The effects had never been studied yet on a lung model for large mammals. Our data showed dose-dependent effects of CsA on gas exchanges, but also on pulmonary hemodynamics, and possibly an aggravation of the IRI due to high doses of CsA. These results constitute an important step toward the use of CsA on humans to reduce lung IRI and consequently, primary graft dysfunction. Within a few years, the EVLP technique has become a reference for the evaluation of lung grafts. Its interest has been demonstrated Selleck Talazoparib on animal

lung preparations, especially on pig [43] and human lungs [12]. This technique can be seen as bench test for lung function, allowing for the assessment of new therapies suppose to limit IRI. Gas exchange capacities and total pulmonary arterial resistance are more commonly studied physiopathological parameters. We also measured other hemodynamics (Pcap,

longitudinal pulmonary resistance) and markers (AFC, RAGE, cytokines, lung permeability) that have showed their pertinence in the evaluation of lung IRI [5, 7]. It has been hypothesized that IRI is mostly related to mitochondrial death as a consequence of MPTP opening. Located in the inner mitochondrial membrane, the MPTP remains unremarkable under normal physiological conditions. Stress can lead to its opening, resulting in the swelling of the matrix due to osmotic forces. It then induces further failure of the mitochondrial outer membrane and the release of the cytosol pro-apoptotic factors [19]. The inhibition of Estrogen antagonist the opening of MPTP is thought to be the main pathway for CsA action. Several in vitro and in vivo animal models showed CsA interests in pre and post-conditioning for the

prevention of IRI on different organs such as heart, kidney, and liver [19, 20, 45, 50]. In humans, CsA administered just before coronary reperfusion (post-conditioning) has been proven to be an efficient way to reduce the size of myocardial infarction [33]. However, few studies have been published on CsA effects on lung IRI. In vitro studies on post hypoxia-reoxygenation injuries showed that alveolar macrophages pretreated by CsA secreted less chemokines than Axenfeld syndrome controls [30]. Moreover, endothelial cells incubated with CsA selectively reduced pro-inflammatory mediator secretion of NFκB and EGR-1 [15]. Nevertheless, some of the pathways involved in IRI can be activated by CsA, such as the metalloproteinase and the TLR [1, 28, 41]. Such insights can explain the increased levels of pro-inflammatory cytokines we measured in our experiments with high doses of CsA (30 μM). In an in vivo ischemic lung model, Krishnadasan et al. showed that rats pre-conditioned with CsA displayed less tissue myeloperoxidase content, leukocyte accumulation, and vascular permeability [25].

6e). To determine if xeno-GVHD resulted

from a loss of peripheral tolerance, we evaluated the levels of human Treg detectable in the blood of standard NSG–BLT mice (with irradiation) over time (Fig. 6f). The percentage of CD25+/CD127dim/FoxP3+ cells in the blood of NSG–BLT mice did not decrease over time. To determine the contribution of irradiation in the development of xeno-GVHD in BLT mice, we compared the survival of NSG–BLT mice that were either irradiated or non-irradiated (Fig. 6g). Overall, there was an increased survival of non-irradiated NSG–BLT mice; however, these animals click here ultimately developed GVHD-like symptoms. The BLT mouse, also referred to as the Thy/Liv mouse, is an ideal model to study human immune and T cell functions, as the implant of human thymic tissues and autologous human HSC enable the efficient development of HLA-restricted human CD4 and CD8 T cells [63]. Following implantation into the subcapsular Pritelivir concentration renal space, the human fetal thymus grows significantly, is populated with a normal distribution of human thymocyte subsets and allows high levels of human T cells to repopulate the peripheral lymphoid tissues [21-23]. The BLT model is based on the severe compromised immunodeficient-humanized

(SCID-hu) mouse described by McCune and colleagues [6]. The original SCID-hu model was created using CB17-scid mice and involved the transplant of human fetal thymic tissues in the renal subcapsular space and i.v. injection of autologous or allogeneic HSC derived from the fetal liver. The SCID-hu mouse enabled the development of human T cells, which required both the implant of thymic tissues and injection of HSC. However, in CB17-scid mice the C59 concentration persistence of human T cells in the peripheral

tissues was transient, as CD3+ cells were not detectable in the peripheral blood at 12 weeks post-implant and the ability of these cells to mediate an immune response was limited [64]. The persistence and functionality of human T cells was improved significantly by the use of NOD-scid mice as recipients of human thymic and liver tissues [22, 23]. However, engraftment of fetal thymic and liver tissues into NSG mice enhances human cell chimerism significantly, including reconstitution of a mucosal immune system, compared to other mouse strains [17, 65]. Continued improvement of the NSG mouse by the transgenic expression of human-specific cytokines and growth factors and expression of HLA that will allow matching with the donor tissues will further augment the development of human immune systems in BLT mice [3, 66]. In an effort to provide an analysis of optimal parameters for establishing the NSG–BLT model, we have assessed the requirement for irradiation to attain high-level human cell chimerism, the optimal implantation sites for thymic tissues, the stability of human cell chimerism and the longevity of engrafted mice.

