Cystatin C was measured AZD1208 using a particle-enhanced nephelometric assay. Results:  CKD stages were more sensitively differentiated by cystatin C compared to sCr, especially in moderate and severe kidney dysfunction. Sex and body mass index did not affect cystatin C level. Pearson’s correlation coefficients of reciprocal of cystatin C, measured and recalibrated sCr compared to systemic inulin clearance (Clin) were 0.757, 0.734 and 0.709, respectively. We derived novel pertinent equations based on cystatin C (model 1: 1.404 × cystatin C−0.895 × age0.006 × weight1.074 × height−1.562 × (0.865; if female); model 2: 43.287 × cystatin C−0.906 × age0.101 × (0.762;

if female)]. Models 1 and 2 showed superior performance in representing systemic Clin than the IDMS Modification of Diet in Renal Disease (MDRD) study equations did (adjusted r2 = 0.76 and 0.72 for models 1 and 2, and 0.64 and 0.65 for 4 and 6 variable IDMS MDRD equations, respectively). Conclusion:  Cystatin C reflects kidney dysfunction sensitively, and thus cystatin C-based estimation of GFR could provide a reliable support for clinical practice. “
“Acute kidney injury Daporinad (AKI) is a frequent complication in critically ill patients and is associated

with a high mortality. Clinicians have limited tools to predict the course of AKI at the time of serum creatinine increase. We evaluated the diagnostic and prognostic utility of urinary cystatin C (uCysC) in patients with AKI. In this study, serum and uCysC and urinary creatinine (uCr) were measured in patients presenting with acute kidney injury. The patients were divided into two groups: those with prerenal AKI and those with an intrinsic AKI. Prerenal AKI was defined as a new-onset

increase in serum creatinine (sCr) that resolved within 72 h and returned to the baseline kidney function level. Patients with intrinsic AKI were defined and classified according to the Acute Kidney Injury Network (AKIN) criteria. Of the total number of patients (n = 213), 40.4% (n = 86) were judged to have prerenal AKI and 59.6% (n = 127) intrinsic AKI. Loperamide uCysC values and the uCysC/uCr ratio were significantly higher in intrinsic AKI versus prerenal AKI. In intrinsic AKI, the uCysC concentration increased with AKI severity. The uCysC/uCr ratio was significantly higher in the RRT group versus the non-RRT group (0.15 vs. 0.08, respectively; P = 0.037). In a multivariate analysis, the uCysC/uCr ratio was associated with in-hospital mortality (P = 0.019). uCysC level and the uCysC/uCr ratio were useful biomarkers of intrinsic AKI, and the uCysC/uCr ratio was predictive of in-hospital death in AKI patients.

7b). Antibody stimulation was used instead of antigen stimulation to demonstrate the direct effect of learn more the inhibitor on

Th1 cells and to discount the indirect effects on APCs. Inhibition of JNK activity by SP600125 was sufficient to suppress the proliferation of the KLH-specific Th1 cells, indicating that Th1 cells used in this model are no different from primary CD4+ T cells in that the inhibition of JNK alone is sufficient to block proliferation. In conclusion, p21Cip1-mediated suppression of JNK activity in anergic Th1 cells is a novel potential mechanism that could account for the proliferative unresponsiveness found in these cells. This manuscript examined the role of p21Cip1 in maintaining the proliferative unresponsiveness found 5-Fluoracil in Th1 cells anergized by exposure to antigen and n-butyrate. The results presented in this work suggest that p21Cip1 functions in these Th1 cells primarily through the inhibition of members of the MAPK family rather than inhibition of its classical interaction partners,

namely cdk. p21Cip1 has long been described as a negative regulator of the cdk-mediated G1 to S phase transition.25 However, based on the association pattern of p21Cip1 and cdk in anergic compared to control Th1 cells, the p21Cip1 inhibition of cdk activity does not appear to be the primary mechanism for cell cycle inhibition. Instead, the results suggest that p21Cip1 specifically interacts with p-JNK and p-c-jun in antigen-restimulated anergic Th1 cells. The role of p21Cip1 in the normal cell cycle has been at

