to facilitate pathogen clearance by recruitment of leukocytes. An excessive inflammatory response, however, is harmful to the host because it causes severe tissue damage (Hersh et al., 1998). Tight control of inflammation is thus critical for host immune defense and can be achieved by balancing the expression of proinflammatory FK506 price cytokines and anti-inflammatory cytokines (Dinarello, 2000). Proinflammatory cytokines such as interleukin-1β (IL-1β) and tumor necrosis factor α (TNF-α) serve to promote inflammation by promoting a diverse

BYL719 order range of activities including the induction of adhesion molecules required for the transmigration of leukocytes to infection sites (Dinarello, 2000). The release of proinflammatory cytokines can be triggered by various bacterial products including LPS of Gram-negative bacteria, peptidoglycan of Gram-positive bacteria or specific molecules from diverse microorganisms (Henderson et al., 1996). Gram-positive bacterium Streptococcus pneumoniae is an important cause of morbidity and mortality in humans, especially among young children (Bluestone et al., 1992). Among the numerous virulence factors identified in PDK4 S. pneumoniae to date, the cell wall plays an important role in initiating inflammation during infection, which is characterized by the production of proinflammatory cytokines and leukocyte influx (Tuomanen

et al., 1985; Bruyn & van Furth, 1991; Cundell et al., 1995). The cell wall components consist of polysaccharides and teichoic acid, which are recognized by Toll-like receptor 2 (TLR2) (Yoshimura et al., 1999). On the surface of the cell wall, there are a range of cell surface-associated proteins involved in the pathogenesis of S. pneumoniae during infection, including autolysin, pneumococcal surface protein A (PspA), PspC, hyaluronidase, neuraminidase, and pneumococcal surface antigen A (PsaA) (Mitchell, 2006). On the other hand, pneumolysin, which is 53 kDa in size, is localized in the cytoplasm and seems to be released during infections by the action of pneumococcal autolysis from virtually all clinical isolates (Canvin et al., 1995; Wheeler et al., 1999). However, it has been reported recently that the pneumolysin is also localized to the cell wall compartment (Price & Camilli, 2009). The upper respiratory tract is the ecological niche for various bacterial species including S. pneumoniae and nontypable Haemophilus influenzae (NTHi) (Faden et al., 1990; Givon-Lavi et al., 2002). NTHi has been identified as a major pathogen causing otitis media (OM) and pneumonia along with S. pneumoniae (Gok et al., 2001; Ozyilmaz et al., 2005).