Specific central memory CD4+ T cells, defined by CCR7 expression, were virtually undetectable 2 months after vaccination. A change to central

memory phenotype may occur at a later post-vaccination time and this will be explored in future studies. In MVA85A-vaccinated subjects from the UK, Ag85A-specific T-cell proliferation peaked 6 months post-vaccination 32. Interestingly, in mice MVA-induced CD8+ T cells mostly convert to a central memory phenotype within weeks of immunization 44, suggesting that the rate of conversion to central memory cells may differ between species. In other human studies, we have also consistently observed predominant effector phenotypes of human mycobacteria-specific CD4+ T cells in infants 33, 45 and adults 20. Mycobacteria-specific CD4+ T cells from children with latent M.tb infection or active TB 16, and chronically HIV-infected adults with latent M.tb infection 46, selleckchem also display this phenotype. Long-lived central memory cells prevail when Ag is cleared after vaccination, e.g. after tetanus toxoid vaccination 42, whereas chronic CMV, EBV or HIV infection is associated with predominance of effector memory cells 47. One might hypothesize

that chronic exposure to mycobacterial Ag is responsible for our observed phenotype. Adolescents with latent M.tb infection, one potential source of such chronic exposure, were not enrolled DCLK1 into our study. Additional studies are required to dissect this further. No serious adverse events were recorded, and mild local reactions at the vaccination site were FK506 manufacturer predominant. These reactions were commonly reported in the first week after vaccination, did not interfere with daily activities and did not persist. Systemic reactions were uncommon and included mild flu-like symptoms. Clinically, there were no major differences between the adolescents’ and the children’s experience in the trial with a slightly increased incidence of non-vaccine-related systemic events reflecting

this younger age group’s increased risk of transient viral illnesses. This complements the good safety profile of MVA85A found in healthy adults from the same region 25, the United Kingdom 36 and The Gambia 24, as well as other recombinant MVA being tested in clinical trials 40, 48. Together these small phase I/II trials demonstrate a very promising safety profile of MVA85A, which is now being assessed in larger groups of participants, in an infant, phase IIb safety and efficacy study. In conclusion, MVA85A was found to be safe and highly immunogenic in TB-naïve, HIV-uninfected adolescents and children. The vaccine induced durable, polyfunctional CD4+ T-cell responses with a CCR7− effector memory phenotype. These data support future studies to evaluate the efficacy of this vaccine to prevent TB.

Antioxidants, free radical scavengers or substances inhibiting I/R injury may reduce bladder damages caused by BOO or overdistention. As any organ in the body, the urinary bladder needs an adequate blood supply to obtain oxygen and nutrition to function normally. Ischemia with the accompanied https://www.selleckchem.com/products/ABT-263.html hypoxia would expect to impair bladder function. Cumulated evidences have demonstrated that ischemia, hypoxia and ischemia/reperfusion (I/R), with the accompanying generation of reactive oxygen/nitrogen species, are important etiologic factors in obstructive

bladder dysfunction.1,2 The present paper reviews and summarizes the effects of ischemia and hypoxia on the energy metabolism and contractile Selleck Everolimus function of the urinary bladder. I/R injury on the bladder and its role in chronic bladder outlet obstruction and acute overdistention are further reviewed. Chronic partial ischemia of the bladder has been shown to impair bladder function. Gill et al. have shown that bladder ischemia induced by ligation of the vesical artery impaired contractile responses of the detrusor strips.3 Lin el al. found that chronic ischemia of the bladder resulted in a decrease of bladder compliance with a reduction in the contractility of the whole bladder.4

Lit et al. further demonstrated that chronic ischemia deranged glucose metabolism of the detrusor with a reduction in glycogen content and an increase ROS1 in anaerobic metabolism, resulting in a much lower production of high-energy molecules.5 Using an atherosclerosis rabbit model, a recent study also demonstrated that chronic ischemia of the urinary bladder resulted in mitochondrial injury, fibrosis, microvasculature damage and neurodegeneration.6 Lin et al. have demonstrated that urinary bladder blood flow was reduced by outlet obstruction

and the reduction in blood flow was associated with decreased tissue level of high-energy phosphates, adenosine triphosphate (ATP) and creatine phosphate.7 They further showed that the BOO-induced blood flow reduction could be recovered gradually after relieving outlet obstruction and was in parallel with the recovery of energy producing-related mitochondrial enzyme activity and energy producing capability of the bladder.8,9 During bladder emptying, the increased intra-wall tension results in blood vessel compression, decreased blood flow and tissue hypoxia. This occurs in normal bladders; nevertheless, this phenomenon is significantly exaggerated in the obstructed hypertrophied bladder.2,10 Under conditions of increased oxidative stress, cellular and subcellular membranes become subject to attacks when the generation of free radicals outweighs the system’s ability to eliminate.

