, 2003). For C. pneumoniae, there was a reduction in chemokine expression only in the absence of TLR2 and TLR4 (Da Costa et al., 2004). Moreover, C. pneumoniae survival was significantly reduced upon double knock out of TLR2 and TLR4 (Rodriguez et al., 2006). Different combinations of antibodies or knock outs against TLRs may thus be useful to dissect the PAMP recognition network. Another very useful approach is to transfect TLRs into HEK cells

(that lack most of these receptors) and to use a reporter system such as luciferin to detect TLR activation (Brightbill et al., 1999). Activation of TLR4 or TLR2 also influences their own expression levels (Wissel et al., 2005), as well as those of cytokine receptors. This allows a more rapid and amplified response

to PAMPs by neighboring cells. Besides TLRs, other PRRs are triggered by C. pneumoniae and C. trachomatis infection. Nod1 not only controls cytokine activation Lapatinib but also induces the production of the bactericidal NO by inducible nitric oxide synthase (iNOS) (Shimada et al., 2009). Failure to activate iNOS allows uncontrolled bacterial growth. CD14 recognizes chlamydial lipopolysaccharide, which is a much weaker inducer than other lipopolysaccharides click here (Heine et al., 2003). Thus, PPRs should be seen as a network that can lead to the activation of the same downstream components. Furthermore, PRRs have very specific effectors and their activation is cell and pathogen dependent. Chlamydiales seem to have effector proteins that counteract TLR-induced immune response (reviewed in Betts et al., 2009). For example, C. psittaci elicits IFN-γ receptor (IFN-γR) expression Afatinib solubility dmso through TLR4 and TLR2, but at the same time its function is impaired (Shirey et al., 2006). How this inhibition is performed is unknown. Other interferons are also induced by C. pneumoniae infection, leading to an IFN-γ response. The interferons were activated by a TLR4/MyD88 signaling pathway (Rothfuchs et al., 2004). IFN-γ induces

tryptophan breakdown by increasing host cell indolamine 2,3 dioxygenase expression. This is detrimental for Chlamydiales because most cannot synthesize tryptophan. Chlamydia trachomatis genital strains can use indole produced by other bacteria of the vaginal flora to synthesize tryptophan. Ocular strains of C. trachomatis have a mutation that prevents correct enzyme activity (Bavoil, 2006). Parachlamydia acanthamoebae also does not encode the tryptophan synthase enzyme and can therefore not circumvent tryptophan depletion. Induction of IFN-γ by chlamydial PAMPs is thus a potent bacterial growth inhibitor, at least for some C. trachomatis strains and P. acanthamoebae. Moreover, recent studies highlighted new IFN-γ-inducible effectors, so-called p47 GTPases. The absence of any of the two members of the p47 GTPases (Igtp[Irgm3] and Irgb10) was linked to an increase in susceptibility to C. trachomatis infection (Bernstein-Hanley et al., 2006).

Fibroblasts play a crucial role in the proliferative phase. They migrate from normal tissue into the wound area from its margins, where they grow and form a new, provisional extracellular matrix by excreting collagen and fibronectin. Due to the crucial role of fibroblasts in the wound healing process, we investigated the effects of different concentrations of local anaesthetics on viability and proliferation of fibroblasts. Based on previous results in an inflammatory model of acute lung injury [13], we hypothesized that local anaesthetics do not have

an adverse effect on fibroblasts. In this study, human osteosarcoma cells (LGC Standard GmbH, Wesel, PR-171 datasheet Germany), osteoblast-like cell types with the morphology of human fibroblasts, were used. According to a study from Jukkola et al. in 1993, these cells have the characteristics of proliferative wound fibroblasts [14]. Cells were cultured in α-modified Eagle’s medium (MEM; LGC Standard GmbH) with 10% fetal bovine serum

