In DC-based immunotherapy, it is occasionally difficult to obtain

In DC-based immunotherapy, it is occasionally difficult to obtain a sufficient number of quality-guaranteed DC for some patient groups, such as: (1) paediatric cancer patients, who are too

small to receive leukapheresis for DC preparation [20], (2) cancer patients with pancytopaenia owing to cachexia or basal disease-related factors such as liver cirrhosis or (3) patients with haematological malignancy, in whom peripheral blood may be contaminated with a large number of viable malignant cells. In such patients, allogeneic DC may be an alternative source. It has been suggested that the host alloresponse to the injected DC may actually facilitate the antitumour response AZD1208 chemical structure and that their alloantigens may work as helper antigens [21]. However, this theory is controversial [22, 23]. Moreover, Daporinad some preclinical studies using murine s.c. tumour models have shown that s.c. immunization using fully allogeneic DC failed to induce antitumour effects [14, 24]; thus, the use of allogeneic DC in DC-based immunotherapy may be limited. When allogeneic DC are used for cancer immunotherapy, three important factors must be considered. First,

the major histocompatibility complex (MHC) incompatibility of the DC used may be the most important factor for priming the MHC-restricted TAA-specific CD8+ T cells [25, 26] because during T-cell development, the host T cells acquire MHC restriction because of positive selection [27] by somatic cells (cortical thymic epithelial cells (cTECs), which are the crucial APC for expressing the MHC), rather than

haematopoietic cells [27]. Second, the survival of injected allogeneic DC may be shortened by T-cell-mediated rejection, and this may have an effect on the resulting antitumour response because DC survival is an important factor in priming antigen-specific T-cell responses. Third, it is not known whether host-derived pAPC can function in an antitumour capacity in DC-based immunotherapy, especially via the i.t. injection route. Until Loperamide now, no experimental model has been developed that assesses these factors individually, and it is unclear which of the factors, and to what degree, will affect the antitumour responses of allogeneic DC. It is also unclear which injection route is most preferable when using allogeneic DC. Here, we aimed at evaluating the availability of allogeneic DC for DC-based immunotherapy and to elucidate the mechanism for the antitumour effect, focusing on the three important factors related to allogeneic DC. We demonstrate that s.c. immunization using semi- or fully allogeneic DC pulsed with tumour lysate has a limited antitumour effect and does not induce a significant number of IFN-γ-producing tumour-specific CD8+ T cells. When semi-allogeneic DC were injected via an i.t. injection route, we observed the induction of an efficient antitumour response and a significant tumour-specific CD8+ T-cell response.

Following incubation with 50% chamber fluid, the CD11b activation

Following incubation with 50% chamber fluid, the CD11b activation epitope was significantly induced compared with cells incubated with the corresponding serum. Furthermore, the expression induced by chamber fluid corresponded to the expression induced by 100 ng/ml recombinant IL-8. The result is in line with previous findings indicating an

increased expression of CBRM1/5 after 10 min of incubation with relatively strong activators such as phorbol 12-myristate 13-acetate (PMA) and N-formylmethionyl leucyl phenylalanine (fMLP) [27], as well as weaker activators such as IL-8, C3a or platelet-activating factor (PAF) [28]. Interestingly, in our model, which is based on mediators released during a physiological response, IL-8 was the sole mediator correlating to CD11b activation. To further examine the correlation between IL-8 and CD11b activation, the expression of CD11b activation epitope was assessed following in vitro incubation with recombinant IL-8 corresponding to the concentration in serum and chamber fluid. The expression of the activation epitope was concentration dependent and increased gradually at levels corresponding to chamber fluid. Interestingly, in a former publication, a single dose of 10 ng/ml IL-8 induced

an almost identical expression of CBRM1/5 as in the Selleck Atezolizumab present article using the same concentration [28]. In this article, we demonstrate for the first time a concentration-dependent induction of the CD11b activation epitope by use of both endogenous and recombinant IL-8. Recombinant IL-8 required 10 times

higher the concentration of IL-8 in chamber fluid to induce a similar activation of CD11b. This could be explained by an increased biological activity of IL-8 in vivo, which has been demonstrated following gelatinase-mediated truncations [29] or by the combined action of other inflammatory mediators, not by themselves correlating to the CBRM1/5 expression. In summary, the concentration of IL-8 was a major determinant for neutrophil transmigration both in vivo and in vitro. One Arachidonate 15-lipoxygenase possible mechanism could be through regulation of the activation epitope on CD11b, and the present data on an IL-8 dose-dependent activation of CD11b support this view. Endogenous IL-8, compared with recombinant, mounted an enhanced response, probably reflecting an increased potency of in vivo IL-8. We, therefore, suggest IL-8 to be a major determinant for neutrophil CD11b activation and extravasation. The authors would like to thank Anette Bygden-Nylander for assistance with the skin blister method. The study was supported by unrestricted grants from Karolinska Institute and Hesselman Foundation. JMP, JL and SHJ wrote the paper; JMP conceived, designed and performed the experiments; JMP and JL analysed the data; and SHJ contributed to reagents.

