Current challenges and opportunities for bacillus Calmette-Guérin replacement vaccine candidates.

Bacillus Calmette-Guérin (BCG) remains the only licensed vaccine against human tuberculosis (TB). BCG is a live-attenuated strain of Mycobacterium bovis, with limitations in efficacy against respiratory TB, the most common form of the disease responsible for transmission. However, continues to be used in the immunization programmes of different countries in the absence of another alternative. In order to improve BCG efficacy against pulmonary TB, in the current clinical TB vaccine pipeline, there are live-attenuated TB vaccines to replace BCG. This review discusses the current status of the development of live vaccine candidates designed to replace BCG from the rational strategies and immunological challenges to its clinical trial and identify key areas in the next years considered essential to confer improved safety and efficacy over BCG.

Novel vaccination strategies and approaches against human tuberculosis.

Tuberculosis (TB) remains one of a major health problem worldwide. Tuberculosis vaccine research has made an extraordinary progress over the past few years. However, there is still no replacement for the Bacillus Calmette-Guérin vaccine, the only TB vaccine licensed for human use. Therefore, the discovery and development of new TB vaccines remains a priority. This article discusses current strategies used to diversify TB vaccines and includes discussion of the status of efforts to improve protection against Mycobacterium tuberculosis (M tb) infection or TB disease by developing new and safe TB vaccines. This article also highlights the current research efforts in immune-enhancing approaches to improve vaccination efficacy. The development of more effective TB vaccines might have significant impact on global TB control.

Development of tuberculosis vaccines in clinical trials: Current status.

Tuberculosis (TB) is an important infectious disease worldwide. Currently, Bacillus Calmette-Guérin (BCG) remains the only TB vaccine licensed for human use. This TB vaccine is effective in protecting children against severe military TB but offers variable protective efficacy in adults. Therefore, new vaccines against TB are needed to overcome this serious disease. At present, around 14 TB vaccine candidates are in different phases of clinical trials. These TB vaccines in clinical evaluation can be classified into two groups including preventive pre- and post-exposure vaccines: subunit vaccines (attenuated viral vectors or adjuvanted fusion proteins), and whole-cell vaccines (genetically attenuated Mycobacterium tuberculosis (M. tb), recombinant BCG, killed M. tb or M. vaccae). Although, over the last two decades a great progress in the search for a more effective TB vaccine has been demonstrated there is still no replacement for the licensed BCG vaccine. This article summarizes the current status of TB vaccine development and identifies crucial gaps of research for the development of an effective TB vaccine in all age groups.

Molecular events by which dendritic cells promote Th2 immune protection in helmith infection.

Helminth parasites are a major cause of global infectious diseases, affecting nearly one quarter of the world's population. The common feature of helminth infections is to skew the immune system towards a T-helper 2 (Th2) response that helps to control disease. Dendritic cells (DCs), which are professional antigen-presenting cells, play a critical role for Th2 skewing against helminth parasites. However, the molecular mechanisms by which helminth antigens activate DCs for Th2 polarization have not yet been clearly defined. This review provides a focused update on the major role of DCs for inducing and/or enhancing Th2 immune responses in helminthic infection and will discuss the main signalling-dependent and independent mechanisms by which helminth antigens activate DCs for Th2 skewing.

Peptidomimetics as a new generation of antimicrobial agents: current progress.

Antibiotic resistance is an increasing public health concern around the world. Rapid increase in the emergence of multidrug-resistant bacteria has been the target of extensive research efforts to develop a novel class of antibiotics. Antimicrobial peptides (AMPs) are small cationic amphiphilic peptides, which play an important role in the defense against bacterial infections through disruption of their membranes. They have been regarded as a potential source of future antibiotics, owing to a remarkable set of advantageous properties such as broad-spectrum activity, and they do not readily induce drug-resistance. However, AMPs have some intrinsic drawbacks, such as susceptibility to enzymatic degradation, toxicity, and high production cost. Currently, a new class of AMPs termed "peptidomimetics" have been developed, which can mimic the bactericidal mechanism of AMPs, while being stable to enzymatic degradation and displaying potent activity against multidrug-resistant bacteria. This review will focus on current findings of antimicrobial peptidomimetics. The potential future directions in the development of more potent analogs of peptidomimetics as a new generation of antimicrobial agents are also presented.

Isolation of IgG antibodies to Toxocara in ankylosing spondylitis patients with acute anterior uveitis.