According to the size of clone, cloning rings are usually used to pick larger size clones. When the cloning ring is sealed firmly on a clone,

add trypsin/EDTA into rings as per normal trypsinization of cells. Trypsin Antiinfection Compound Library datasheet needs 5 min at 37°C. Then transfer cloning cells or discs into individual flasks or culture plate at 33°C. Leave discs in for at least 48 h. Keep culturing cells until they are confluent and then freeze cells, make sure there are plenty of stocks all the time (Fig. 4). Experimental procedures are performed on the clonally selected cells by growing cells at 40% confluence on cover slips in Petri dishes at 33°C followed by differentiation for 10–14 days at 37°C. Fix cells before staining with 2% paraformaldehyde solution adding 2% sucrose. Immunofluorescence staining for podocyte markers, protein extraction BVD-523 order from culture flasks or plates is performed after differentiation for 14 days at 37°C. We detect podocyte proteins, such as nephrin, podocin, CD2AP, and synaptopodin, and known molecules of the slit diaphragm

ZO-1, alpha-, beta-, and gamma-catenin and P-cadherin (Fig. 5). Incubators kept at 33°C and 37°C, 5% CO2, RPMI-1640 Sigma R-8758 Use of antibiotics (Pen/Strep) is optional for cell lines. Use standard tissue culture-treated flasks or plates. We do not use special coatings such as collagen routinely as we have concluded that they do not offer any further benefit to cell culture. We do not specially treat flasks or plates

ourselves. Let immortalized podocytes grow at 33°C to 100% confluence, then freeze 40% and split the rest 1:3. For subsequent passages, split cells 1:3 to 1:5 when at 80% confluence. Use low concentrations of trypsin/EDTA (Sigma T3924 or equivalent with trypsin 0.05%) and expose the cells for as short a time as possible. Ensure freezing of at least 30% of each passage for long-term storage (liquid nitrogen) and availability of low passage numbers for the future. Move cells from 33°C to 37°C when cells are 40–60% confluent. Change medium three times per week. Usually it takes 14 days for full differentiation. They proliferate abundantly at 33°C, and Carbohydrate after thermoswitching to 37°C, usually take 1–3 days before cell division fully ceases. The transgene is actually designed to inactivate fully at 39.5°C but we normally see complete quiescence at 37°C for most human podocytes (sometimes with mouse podocytes it is necessary to go up to 38.5°C or above for full differentiation). We would like to finish with a word about cell co-culture. We restate the view2 that the glomerular capillary wall should be seen as a tripartite structure in which the three components (podocytes, glomerular basement membrane and glomerular endothelial cells) are interdependent and each of crucial significance, such that a focus on any one component of that structure might be inappropriately simplistic.

Appropriate regulation of gut immunity thus depends upon a complex three-way interplay between host cells, commensals and pathogens, and can exert a major impact on systemic responses including allergy and autoimmunity. In the gastrointestinal (GI) tract, the immune GW-572016 clinical trial system is faced with the most demanding of all decision-making, with little room for error. It is imperative at all times to discriminate between, and respond correctly to, beneficial symbionts, harmless food antigens and potential pathogens [1]. There is increasing appreciation that regulatory T cells (Tregs)

play a prominent and essential role in maintaining appropriate responsiveness in the gut [2,3], actively enforcing homeostasis and preventing untoward immune responses occurring. While stimulated by specific antigens, of both self and non-self origin, Tregs can transcend antigen specificity, mediating bystander suppression in a manner likely to modify systemic immune status as suggested by the ‘hygiene hypothesis’. Recent studies have changed our perspective of commensal microbes from benign but inert passengers to active participants in both the postnatal development of mucosal immunity and in its long-term steady-state function. Germ-free mice show extensive deficiencies

in intestinal immune system development, with reduced lymphoid tissue and fewer Acalabrutinib in vivo lymphocytes [4]. The CD4+ T cell population is diminished, affecting T helper type 1 (Th1) cells disproportionately although, remarkably, Treg frequencies are maintained or increased in germ-free mice. These and other data have established that defined components of the gut flora can play a major role in intestinal homeostasis, including protection against gut injury and mediating oral tolerance against dietary