Then, the T I can vary from 0 (normal) to 45 (most abnormal) T

Then, the T.I. can vary from 0 (normal) to 45 (most abnormal). T.I. < 10 is considered normal.[9, 10] Surgical approach included complete excision of lymphocele with its capsule and microsurgical lymphatic-venous anastomoses (LVA) between afferent lymphatics and venous branch of great saphenous vein (Fig. 1). LVA were performed using microsurgical technique at the operating microscope (25–30× magnification) with an arm-sleeve technique. A U-shaped stitch was used to pull the lymphatics inside the vein all together, anastomosing several lymphatics to the same vein, due to the higher caliber of the vein LGK-974 research buy (2–3 mm), compared to the lymphatic one (0.1–0.2 mm). The segment of the

vein used for anastomosis was usually collateral branch of the main vein with a competent valvular system, so that there was no blood reflux into the anastomosis, thus preventing the closure of anastomosis with time. Six to eight stitches were used to fix adventitial lymphatic tissues to the venous cut-end (Fig. 2).[11] Patent Blue dye injection was used to identify lower limb lymphatics intraoperatively. The surgeon could find a technical difficulty to find out a proper venous segment for microanastomosis, if great saphenous vein had been previously ligated during nodal dissection. In this case, a preoperative venous ultrasound-guided

INK 128 mapping was indispensable to look for a sound venous branch to use for lymphatic-venous bypass. It was, furthermore, important to use a competent vein in order not to have any blood reflux into the shunt, thus avoiding its closure with time. In case there were no superficial veins, deep collateral branch of femoral vein could be prepared for anastomosis. Two suction drains, one round and one flat, were placed and removed averagely after 3–5 days with leg

bandaging in case of associated lymphedema. Drains were usually removed separately (before round drain) when 24-hour output was less than 30 ml. Patients were followed up clinically and by ultrasounds as Obatoclax Mesylate (GX15-070) concerns lymphocele and by volumetry for lower limb lymphedema (at 3 months and 1-year postoperative). Postoperative LS was performed after 1 year from operation. In nine patients with GL without LL, lymphocele completely disappeared and no appearance of lower limb postoperative lymphedema occurred. The other seven patients with associated secondary lymphedema had complete disappearance of lymphocele and a remarkable reduction of leg volume (averagely 80% excess volume decrease). Four of them completely recovered without the need of any compression garment, after the first year postoperative. After 3 months, either there were no clinical or instrumental signs of lymphocele and a significant reduction of limb excess volume. After 1 year, there was an almost complete decrease of this volume (Table 1). The preoperative volume difference between both legs was 2123 cc averagely. After 1 year, the mean volume difference was 265 cc (157–447).

Ultrapure LPS was purchased from Invitrogen and used at a concent

Ultrapure LPS was purchased from Invitrogen and used at a concentration of 10 μg/mL. selleck compound ATP was from Sigma and used at a final concentration of 3 mM. For immunoblotting, cells were washed twice with sterile PBS and lyzed in buffer (150 mM NaCl, 10 mM Tris, pH

7.4, 5 mM EDTA, 1 mM EGTA, 0.1% Nonidet P-40) which was supplemented with a Roche protease inhibitor cocktail tablet. After clarification and denaturation with SDS buffer, samples were boiled for 5 min. Separation of the proteins was done by using SDS-PAGE and thereafter transferred into a nitrocellulose membrane. These membranes were coated with primary antibodies and active caspase-1 was detected using secondary anti-rabbit antibody conjugated to horseradish peroxidase followed by enhanced chemiluminiscence. Sotrastaurin manufacturer Peritoneal macrophages were isolated by injecting