This is of interest for diseases, such as systemic infections, rheumatoid arthritis and osteoarthritis, which are associated with an increased activation of coagulation and the presence of physiological concentrations

of coagulation proteases, which may contribute to pro- or anti-inflammatory responses in a PAR-dependent manner. Therefore, in this study, it was investigated whether coagulation proteases (FVIIa, the binary TF-FVIIa complex, the binary TF-FVIIa complex with free FX, free FX, free FXa and thrombin) in physiological concentrations can elicit pro- or anti-inflammatory responses in a PAR-dependent manner in naïve (non-preactivated) human monocytes and PBMCs. Ficoll-Paque was purchased from Pharmacia (Uppsala, Sweden) and CD14 microbeads from Miltenyi Biotec (Bergisch Gladbach, buy PD-0332991 Germany). Dulbecco’s modified Eagle’s medium (DMEM) was obtained from Invitrogen (Carlsbad, CA, USA). Heat-inactivated human male AB serum was from

Sigma-Aldrich (St. Louis, MO, USA). Allophycocyanin (APC)-conjugated monoclonal mouse anti-human CD14 antibody and APC-conjugated isotype control antibody were from BD Biosciences (Franklin Lakes, NJ, USA). Phycoerythrin (PE)-conjugated monoclonal mouse anti-human PAR-1 (ATAP2) antibody, FITC-conjugated monoclonal mouse anti-human PAR-2 (SAM11) antibody, GS-1101 PE-conjugated monoclonal mouse anti-human PAR-3 (8E8) antibody, and APC-, PE- and FITC-conjugated isotype control antibodies were from Santa Cruz Biotechnology Inc. (Santa Cruz, CA, USA). FITC-conjugated polyclonal rabbit anti-human PAR-4 (APR-034-F) antibody was obtained from Alomone Labs (Jerusalem, Israel). PE-conjugated monoclonal mouse anti-human TF (HTF-1) antibody and PE-conjugated isotype control antibody were from BD Biosciences. Recombinant human FVIIa was kindly provided by Novo Nordisk A/S (Maaloev, Denmark). Recombinant human tissue factor (4500L), human factor X (527) and human activated factor X (526) were purchased from American Diagnostica

Inc. (Stamford, CT, USA). Human alpha thrombin factor IIa (IHT; activity ≥2700 NIH units/mg) was obtained from Innovative Research (Novi, USA). The GBA3 activity of the purchased coagulation proteases was tested positive in coagulation assays before use. Purified LPS was purchased from Sigma-Aldrich. PAR-1 antagonist FR171113 was obtained from Tocris Bioscience (Bristol, UK). FR171113 is a highly purified (>98%) specific PAR-1 antagonist which is able to inhibit thrombin-induced platelet aggregation. Interleukin-1β (IL-1β), Interleukin-10 (IL-10) and tumour necrosis factor-alpha (TNF-α) enzyme-linked immunosorbent assay (ELISA) kits were from Invitrogen. Interleukin-6 and IL-8 ELISA kits were obtained from eBioscience (San Diego, CA, USA). All other chemicals were from Sigma-Aldrich. Peripheral blood was obtained from five different healthy donors after informed consent (age 37.2 ± 4.9 years; 2 males and 3 females). PBMCs were isolated by Ficoll-Paque (Pharmacia) according to standard procedures.