antigens. ADP ribosylation factor In mice which acquire a conventional microbiome, the immune system develops normally while maintaining a continuing dialogue with the commensal population. Here, one of the dominant roles of Tregs is to prevent exuberant responses against gut flora, with which the intestinal tract is in intimate contact. Nevertheless, how commensals communicate with cells to ensure immune homeostasis is still unclear. One critical factor in this interaction at the molecular level is the host Toll-like receptor (TLR) system, as demonstrated by spontaneous colitis in TLR-5-deficient mice [5]. Where colitis is induced experimentally (e.g. by dextran sulphate administration), the absence of TLR signalling then results in greatly aggravated pathology, again indicating that TLR-mediated recognition of commensal molecules contributes to dampening immune reactivity [6]. The requirement for TLR signalling in induction of oral tolerance to dietary antigens [7] also speaks to the bimodal participation of the TLR system in both stimulatory and regulatory arms of the immune response. Recent evidence suggests that TLR signalling can impact Treg homeostasis and that Tregs themselves express TLRs selectively.

For example, the cathelicidin-derived peptide, LL-37, can enhance IL-1β release from lipopolysaccharide-primed monocytes via a P2X7-dependent mechanism and can also induce the production of monocyte chemoattractant protein-1 (MCP-1) chemokine from these cells.[9] LL-37 is also reported to influence monocyte maturation, potentially resulting in cells with more pro-inflammatory characteristics.[10] To further assess the effects of

antimicrobial peptides on monocytic cells, we examined the induction of co-stimulatory molecules, CD80 and CD86, as well as an array of chemokines by hBD-3, LL-37 and a well-defined Toll-like receptor 1/2 (TLR1/2) agonist, PAM3CSK4. In addition, we asked if chemokine induction by hBD-3 might be diminished in cells selleck inhibitor from HIV+ donors because we have previously found evidence for decreased induction of CD80 in cells from HIV+ donors compared with cells from healthy controls.[11] Our results suggest that hBD-3 activation of monocytic cells could play an important role in orchestrating inflammatory microenvironments by inducing chemokine expression and this activity may be modified in HIV disease. Cells were obtained from healthy adult volunteers and HIV+ Alisertib ic50 donors with IRB-approved

protocols and informed consent. For chemokine production studies, the HIV+ donors consisted of three viraemic and six aviraemic subjects. Purified monocytes were prepared with EasySep monocyte isolation kits (STEMCELL Technologies) and achieved > 85% purity. Monocyte-derived macrophages were generated by incubating cells with 100 ng/ml macrophage colony-stimulating factor (M-CSF) for 7 days. Cells were incubated in complete medium consisting of RPMI 10% fetal calf serum plus l-glutamine. For studies of chemokine receptor expression, freshly isolated peripheral blood mononuclear cells (PBMC) were stained with anti-CD14 Peridinin chlorophyll protein (PerCP; Janus kinase (JAK) BD Biosciences, San Jose, CA), anti-CD16 allophycocyanin-chychrom

7 (APC-Cy7; Biolegend, San Diego, CA), anti-CCR5 APC (BD Pharmingen), anti-CCR2 PerCP Cy5.5 (Biolegend), anti-CXCR2 FITC (Biolegend) and anti-CCR4 phycoerythrin-Cy7 (BD Pharmingen, Franklin Lakes, NJ). Cells were incubated for 10 min at room temperature, washed in PBS/BSA buffer, fixed in 1% paraformaldehyde and analysed by flow cytometry. Subjects for these studies included 27 HIV+ donors and 18 healthy control donors. The HIV+ donors had a median CD4 cell count of 589 cells/μl and a median plasma HIV RNA of 33 copies/ml. All but three HIV+ donors were receiving anti-retroviral therapy at the time of the study and all but four of the HIV+ donors had a viral load below 500 copies/ml. The age of the HIV+ donors (median = 47 years) and HIV– donors (median = 38 years) was not significantly different.