variance in different studies. Eventually, a dual role has been suggested for p21Cip1 in which low levels of p21Cip1 facilitated the cell cycle by promoting cdk–cyclin complex assembly whereas high levels inhibited cdk activity.25–27 The role of p21Cip1 in normal T-cell activation is not clear. T cells from the p21Cip1-deficient mice exhibited enhanced homeostatic proliferation and increased the the frequency of cycling T cells.28 Another study using p21Cip1-deficient mice reported that p21Cip1 did not affect primary proliferation of naïve T cells, but was required for the regulation of activated/memory T-cell proliferation.29 In the present study, control Th1 cells stimulated for 36 hr with antigen contained appreciable amounts of p21Cip1, much of which associated with cdk2, cdk4 and cdk6. It would therefore seem likely that at least some of the regulatory effect of p21Cip1 in stimulated control Th1 cells in our system involves interaction with cdk. The amount and timing of p21Cip1 induced in activated T cells may be sufficient to promote cdk–cyclin assembly but not enough to block cdk activity. Alternatively, p21Cip1 may be up-regulated in activated T cells as a fail-safe mechanism in case some kind of cellular stress necessitates regulation of DNA replication or repair.

Serum samples from patients with TB reacted more strongly with MPB64 antigen than did those from uninfected individuals. In addition, serum samples from TB patients

with active infection reacted more strongly with the antigen than did samples from patients with inactive TB. When urine samples were assessed using this assay, similar results were obtained. Correlations between the data obtained from serum and urine samples were analyzed for all subjects, including uninfected individuals, and a strong positive correlation between the results of serum and urine tests (n = 36, r = 0.672) was found. The sensitivity and specificity of this assay for serum samples was 85.7 % and 85.0 %, and for urine samples 75.0 % and 85.0 %, respectively. These results suggest that dot-blot assay with MPB64 Fulvestrant antigen could be a useful screening test for active

TB. Because urine samples can be obtained more easily than serum samples and because urine is less contagious, urine testing should probably be employed for screening purposes. selleck According to the World Health Organization, about two billion people, approximately one third of the world’s population, are infected with M. tuberculosis. In 2011, around 8.8 million new cases of TB and 1.1 million deaths from this disease were reported (1). This is the greatest number of deaths caused by any single pathogen. From sub-Saharan Africa to Asia, the annual incidence of TB now exceeds 300 per 100,000. In Japan, the number of new cases of TB and its incidence has been increasing since 1997. In 2007, the number of new TB patients reached 25,311, with the total incidence rising to 19.8, which is higher than in many other developed countries (1). In Japan, a high percentage of infected elderly patients develop active TB and, in urban areas, the percentage of immigrants from Southeast

Asian countries with TB is not negligible. The diagnosis of pulmonary TB is based on the presence of respiratory symptoms (cough, sputum, and hemoptysis) and systemic symptoms (fever, malaise, and weight loss), and the findings on chest X-ray films and computed tomography scans. Examination of the patient’s sputum and gastric C-X-C chemokine receptor type 7 (CXCR-7) juice, as well as auxiliary diagnostic tests such as the QuantiFERON test, tuberculin skin test, and bronchoscopy, can also be performed (2). For many years, the tuberculin skin test was the standard test for TB infection. However, this test does not become positive until 4–6 weeks after establishment of infection and prior BCG vaccination can influence its results. Accordingly, the QuantiFERON-TB Gold In-Tube, which is based on three tuberculosis-specific antigens (ESAT-6, CFP-10 and TB7.7 proteins), is now recommended as a more specific test for TB (3, 4). There have been many attempts to develop serodiagnostic tests for TB that detect antibodies targeting various structural components of M. tuberculosis.

We also developed a bioinformatics method to predict pMHC-I stability, which suggested that 30% of the nonimmunogenic binders hitherto classified as “holes in the T-cell repertoire” can be explained as being unstably

bound to MHC-I. Finally, we suggest that nonoptimal anchor residues in position 2 of the peptide are particularly prone to cause unstable interactions Erlotinib manufacturer with MHC-I. We conclude that the availability of accurate predictors of pMHC-I stability might be helpful in the elucidation of MHC-I restricted antigen presentation, and might be instrumental in future search strategies for MHC-I epitopes. Major histocompatibility complex class I (MHC-I) plays a pivotal role in the generation of specific immune responses mediated

by cytotoxic T lymphocytes (CTLs). MHC-I molecules sample peptides derived from intracellular proteins, translocate them to the cell surface, and display them to CTLs, allowing immune scrutiny of the ongoing intracellular metabolism leading to the detection of the presence of any intracellular pathogens. To fulfill this crucial antigen presenting function, MHC-I molecules must be endowed with the ability to retain bound peptides at the cell surface while waiting for the arrival of rare circulating CTL clones of the appropriate specificity. Sustained presentation at the cell surface and induction of specific immune T-cell responses therefore requires