Having analyzed the very early stages of this differentiation process we next looked at the long-term development of memory cells by phenotypically analyzing cell surface marker expression profiles on WT and IFNAR−/− P14 cells in the blood of LCMV8.7 and VVG2 co-infected mice (Fig. 3C). This longitudinal analysis revealed that IFNAR−/− P14 cells initially begin

to down-regulate surface CD62L expression but after day 3 the level of CD62L is gradually regained on the population of IFNAR−/− P14 cells. This same trend is seen for the expression of CD127, and the opposite is seen for KLRG1 and CD25 expression (Fig. 3C). Of note, a comparable MPEC phenotype of IFNAR−/− P14 cells could be observed upon single HM781-36B nmr KU-60019 concentration LCMV-WE infection (Fig. 4A), indicating that although the antigen load seen by P14 cells profoundly differs between an infection with VVG2 or LCMV, type-I IFN is the main regulator of the fate decision toward the SLEC subset. Importantly, SLEC differentiation of IFNAR−/− P14 was similar to that of WT P14 cells in the context of a VVG2 only infection (Fig. 4B) 22, where high levels of IL-12 are produced at the expense of type-I IFN 17. These

results strongly suggest that depending on the type of infection and the predominant cytokines induced, different inflammatory signals instruct effector phenotype differentiation. Thus, in the context of VV infection, the high levels of IL-12 induced upon infection are sufficient to drive the differentiation of IFNAR−/− P14 cells into SLECs 23 and type-I IFN is not required for this process.

Furthermore, this finding shows that CD8+ T cells lacking type-I IFN signaling are not inherently impaired in their capacity to gain an SLEC phenotype 22. Based on these phenotypic results we reasoned that the amount of T-bet, an important transcription Oxymatrine factor that is more abundantly expressed in SLECs compared with MPECs 4, 24, might also differ in WT and IFNAR−/− P14 cells. Upon in vivo activation, WT and IFNAR−/− P14 cells upregulated T-bet expression independent of their phenotype (Fig. 5A). However, WT P14 cells expressed significantly higher T-bet levels than IFNAR−/− P14 cells at day 3 and even more pronounced at day 6 post-infection (Fig. 5A and B). As terminal effector differentiation is accompanied by high levels of T-bet whereas low amounts of T-bet rather promote MPEC development 4, we reasoned that in a type-I IFN biased cytokine milieu direct signaling via the type-I IFN receptor might regulate T-bet expression and thereby drive the fate decision toward an SLEC phenotype. We therefore examined the ability of type-I IFN to directly regulate the expression of T-bet. To this end, IFN-β was added to CD8+ T cells during in vitro activation with anti-CD3/CD28 and the relative expression levels of T-bet mRNA were monitored after 24 and 48 h (Fig. 5C).

The S100 proteins are thought to play a role in inflammatory conditions and tumorigenesis [8]. MRP14 was thought initially to occur only as a heterodimer complex with MRP8, but recently MRP14 is more often found to act on its own [9–12]. It is expressed in healthy skin and lung, while Navitoclax chemical structure MRP8 is undetectable in these tissues [12]. Although the exact function of MRP14 is not known, it may

be associated with disease severity in chronic inflammatory diseases and it was found to stimulate fibroblast proliferation in vitro[11,13,14]. MRP14 is expressed in affected tissue of gingivitis, rheumatoid arthritis, tuberculosis and sarcoidosis patients [12,14,15]. In sarcoidosis, MRP14 is expressed in epitheloid cells and giant cells composing the granuloma, whereas MRP8 is expressed only weakly or is even absent [15]. Using 2D electrophoresis, Bargagli et al. recently found MRP14 to be expressed

differentially in the BALF of sarcoidosis and IPF patients [16], but it was not possible to assess quantitatively the relationship of MRP14 with patient characteristics. In this study, we quantified BALF MRP14 levels in sarcoidosis and IPF patients using enzyme-linked immunosorbent assay (ELISA), and investigated whether MRP14 levels are associated with clinical parameters and disease severity. This is the first step towards understanding the role of MRP14 in fibrosing interstitial lung diseases. In this study, click here 74 sarcoidosis patients (54 male, 20 female) and 54 IPF patients (44 male, 10 female) were included retrospectively (Table 1). IPF patients were diagnosed at the Department of Pulmonology of the St Antonius Hospital Nieuwegein in the Netherlands and included when current American Thoracic Society/European Respiratory Society (ATS/ERS) criteria were met [4]. All Coproporphyrinogen III oxidase patients who underwent bronchoalveolar lavage (BAL) within 3 months from diagnosis were included. Eight IPF patients were treated with low-dose steroids at the time of diagnosis and BAL; the other IPF patients did not use

immunosuppressants. Sarcoidosis patients were diagnosed in accordance with the consensus of the ATS/ERS/World Association of Sarcoidosis and Other Granulomatous Disorders (WASOG) statement on sarcoidosis [17]. Sarcoidosis patients were classified based on chest radiographic stages according to Scadding [18]. Stage I showed bilateral lymphadenopathy (12 patients), stage II lymphadenopathy with parenchymal abnormalities (11 patients), stage III showed no lymphadenopathy but parenchymal abnormalities (19 patients) and stage IV showed fibrosis (32 patients, 16 non-steroid users and 16 steroid users). We first selected patients who had BALF and a clear classifying chest radiograph at presentation and were not treated with steroids at that time (12/11/12/eight per stages I, II, III and IV, respectively).