5 mL of ice-cold sterile PBS (pH 7.4) in the peritoneal cavity. After centrifugation and washing, cells were resuspended in RPMI 1640 containing 1 mM pyruvate, 2 mM L-glutamine and 100 μg/mL gentamycin (culture medium). Cells were counted using a Z1 Coulter Particle Counter (Beckman Coulter, Woerden, The Netherlands) and adjusted to 1×106 cells/mL. Cells were cultured in 96-well round-bottom microtiter plates (Costar, Corning, The Netherlands) at 1×105 cells/well, at a final volume of 200 μL. The cells were stimulated with RPMI or two different heat-killed Borrelia strains. After 24 h of incubation at 37°C in air and 5% CO2, the plates were centrifuged at 500×g for 10 min, and the supernatant was collected and stored at −80°C until cytokine assays were performed. Investigating the role of IL-33 was not performed by incubation of peritoneal macrophages that were stimulated with RPMI or Borrelia in the presence or absence of 10 μg/mL anti-mouse

IL-33 antibody (R&D Systems). After 48 h of incubation, IL-4 and IL-5 levels were measured using ELISA kits (eBioscience, San Diego, CA, USA). ELISA were performed according to the manufacturer’s instructions. Spleen cells were isolated by gently squeezing spleens in a sterile 200 μm filter chamber. After washing with sterile PBS and centrifugation at 4°C (1200 rpm, 5 min), cells were resuspended in 4 mL RPMI 1640 in presence of 20% FCS. Cells were counted and concentrations were adjusted to 1×107 cells/mL. Cells were cultured in 24-well plates (Greiner, Alphen a/d Rijn, The Netherlands) at 5×106 cells/well, at a final volume of 1000 μL. After 5 days of incubation, supernatant was collected and stored at −80°C until cytokine assays were performed. Concentrations of mouse IL-1β were determined by specific radioimmunoassay (RIA; detection limit is 20 pg/mL) as described by Netea et al52. Mouse IL-6, IL-17 and IFN-γ, concentrations were measured by a commercial ELISA kit (Biosource, Camarillo, CA; detection limits 16 pg/mL), according to the instructions of the manufacturer.

tuberculosis DNA in all the pleural TB samples, thus demonstratin

tuberculosis DNA in all the pleural TB samples, thus demonstrating that the DNA extraction method could affect the performance of real-time PCR. Because of extremely high sensitivity of PCR, the carry-over contamination of amplicon, previous infection

or asymptomatic EPTB infection at another site could result into false positivity (Honore-Bouakline et al., 2003; Chakravorty et al., 2005; Sun et al., 2011). The false positivity of PCR reports in the absence check details of clinical findings poses serious challenges these days in diagnosing EPTB cases (Thangappah et al., 2011). The lack of proper gold standard remains the major hindrance for evaluating new diagnostics in EPTB-infected individuals (Sun et al., 2011; Vadwai et al., 2011). The true accuracy of PCR tests may actually be different than the reported one when using an imperfect gold/reference standard (Abbara & Davidson, 2011; Tortoli et al., 2012). Culture (on solid and liquid media) is the most widely used gold standard for validating PCR results in diagnosing EPTB specimens although it is suboptimal gold standard with varying sensitivities and leads to inaccurate PCR

results (Negi et al., 2005a; Hillemann et al., 2011; Sun et al., 2011). MLN0128 cell line The other gold/reference standards include BACTEC culture, histopathology and response to anti-tubercular therapy (ATT) and also the combination of these methods (Negi et al., 2005b; Kulkarni et al., 2006; Abdalla et al., 2009; Noussair et al., 2009). Chakravorty et al. (2005) as well as Vadwai et al. (2011) have used smear, culture, histology/cytology, clinical findings and response to ATT, all together as the gold/reference standard for validating

their PCR results in diagnosing EPTB specimens. There are several potential commercial kits Farnesyltransferase devised to diagnose TB such as Amplicor M. tuberculosis test (Roche Molecular Systems Branchburg, NJ), Gen-probe Amplified M. tuberculosis Direct Test (AMTD; Gen-Probe, CA), COBAS TaqMan M. tuberculosis (Roche Molecular Systems Branchburg) and LightCycler (Roche Molecular Diagnostics, Mannheim, Germany; Ritis et al., 2005; Causse et al., 2011; Parrish & Carroll, 2011) Among these, Amplicor M. tuberculosis test and AMTD based on 16S rRNA gene have been approved by the US Food and Drug Administration (FDA) for the diagnosis of PTB only (Brodie & Schluger, 2009), and none of these commercial tests have been approved by FDA for the diagnosis of EPTB (Parrish & Carroll, 2011). However, the utility of these commercial tests has been extensively explored in the diagnosis of EPTB (Honore-Bouakline et al., 2003; Causse et al., 2011). Moreover, the meta-analyses of PCR tests have suggested that the commercial tests yielded high specificities but variable sensitivities for the diagnosis of EPTB, while heterogeneous sensitivities and specificities were observed with in-house PCR tests (Pai et al., 2004; Daley et al., 2007).