AFLP was a useful tool for identification to species-level and for the ABT-888 supplier discrimination of inter- and intra-patient isolates. Scedosporium prolificans represents the most prevalent species in the respiratory tract of CF patients and immunocompromised patients in Northern-Spain, followed by Pseudallescheria boydii, P. apiosperma, and P. ellipsoidea. CF patients were exclusively colonised with either P. boydii or S. prolificans. Patients were colonised over years exclusively with isolates affiliated to one species, but some patients were colonised with multiple strains with different AFSP. The sum of those

co-colonising strains in one patient, may appear in vitro and in vivo as a multi-resistant S. prolificans isolate, as strains are morphologically identical and might therefore be regarded as only

one strain. A majority of Scedosporium strains (with exception of S. prolificans) were found susceptible for voriconazole and micafungin. Pseudallescheria/Scedosporium Selleck Z-IETD-FMK species are the second most frequently cultured filamentous fungi from the lungs of patients with cystic fibrosis (CF).1 Until 2005, only two clinically relevant species of Scedosporium were known: Scedosporium apiospermum (teleomorph: Pseudallescheria boydii) and S. prolificans. During the last 5 years, several sibling species have been introduced, 1–5 which has led to the subdivision of P. boydii into the following species: S. apiospermum (teleomorph P. apiosperma), S. aurantiacum, S. boydii (teleomorph: P. boydii), S. dehoogii, P. fusoidea, P. ellipsoidea, P. angusta, and P. minutispora. Pseudallescheria Tenoxicam and Scedosporium infections are difficult to treat because of their therapy-refractory nature.6,7 Several infections by multi-drug resistant strains of Scedosporium species have been reported.8–11 Among these, S. prolificans is the most frequently encountered pathogen.12 Delayed diagnosis of the causative agent and ineffective antifungal therapy may have a negative impact on mortality rates of

patients suffering from systemic Scedosporium infections.13,14 Since the segregation of these sibling species, no comprehensive studies on species-specific antifungal susceptibilities and clinical epidemiology have been published. The aim of this study was to provide antifungal susceptibility patterns of isolates identified according to the taxonomy proposed by Gilgado et al.2–5 Strains were identified using AFLP analysis. Moreover, this study provides qualitative molecular epidemiology data on Northern Spanish patients colonised or infected with Scedosporium strains. In total, 60 clinical isolates from 21 patients isolated at the University Hospital Miguel Servet, located in Zaragoza (Northern-East Spain) were included in this study. The University Hospital has an adherence of more than 500 000 persons.

Four of the nine mutations (9%) that were detected in embB306 indicating resistance to ethambutol were not detected by the DST method, giving the lowest rate of concordance (44.4%) of the PCR with the DST method. One of the greatest concerns of national tuberculosis control programs in several countries

is the emergence and spread of drug resistant and MDR-TB. The actual extent and type of drug-resistant tuberculosis in Jordan is unknown. To determine this, the present study characterized 100 M. tuberculosis strains by PCR that were identified as drug resistant in the reference laboratory for mycobacteria. This is the first investigation involving the molecular characterization of drug resistance of M.

tuberculosis clinical isolates from Jordan. It was initiated as a result of the growing demand for rapid molecular characterization click here of Mycobacterium ZD1839 datasheet strains isolated from patients whose clinical details and history suggested the presence of drug-resistant M. tuberculosis, i.e. previous tuberculosis, recent immigration from or travel to an area with a high prevalence of MDR-TB, failure to respond to therapy, or contact with a known MDR-TB patient (Watterson et al., 1998). In this study, 34 isolates resistant to one or more of the tested drugs were identified. This is comparable to what has been reported in the neighboring countries, with resistance to isoniazid and rifampicin being more common than resistance to ethambutol. The World Health Organization has estimated the prevalence of MDR-TB in several Mediterranean and neighboring from countries as follows: Bahrain 3.5%, Egypt 5%, Iran 7.1%, Iraq 5.6%, Israel 5.6%, Kuwait 2.4%, Lebanon 2.4%, Oman 1.8%, Qatar 1.1%, Saudi Arabia 3.4%, United Arab Emirates 3.8%, and Yemen 3.2% (WHO, 1997, 2000a, b, 2003). In Jordan, there is very limited documentation of MDR-TB cases. Previous studies reported that over 90% of the M. tuberculosis rifampicin-resistant

clinical isolates harbored mutations in the 81-bp core region of the rpoB516, rpoB526, and rpoB531, the most frequent (70–95%) worldwide (Bártfai et al., 2001; Mokrousov et al., 2003). The discrepancies between the molecular and phenotypic drug resistance reported in this study have been reported by others previously (Baldeviano-Vidalon et al., 2005; Chan et al., 2007; Plinke et al., 2009). These discrepancies are most likely caused by problems with conventional susceptibility testing (Plinke et al., 2009) or by a single base substitution of a silent point mutation. Another possibility is the presence of heterogeneous isolates or mixed populations of resistant and susceptible M. tuberculosis bacilli in the initial sputum specimens with mutant genotypes being recognized by the molecular assay and therefore masking or dominating the susceptible genotypes.