PURPOSE: Since few reports had been published on the prevalence of toxocariasis in ankylosing spondylitis (AS) patients with acute non-granulomatous anterior uveitis (ANGAU), the aim of this work was to determine the presence of antibodies against Toxocara canis in AS patients with ANGAU. METHODS: Thirty-six patients (14 female and 22 male) with AS were enrolled in the study. The history of ANGAU was accepted only if diagnosed by an ophthalmologist. The detection of IgG antibodies to T. canis was determined by enzyme-linked immunosorbent assay. In addition, antibodies to Ascaris lumbricoides were also tested to verify non-specific reactions. RESULTS: The prevalence of ANGAU in the AS patients was 58% (21 / 36), and 38% (8 / 21) of the patients with ANGAU were positive for antibodies to Toxocara, while 7% (1 / 15) of AS patients without ANGAU were positive for T. canis (p = 0.038, two tails; mid-p exact). No antibodies were detected to A. lumbricoides antigens in the serum samples of patients with AS. CONCLUSIONS: These data suggest that the seroprevalence of antibodies to T. canis is high in Mexican patients with AS-associated uveitis, suggesting a chronic asymptomatic toxocariosis, which could be associated with the pathogenesis of ANGAU; however, further larger-scale studies are needed to confirm this observation.

Recent advances in the field of antimicrobial peptides in inflammatory diseases.

Antimicrobial peptides are cationic molecules, which participate in multiple aspects of the immune response including the control of inflammatory diseases, characteristic that make these molecules attractive as therapeutic tools. These peptides are produced in bacteria, insects, plants and vertebrates, and are classified together due to their capacity to directly inhibit the growth of microorganisms, and to regulate the immune response by inducing the secretion of chemokines and cytokines. Various families of antimicrobial peptides have been identified including the cathelicidins and defensins, the most investigated human antimicrobial peptides. This review will cover the main biological functions of antimicrobial and cell-penetrating peptides in inflammation, and describe the importance and utility of antimicrobial peptides as therapeutics for inflammatory diseases.

Immunological mechanisms by which concomitant helminth infections predispose to the development of human tuberculosis.

Helminthic infections afflict over 1.5 billion people worldwide, while Mycobacterium tuberculosis infects one third of the world's population, resulting in 2 million deaths per year. Although tuberculosis and helminthic infections coexist in many parts of the world, and it has been demonstrated that the T-helper 2 and T-regulatory cell responses elicited by helminths can affect the ability of the host to control mycobacterial infection, it is still unclear whether helminth infections in fact affect tuberculosis disease. In this review article, current progress in the knowledge about the immunomodulation induced by helminths to diminish the protective immune responses to bacille Calmette-Guerin vaccination is reviewed, and the knowledge about the types of immune responses modulated by helminths and the consequences for tuberculosis are summarized. In addition, recent data supporting the significant reduction of both M. tuberculosis antigen-specific Toll-like receptor (TLR) 2 and TLR9 expression, and pro-inflammatory cytokine responses to TLR2 and TLR9 ligands in individuals with M. tuberculosis and helminth co-infection were discussed. This examination will allow to improve understanding of the immune responses to mycobacterial infection and also be of great relevance in combating human tuberculosis.

The human cathelicidin hCAP18/LL-37: a multifunctional peptide involved in mycobacterial infections.

Antimicrobial peptides are predominantly small cationic polypeptides that are classified together on the basis of these molecules to directly kill or inhibit the growth of microorganisms including mycobacteria, and to activate mechanisms of cellular and adaptive immunity. Various families of antimicrobial peptides have been identified, including the cathelicidins. The cathelicidin family is characterised by a conserved N-terminal cathelin domain and a variable C-terminal antimicrobial domain that can be released from the precursor protein after cleavage by proteinases. LL-37 is the C-terminal part of the only human cathelicidin identified to date called human cationic antimicrobial protein (hCAP18), which is mainly expressed by neutrophils and epithelial cells. The cathelicidin hCAP18/LL-37 is a multifunctional molecule that may mediate various host responses, including bactericidal action, chemotaxis, epithelial cell activation, angiogenesis, epithelial wound repair and activation of chemokine secretion. The antimicrobial peptide LL-37 is produced from human cells during infection of mycobacteria and exerts a microbicidal effect. The discussion will (1) describe recent work on the antimicrobial and immunomodulatory functions of the cathelicidin hCAP18/LL-37, (2) highlight the effectiveness of the cathelicidin hCAP18/LL-37 as a potent component in antimycobacterial immune responses and (3) summarise current progress in the understanding of the therapeutic application of hCAP18/LL-37 and its derivates antimicrobial peptides in mycobacterial infection.

Reactive oxygen species-activated p38/ERK 1/2 MAPK signaling pathway in the Mycobacterium bovis bacillus Calmette Guérin (BCG)-induced CCL2 secretion in human monocytic cell line THP-1.