(FBS; LGC Standard GmbH) and 10 000 U/l penicillin/streptomycin (LGC Standard GmbH) at 37°C and 5% CO2. Lidocaine (Lidocain CO2 2% Sintetica®) was purchased from Sintetica AG, Mendrisio, Switzerland, bupivacaine (Bucain®) from DeltaSelect GmbH, Munich, Germany and ropivacaine (Naropin®) from AstraZeneca, Wedel, Germany. Serial dilutions were chosen with lidocaine, bupivacaine and ropivacaine resulting in concentrations AZD9668 cell line of 0·3 mg/ml and 0·6 mg/ml, representing comparable tissue concentrations measured in clinical practice [15]. In group 1, cells were exposed to the LA for 2 days followed by another incubation time of 1, 4 or 7 days with normal medium without LA. In group 2, cells were exposed permanently to local anaesthetics for 3, 6 or 9 days. The LA-containing medium was changed every second day to provide stable and constant drug concentrations. Control cells were incubated with medium only for the ADP ribosylation factor according period of time. All changes

of medium performed in the treated group were performed similarly in control cells. On days 3, 6 and 9, living cells were counted manually in the Neubauer chamber, using trypan blue [16,17]. The tetrazolium bromide (MTT) assay is a well-known and recognized method to measure cell viability in vitro[18]. The method is based on the reduction of yellow tetrazoliumsalt 3-[4, 5-dimethylthiazol-2-yl]-2, 5-diphenyl tetrazolium bromide into purple formazan crystals by mitochondrial dehydrogenases. Dehydrogenases are active only in living cells. Conversion of MTT is therefore related directly to cell viability. Proliferation tests were performed with the help of the colorimetric bromodeoxyuridine (BrdU) assay (Roche, Basel, Switzerland). The test analyses the proliferation of cells by utilizing BrdU as an analogue of the DNA nucleotide thymidine, which is incorporated into the synthesized DNA of actively dividing cells.

5a–c). In relation to the maturation profile of T lymphocytes in skin lesions, the number of CD4+ T cells co-localizing with CD45RA was similar in both groups of patients (25%)

(Fig. 5d). In contrast, analysis of CD8+ T-cell maturation revealed a higher number of CD8+ T cells co-localizing with CD45RA in the RR/HIV lesions (20%), a result not observed in the RR lesions (< 5%), which only exhibited a few double-positive cells. This profile may indicate the presence of a TEMRA phenotype in the granuloma of RR/HIV despite it being impossible to evaluate the CCR7 marker in these biopsies. Analyses were performed of CD38 activation cell marker expression in different maturation phenotypes of CD8+ T cells in the RR and RR/HIV groups after ML stimulation. Selleck Navitoclax CD38 was significantly

up-regulated among RR/HIV patients in the TCM CD8+ T-cell subset [Fig. 6a,b; NS = 12·18 (9·8–12·9) versus ML = 22·32 (17·5–26·1); P < 0·05] and the TEM CD8+ T-cell subset [Fig. 6a,b; NS = 12·6 (7·1–20·5) versus ML = 28·3 (21·6–36·9); P < 0·05]. These data suggest that TEM and TCM CD8+ in RR/HIV patients preserve an activated phenotype in response to ML. The double-immune labelling of CD38 with CD45RO revealed a similar pattern in both groups under study, with many CD38+ cells co-localizing with CD45RO (30–40%). This result indicates the presence of a maturation-activated phenotype in Everolimus chemical structure RR and RR/HIV patients (Fig. 6c). It has been recently discovered that among human PBMCs, most of the perforin and granzymes are expressed in CD8+ T cells and that the high cytolytic granular content is related to cellular maturity.[28] ROS1 In light