We recently observed immunostimulatory properties in the root extracts of chemotypes NMITLI-101,

NMITLI-118, NMITLI-128 and pure withanolide, Withaferin A. In the present study, we evaluated the potential immunoprophylactic efficacies of these extracts against an infective pathogen. Our results show that administration of aqueous ethanol extracts (10 mg/kg) and Withaferin A (0.3 mg/kg), 7 days before and after challenge with human filarial parasite Brugia malayi offer differential protection in Mastomys coucha with chemotype 101R offering best protection (53.57%) as compared to other chemotypes. Our findings also demonstrate that establishment of B .malayi larvae was adversely affected by pre-treatment with Withaferin A as evidenced Selleck Selumetinib by 63.6% reduction in adult

worm establishment. Moreover, a large percentage of the established female worms (66.2%) also showed defective embryogenesis. While the filaria-specific immunological response induced by Withaferin A and NMITLI-101 showed a mixed Th1/Th2 phenotype, 118R stimulated production of IFN-γ, and 128R increased levels of IL-4. Taken together, our findings reveal potential immunoprophylactic properties of Withania somnifera and further studies are needed to ascertain the benefits of this plant against other pathogens as well. 2011 Blackwell Publishing Ltd “
“Over the last decade, live cell imaging has revealed the surprisingly complex orchestration of antigen receptor Topoisomerase inhibitor signalling at the immunological synapse. The imaging studies showed that one of the earliest steps in antigen receptor activation Erlotinib solubility dmso is the formation of submicroscopic clusters, which regulate the early signalling events. However, the molecular mechanisms operating inside these microclusters have remained beyond the resolution of optical microscopy. Recent development of imaging techniques that approach molecular resolution in intact cells offers a first view of the molecular processes inside these structures. Here I review the contributions

of molecular imaging of the immunological synapse to our understanding of antigen receptor clustering, binding to antigens, and recruitment of signalling molecules. Finally, I provide an outlook on the future prospects of this rapidly advancing technology. Activation of antigen receptors, the T-cell receptor (TCR) and the B-cell receptor (BCR), is a highly regulated process that sets in motion the adaptive immune response. In accord with their pivotal role in immune responses, antigen receptors are tuned to an unusually high degree of ligand discrimination and sensitivity. Each lymphocyte clone responds specifically to high-affinity interactions with the cognate antigen, which potentially signifies an infection, but disregards low-affinity interactions, which occur with self structures.

The estimated efficiency of peripheral B cells producing buy MAPK Inhibitor Library CD4-reactive Ab was ∼0.0013% (three clones/2.4×105 estimated screened B cells×100 (%), given that the B cells compose 10% of PBMC and that EBV immortalization is 30% efficient on average) 14. According to the ELISA data, the Fab concentrations that yielded 50% maximal binding were ∼8 μg/mL for HO538-213, and ∼1 μg/mL for HO702-001 and HO702-016 (Fig. 1B). Consistent with these data, the BIACORE

assay revealed that the dissociation constant (Kd) of HO538-213, HO702-001, and HO702-016 to rhCD4 was 6.5×10−8, 7.7×10−9, and 2.7×10−9 M, respectively (Fig. 1C), which is relatively weak compared with average Ab–Ag interactions (e.g. the Kd of mouse mAb Leu-3a to rhCD4 is 2.2×10−10 M). The Fab sequences were analyzed by the Kabat database (http://www.ncbi.nlm.nih.gov/igblast/) in GenBank, as previously described 15, 16. The Ig gene family of each gene and the most homologous germline are indicated (Fig. 2A). All the three clones were of the IgM class and

had a κ-chain for VL. Comparison of the heavy chain with the germlines revealed that the μ-chains of HO538-213 and HO702-001 were 95 and 97% homologous to germ line VH3-33, respectively, while HO702-016 was 96% homologous to germline VH4-4 17. For the light chains, the κ-chain Vkappa3 of HO538-213 was 97% homologous to germline L6 6, 18, 19, and κ-chain Vkappa1 Selleck Sirolimus of both HO702-001 and HO702-016 was 97% homologous to the germline L12 6, 18, 19. These data suggest that there is a preferential use of VH and VL genes to develop CD4-reactive Ab, considering the number of VH and VL genes present before the Ig gene rearrangement. According to the sequence analysis, the VH amino acid sequences of HO538-213

