Supernatants were harvested, centrifuged to pellet cells and insoluble debris and assessed for cytokine levels by ELISA or cytokine array. For differentiation experiments, monocytes grown in OptiMEM were treated with dibutyryl-cAMP (db-cAMP, 100 μm), macrophage colony-stimulating factor (M-CSF; 5 ng/ml) or granulocyte–macrophage colony-stimulating factor (GM-CSF; 2 ng/ml) for 4 days before analysis by flow cytometry or assay of cytokine release. For flow cytometric analysis, 100-μl aliquots of cells (5 × 106/ml) were stained with the mAb for individual integrins for 30–60 min on ice before washing in PBS; if required, a learn more fluorophore-conjugated secondary

reagent was added and a further 30–60 minutes of incubation was conducted before washing and analysis. Appropriate isotype controls were included. Data were collected from a minimum of 104 cells using a FACScan instrument (BD Biosciences) and analysed using CellQuest software (BD Biosciences). Human monocytes release cytokines following stimulation by a range of stimuli. Other groups have demonstrated that exposure of human PBMC to sCD23 promoted TNF-α release, via ligation of the αVβ3 integrin,18 and other cytokines via ligation of β2 integrins.17,35Figure 1(a) illustrates that normal PBMC released TNF-α following stimulation with

lipopolysaccharide (LPS) or sCD23 but not when treated with the extracellular matrix proteins vitronectin (Vn) or fibronectin (Fn), which are additional ligands for αVβ3 and αVβ5. However, these

Rapamycin order cells expressed high levels of three of the four integrins that are known to bind sCD23; namely αVβ3, αVβ5 and αXβ2 (Fig. 1b). Therefore, it is not clear which of the four possible sCD23-binding integrins would be responsible for acute regulation of release of one or more discrete cytokines or groups of cytokines (Fig. 1c), or whether these integrins generate synergistic or mutually inhibitory signals. To test the broad hypothesis that individual sCD23-binding integrins differentially regulate acute cytokine release Selleck CHIR-99021 from monocytic cells, an antibody array approach was employed to determine the qualitative patterns of cytokine release from THP-1 cells following stimulation with antibodies directed against individual sCD23-binding integrin isoforms (Fig. 1c). The general principle of the assay is shown in Supplementary material, Fig. S1A and the patterns of pairs of anti-cytokine antibodies printed on the array are shown in Supplementary material, Fig. S1B. The pattern of release of cytokines driven by sCD23 in monocytic cells is complex and may reflect the fact that up to four distinct sCD23 binding integrins can be ligated on the same cell, with each potentially giving rise to a distinct effect on cytokine synthesis and release.

Of the 47 patients who received anakinra (25 anakinra with dexamethasone), progression-free survival ensued for more than three years and in 8 patients for

more than 4 years 100. Patients with a decrease in serum CRP of 15% or greater after 6 months of anakinra monotherapy resulted in progression-free survival times greater than 3 years as compared with 6 months in patients with less than a 15% fall during anakinra therapy (p<0.002). Thus, an effective reduction in IL-1β activity using CRP as the marker for IL-1β-induced IL-6 halts progression to active myeloma. Anakinra results in resolution of all signs and symptoms within hours after the first injection. However, approximately 20% of patients with Schnitzler's syndrome develop a lymphoproliferative disorder, mostly lymphoma or Waldenstrom

disease, which is similar to patients with IgM MGUS. This latter point and its consequences have been already been addressed in the find more literature 101. Blocking IL-1β may reduce the progression to a lymphoproliferative disorder in patients with Schnitzler’s syndrome. Similar to smoldering myeloma, the concept that IL-1β drives IL-6 production was tested in a patient with another lymphoproliferative disorder, Castleman’s https://www.selleckchem.com/products/byl719.html disease, which is usually treated with anti-IL-6 receptor antibodies 102. The patient failed to respond to cladribine, rituximab, steroids, etanercept and anti-IL-6 antibody but within 1 wk of anakinra treatment, the constitutional symptoms markedly improved, and anemia, thrombocytosis, leukocytosis, and elevated markers of systemic inflammation reverted to normality 103. In cytokine biology as applied to the treatment of disease, associations of elevated circulating levels of a particular cytokine with a disease do not allow for a conclusion of causation by that cytokine for the pathological process. Rather, only specific