of the observed increase in TCM and TEM CD8+ cell frequencies in RR/HIV patients and given the key roles played by perforin and granzyme B in the cell death pathway, the expression of these proteins in the PBMCs of RR and RR/HIV patients in conjunction with the expression of CD8, CD45RA and CCR7 markers was investigated. The ML increased granzyme B+ TEM CD8+ T-cell frequencies in PBMCs of the RR/HIV patients compared with the NS culture [Fig. 7a; NS = 8·7 (1·1–10·3) versus ML = 21·3 (7·8–25·7); P < 0·01], which was not observed in the HC and RR groups. ML also led to an increase in perforin+ cell frequency in the naive CD8+ T-cell population among RR and RR/HIV patients but with no significant difference. Based on the elevated expression of granzyme B and perforin, the cytotoxicity capacity of T cells isolated from RR/HIV patient blood was investigated. Purified lymphocytes led to an increase in the percentage of cell death (Propidium iodide-positive cells) in ML-stimulated RR/HIV monocytes [Fig.

abortus rough strain RB51 and smooth strain 2308 to stimulate murine bone marrow-derived DC (BMDC) activation and function based on the cell surface expression of costimulatory molecule and cytokine production. This study assessed simultaneously, for the first time, the differential ability of live, HK and IR rough and smooth strains of B. abortus YAP-TEAD Inhibitor 1 research buy at the same doses to stimulate DC activation and function. Female 6–8-week-old BALB/c mice were obtained from Charles River Laboratories Inc. (Wilmington, MA). Mice were used under animal care protocols approved by Institutional Animal Care and Use Committee at Virginia Tech. BMDCs were generated,

as described previously (Inaba et al., 1992). Briefly, tibias and fibulas of 7–8-week-old BALB/c mice were incised and bone marrow (BM) cells were removed. Following red blood cell lysis and filtration, the cells were resuspended and plated in RPMI 1640 complete media with 10% non-heat-inactivated fetal bovine serum and 20 ng mL−1 rGM-CSF (recombinant Granulocyte colony stimulating factor; Invitrogen, Carlsbad, CA). The cells were incubated at 37 °C in 5% CO2. Fresh media containing rGM-CSF was added at days 2, 4 and 5 and harvested on day 6. The cells harvested on day 6 were typically 70% CD11c+ and displayed low levels of major

histocompatibility complex (MHC) class II, CD40 and CD86 expression, consistent with immature DCs. Flow cytometry was performed to confirm DC activation status (Inaba et al., 1992). Stock cultures of live-attenuated rough B. abortus vaccine strain RB51 and virulent smooth this website strain 2308 from our culture collection (Schurig et al., 1991; Vemulapalli et al., 2000) were stored at −80 °C. An aliquot each of strain RB51 and strain

2308 were subjected to γ-irradiation using a 60Co source irradiator with a radiation output of 2200 rads min−1 (Model 109-68R by J.L. Shepherd and Associates, San Fernando, CA) for 3 h (396 krads Mirabegron of γ-radiation). Aliquots of strain RB51 and strain 2308 were subjected to heat killing by incubating in an 80 °C water bath for 60 min. IR and HK bacterial preparations were confirmed to be nonviable by plating aliquots on TSA plates and confirming lack of growth following 4 days of incubation. All experiments with Brucella were performed in our CDC-approved (C2003 1120-0016) Biosafety Level-3 facility. On day 6, DCs were harvested and plated at 5 × 105 cells per well in 24-well plates and stimulated with live, IR or HK strain RB51 or strain 2308 at 1 : 10 (DC : Brucella) or 1 : 100 CFUs per well (i.e. 5 × 106 or 5 × 107 CFU equivalents per well of IR or HK B. abortus). Stimulation was enhanced by a short spin at 400 g for 5 min at room temperature. The stimulated cells were incubated for 4 h at 37 °C in 5% CO2. Then cells were washed with media containing gentamicin (Sigma, St. Louis, MO) 30 μg mL−1. The stimulated cells were incubated for an additional 20 h in complete media with 10 ng mL−1 rGM-CSF and 30 μg mL−1 gentamicin.