Conflict of interest: A -L I , M J O , M B , R B and I A

Conflict of interest: A.-L. I., M. J. O., M. B., R. B. and I. A. have potential conflict of interests that include stock options, salaries or consulting fees

from OMT. G. J. C. has potential conflict of interest that includes salary fees from Sangamo BioSciences. Detailed facts of importance to specialist readers are published Small molecule library in vitro as ”Supporting Information”. Such documents are peer-reviewed, but not copy-edited or typeset. They are made available as submitted by the authors. “
“Citation McDonald EA, Wolfe MW. The pro-inflammatory role of adiponectin at the maternal–fetal interface. Am J Reprod Immunol 2011; 66: 128–136 Problem  A successful pregnancy is contingent on maternal tolerance of the immunologically foreign fetus. Prevalent diseases such as preeclampsia arise in part due to an inappropriate immune response by the placenta. A number of molecules have been proposed to temper cellular response to pro-inflammatory mediators, including CD24 and Siglec10. Methods  Cytotrophoblast cells from

healthy term placentas were treated with adiponectin in vitro and analyzed with qPCR and ELISA-based assays. Immunohistochemistry was performed on term villous sections and cultured trophoblasts. Results  Treatment with adiponectin increased expression of IL-1β and IL-8. Term villi express CD24 in cytotrophoblasts and the syncytiotrophoblast, and Siglec10 by the syncytiotrophoblast. Treatment of trophoblast cells with adiponectin increased Siglec10 expression. Conclusion  These data describe a role for adiponectin in enhancing pro-inflammatory signals in in vitro click here syncytialized trophoblasts. Additionally, this represents the first time the CD24/Siglec10

pathway has been implicated in a trophoblast response to a pro-inflammatory mediator. “
“Myeloid-derived suppressor cells (MDSC) are a heterogeneous population of cells that negatively regulate the immune response during tumour progression, inflammation PtdIns(3,4)P2 and infection. Only limited data are available on human MDSC because of the lack of specific markers. We have identified members of the S100 protein family – S100A8, S100A9 and S100A12 – specifically expressed in CD14+ HLA-DR−/low MDSC. S100A9 staining in combination with anti-CD14 could be used to identify MDSC in whole blood from patients with colon cancer. An increase in the population of CD14+ S100A9high MDSC was observed in the peripheral blood from colon cancer patients in comparison with healthy controls. Finally, nitric oxide synthase expression, a hallmark of MDSC, was induced in CD14+ S100A9high upon lipopolysaccharide/interferon-γ stimulation. We propose S100 proteins as useful markers for the analysis and further characterization of human MDSC. Myeloid-derived suppressor cells (MDSC) have been characterized as a population of cells that can negatively regulate T-cell function.

Our current data support previous clinical studies in suggesting

Our current data support previous clinical studies in suggesting a role of E. coli in human PBC. Hopf et al. [63] reported an association between PBC and the presence of rough-form mutants of E. coli in the patients’ fecal BGB324 mouse samples. In addition, Butler et al. reported reactivity to PDC-E2 in 52% of sera from patients with chronic UTIs [7, 64]. In the first controlled epidemiological analysis for the relationship between

E. coli and PBC, Parikh-Patel et al. showed a positive association between PBC and recurrent UTI [65]. A recent epidemiological study on 1032 PBC patients followed-up in 20 tertiary referral centres in the United States and 1041 demographically matched controls confirms earlier studies indicating a connection buy Trichostatin A of UTI with PBC [66]. The discovery of E. coli infection-triggered autoimmunity and liver pathology warrant further consideration in the elucidation of aetiological mechanisms of autoimmune syndromes and may suggest new and simpler ways to diagnose and treat these debilitating diseases. Our data also highlight the importance of microbial

infections in autoimmunity either as primary or co-existing secondary inciting events. This work was supported in part by National Institutes of Health grants DK39588 (M. E. G.) DK067003 (M. E. G.), AI71922 (M. K.) and AI083029 (J. L. V.) The authors have no financial conflicts of interest. “
“Bidirectional signals via Eph receptors/ephrins have been recognized as major forms of contact-dependent cell communications such as cell attraction and repulsion. T cells express EphBs, and their ligands, the ephrin-Bs, have been