blockade or neutralization provides the evidence. This is especially Fossariinae the case with IL-1β, as circulating levels, even in severe systemic inflammatory diseases, are undetectable and yet the disease manifestations are dramatically reduced upon blockade of IL-1 activity. This commonly observed therapeutic response is due to the high specific activity of IL-1β, which can be in the picomolar range in humans. Therefore, establishing a role for IL-1β in inflammatory diseases has succeeded by using short-term IL-1β-blockade and its role and usefulness will likely increase with clinical testing, facilitated by the safety of short-acting anakinra and the availability of neutralizing anti-IL-1β antibodies. Supported by NIH Grants AI-15614, CA-04 6934 and JDRF 26-2008-893. The author thanks Antonio Abbate, Mihai Netea, Leo Joosten, Anna Simon and Jos van der Meer for many helpful suggestions in the preparation of this MS. Conflict of interest: The author declares no financial or commercial conflict of interest. This article is editorially independent of Novartis.

Interestingly, the marked differences between WT and CD68TGF-βDNRII mice were primarily associated with the resolution of colitic inflammation. Impairment of TGF-β responsiveness in Mϕs delayed the reduction of granulocytic inflammation, impaired IL-10 release, but increased the production of IL-33, a type 2 cytokine that is produced at high levels in the mucosa of UC patients. selleck chemicals llc Hence, TGF-β promotes the normal resolution of intestinal inflammation at least in part, through limiting the production of type 2 cytokines from colonic Mϕs. CD68

(macrosialin) encodes a type 1 transmembrane protein in mononuclear phagocyte endosomes and its promoter drives Mϕ-specific transgene expression in mice 27, 37. We demonstrate that the CD68 promoter drives transgene expression in colonic F4/80+ and F4/80+ CD11c+ populations, but is only marginally expressed in CD11c+ (specific for dendritic cells) or Gr-1+ cell populations (specific for neutrophils/granulocytes) (Fig. selleck chemicals 2) (data not shown). This is distinct from all other myeloid-specific promoters such as human CD11b, c-fms, and lysozyme that confer dendritic cell- and neutrophil-specific expression 38–40. Neutrophils promote oxidative tissue injury during DSS-induced colitis 41 and

TGF-β is known to directly modulate neutrophil function in vivo 42, which makes the lack of transgene expression in granulocytes an important issue in this model system. Our data are consistent with prior evidence that the human CD68 promoter is primarily active in mature tissue-resident Mϕ populations 43, 44. Prior to colitis induction, CD68

TGF-βDNRII mice do not have signs of overt inflammation or tissue injury. On the contrary, mice that lack STAT-3 responsiveness in Mϕs and neutrophils develop spontaneous colitis by 20 wk of age 45. As STAT-3 is an important transcription factor for IL-10 responses 46, this may suggest distinct roles for IL-10 and TGF-β in the regulation of gastrointestinal inflammation. Exacerbated intestinal immunopathology following the cessation of DSS administration in CD68 TGF-βDNRII mice was associated with an extended period of granulocyte infiltration, G-CSF production, chemokine release, and myeloperoxidase (MPO) production (data not shown). This is consistent Buspirone HCl with prior evidence in this model that excess accumulation of activated Mϕs, neutrophils, eosinophils causes irreparable mucosal damage and lethality 47, 48. Insufficient IL-10 production may partially explain the increased inflammation in CD68TGF-βDNRII mice, as IL-10-mediated suppression of colitis can be TGF-β dependent 49 and TGF-β induces Mϕs to produce IL-10 34. Furthermore, Mϕs from CD68TGF-βDNRII mice produced significantly less IL-10 following TGF-β stimulation in vitro (Fig. 1E) and in vivo (Fig. 5B and C). This link between TGF-β responsiveness in Mϕs and IL-10 production is consistent with evidence that TGF-β suppresses intestinal inflammation via regulatory Mϕs that produce IL-10 50.