Approximately, half of the CD56bright in peripheral blood express CD27, a marker virtually absent from CD56dim13–15. Hence, CD56bright in peripheral blood are identical or closely related

to the NK cells residing in secondary lymphoid organs (SLO) that produce cytokines to guide the adaptive immune response 4–6, 10, 11. It is worth noting that the precise relationship between NK cells in SLO, CD56bright in peripheral blood and CD56dim is not completely understood. Although some of the NK cells in SLO might be CD56bright recirculating from the blood, others could represent early maturation stages of NK cells developing from Selleckchem Doxorubicin hematopoietic precursor cells that repopulate extramedullary tissues and retain multi-lineage reconstitution capacity 16. Caligiuri’s group has identified lymphocytes in tonsils and lymph nodes representative of distinct stages of NK-cell development that differentiated into NK cells expressing high levels of CD56 17–19. NK-cell lineage commitment occurred in cells referred to as immature NK cells (iNK) that expressed no, or only very low levels of the NK-cell-associated markers NKp46, CD94, KIR and CD16. Furthermore, they lacked the characteristic attributes of mature NK cells: a high expression of the complement receptor CD11b as well as the ability to mediate cytotoxicity against MHC class I-negative targets and

to produce IFN-γ. iNK acquire all these features during the transition to the next maturation stage after which they closely resemble CD56bright GPCR Compound Library supplier in peripheral blood and are considered to be mature. The proof of progression from CD56bright to CD56dim Nutlin3 has remained elusive for a long time. CD56bright isolated from peripheral blood start to express KIR and CD16, downregulate c-kit and acquire cytolytic activity upon activation by IL-2 or IL-15 5, 12. However, IL-15 induces only CD56dim-like

levels of CD56, CD16 and KIR in CD56bright in contact with fibroblasts 20 or after infusion of CD56bright into immune-deficient mice 20, 21. Furthermore, skewing NK-cell differentiation toward CD56dim is far superior when IL-15 is trans-presented by IL-15Rα-Fc 21, which mimics the way IL-15 is presented by dendritic cells to NK cells in lymph nodes 22. Although these results provide direct evidence that a transition of CD56bright to CD56dim may occur, it remains unclear to what extent this transition represents the typical differentiation pathway in vivo. Furthermore, the fact that CD56bright may acquire many features of CD56dim may not be ground enough to denote them as less mature or as CD56dim precursors, not only because they largely outnumber CD56dim5, 6 but also because most CD56bright probably exert their effector functions without ever “maturing” into CD56dim.

, 2007). Such strains may possibly be able to form a biofilm in vivo without PNAG. Testing of other S. epidermidis

from the same collection (Table 1) indicates the presence of two B+, I+, P+ strains that are completely unable to develop an infection in spite of possessing the ica locus and forming a biofilm in vitro. This result indicates that in the TC-GP model, not all the clinical strains are able to develop and maintain an infection. Three negative B−, I−, P− clinical and commensal strains showed, to some extent, a capacity to develop and maintain an infection. Such strains may form a biofilm in vivo without PIA. The presence of a significant amount of bacteria after sonication AZD2281 research buy in the implants infected by these strains could indicate their presence in a biofilm form. It is also conceivable that these negative strains may develop and maintain an infection without a biofilm. Further experiments are needed to evaluate the capacity of the different strains to form a biofilm in vivo. However, the fact that the strains belonging to the ‘B+, I+, P+’ type showed a high capacity to cause persistent infections, compared with the opposite ‘B−, I−, P−’ type, emphasized the potential role of PNAG and the ica locus in the pathophysiology of strains. Whatever the strains, the exact mechanism responsible

buy Ixazomib for virulence remains to be determined, and it can be assumed that subspecies-specific differences exist in the abilities of S. epidermidis isolates to form a biofilm