known as costimulatory molecules for T-cell proliferation. Recently, another remarkable feature of ephrin-As has emerged in the form of a concentration-dependent transition from promotion to inhibition in axon growth. Here we examined whether this modification plays a role in ephrin-B costimulation in murine primary T cells. Low doses of ephrin-B1 and ephrin-B2 costimulated T-cell proliferation induced by anti-CD3, but PLEKHB2 high concentrations strongly inhibited it. In contrast, ephrin-B3 showed a steadily increasing stimulatory effect. This modulation was virtually preserved in T cells from mice simultaneously lacking four genes, EphB1, EphB2, EphB3, and EphB6. High concentrations of ephrin-B1/B2, but not ephrin-B3, inhibited the anti-CD3-induced phosphorylation of Lck and its downstream signals such as Erk and Akt. Additionally, high doses of any ephrin-Bs could phosphorylate EphB4. However, only ephrin-B1/B2 but not ephrin-B3 recruited SHP1, a phosphatase to suppress the phosphorylation of Lck. These data suggest that EphB4 signaling could engage in negative feedback to TCR signals. T-cell activation may be finely adjusted by the combination and concentration of ephrin-Bs expressed in the immunological microenvironment.

70F, CBS 700 71 and CBS 700 68 were used as negative controls in

70F, CBS 700.71 and CBS 700.68 were used as negative controls in tests with non-orthologous taxa. The concordance of RCA results and identification by multilocus

sequencing was 100%. Products of the RCA reaction were visualised by electrophoresis on 1% agarose gels. With exonucleolysis, positive responses showed ladder-like patterns after RCA, whereas with negative results the background remained clean. When exonucleolysis was omitted, a single, weak or strong band was visible on negative lanes, representing a non-specific band that did not interfere with the RCA reaction. Sensitivity testing showed that RCA yields positive results in wide ranges of amplicon concentrations down to 3.2 × 105 copies of amplicon (Fig. 2). rDNA ITS is a sufficient barcoding marker in Mucorales, because interspecific distances tend to be relatively large compared to e.g. more recently evolved BAY 80-6946 mouse ascomycetes.[11] In addition, the majority of clinically relevant taxa are located in distantly related clades. The main exception is with R. arrhizus var. arrhizus and R. arrhizus var. delemar which differ in 3 bp in ITS, show occasional interbreeding and have been considered to be varieties of a single species rather than separate species.[21] RCA

reportedly has a specific detection limit of single nucleotide [17] and thus should be able to differentiate between these groups. Our results showed that this was indeed the case (Fig. 1). The purpose of the present BAY 73-4506 study was to establish a screening method based on the RCA enabling rapid detection, with specificity down to few nucleotide differences and assess the limits Fluorouracil mouse of this molecular method. We found specificity of 100%, and high sensitivity. RCA is a robust

and simple isothermal DNA amplification technique allowing rapid detection of specific nucleic-acid sequences with no need of sequencing and can be performed within 2 h, and is therefore applicable for rapid and economic screening purposes.[16, 17] Specially designed padlock probes hybridise to a target DNA or RNA and permit the detection of single nucleotide mismatch and prevent non-specific amplification, a common risk factor in conventional PCR. To date, RCA has been used for different fungi, such as Cryptococcus, Trichophyton, Candida, Aspergillus, Talaromyces marneffei, Scedosporium and black yeast.[17] In Mucorales no cross reactivity was observed within tested strains. RCA is particularly suited for high throughput applications. Wide ranges of amplicon concentrations yield positive results. The amplification product can be visualised by agarose gel electrophoresis, but also in gel-free systems using fluorescence staining of amplified product by SYBR Green in combination with UV-transillumination, and this can add to the speed and ease of the test. RCA is practical for detection of low copy number DNA. The method can be performed with a variety of DNA polymerases compared to direct PCR.