High molecular weight chaperone complexes, hsp110- or grp170-tyrosinase-related protein 2 peptide (TRP2175–192), were superior to conventional chaperones as a vaccine platform to deliver tumour-derived antigens.[74] In addition, the immunization with chaperones combined to two different melanoma antigens (gp100, TRP2) significantly improved anti-tumour efficacy compared with either of the single antigen vaccines,[74] demonstrating that hsp combination vaccines can offer increased efficacy. In a Phase II clinical

trial, vaccination with autologous tumour-derived gp96–peptide complex vaccine (hsp complex-96) together with granulocyte–macrophage colony-stimulating factor and interferon-α was associated with mild local and systemic toxicity.[75] Vaccination was proven to instigate both tumour-specific T-cell-mediated and natural killer cell responses in some check details patients. However, neither immunological nor clinical responses were improved compared with those recorded in a previous study investigating hsp complex-96 monotherapy. A recent study has provided the first evidence

in man of patient-specific immune responses against autologous tumour-derived peptides bound to gp96.[76] Over-expression of hsp70 increases significantly the immunogenicity of cancer cell extracts; with the mechanism of cell death influencing both hsp70 expression levels and the immunogenicity of cell extracts.[77] In addition PAK5 to hsp complex from hsp70 (hsp70C), synthetic peptide-mimics of hsp70C can modulate positively mTOR inhibitor the immune response against tumours[78] and therefore provide an additional approach for therapeutic intervention. Heat shock protein 70 derived from tumours of characterized antigenic makeup could be used as a generic subunit tumour vaccine.[73] Vaccines derived from tumours or cell lines that have undergone heating to increase the abundance of hsp

may provide an innovative approach. For example, vaccination with heated autologous prostate cancer cells elicits protection against tumour challenge in 60% of vaccinated rats, compared with 0% protection in control rats receiving vaccines from non-shocked cells, together with an increase in the T helper type 1 (interferon-γ) response.[79] Heat shock protein 70 extracted from DC fused to patient-derived ovarian cancer or breast cancer cells (hsp70.PC-F) were tested as tumour vaccines.[80] The hsp70.PC-F induced T-cells expressing higher levels of interferon-γ and with increased killing capacity for tumour cells, compared with those induced by hsp derived from tumour cells, although these were characterized by a higher content of tumour antigens and the detection of hsp such as hsp90 and hsp110.

Two hundred and twenty-five patients have been recruited within a collaborative project (GenHomme, Research French ministry) involving the Nantes Institute of Transplantation,

the Center for Adult Transplantation of the Necker Hospital (Paris, France) and the Biotechnology Company, TcLand Expression (Nantes, France). Sixty-one additional patients were recruited in the framework of the European “Indices of Tolerance” Network. Selleckchem 3-deazaneplanocin A The protocol of the study was approved by the Ethical Committees of Nantes and Paris Universities and of the European Commission. All patients signed a written informed consent before inclusion. Several different clinical groups were studied (Table 1). Operationally tolerant patients (TOL, n=14) are defined by a stable kidney graft function (Creatininemia<150 μmol/L, Proteinuria<1 g 24 h−1) off immunosuppressive drugs for more than 1 year (mean drug-free duration=8.3±5.7 years) at the time of testing. This definition fulfills click here EU criteria for operational tolerance (for review, see 4). Immunosuppressive treatment, including corticosteroids, was

stopped on account of non-compliance (n=11), calcineurin inhibitor toxicity (n=1), post-transplant lymphoproliferative disorder (n=1) or cancer (n=1). Patients with the “suspicious” form of chronic humoral rejection (CHR, n=21) all had a progressive degradation of their renal function (Creatininemia >150 μmol/L and Proteinuria >1 g 24 h−1). In all cases, transplant renal biopsies documented histological signs of chronic humoral rejection at the time of the blood test (Banff 05 grade II or IIIb) with either C4d deposition (in 14 patients out of 21) or circulating anti-donor class II Ab in 11 out of 21 patients. Because the patients had not necessarily both circulating anti-donor class II Ab and

Bay 11-7085 C4d deposits, they were referred to as “suspicious” of chronic humoral rejection, as suggested by Banff ’07 classification 2. Long-term stable patients (n=229) comprised patients who had stable kidney graft function on immunosuppresants (either mycophenolate mofetil or azathioprine), supplemented with calcineurin inhibitors treatment in some (n=209 referred as STA) but not in others cases (n=8, referred as STN). Patients also received corticosteroids. The cohort of 209 STA patients is composed of 182 patients recruited from the GenHomme study (patients who have been transplanted at least 5 years previously) and 27 patients from the “Indices of Tolerance” network. Patients were included based on the function of their kidney graft assessed at least 5 years after transplantation (Creatininemia <150 μmol/L, Proteinuria <1 g 24 h−1). Ongoing infection and episodes of rejection defined the exclusion criteria.