BACKGROUND AND AIMS: CCL2 plays an important role in mycobacterial infection by inducing leukocyte recruitment and activation. Here we assess the role of reactive oxygen species (ROS) in the secretion of the CCL2 and the activation of mitogen-activated protein kinases (MAPKs) by human monocytic cells infected with Mycobacterium bovis bacillus Calmette Guérin (BCG). METHODS: CCL2 mRNA and protein expression were measured by reverse transcriptase polymerase chain reaction (RT-PCR), quantitative PCR and ELISA. Kinase phosphorylation was determined by immunoblotting. RESULTS: Treatment of human monocytic cells with M. bovis BCG activated rapid superoxide generation. mRNA expression of CCL2 was increased in M. bovis BCG-infected monocytic cells, and this increase was abrogated by administration of the nicotinamide adenine dinucleotide phosphate (NADPH) oxidase inhibitor diphenyleneiodonium (DPI). Importantly, M. bovis BCG-induced CCL2 protein secretion was also inhibited by the NADPH oxidase inhibitor DPI, the selective inhibitor of NADPH oxidase apocynin, the mitochondrial electron transfer chain subunit I inhibitor rotenone and H(2)O(2) scavenging enzyme catalase, indicating that the inhibition is through the NADPH/ROS pathway. Analysis of downstream signals showed that inhibition of NADPH oxidase inhibited M. bovis BCG-induced phosphorylation of MAPK (extracellular signal-regulated kinase (ERK) 1/2 and p38). CONCLUSIONS: These results strongly suggest that NADPH oxidase-derived ROS-mediated activation of p38 and ERK 1/2 is essential for the M. bovis BCG-induced CCL2 production.

Role of interleukin-12 family cytokines in the cellular response to mycobacterial disease.

Interleukin (IL)-12 is a multifunctional cytokine acting as a key regulator of cell-mediated immune responses through the differentiation of naïve CD4+ T cells into type 1 helper T cells (Th1) producing interferon-gamma. As our knowledge of IL-12 family members is rapidly growing, it will be important to specify their involvement in the regulation of mycobacterial infection. This article is a review of the current knowledge regarding the functions of the IL-12 family cytokines in the immune host defense system against mycobacteria. Specifically, this review aims to describe recent scientific evidence concerning the protective role of some members of the IL-12 family cytokines for the control of mycobacterial infection, as well as to summarize knowledge of the potential use of the IL-12 family members as potent adjuvants in the prevention and treatment of mycobacterial infectious diseases. In addition, recent data supporting the importance of the IL-12 family members in mycobacterial diseases in relation to Th17 function are discussed. This examination will help to improve our understanding of the immune response to mycobacterial infection and also improve vaccine design and immunotherapeutic intervention against tuberculosis.

Role of reactive oxygen species (ROS) in Mycobacterium bovis bacillus Calmette Guérin-mediated up-regulation of the human cathelicidin LL-37 in A549 cells.

The human cathelicidin LL-37 is one of the major antimicrobial peptides of the non-specific innate immune system in Mycobacterium tuberculosis infection. Its expression has been reported in epithelial cells infected with mycobacteria. However, the underlying molecular mechanisms by which Mycobacterium bovis bacillus Calmette-Guérin (BCG) triggers gene transcription of cathelicidin have not been elucidated. The objective of this study was to investigate the role of reactive oxygen species (ROS) in the M. bovis BCG-mediated up-regulation of the antimicrobial peptide cathelicidin LL-37 in human epithelial cells. Infection of A549 cells with M. bovis BCG led to a rapid ROS production. Importantly, blockade of ROS by preincubation of cells with the general ROS scavenger N-acetyl-l-cysteine (NAC) or the NADPH oxidase inhibitor DPI significantly reduced M. bovis BCG-induced up-regulation of cathelicidin LL-37 mRNA expression as determined by semi-quantitative RT-PCR or real-time PCR. In contrast, the xanthine oxidase inhibitor allopurinol did not affect M. bovis BCG-mediated up-regulation of cathelicidin LL-37 mRNA. Moreover, M. bovis BCG-mediated cathelicidin LL-37 mRNA expression was significantly blocked by the effect of the mitochondrial electron transfer chain subunit I inhibitor rotenone and H(2)O(2) scavenging enzyme catalase. In addition, M. bovis BCG-induced cathelicidin LL-37 protein secretion was inhibited by the addition of NAC, DPI, and the selective inhibitor of NADPH oxidase apocynin. Our results collectively indicate that M. bovis BCG-mediated up-regulation of cathelicidin is influenced by NADPH/ROS signaling pathways. In conclusion, these findings demonstrate a novel regulatory mechanism for the expression of cathelicidin LL-37 in human epithelial cells stimulated with M. bovis BCG.

Mycobacterium bovis Bacillus Calmette-Guérin (BCG)-induced activation of PI3K/Akt and NF-kB signaling pathways regulates expression of CXCL10 in epithelial cells.