some selleck degree of pMHC-I stability. Indeed, it has been claimed that stability, rather than affinity, of pMHC-I complexes is the better correlate of immunogenicity and immunodominance [[1-5]]. Experimentally, however, affinity remains the most frequently Atezolizumab established correlate of immunogenicity. Thus, when Assarsson et al. [[6]] recently conducted a quantitative analysis of the variables affecting the repertoire of T-cell specificities recognized after vaccinia virus infection, they found that the vast majority of epitopes (85%) bound their restricting allele with an affinity of 500 nM or better, and most (75%) bound with an affinity of 100 nM or better. Investigating the stability of pMHC-I complexes for a small sample of immunogenic and nonimmunogenic peptides, they found a suggestive, but not statistically significant, trend for off-rates and immunodominance being correlated. The authors concluded that “in our hands, peptide stability did not correlate significantly better with immunodominance than did equilibrium binding measurements”. One reason why pMHC stability has not been addressed more extensively undoubtedly relates to the cumbersome and/or low-throughput nature of current biochemical methods used to measure the dissociation of pMHC complexes [[6-12]]. A particularly interesting dissociation assay developed by Parker et al.

Rabbit monoclonal Ab against GAPDH was obtained from Cell Signaling Technology MK-2206 chemical structure (Danvers, MA). Western blotting of lung homogenates was performed as described previously [[46, 47]]. RNA was extracted from lung homogenates and cells with Trizol (Invitrogen Life Technologies, Carlsbad, CA) according to the manufacturer’s instructions. Reverse transcription was performed using 1.5 μg of RNA and cDNA was amplified using gene-specific primers [[48, 49]]. The results were normalized with GAPDH. MPO assay was performed as described previously (15). Samples were homogenized in 50 mM hexadecyltrimethylammonium bromide

(HTAB) and assayed as previously described [[45, 50]]. H2DCF dye (Molecular Probes) does not normally AZD6738 chemical structure fluoresce under resting conditions, but emits green fluorescence upon reaction with superoxide inside cells. Cells were treated as above and equal amounts of dye added [[16]]. This assay measures color change of MTT upon reduction by enzymes to assess the viability of cells. After infection of MLE-12 cells with K. pneumoniae, MTT dye was added at a final concentration of 1 μg/mL as described previously [[47]]. We used LipofectAmine2000 to transfect cells at 60% confluency and achieved high efficiency in transfection [[22, 51]]. The yellow fluorescent protein (YFP)-Cav-1, YFP-Cav-1Δ51-169 dominant negative (DN) plasmids were generated as described previously [[18]].

MLE-12 cells were infected with K. pneumoniae at MOI 10:1 for 1 h and the free bacteria were removed by washing three times with PBS. The surface bacteria were killed by incubation with 100 μg/mL polymyxin B for 1 h and intracellular bacteria were enumerated to determine CFU. Transfection with cav1 DN plasmid did not affect survival of MLE-12 cells prior to incubation with K. pneumoniae. WP1066 (a novel STAT5 inhibitor from Sigma) was dissolved in 1% DMSO solution and used at a final concentration of 2 μM in culture medium. No adverse effect of the vehicle control was observed in the assays. The differences

in outcomes between cav1 KO and WT control animals after K. pneumoniae infection were calculated by Kaplan–Meier survival curve comparisons, and the Liothyronine Sodium p values were derived from a log-rank test. Most experiments were performed three times in triplicate. Comparison of experimental groups with controls was done with one-way ANOVA (Tukey’s post-hoc) [[16, 52]]. This project was supported by NIH ES014690, Flight Attendant Medical Research Institute (FAMRI, 103007), and American Heart Association Scientist Development Grant (MW); and by NIH 5R01HL092905-04 and 3R01HL092905-02S1 (HG). We thank S. Rolling of UND imaging core for help with confocal imaging. The authors declare that they have no competing financial interests. Disclaimer: Supplementary materials have been peer-reviewed but not copyedited.

In this sample of patients, there was a predominance of middle-aged male patients, who were primarily rural workers. Chronic multifocal disease was prevalent, with lesions also detected in the lungs, lymph nodes, skin or adrenal glands.