For example, phosphorylation

of JIP-1 enhances its ability to bind JNK indicating a feedback mechanism that may be responsible for the changes [46, 47]. Moreover, modifications by AKT1/2 [48] and Siah1 [49] can regulate POSH function and change the composition of the complex. Our data suggests that the POSH SH3.3 domain is dispensable for TCR-mediated NF-κB Selleckchem Ku-0059436 activation in CD8+ T cells [26]. Curiously though, POSH binds TAK1, a MAP3K responsible for IKKα/β phosphorylation and NF-κB (and JNK) activation [50]. When this is considered with the role of the Carma1/Bcl10 complex in the regulation of JNK2 (and NF-κB) [28], these data suggest the intriguing possibility that POSH could have a role in regulating JNK2 and NF-κB activity through the sequestration of TAK1. Considered together, these findings provide insight into the complex mechanisms that regulate the JNK pathway. The defect in POSH/JIP-1/JNK1-dependent Eomes expression may be indicative of impaired T-cell memory Luminespib research buy [45]. The loss of Tat-POSH-treated cells between days

9 and 20 supports this idea. Eomes−/− CD8+ effector T cells are both impaired in survival and the ability to re-expand upon rechallenge [41]. Interestingly, CD8+ T cells lacking both Eomes/T-bet acquire effector functions but are unable to mount an effective antitumor response [40]. In apparent contradiction, memory numbers and function were normal in both JNK1−/− and JNK2−/− mice after infection with LCMV [16]. However, the presence of high levels of proinflammatory cytokines in the LCMV-infected mice may have compensated for the lack of JNK activation. Therefore, whether the difference in these outcomes (LCMV versus tumor) is due to the nature of the “pathogen,” the inflammatory milieu, or POSH (and or TCR) independent signals in vivo remains to be determined. Regardless, the differential expression of T-bet and Eomes strongly suggests a mechanism

to explain how the POSH/JIP-1/JNK1 complex contributes to the T-cell effector differentiation program. In summary, our data indicate that the POSH/JIP-1 scaffold complex regulates JNK1 signaling and the development of T-cell effector function. This study highlights a mechanism by which unique scaffold complexes specifically regulate different isoforms of the same protein; JNK1 uses POSH/JIP-1 while JNK2 uses the Carma1/Bcl10 scaffold complex [28]. This provides the cell Isotretinoin with multiple points of control over the JNK signal pathways. How the POSH/JIP-1 scaffold complex regulates the unique role of JNK during thymic selection, CD4+ T-cell differentiation and the role of POSH in TCR-independent activation of JNK remains an open question. Together, given the various roles of JNK in T cells, inflammatory cells, neurons, and numerous cancers, these data identify POSH as a promising therapeutic target for manipulating cell fates and function. C57BL/6, OT-I, and OT-I Rag−/− mice were maintained in our animal facilities at the University of Missouri.

5 T cells into Foxp3+ Treg cells in the PLNs accumulating in the pancreatic islets. Furthermore, tolerogenic plasmacytoid DCs (pDCs) characterized by low MHC class II molecule expression and TGF-β production are critical in the PLNs for the recruitment of Treg

cells into the pancreatic islets by inducing CXCR3 expression. PI3K inhibitor Accordingly, pDC depletion in α-galactosylceramide-treated proinsulin 2−/− NOD mice abrogates the protection against T1D. These findings reveal that upon repetitive iNKT-cell stimulation, pDCs are critical for the recruitment of Treg cells in the pancreatic islets and the prevention of T1D development. “
“Immunoproteasomes containing the IFN-inducible subunits β1i (LMP2), β2i (MECL-1) and β5i (LMP7) alter proteasomal cleavage preference and optimize the generation of peptide ligands of MHC class I molecules. Here, we report on an unexpected new function of immunoproteasome subunits

for the survival and expansion of CD4+ and CD8+ T cells during viral infection of mice. The effect of immunoproteasome subunit deficiency on T-cell survival upon adoptive transfer was most prominent for the lack of LMP7 followed by MECL-1 and LMP2. The survival of T cells in uninfected mice or the homeostatic expansion after transfer into buy BAY 73-4506 RAG-2−/− mice was not affected by the lack of the immunosubunits. Lymphocytic choriomeningitis virus (LCMV)-specific CD8+ T cells lacking LMP7 or MECL-1 started to divide after transfer into LCMV-infected mice but experienced a considerable cell loss within 2 days after transfer. We provide strong evidence that the loss of immunoproteasome-deficient T cells after transfer is not a consequence of graft rejection by the host, but instead is based on the requirement for immunoproteasomes for the survival of T cells in LCMV-infected mice. Therefore, the immunoproteasome