CXCL10 production is a critical step in limiting mycobacterial infection. Although induction of this chemokine by mycobacteria in epithelial cells has been reported, it is still unclear how CXCL10 is regulated in Mycobacterium bovis BCG-infected epithelial cells. In this study, we demonstrate that phosphatidylinoditol 3-kinase (PI3K)/Akt and the nuclear factor kB (NF-kB) signaling pathways play an important role in CXCL10 expression at the protein and mRNA level in A549 cells. We demonstrate that treatment of A549 cells with LY294002 and wortmannin, two PI3K inhibitors, inhibited M. bovis BCG-induced CXCL10 expression. In addition, treatment of A549 cells with an Akt inhibitor significantly blocked M. bovis BCG-induced CXCL10 production. Moreover, our data show that treatment of epithelial cells with CAPE, BAY 11-7082, and PDTC three selective inhibitors of NF-kB, significantly reduced the effect of M. bovis BCG on induced CXCL10 mRNA expression (74%, 69% and 83% inhibition by 8microM CAPE, 10microM BAY 11-7082 and 3microM PDTC as assessed by real-time PCR, respectively). In accordance with the gene induction, the production of CXCL10 was also significantly reduced by these inhibitors. Finally, the inhibition of PI3K affect NF-kB activation in M. bovis BCG-infected cells, indicating that PI3K activity is required for the M. bovis BCG-induced activation of NF-kB. The functional association between PI3K/Akt and NF-kB demonstrates another mechanism in the regulation of M. bovis BCG-induced CXCL10 in A549 cells.

Expression and regulation of chemokines in mycobacterial infection.

Chemokines are the key molecules that recruit immune cells by chemotaxis and act in leukocyte activation during mycobacterial diseases. Currently, tuberculosis is a leading infectious disease affecting millions of people worldwide. The purpose of this review is to describe a series of recent scientific evidence concerning to the protective role of some members of the CC- and the CXC chemokine subfamilies for the control of mycobacterial infection. The discussion will (1) highlight the effectiveness of some chemokines as potent immunoprophylactic tool for controlling the mycobacterial establishment within the host, (2) describe recent work on the relevance of cellular signaling pathways by which mycobacterial antigens mediate chemokine induction, and (3) summarize current progress in the understanding of the potential use of chemokines as potent adjuvants in antimycobacterial immune responses.

Regulation of human beta-defensin-2 by Mycobacterium bovis bacillus Calmette-Guérin (BCG): involvement of PKC, JNK, and PI3K in human lung epithelial cell line (A549).

Human beta-defensin (HBD)-2 is an inducible antimicrobial peptide that plays an important role in innate immunity. Induction of this peptide by mycobacteria in epithelial cells has been reported. However, the mechanism(s) by which Mycobacterium bovis bacillus Calmette-Guérin (BCG) triggers gene transcription of HBD-2 remains poorly understood. In the present work we found that treatment of human epithelial cells with Ro32-0432 or Gö6976, two selective inhibitors of protein kinase C (PKC), significantly reduced the effect of M. bovis BCG on induced HBD-2 mRNA expression (65 and 80% inhibition by 10microM Ro32-0432, and 1microM Gö6976 as assessed by real-time PCR, respectively). Moreover, there was increased activation of c-Jun N-terminal kinase (JNK) and phosphatidylinositol-3-kinase (PI3K)/Akt in A549 cells infected with M. bovis BCG, and this JNK and PI3K activation was mediated through PKC. Finally, we found that M. bovis BCG-induced HBD-2 mRNA gene expression in A549 cells was dependent on JNK, and PI3K determined by real-time PCR analysis, which was attenuated by inhibitors of JNK (SP600125 and AG126), and PI3K (wortmannin and Ly294002). These studies are the first to show that M. bovis BCG-induced HBD-2 mRNA expression in A549 cells is regulated at least in part through activation of signaling proteins of PKC, JNK and PI3K.

Role of antimicrobial peptides in host defense against mycobacterial infections.

Worldwide, tuberculosis remains the most important infectious disease causing morbidity and death. Currently, at least one-third of the world's population is infected with Mycobacterium tuberculosis. In addition, the World Health Organization estimates that about 8-10 million new tuberculosis cases occur annually worldwide and this incidence is currently increasing. Moreover, multidrug-resistant tuberculosis has been increasing in incidence in many areas during the past decade. These situations underscore the importance of the development of new therapeutic agents against mycobacterial infectious diseases. In this article, it is review current progress in the understanding of antimicrobial peptides as potential candidates to develop an alternative/adjunct therapeutic strategy against tuberculosis. This immunoadjunctive therapy might be evaluated in the context of possible drug resistance. This review also summarizes the knowledge about the functions of antimicrobial peptides in the pulmonary innate host defense system and their role in mycobacterial infection, and at the same time outlines recent advances in our understanding of the combined effect of antimicrobial peptides and anti-tuberculosis drugs against intracellular mycobacteria. A concerted effort should now focus on the clinical application of antimicrobial peptides for their practical use.