Most of the cases presented with lesions at the gingival mucosa followed by the palate and lips; these conditions occurring in the oral cavity were frequently associated with pain. Importantly, most of the patients sought professional care for oral lesions. The diagnosis was obtained through exfoliative cytology and/or biopsy of the oral lesions. Medical treatment was effective, and there were no mortalities in the sample. The present findings not only confirm the importance of oral lesions in the diagnosis and management of PCM but also illustrate that questions still remain unclear, such as the possibility of Selleckchem PI3K Inhibitor Library direct inoculation of the fungus onto oral tissues. “
“To report an outbreak of Fusarium solani endophthalmitis after uneventful cataract surgeries performed on the same day in the same operating room. Nine patients underwent GPCR Compound Library order phacoemulsification at 4th Clinic of Beyoglu Eye Training and Research Hospital in Istanbul. Cefuroxime axetyl

was injected intracamerally from the same vial to all patients at the end of surgery. All patients developed acute postoperative endophthalmitis. Presentation, cultural studies, treatment, clinical responses and risk factors were evaluated. Cultural and DNA sequence findings revealed F. solani. Antifungal therapy was begun and pars plana vitrectomy, intraocular lens and capsule extraction were performed. Corneal involvement was correlated with old age and systemic disease. Fusarium solani should be considered in acute postoperative endophthalmitis. This infection can be controlled with early and aggressive combined antifungal and surgical treatment. The patients with corneal involvement had poor prognosis. It is important to use solutions prepared separately for each patient. “
“Mucormycoses are life-threatening infections with fungi

from the order Mucorales (Mucoromycotina). Although mucormycoses are uncommon compared to other fungal infections, e.g. N-acetylglucosamine-1-phosphate transferase aspergillosis and candidiasis, the number of cases is increasing especially in immunocompromised patients. Lichtheimia (formerly Absidia) species represent the second to third most common cause of mucormycoses in Europe. This mini review presents current knowledge about taxonomy and clinical relevance of Lichtheimia species. In addition, clinical presentation and risk factors will be discussed. Proper animal infection models are essential for the understanding of the pathogenesis and the identification of virulence factors of fungal pathogens. To date, several animal models have been used to study Lichtheimia infection.

TNF-α treatment induced a decrease in TNF-α, IL-12p40 and IL-10 mRNA levels in peritoneal cells following PPD stimulation while live M. tuberculosis caused an increase in TNF-α mRNA and a decrease in the IL-10 mRNA expression. TNF-α injection also induced an increase in the infiltration of mononuclear cells and in the proportions of CD3+ T cells in the lymph nodes. These Palbociclib order results indicate that rgpTNF-α enhances some aspects of T cell immunity and promotes control of mycobacteria in the tissues. Future studies will address the role of TNF-α in BCG-vaccinated guinea pigs following low-dose pulmonary challenge with virulent M. tuberculosis. Among the many cytokines that contribute to a protective immune

response against Mycobacterium tuberculosis, tumour necrosis factor (TNF)-α is known to play an essential role in the formation and maintenance of granulomas [1,2]. Resistance against M. tuberculosis

is mediated by T cells and macrophages [3–5]. Several cytokines, including interleukin (IL)-12, IL-17 and IL-23, contribute to the host-response to mycobacteria by enhancing the development of T helper type 1 (Th1) immunity [6,7]. Among the Th1 cytokines, interferon (IFN)-γ and TNF-α have been identified as the most important players in the cytokine cascade for anti-mycobacterial immunity because the formation as well as the maintenance of the granuloma are mediated by TNF-α, and it synergizes with IFN-γ in activating macrophages for the production of effector molecules [2,8]. It is known that susceptibility to tuberculosis

occurs with defects in the type-1 cytokine pathway in humans [9,10]. The importance Selleckchem Lorlatinib of IFN-γ has been well established in mouse models, as disruption of IFN-γ, the IFN-γ receptor gene or components of the IFN-γ receptor signal-transducing chain resulted in an exacerbation of disease after M. tuberculosis or M. bovis infection [9,11]. Neutralization of TNF-α in mice resulted Tolmetin in the reactivation of latent M. tuberculosis infection, disrupted granuloma formation and rapid death [12]. In another study, neutralization of TNF-α resulted in marked disorganization of granulomas and an increase in proinflammatory cytokine and chemokine expression in mice given an aerosol infection with M. tuberculosis[13]. Mice deficient for TNF-α or TNF-R1 showed disruption in granuloma formation and succumbed to infection with M. tuberculosis[14]. The importance of TNF-α in anti-mycobacterial immunity has been reinforced by reports that the use of TNF-α neutralizing antibody in the treatment of chronic inflammatory diseases resulted in the reactivation of latent tuberculosis in humans [15], [10]. Several reports also indicate that injection of mice with recombinant TNF-α or IFN-γ alone or in combination was associated with decreased microbial growth and increased survival after infection with disseminated M. avium complex or M. tuberculosis[16,17].