Biofilms of Candida spp. may be associated with increasing candidemia cases and treatment failure, as mature biofilms can become reservoirs of cells resistant to antifungal agents.[115] C. albicans

secretes higher amounts of Sap when grown in the form of biofilms, suggesting a relationship between secretion of Sap and the maintenance of biofilms on surfaces.[104, 116] Mores et al. [104] observed that secretion of Sap by sessile cells is greater than by planktonic cells and tends to increase if they grow in the presence of sub-MIC concentrations of fluconazole. Several studies have pointed out differences in patterns of secretion and in Sap activity in the presence of antifungal agents, but these can be related to differences in the sensitivity of the methods used to evaluate the proteolytic activity of Sap. Contrasting

FDA-approved Drug Library in vivo results were seen in the levels of Sap activity in the presence of antifungal buy JQ1 agents.[100] Most of the studies included in this review observed an increased expression of Sap in resistant isolates in the presence of sub-MIC concentrations of antifungal agents.[100, 101, 107, 108, 111] However, in a study by Copping et al. [113], the increase in Sap activity was mainly observed in susceptible isolates, whereas in resistant isolates there was a reduction in activity. Schulz et al. [110] observed a single isolate before and after exposure to fluconazole and despite not having found significant differences in Sap activity, they observed alterations in other factors associated with virulence, such as the ability to form biofilms. Induction of SAP gene expression by exposure Palmatine to antifungal agents is generally done using sub-MIC concentrations. However, in work by Ripeau et al.

[112], caspofungin was tested at fungicide concentrations and no induction or suppression of SAP gene expression was observed. Our review suggests that naturally resistant Candida spp. isolates or isolates that have developed resistance after prolonged exposure to drugs may present an increase in the secretion pattern and proteolytic activity of Sap. However, discrepancies in the results from different studies conducted under similar conditions may be due to the fact that virulence-associated factors are correlated to ensuing pathogenicity. Currently, there are very few studies on SAP gene expression and they are predominantly carried out on the more common species, such as C. albicans. The role of Sap in the virulence and pathogenesis of Candida spp. has been studied in detail, but more studies are needed to elucidate its relation to antifungal resistance fully. The Fundação de Amparo à Pesquisa do Estado de Minas Gerais (FAPEMIG) (APQ-01684/08; 02782/10, 01413/12) and Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES). All authors report no conflicts of interest relevant to this study.

2, black bars, right Y-axis), and that led to decreasing expression of endogenous miR-221 (Fig. 2A, white bars, left Y-axis) and miR-222 (Fig. 2B, white bars, left Y-axis). We conclude that Pax5 downregulates, either directly or indirectly, the expression of miR-221 and miR-222. We retrovirally introduced a doxycycline-inducible system of overexpression of miR-221 and miR-222 in Pax5+/+ pre-B-I cells to test whether miR-221 or miR-222 has

a modifying effect on the differentiation or migration of pre-B-I cells. In this system GFP becomes expressed when mature miRNA is formed by splicing [21, 22] (Supporting Information Nutlin3a Fig. 2A and B). We assayed the overexpression

of miR-221 by quantitative real-time PCR with a probe specific for the mature miR-221 and confirmed its time-dependent overexpression (Supporting Information Fig. 2C). The highest upregulation of miR-221 (14- and 18-fold, compared with that of the empty vector control) was detected 24 and 72 hours after addition of 1 μg/mL doxy-cycline. We used a luciferase reporter assay to test the function of miR-221 to downregulate gene expression (Supporting Information Fig. 3). The results show that expressed, mature miR-221 functions to reduce luciferase activity by inhibiting the translation of the luciferase gene. To test whether single overexpression of miR-221 would revert the B cell-monopotency of the pre-B-I cell line back to the multi-myeloid/lymphoid potency of the miR-221/miR-222 expressing MPPs and

