Characteristic PML lesions have been described as large, subcortical, grey-matter-sparing lesions appearing hyperintense on T2 and fluid-attenuated inversion recovery and hypointense

on T1 scans; contrast enhancement may occur [47]. The anti-CD52 mAb alemtuzumab (Lemtrada®) has been shown to be highly effective and is approved for active relapsing MS in Europe [10-12, 69]. Disease activity is defined as clinical or radiological deterioration [70]. Mechanisms of action include depletion of CD52-expressing T/B lymphocytes, natural killer (NK) cells, dendritic cells and monocytes/macrophages with skewed repopulation leading to a reprogramming of the immune repertoire [71, 72]. Already in earlier studies, patients especially with an early relapsing disease course appeared

to benefit most from alemtuzumab MEK inhibitor treatment, leading to the concept of a therapeutic window relatively early during the disease, when highly active immunotherapy may exert most profound effects [72]. This was reflected in the inclusion criteria for the pivotal Phase III trials selleck compound CARE-MS I and II (Comparison of Alemtuzumab and Rebif® Efficacy in Multiple Sclerosis, Studies One and Two). CARE-MS I included active relapsing, therapy-naive MS patients, whereas CARE-MS II focused on relapsing MS refractory to first-line therapy [10, 12]. Especially in terms of disease progression, the latter patient group appeared to benefit most. Whereas current EMA approval is relatively broad [70], careful patient selection

is mandatory, as SADRs have been reported and thorough adherence to safety assessments is necessary. This is stressed by long-term data from the Phase II trial CAMMS223, with one additional SADR (Goodpasture syndrome), but also sustained reduction of disability accumulation and relapse rates compared to active comparator [73], revealing the dilemma of long-lasting efficacy versus potential SADRs. Alemtuzumab is applied intravenously with a first treatment cycle of 12 mg over 5 days, followed by a second therapy cycle over 3 days after 12 months [10, 12, 69]. Further cycles are not intended, but the question of when and how to continue DMD treatment after two cycles is unanswered. There is no class I evidence for different treatment protocols in this indication. During and for 1 month after treatment, acyclovir (200 mg twice daily) has to be administered prophylactically. Florfenicol Therapy surveillance with large treatment intervals, but necessarily close safety monitoring, will be a challenge in clinical practice [74] and emphasizes even more the importance of patient education, counselling and informed consent to assure adherence to safety measures. These include differential blood count, serum creatinine and urine analysis before first administration and monthly afterwards; regular testing of thyroid stimulating hormone (TSH) levels has to be performed before treatment initiation and every 3 months up to 4 years after the last administration [70].

Sections from lungs were excised and stained by the Ziehl–Neelsen technique for identification of acid-fast bacilli in the tissue. AMM + AMH vaccine resulted in the lowest number of acid-fast bacteria (data not shown), which is consistent with the CFU data. HE stain showed that the granuloma areas per section of the lungs from mice immunized with BCG or boosted with fusion proteins AMM, AMH or AMM + AMH

were smaller (P < 0.05) compared with PBS. There was no difference 3-deazaneplanocin A in vivo between the three fusion protein boosting groups and BCG-immunized group (Fig. 5), indicating that boosting with the fusion protein vaccines did not aggravate pathology. In this research, we constructed a fusion protein AMH, which included the protective antigen HspX highly expressed in dormant stage of bacteria. Mice immunized with AMH subunit vaccine generated high levels of antigen-specific antibodies and IFN-γ-producing lymphocytes. AMH combined with AMM could enhance the BCG-primed immune protection against M. tuberculosis infection in mice. Dormant bacteria exist