may qualify as a potential new target for the suppression of undesired proinflammatory T-cell responses. The proteasome Selleck Fluorouracil core complex, referred to as 20S proteasome, is a cylinder-shaped structure consisting of 28 subunits, arranged in four stacked rings. The two outer rings, each made up of seven α-type subunits (α1–α7) are framing the two inner rings, each composed of seven β-type subunits (β1–β7). The catalytic activity is performed by three β-subunits of each inner ring: β1 (δ), β2 (MC14) and β5 (MB1). In the course of an immune response, the constitutive β-subunits are replaced in newly assembled proteasomes by the IFN-γ- and TNF-α-inducible subunits β1i (LMP2), β5i (LMP7) and β2i (MECL-1) 1, thereby building so-called immunoproteasomes 2. Immunoproteasomes cleave Ag with a different cleavage preference 3, 4, thus optimizing the quantity and quality of the generated peptides for presentation by MHC class I molecules 5–8.

In this review, we aim to discuss current knowledge of intestinal (butyrate-producing) microbiota composition in obesity as well as the use of faecal transplantation using different donors to mine for beneficial intestinal bacterial strains to treat obesity and subsequent type 2 diabetes mellitus. The intestinal microbiota of the newborn human was thought to be essentially sterile, but recent data suggest that modest bacterial translocation via placental circulation antenatally is likely to provide a primitive bacterial

community to the meconium [8]. Although the new concept of fetal intestinal colonization remains controversial, recent ongoing studies using 16S rRNA gene pyrosequencing to characterize the bacterial population in meconium of preterm infants suggest that the bacteria of maternal intestine are able to cross the LY294002 chemical structure placental barrier and act as

the initial inoculum for the fetal gut microbiota [8, Poziotinib 9]. Nevertheless, the infant’s gut is only colonized fully by maternal and environmental bacteria during birth. Whereas the vaginally delivered infant’s intestinal microbial communities resemble their own mother’s vaginal microbiota (dominated by Lactobacillus, Prevotella or Sneathia spp.), newborns delivered by caesarean section harbour intestinal bacterial societies similar to those found on maternal skin surface, dominated by Staphylococcus, Corynebacterium and Propionibacterium spp. [9]. In this regard, it is interesting to note that mode of delivery (caesarean) is associated with increased risk of obesity later in life [10]. Other than the delivery mode, gestational age

at birth, diet composition and antibiotic use by the infant may have significant impacts to determine the composition of the infant’s intestinal microbial communities and body mass index (BMI) [11]. With respect to feeding pattern, the composition of intestinal bacteria differs substantially between breast-fed and formula-fed infants, which is thought to be due to the breast milk containing (prebiotic) oligosaccharides [12, 13]. The subsequent transformation of the intestinal microbiota from infant- to adult-type is triggered via bidirectional cross-talk between Janus kinase (JAK) host and predominantly dietary and environmental factors [12, 14], but remains relatively stable until the 7th decade of life [15]. It is thus likely that host (immunological) responses to inhabitant commensal bacteria differ from those elicited towards pathogens that do not belong to the indigenous microbiota [16, 17]. The precise mechanisms of how intestinal microbes affect and protect host immune physiology, however, are yet to be revealed. There is now solid evidence that composition of the intestinal microbiota is altered in obese people on a western diet compared to lean [18, 19]. Moreover, dietary composition seems to be one the most important determinants of intestinal microbiota diversity driving obesity [20, 21].

2a). Interestingly, no production or secretion of FhaB was detected Torin 1 ic50 under the iron-starved conditions (Fig. 2b). On the other hand, production and secretion of CyaA, Prn, and DNT were not significantly affected by the iron concentration (Fig. 2b). These results clearly indicate that BvgAS-regulated gene expression is not always enhanced by iron-starved conditions. To further investigate BvgAS-regulated gene expression

under iron-starved conditions, total RNA was prepared from B. bronchiseptica cultured under iron-replete or -depleted conditions. The cDNA samples reverse-transcribed from the total RNA samples were subjected to quantitative RT-PCR analysis to quantify the relative amounts of bsp22 and fhaB mRNA as a hallmark of the BvgAS-regulated