and to cause infection in vivo. The early detection of the medical device-related staphylococcal infections is difficult using the classical tools of microbiological analyses. During an implant-related biofilm infection, the quantity of bacteria in the bloodstream is very low, and their direct detection is nearly impossible. The diagnosis is often made only at advanced stages of infection, when severe complications occur: formation of abscesses, pain, and unsealing of the prosthetic devices. Specific and noninvasive laboratory tests to diagnose these infections are not yet available. Because the pathogenicity of S. epidermidis is mostly due to its ability to colonize others indwelling polymeric devices and form a biofilm, a diagnostic test could be based on the detection of antibodies specific for biofilm components of CoNS, particularly S. epidermidis. A detection of specific ‘antibiofilm’ antibodies in the blood serum of patients could serve as a convenient noninvasive and inexpensive diagnostic tool for the detection of foreign body-associated infections. However, no antigens specific for staphylococcal infection have been identified. Different extracellular antigenic preparations have been proposed by different authors as candidates for immunological tests: an extracellular extract of a clinical S. epidermidis strain (staphylococcal slime polysaccharide antigen, Selan et al., 2002), a ‘20-kDa sulphated polysaccharide’, an ‘80-kDa peptidoclycan’ (Karamanos et al.

8,9 Herein we report the first case of a PJI due to Pseudallescheria apiosperma in an immunocompetent patient. A 61-year-old male immunocompetent farmer had a car accident in November 2000, ending up with his car in a fresh water canal. Besides a whiplash the patient had no injuries after the accident and water was not aspirated. Two months after the car accident he was find more first seen suffering from increasing knee pain. Previous to the car accident, the patient had an unremarkable medical history. Orthopaedic investigation in January 2001 disclosed a gonarthrosis of his left knee which was probably not the result of the car accident (Fig. 1). Since the patient

had a severe form of osteoarthrosis, a hemi-prosthesis was implanted in March 2001, 1 month after his first clinical presentation. Five weeks after implantation the patient was admitted with pain in his left knee. The radiograph showed a well-positioned and well-fixed hemi prosthesis (Fig. 2). The initial blood test found a white blood cell count of 12.5 × 109 l−1, an erythrocyte sedimentation rate (ESR) of 102 mm h−1, and a C-reactive protein (CRP) of 200 mg l−1, suggesting inflammation of the

patient’s left knee. A drainage system was installed, but all routinely taken microbiological Ibrutinib nmr cultures remained negative. The patient was treated with empirical antibiotics (3 dd 1000 mg cefazoline) for 2 weeks. One month later, in May 2001, he was re-admitted with fever (38.6 °C) and a red, swollen

and aching knee. Surgical drainage of the knee was started immediately with evacuation of approximately 100 ml foul-smelling, brownish-greyish pus. No bacteria were found using Gram staining, but in the blankophor preparation fungal elements were clearly visible. Cultures became positive with a fungus and routine morphological identification revealed a member of the Scedosporium/Pseudallescheria complex as causative agent. Based on good outcomes of itraconazole (ITZ)-treated Scedosporium-infections10–13 our patient started initially with ITZ 200 mg day−1 oral solution also which was increased to 400 mg day−1 when the prosthesis remained in situ, as patient refused removal. Molecular identification was performed with ITS-sequencing. By BLAST analysis of the obtained sequence vs. a validated in-house Centraalbureau voor Schimmelcultures (CBS) research database the isolate was identified as P. apiosperma.14 The isolate has been deposited in the CBS collection under CBS 129357. The sequence of the ITS/D1D2 region of the isolate has been submitted to Genbank with accession number JF906010. During the next two months several pus-filled pockets and fistulas around the knee were drained and mycological cultures grew Pseudallescheria despite ongoing ITZ administration. Notably no grains were observed in the pus collections.