HSCs, transduced signaling pathway cells were cultured under conditions that allow Pax5−/− cells to develop to CD4/CD8 double Mannose-binding protein-associated serine protease negative, and to CD4+CD8+ T-lineage cells in vitro [23]. The different transduced pre-B-I-cell lines failed to develop to T-lineage cells (Supporting Information Fig. 4). In addition, none of the miRNAs downregulated the expression of CD19 (Supporting Information Fig. 2B). We conclude that overexpression of these miRNAs did not induce dedifferentiation of pre-B-I cells to the earlier, CD19−flt3+ multipotent CLP-like pro-/pre-B cell stage of B cell differentiation. To test the in vivo differentiation and migration potential of the rtTA/tetO-miRNA-double-transduced pre-B-I cells we established a series of pre-B-I-cell lines from 18 day-old CD45.1+C57BL/6J fetal liver. These CD45.1+ pre-B-cell lines can be detected in the CD45.2+ host also by their GFP expression in the presence of doxycycline, when they express mature miRNA. It also allows the capacity of these cell lines to mature in the host to CD45.1+CD19+sIgM+ B cells to be measured, as long as they still express doxycycline-induced miRNA/GFP, or after doxycycline removal when they no longer express miRNA/GFP.

vulnificus components with pattern recognition receptors (PRRs) (Espat et al., 1996; Powell et al., 1997, 2003; Shin et al., 2002; Lu et al., 2009). Recent studies showed that recombinant-produced V. vulnificus lipoprotein (Ilpa) and flagellar filament protein (FlaB) are recognized by Toll-like receptor 2 (TLR2) and TLR5, respectively (Lee et al., 2006; Goo et al., 2007). TLRs are a family of PRRs that are among the first line of host defense (Takeda & Akira, 2005; Gerold et al., 2007). Upon recognition of agonists, TLRs associate with central adapter

molecules such as myeloid differentiation factor 88 (MyD88). This interaction initiates a signaling cascade that results in production of TNFα and other proinflammatory cytokines. Although ABT-199 molecular weight TLR signaling is usually essential for activating an effective host immune response, it also plays a lead role in induction of the systemic inflammatory response that causes septic shock (Leaver et al., 2007). Thus, TLRs have attracted attention as Selleck Y27632 targets for treatment of sepsis. However, blockade of harmful TLR signaling requires knowledge of the TLR repertoire activated by a pathogen and the effect of TLR signaling on the host response and the outcome of infection (Gao et al., 2008). In addition to TLR2 and TLR5 agonists, V. vulnificus synthesizes lipopolysaccharide, which elicits a proinflammatory

cytokine response (e.g. TNFα secretion and cytokine mRNA expression) from human peripheral blood monocytes (Powell et al., 1997). Many Gram-negative bacteria activate TLR signaling due to recognition of their lipopolysaccharide via TLR4 (Takeda & Akira, 2005; Gerold et al., 2007). However, there was no information concerning whether V. vulnificus activates TLR4.

The goal of this study was to investigate the role of TLR4 in the host response to V. vulnificus using mice that are genetically deficient for this receptor. Wild-type (WT) male C57BL/6 mice were purchased from the Jackson Laboratory (Bar Harbor, ME). Homozygous TLR4 knockout (KO) (Hoshino et al., 1999) and MyD88 KO (Adachi Inositol monophosphatase 1 et al., 1998) mice that had been backcrossed for eight generations to WT mice were obtained via S. Akira (Osaka, Japan). Homozygous TNFα KO mice generated on a C57BL/6 background were obtained via L. Old (New York, NY). All mice were housed under specific pathogen-free conditions. MyD88 KO mice were reared without antibiotics and received sterile water and food. Animal procedures were approved by the University of North Carolina at Chapel Hill (UNC-CH) Institutional Animal Care and Use Committee. Vibrio vulnificus type strain ATCC 27562, a clinical (blood) isolate, was purchased from Remel (Lake Charles, LA) and grown in Bacto heart infusion (HI) broth (Becton Dickinson and Co., Sparks, MD) or on HI agar. Stocks were prepared by addition of glycerol (10% final concentration) to broth cultures and stored at −70 °C. Inactivated V.