and HO702-001 carried distinct mutations, although both were derived from the same germline VH3-33. The mutations were more frequent in the CDR regions (Fig. 2B and C, Supporting Information Fig. 3), which is characteristic of somatic hypermutation (SHM) associated with affinity maturation. Unlike most SHM, however, mutations involving G/C were not dominant. We next examined the potential impact of these CD4-reactive Fab Ab on HIV replication. Viral replication was monitored in PBMC by measuring p24CA viral Ag levels in the culture supernatant. Among the three IgM Fab clones, HO538-213 suppressed Cepharanthine R5-tropic virus HIV-1JR-FL replication by 3.5±1.5-fold at 1–2.5 μg/mL (average±SD from four independent experiments, Fig. 3A). There was a modest but consistent suppression of X4-tropic virus HIV-1HXB2 replication (1.4±0.2-fold, average±SD from three independent experiments). BIACORE and ELISA revealed that HO538-213 did not compete with the anti-CD4 mAb Leu-3a 20, 21 for CD4 binding. Leu-3a restricts HIV-1 replication by physically blocking the Env-CD4 interaction (data not shown), suggesting that the epitope recognized by HO538-213 is distinct from the Env-interacting domain of CD4 7, 22, 23.

Interestingly, the ability of Lcn2 to

induce neutrophil migration was not affected TSA HDAC datasheet by the binding of a bacterial siderophore, such as enterobactin, to the peptide. The physiological relevance of Lcn2 as a chemoattractant was confirmed by in vivo studies in mice. Consistently, i.p., i.v. injection, and intradermal administration of Lcn2 resulted in increased leukocyte migration, mobilization, or infiltration. In addition, we found that Lcn2 plays an important role for PMN migration because PMNs from Lcn2−/− mice had a significantly reduced adhesion capacity, which we could link to reduced expression of adhesion associated surface proteins and the chemokine receptor CXCR2 on these cells. Similar biological effects as observed herein for Lcn2 were previously reported for several myeloid-related proteins (MRPs), such as S100A9 ABT-263 manufacturer (MRP14), S100A8 (MRP8), and S100A8/A9 [33-36]. These proteins have been reported to be, at least in part, expressed and stored in secondary granules such as Lcn2 and to act as chemotactic agents and modulators of neutrophil transmigration, which has been referred to stimulation of CD11b/CD18 integrin receptor expression [33]. Interestingly, MRPs can induce shedding

of CD62L and expression of CD11b on human PMNs [37]. Importantly, the expression of these adhesion molecules was significantly impaired on PMNs from Lcn2−/− mice as compared to Lcn2+/+ mice following an inflammatory stimulus. Moreover, the reduced expression of CXCR2 on PMNs of Lcn2−/− mice may negatively impact on the induction of chemotaxis by KC [38]. As we wanted to understand by which pathways Lcn2 exerts its chemoattractant activity, we analyzed the expression of the two previously described receptors of Lcn2, namely megalin and 24p3R [17]. We were able to show that primary PMNs express 24p3R but not megalin. Moreover, we found that the pharmacological blockage of Erk1/Erk2 signaling, a pathway that is induced

upon 24p3R/Lcn2 interaction [17], inhibited the Lcn2-inducible migration of neutrophils, whereas blocking of IL-8-inducible signaling cascades via DIC, PI3, and PKC did not affect Lcn2-dependent chemotaxis. We then employed Lcn2+/+ and Lcn2−/− mice to compare their PMN function. According to our previous results, the reduced in vitro migration of PMNs from Lcn2−/− Phloretin as compared to Lcn2+/+ mice was not unexpected. Surprisingly, we observed, that the addition of rmKC or rmLcn2 could not ameliorate the diminished migration of Lcn2−/− PMNs. However, this could not be traced back to reduced expression of the Lcn2 receptor 24p3R, which was comparable on PMNs from Lcn2−/− and Lcn2+/+ mice. We could then demonstrate that the impaired PMN migration and mobilization in Lcn2−/− compared to Lcn2+/+ mice is also seen in vivo in the very early phase of host responses to bacterial infection. Such differences — although in different experimental approaches — have not been observed by Flo et al.