brasiliensis. For analysis

of bystander activation 2 × 106 leucocytes from DO11/4get/rag−/− mice were transferred by intravenous injection into naive 4get recipient mice. One day after cell transfer, recipients were infected with N. brasiliensis alone (Nb) or with a mixture of N. brasiliensis and 100 μg of ovalbumin (Nb-OVA). The Nb-infected mice were analysed on day 9 after infection. The Nb-OVA-infected mice received an intranasal challenge with 500 μg OVA in 50 μl PBS on day 3 after infection and were then analysed on day 6. DAPT molecular weight For reconstitution of Smarta/4get mice 107 MACS-purified (Miltenyi-Biotec) CD4 T cells from 4get mice were transferred 3 days before infection and mice were analysed on day 9 after infection. The Mann–Whitney-U-test was used to calculate P-values. Single asterisks indicate P < 0·05, double asterisks indicate P < 0·01. Infection of mice with the helminth N. brasiliensis induces a strong Th2 response, which can be visualized by using IL-4/eGFP reporter mice (4get mice).2 At the peak of the response, on EPZ-6438 day 9 after infection, 30–50% of CD4 T cells in the lung express IL-4 transcripts, which marks them as Th2 cells (Figs 1a and 2). The Th2 cells display an activated phenotype with low expression

of CD62L and high expression of CD44, CD29 and CD11a. However, the majority of Th2 cells appear negative for expression PD184352 (CI-1040) of early activation markers like CD25 or CD69 (Fig. 1b). To determine whether N. brasiliensis infection leads to biased expansion of T-cell populations with certain TCR-Vβ chains, we compared the TCR-Vβ repertoire of resting (CD62Lhi) or activated (CD62Llo) CD4 T cells from naive and N. brasiliensis-infected mice. No major differences between naive and infected

mice could be observed (Fig. 2a). As IL-4 protein is only detectable in Th2 cells with the highest expression level of IL-4/eGFP, we performed IL-4 cytokine capture assay after brief re-stimulation of ex vivo isolated Th2 cells and directly compared the TCR-Vβ repertoire of IL-4 protein-expressing or IL-4/eGFP-expressing T cells. No difference in TCR-Vβ usage was observed between the two populations (Fig. 2b). Furthermore, we found no alterations of the TCR-Vβ repertoire when we compared IL-4/eGFP+ and IL-4/eGFP− cells among the CD62Llo population, suggesting that the N. brasiliensis-induced Th2 response was polyclonal and not biased toward expansion of certain TCR-Vβ families (Fig. 2c). It remains unclear whether all Th2 cells in N. brasiliensis-infected mice are indeed parasite-specific T cells or whether early release of cytokines by antigen-specific T cells or cells of the innate immune system could induce differentiation of Th2 cells by bystander activation.

Several pathogenic bacteria including Staphylococcus aureus, Klebsiella pneumonia and Streptococcus pyogenes also activate caspase-1 via NLRP3 46–48. Exotoxins acting as pore-forming or membrane-damaging factors are important in mediating activation of the NLRP3 inflammasome 49, 50. For example, S. aureus hemolysins and Deforolimus S. pyogenes streptolysin O are critical for NLRP3 activation 46, 47. Although TLR stimulation contributes to NLRP3 activation via priming, S. aureus and S. pyogenes can activate caspase-1 independently of MyD88/TRIF, the critical adaptors required for all TLR signaling 46, 47. One possibility

is that pathogenic bacteria induce priming of the NLRP3 inflammasome via TLR-independent mechanisms. Alternatively, exotoxins may mediate the delivery of microbial molecules for NLRP3 activation. Unlike that triggered by TLR ligands, NLRP3 activation induced by bacterial or fungal infection is independent of the P2X7R 46, 47. Thus, the role of ATP-induced P2X7R signaling in microbial

activation of the NLRP3 inflammasome in vivo is unclear. Recent studies suggest a model of NLRP3 activation that is mediated by two signals. The first, signal one, is provided by microbial molecules such as TLR ligands or by certain cytokines that induce priming of the inflammasome at least in part by NF-κB and NLRP3 induction (Fig. 1) 29, 30. The second signal selleck chemicals directly triggers caspase-1 activation, and can be mediated by at least four separate pathways that include ATP-P2X7R-pannexin-1, Syk signaling,