The see more authors thank Rosario Cerrato for her excellent technical assistance in running the cAMP and GTPγ binding assays in FPR2/ALX recombinant cells and membranes. The authors also thank Sonia Pascual and Vicente García for technical and scientific support. The authors have no conflicts of interest to declare. “
“Treg homeostasis

is disturbed in multiple sclerosis (MS). Frequencies of recent thymic emigrant (RTE)-Treg are reduced and the disparity between RTE-Treg and long-lived memory Treg coincides with the MS-associated Treg defect, as shown previously. Recent studies demonstrate that IL-7 and thymic stromal lymphopoietin (TSLP) are critical for Treg maturation. Therefore, altered signaling through their receptors (IL-7R, check details TSLP receptor (TSLPR)), sharing the IL-7Rα-chain (IL-7Rα), might contribute to impaired Treg development. Using blood samples from 56 patients with MS and 33 healthy controls, we assessed IL-7Rα-expression on conventional T cells; frequencies, phenotypes and suppressive activities of Treg, plasma levels of IL-7 and soluble IL-7Rα; and screened for MS-associated IL-7RA gene polymorphism rs6897932. Moreover, we determined

Treg expressing two different TCR Vα-chains designating thymus-originated cells. As TSLP/TSLPR signaling in thymic myeloid dendritic cells (MDCs) promotes Treg differentiation, we measured TSLPR expression on peripheral MDCs to indirectly test whether altered TSLPR expression might add to compromised Treg neogenesis. We found reduced IL-7Rα expression on conventional T cells and upregulated IL-7 plasma levels together with reduction of RTE-Treg frequencies and Treg function in MS, without clear genetic influence. Decreased IL-7Rα expression in MS correlated with declined dual-receptor-Treg and reduced MDC TSLPR expression, indicating contracted

thymic Treg output. Tenoxicam We suggest that altered IL-7R/TSLPR signaling contributes to impaired Treg neogenesis in MS, which is compensated by expanded memory-Treg and finally results in dysfunctional Treg. Treg of CD4+CD25highCD127lowFOXP3+ phenotype are a small sub-group of thymus-derived T lymphocytes that protect peripheral organs from excess and autoimmune inflammation. Treg are defective in various human autoimmune diseases, including multiple sclerosis (MS), an inflammatory demyelinating disorder of the central nervous system 1. In patients with MS, this functional impairment relates to reduced frequencies of naïve Treg of recent thymic origin (CD45RA+CD31+) among circulating CD4+CD25highFOXP3+CD127low cells, along with compensatory expansion of Treg exhibiting a memory phenotype as we and others have shown previously 2, 3.

This was similarly seen in P. aeruginosa-infected cav1 KO mice [[9]]. Interestingly, IL-6 was also elevated not only in cav1 KO mice challenged with K. pneumoniae, but also in those exposed to P. aeruginosa. IL-6 plays disparate roles in inflammatory responses during bacterial infections [[25]]. IL-6 protects the host from death following K. pneumoniae infection; however, IL-6 neutralizing antibodies improve survival in

polymicrobial septic peritonitis [[26]]. Since IL-17R-deficient mice were shown to be more susceptible to check details K. pneumonia infection [[27]], we measured IL-17 levels and found an increase in cav1 KO mice compared with WT mice lungs. In fact, the susceptibility of IL-17-deficient mice to K. pneumoniae has been directly associated with delayed neutrophil recruitment and reduced G-CSF [[28]]. IL-17 has also been documented to induce secretion

of TNFα, IL-1β, and IL-6 [[29]]. The proinflammatory response to K. pneumoniae may not improve survival rates, but it aggravates existing disease conditions as shown in cav1 KO mice infected with P. aeruginosa [[9, 11]]. Despite the elevated levels of TNF-α, IL-1β, IL-6, and IL-17 in BAL fluid, the overall survival of cav1 KO mice with K. pneumoniae infection deteriorated rapidly. Interestingly, IL-27p28, a novel cytokine, was also increased in infected cav1 KO mice. p28, a subunit of IL-27, has broad inhibitory effects on Th1, Th2, and Th17 subsets