gene that is positively or negatively regulated by iron-starved conditions (Fig. 3). The Bsp22 gene was transcriptionally activated by iron starvation. In contrast, the fhaB gene was repressed in response to iron starvation, demonstrating that the relative amounts of mRNAs are correlated with protein production, as shown in Fig. 2b. It has been reported that B. bronchiseptica induces necrotic cell death of various mammalian cultured cells in a T3SS-dependent manner (6, 8). To examine whether this phenotype is affected by iron-depleted conditions, L2 rat lung epithelial cells infected with B. bronchiseptica precultured under iron-replete or -depleted conditions https://www.selleckchem.com/products/Neratinib(HKI-272).html were fixed and stained with Giemsa solution to analyze cell morphology (Fig. 4a). Approximately 60–70% of cells infected with B. bronchiseptica under iron-replete conditions were detached from the substrata and the remainder of adherent cells

exhibited shrunken cytoplasm and condensed nuclei (Fig. 4a). The L2 cells exposed to the T3SS mutant strain showed normal morphology that was identical to that of uninfected cells. In contrast, more than 90% of cells infected with B. bronchiseptica under iron-depleted conditions were detached, and their morphological changes were more pronounced than those of bacteria cultured under iron-replete conditions. Furthermore, HeLa cells were infected with B. bronchiseptica and the relative amounts of LDH released into the extracellular medium measured (Fig. 4b). The cytotoxicity evident in host see more cells infected with B. bronchiseptica under iron-depleted conditions was statistically greater than that of those infected with B. bronchiseptica under iron-replete conditions. T3SS-dependent hemolytic activity was also evaluated using RBCs (Fig. 4c). Again, hemolytic activity of B. bronchiseptica grown under iron-depleted conditions was statistically greater than that of B. bronchiseptica grown under iron-replete conditions. Collectively, these results suggest that B. bronchiseptica is able to recognize iron-starved conditions and exert the T3SS function in response to them.

Regarding protein homogeneity, the preparations of rK9 and rK26 showed at least one significant protein impurity as verified by SDS-PAGE, and such recombinant antigens were assayed by immunoblotting against a Leishmania infected human panel. The proteins K9 + K39 were analysed by ELISA using a canine serum panel (20 positive and 20 negative sera), and the values of SP (100%) and SE (95%) https://www.selleckchem.com/products/epz-6438.html obtained were identical

to those found for rLci2B. ELISA performed with rLci2B employed a higher number of canine serum samples (138 positive and 119 negative sera) than that used in K9 + K39 immunological assay. The comparison between chimera K9-K39-K26 and rLci2B, in respect to ELISA values, shows that for rLci2B, the SE values were superior (100% vs. 95%), while the SP values were inferior (95% vs. 100%). However, it should be noted that the construction of the chimera K9-K39-K26 with two tags

is a difficult task, and the chimera recovery was low and estimated at approximately 10 mg/L bacterial culture Ganetespib (34). Considering the number of serum samples tested using rLci2B and the chimera K9-K39-K26 as being statistically consistent, the values obtained in this study are significant especially those related to the SE parameter (100%) that eliminates the false negative cases. On the other hand, the value of SP equal to 100% obtained for the chimera protein minimizes the false positive nearly cases. Therefore, the ELISA results obtained for both proteins, mainly rLci2B and the chimera K9-K39-K26, can be considered excellent as commented by Chappuis et al. (20). The recombinant proteins rLci2B and rLci1A did not show cross-reactivity with serum samples of dogs infected with T. caninum, B. canis and E. canis, although cross-reactivity has been observed in serum samples obtained from dogs infected with L. brasiliensis, a parasite responsible for American Cutaneous

Leishmaniasis (ACL) (Table 1). The cross-reactivity for rLci2B (11·7%) and rLci1A (2·9%) observed with L. brasiliensis (n = 34) infected sera is probably due to the fact that this parasite belongs to the same genus of L. chagasi. For canine VL, the sacrifice of dogs positive for ACL is also recommended because there is no effective treatment and the animal also constitutes an important reservoir of this disease (35). In conclusion, based on data obtained from protein recovery (rLci2B: 105 mg/L and rLci1A: 225 mg/L bacteria cultures), protein purity and sensibility/specificity values, both proteins can be proposed as alternative antigens for Leishmania serological assay. We thank the researchers of Centro de Pesquisa Aggeu Magalhães, Pernambuco and Centro Gonçalo Muniz, Bahia, Brazil, especially to Dr. Geraldo G. Oliveira, for the donation of the modified E. coli plasmids containing the genes concerning the recombinant proteins rLci2B and rLci1A. We would also want to thank Dr.