Adenosine lysosomal membrane rupture and bacterial exotoxins (Fig. 1). It is likely that these different pathways culminate in a common step that leads to NLRP3 activation. However, the identification of a unifying mechanism of NLRP3 activation remains elusive. The mechanisms regulating NLRP3 activation are discussed in more detail in accompanying articles of this issue 51, 52. A possible common link is provided by the ROS because NLRP3 activation is blocked by ROS inhibitors 27. However, most of these studies rely on pharmacological inhibitors that are used at high concentrations and exhibit variable effects or RNA interference, which is artifact prone. Nonetheless, Tschopp and colleagues have identified thioredoxin-interacting protein (TXNIP) as an NLRP3-interacting protein 53. Although, it remains to be determined whether TXNIP is an essential activator or just a regulator of the NLRP3 inflammasome. There has been a remarkable growth in our knowledge about the regulation, activation and biological role of the inflammasome. However, many important questions remain. They include identifying the link between microbial stimulation and inflammasome activation given that recognition of NLRC4/NLRP3 appears indirect. The identification of TXNIP as a possible link between ROS and NLRP3 is important, but more work is needed to understand its precise role in inflammasome activation.

To do this, transient transfection assays were performed using either one of the two human CCL20 promoter-luciferase constructs: pCCL20 c/EBPmut, containing the full-length human CCL20 promoter bearing the mutated c/EBP site; and the pCCL20 NF-κBmut, containing the full-length CCL20 promoter bearing the mutated NF-κB site 14. As shown in Fig. SCH 900776 in vivo 4, site-specific mutation of the single NF-κB responsive motif almost completely blocked the ability of IFI16 to trigger luciferase activity. In contrast,

mutation of the single C/EBP site only slightly decreased luciferase activity compared with the wild-type CCL20 promoter. In order to provide definitive evidence supporting the role of NF-κB as the mediator of CCL20 promoter activation by IFI16, HUVEC were transfected with the indicator plasmid 5× NF-κB luc 15, infected thereafter with AdVIFI16 or AdVLacZ and reporter gene activity subsequently measured 24 h later. As shown in Fig. 4, overexpression

of IFI16 significantly increased NF-κB transactivation of the reporter gene although at levels lower than those observed with the endogenous CCL20 promoter. Altogether, these results demonstrate that IFI16 interacts with NF-κB in order to trigger CCL20 promoter activity, in line with the results obtained from the ICAM-1 promoter analysis. However, NF-κB does not appear to be the only transactivator GSK126 research buy stimulated by IFI16 in order to trigger CCL20 promoter. The ligand–receptor pair CCL20-CCR6 is believed to be responsible for the chemoattraction of CD34-derived immature DC, Langerhans DC (L-DC), effector/memory T cells and B cells, and it plays a role at skin and mucosal surfaces under homeostatic and inflammatory conditions 16, 17. If this is the case, it is important to verify a functional link between the ability of IFI16 to trigger CCL4, CCL5 and CCL20 release by HUVEC

Dimethyl sulfoxide and DC and B-lymphocyte chemoattraction. Using a transwell migration assay, we demonstrate that both L-DC and B cells migrate to a significantly greater degree in response to the supernatants from IFI16-infected HUVEC compared with the supernatants from LacZ-infected HUVEC (Fig. 5). This migration was significantly reduced by pre-incubation with the anti-CCL4, anti-CCL5 and anti-CCL20 mAb, but only when added to the supernatants from IFI16-infected HUVEC. In contrast, addition of an unrelated mAb of the same isotype, used as an internal control, did not influence cell migration (data not shown). These results confirm that the secretion of CCL4, CCL5 and CCL20 by IFI16-infected HUVEC is functional and important for inducing L-DC and B-cell migration into the mucosa and skin where these cells are particularly abundant.