Earlier reports showed that mfVSG triggers macrophage activation through a MyD88-dependent signaling cascade 38, 39. Heating VSG antigens for 15 min at 95°C did not abrogate the DC maturation activity (data not shown), indicating that the glycosyl-inositol-phosphate Panobinostat in vitro (GIP) moieties of the GPI anchor are the DC-activating factors as suggested previously for macrophages 38. In analogy with other parasitic protozoa such as Leishmania major, Plasmodium falciparum, and T. cruzi, GPI anchors of T. brucei are believed to form the most prominent inflammation and disease-inducing component 48. Indeed, recent reports

showed that the GPI anchors of P. falciparum mainly trigger MyD88-dependent TLR2 and to a lesser extent TLR4 signaling in macrophages 49. DCs sense different pathogens and respond by upregulating MHC and costimulatory molecules as

well as cytokine production to mount an appropriate T-cell response. However, it appears that DCs release substantial amounts of cytokines only upon strong TLR activation 23, 27, 29, 50. We found that mfVSG and Mitat1.5 sVSG act on DCs through MyD88 to mediate maturation but result in a TNF-like inflammation-induced partial maturation profile, leading to Th2-cell polarization. Although we also detected some IL-9-producing T cells in vitro, this was Kinase Inhibitor Library nmr not observed after injection. Since there is ongoing debate whether IL-9 is part of Th2 cells or belonging to an own Th9 subset 51, we did not further address IL-9 in our Th2-cell studies. Inflammatory mediators can activate DCs also in vivo, which are similarly unable 3-oxoacyl-(acyl-carrier-protein) reductase to produce IL-6 or IL-12p40 27, 50, 52. Sporri and Reis e Sousa have shown that DCs activated by inflammatory mediators in vivo induced Th cells but these were unable to support immunoglobulin isotype switching 27. Similarly, in this study all partially matured DCs types were unable to alter IgG1 and IgE levels in the asthma model. Recent reports also suggest that IL-6 by triggering IL-21 secretion in T cells drives the differentiation of Th cells that acquire the ability to provide B-cell help for isotype switching

53. Here, DCs matured with MiTat1.5 sVSG showed substantial production of IL-6, but DC treatment did not modify the isotype switch compared with other maturation stimuli in the allergic asthma model. However, it remains to be determined whether DCs conditioned by MiTat1.5 sVSG can induce B-lymphocyte helper T cells in the absence of any additional adjuvant activity. The capacity to provide efficient B-cell help might further delineate distinct functions of the Th2-cell subsets induced by inflammatory mediators or TLR agonists as identified in this study. In our study, the nonpathogen-derived inflammatory stimulus TNF and type 2 pathogen-derived antigens show remarkable similarities for the maturation of BM-derived DCs, i.e. the in vitro counterpart of the so-called TNF/iNOS-producing DCs (Tip-DCs) 54.

Although there was no statistical difference in reported history of bronchial asthma between the two groups in this study, several investigators have suggested that a history of bronchial asthma is a significant risk factor for pneumonia associated with pandemic A/H1N1/2009 influenza virus infection (13, 2, 15). Thus, the present data, together with previously reported findings, suggest that allergic responses might have important roles in the pathogenesis of such pneumonia. Pneumonia associated with pandemic A/H1N1/2009 influenza virus infection has attracted considerable attention (1), many studies to elucidate its pathogenesis having been carried out (5, 6). However,

no studies have sought to elucidate the mechanism of leukocytosis, another remarkable finding

that was not seen in previous seasonal influenza virus infections. Therefore, in this study an attempt Small molecule library order was made to Y-27632 ic50 identify the differences between pneumonia patients with and without leukocytosis. To our knowledge, this is the first study to elucidate an association between leukocytosis in patients with pneumonia and the host immune response. An increase in proinflammatory and/or inflammatory cytokines has been demonstrated in critical clinical conditions, including severe pneumonia (12, 9, 4); opposite findings have also been demonstrated by several investigators (8, 10, 7, 3). It has been suggested that not only innate immune responses, but also acquired immune responses, are impaired in critically ill patients (8, 10). Giamarellos-Bourboulis et al. (10) demonstrated that significantly fewer CD4 positive T cells and B cells are present in critically ill patients. oxyclozanide In the present

study, both Th1 and Th2 types of cytokines were down-regulated in pneumonia patients with leukocytosis. These findings suggest that if patients with pneumonia do not receive early treatments such as antiviral drugs and steroids, those with leukocytosis might manifest a more severe clinical course than those without leukocytosis. Such immunological impairment can be associated with exacerbation due to secondary bacterial infection (10); however, no statistical differences were observed in detection rates of bacteria in throat swabs obtained from pneumonia patients with and without leukocytosis. Unfortunately, because none of the patients expelled sufficient sputum or needed endotracheal intubation, no specimens from the lower respiratory tract were obtained for bacterial cultures in the present study. Therefore, any association between leukocytosis and secondary bacterial infection of the lower respiratory tract could not be precisely analyzed. Contrary to expectations, the concentration of IL-8, which is strong neutrophil chemotactant, was significantly decreased in pneumonia patients with neutrophilic leukocytosis.