Moreover, other proteases have been indentified in chromaffines g

Moreover, other proteases have been indentified in chromaffines granules, including the neuroendocrine-specific carboxypeptidase E/H and the Lys/Arg-amino peptidases [55]. These data suggest that Cgs might serve as a prohormone for a shorter fragment having regulatory properties [56]. In the rat and human GI tract, the presence of cell- and tissue-specific processing of CgA has been shown [57–59], but very little is known about the functional role of Cgs in GI pathophysiology. Herein we will discuss the several

data related to the role of Cgs in immune function and inflammation. Due to the similarity click here of sequence with the cell-penetrating peptides family [60], Cgs-derived peptides such as chromofungin (CHR, bCgA 47–66) and vasostatin-I (VS-I, bCgA 1–76) are able to penetrate into

polymorphonuclear neutrophils (PMNs), inducing an extracellular calcium entry by a CaM-regulated iPLA2 pathway. This study highlights the role of CgA-derived peptides in active communication between the neuroendocrine and immune systems [61]. Keeping within the endocrine–immune context, not only can the PMN be regulated by Cgs-derived peptides, but catestatin (CAT; bCgA 344–364) stimulates chemotaxis of human peripheral blood monocytes dose-dependently, exhibiting its maximal effect at a concentration of 1 nM comparable to the established chemoattractant-formulated peptide Met-Leu-Phe (fMLP) [62], suggesting a role of this

peptide as an inflammatory mediator. In the same inflammatory context, secretoneurin reduces IL-16 release from eosinophils; this effect is in addition to that observed Transferase inhibitor with granulocyte–macrophage colony-stimulating factors or IL-5. Results suggest that distinct neuropeptides are able to reduce the number of lymphocytes at inflammatory Parvulin sites during existing eosinophilia by inhibiting the relaease of IL-16, thus attenuating the proinflammatory action of lymphocytes and monocytes. It has also been demonstrated that secretoneurin stimulates migration and cytokine release from human peripheral blood NK cells, implying that activation of this cell type by secretoneurin could affect the accumulation of these cells at loci of neurogenic inflammation [63]. A role for the neuropeptide on neutrophil adhesion and transmigration through a lung fibroblast barrier in vitro has also been shown [64]. Cgs-derived peptides can not only regulate the immune system during inflammation, but can also modulate the endothelial permeability during the inflammatory process, but the actual role of Cgs and derived peptide are not really clear. CgA prevents the vascular leakage induced by tumour necrosis factor (TNF)-α in a mouse model [65]. Studies of the mechanism of action show that CgA and its NH(2)-terminal fragments inhibit TNF-α-induced vascular permeability by preventing endothelial cytoskeleton rearrangements.

Bronchiolitis obliterans syndrome (BOS)

Bronchiolitis obliterans syndrome (BOS) selleck inhibitor is the single most important factor that limits long-term survival following lung transplantation [1]. We have shown that BOS is associated with lack of immunosuppression of T cell T helper

type 1 (Th1) cell proinflammatory cytokines and increased T cell granzyme B by peripheral blood T cells [2, 3]. Current immunosuppressive therapies target Th1 proinflammatory cells [4]; however, they are relatively non-specific and, as we have shown, ineffective at reducing proinflammatory mediators produced by major lymphocyte subsets in the peripheral blood of lung transplant patients undergoing and preceding diagnosis of BOS [2, 3, 5]. Hence, there is an urgent need for new targeted therapy to prevent BOS. Following Selleck Z-VAD-FMK adhesion and antigen presentation, T cells require co-stimulatory

signals from professional antigen-presenting cells through surface receptors for T cell proliferation and cytokine production [6]. Repeated antigen-driven proliferation down-regulates T cell CD28 and expansion of late-differentiated, antigen-specific, oligoclonal T cells [7]. Recently, we have shown CD28 down-regulation on CD8+ T cells, the main effector T cells in patients with chronic obstructive pulmonary disease (COPD), another important

chronic pulmonary disease [8]. We hypothesized that down-regulation of CD28 (to a ‘CD28null’ phenotype) and corresponding up-regulation of alternate co-stimulatory molecules Tyrosine-protein kinase BLK may play an important role in the generation of steroid-resistant cytotoxic molecules such as granzymes/perforin and proinflammatory cytokine production by T cells in BOS. Down-regulation of CD28 expression following persistent antigenic stimulation has also been shown to be associated with up-regulation of CD57 expression, a terminally sulphated carbohydrate determinant found on subsets of natural killer (NK) cells and NK T-like cells associated with ageing [9]. Interestingly, we have shown recently that there are increased peripheral blood CD56+CD3+ NK T-like cells in blood from stable lung transplant patients and that these cells exhibit increased production of proinflammatory cytokines interferon (IFN)-γ and tumour necrosis factor (TNF)-α and expression of cytotoxic molecules, perforin and granzymes [10]. We hypothesized that dysregulated expression of T cell co-stimulatory molecules may be associated with steroid resistance and BOS, and identify potential new therapeutic targets that are needed urgently to improve the morbidity and mortality rates following lung transplantation.