as well as the expansion of regulatory T cells [[30]]. Hence, we Z-VAD-FMK clinical trial Nintedanib (BIBF 1120) propose that the elevated IL-27 may provide a passive regulatory mechanism during acute infection. Given that MIP2 is a chemokine primarily produced by macrophages, our finding that MIP2 levels were not elevated in the lung indicates an impaired alveolar macrophage population. This in turn suggests that distinct compartmental immunity occurs in K. pneumoniae infection [[31]]. In addition, the phagocytic ability of AMs was found to be downregulated in K. pneumoniae-infected cav1 KO mice (data not shown). It has been suggested that Cav1 is an immune-modulatory effector on cytokine production through the MKK3/p38 MAPK pathway [[32]]. We found that ERK1/2 was activated in cav1 KO mice. We also noted a decreased TLR-4 response that was previously linked to gram-negative bacteria, suggesting a troublesome lack of innate immunity in cav1 KO mice. We also observed that GSK3β−β-catenin−Akt pathway may be involved in this infection, with both Akt and β-catenin being downregulated by Cav1 deficiency. By contrast, GSK3β expression and phosphorylation are significantly increased following loss of Cav1. This is consistent with the previous studies that show that GSK3β can destabilize β-catenin [[17]]. Although Akt is usually an upstream signal for GSK3β [[33, 34]], in this case the Akt changes may result from the effects of GSK3β [[35]].

86.115), the Gisela Thier foundation of the Leiden University Medical Center, and the Netherlands Leprosy Foundation. The funders had no role in study design, data selleck chemicals llc collection and analysis, decision to publish, or preparation of the manuscript. Jérémy Bastid is chief operating

officer at OREGA BIOTECH and provided the anti-CD39 monoclonal antibody BY40/OREG-103. Dr. Bastid was not involved in design and execution of experiments or in data analysis. As a service to our authors and readers, this journal provides supporting information supplied by the authors. Such materials are peer reviewed and may be re-organized for online delivery, but are not copy-edited or typeset. Technical support issues arising from supporting information (other than missing files) should be addressed to the authors. Supporting Information Fig. 1. Gating strategy. Supporting Information Fig. 2: Expression of regulatory T cell markers in restimulated CD8+CD39+ T-cell lines. Supporting Information Fig. 3: Inhibition of Th1-responder cell proliferation is

not the result of lysis by CD8+ T cells. “
“Four genotypically distinct strains of L. major collected from persons residing in different endemic areas of cutaneous leishmaniasis in Iran were evaluated in BALB/c selleck chemicals mice. Parasite virulence was evaluated by measuring the parasite burden

in the lymph nodes. Immunogenicity of the strains was assessed by analysis of many cytokines mRNA expression levels in popliteal lymph nodes of the mice in early (3, 16, 40 h) and late (week 1, W3, W5 and W8) time periods after infection. The expression of cytokines mRNA, namely Ifng, Il2,Il4,Il10 and Il12, was quantitated by real-time PCR. The lowest and the highest parasite loads were induced by Damghan (2·15 × 107) and Shiraz (9·59 × 109) strains, respectively. Moreover, Damghan strain elicited higher expression levels of Ifng and Il2 mRNA and the highest ratio of Ifng/Il4 mRNA expression compared with the other strains at 40 h and 8 weeks post-infection. The results indicate that the inoculation of BALB/c mice with different strains induced high diversity in parasite burden and cytokines gene expression. Amongst the four strains, Damghan strain showed the lowest parasite load and the highest tendency to induce expression of Th1 cytokines gene and might be considered as a safe and immunogenic strain. Leishmania major parasites are intra-macrophage organisms and the causative agent of the Old World zoonotic cutaneous leishmaniasis (ZCL) [1]. ZCL is endemic in North Africa, Central Asia and Middle East [2], including Iran and is a major health problem in different parts of the country.