together with replicating bacteria in vivo in human and animal infection [2–4] (Fig. 6). Central to the success of M. tuberculosis as a pathogen is its ability to persist within humans for long periods of time in a latent state [2–4]. BCG is the most widely used vaccine, but it is not sufficient to prevent latent TB or prevent reactivation in adult life [18]. The antigens selleck chemicals of subunit vaccines which were aimed to boost BCG-primed immunity were chosen frequently from secreted proteins in early and log growth phase of bacteria based on in vitro culture and are insufficient to impart sufficient immunity against the latent infection where some bacteria are in dormant state [19]. Therefore, it is potentially important for the subunit vaccines to consist of antigens in multiple stages from active multiplication to non-replicating

dormancy so as to have a maximum impact on all stages of M. tuberculosis infection [2]. Different antigens ioxilan are expressed in different growth stages. Ag85B, Mtb8.4 and MPT64 are main antigens of bacteria in replicating stage, whereas HspX is the protein that is mainly expressed in dormant phase (Fig. 6). Some latency antigens were detected up-regulated in non-replicating conditions [10]. For example, DosR, the hypoxia-related transcriptional regulator, and its genes are up-regulated under conditions closer to in vivo infection and prepare M. tuberculosis for dormancy [2]. Among them, HspX is the first gene to be identified as being induced by hypoxia and has been identified as an important latency antigen [10–13] (Fig. 6). HspX was a major membrane protein in virulent M. tuberculosis [20]. M. tuberculosis and M. bovis have increased thickness of their cell walls which contain large amounts of HspX under low oxygen conditions.

Cells were analyzed by a FACScan equipped with Cell Quest software (Becton Dickinson,

Mountain View, CA, USA). CXCL10, CCL2, and CXCL8 were measured with OptEIA™ kits (BD Pharmingen), TGF-beta inhibitor in cell-free supernatants [sups] from resting or stimulated keratinocyte cultures, according to the manufacturer’ protocols. The plates were analyzed in an ELISA reader mod. 3550 UV Bio-Rad. Results are given as ng/106 cells ± SD. Skin biopsies were minced with a scalpel and placed in culture in complete medium plus 60 U/mL IL-2. After 2–5 days, T cells emigrated from tissue samples were collected for phenotypic and functional characterization and T-cell cloning by limiting dilution (0.6 cells per well), in the presence of irradiated allogeneic feeder cells plus 1% PHA. Subconfluent keratinocytes seeded in culture slides (BD Biosciences) were pretreated with the indicated concentrations of peptides and, then, stimulated with IFN-γ. After 24 h, cells were cocultured with 5 × 105 CFSE-stained autologous T cells clones. In blocking experiments, keratinocytes were incubated for 1 h with 10 μg/mL anti-CD54 prior to cocultures with effector T cells. After 6 h, cocultures were extensively washed in PBS, fixed in 4% paraformaldehyde, and counterstained with hematoxylin. T cells

that adhered to keratinocytes were counted in 20 casual fields for each LY2606368 datasheet condition, as fluorescent dots using a fluorescent microscope (Zeiss, Oberkochen, Germany), and average T-cell number per square millimeter ± SD was calculated. Complete RPMI with 0.5% BSA alone or supernatants (sups) from untreated or 24 h IFN-γ-stimulated keratinocyte cultures

(0.6 mL total amount) were added to the bottom chamber of 24-well Transwell chambers with uncoated 5 mm pore polycarbonate filters (Corning Chlormezanone Costar, Cambridge, MA). T autologous cells were resuspended in complete RPMI with 0.5% BSA, and 0.1 mL cell suspension (106 cells/mL) was added to the top chamber. Transwells were then incubated for 1 h at 37°C with 5% CO2. The number of cells transmigrated to the lower chamber relative to the input was measured with a FACScant by 60 s acquisition at a flow rate of 100 mL/min. The experiments were carried out in triplicate for each condition and the results are given as ng/106 cells ± SD. Five-millimeter punches of normal human skin from three healthy donors undergoing to plastic surgery. Biopsies were taken after informed consent and the study was approved by the Ethical Committee of the Istituto Dermopatico dell’Immacolata (IDI)–IRCCS (Rome, Italy). Biopsies were placed in Keratinocye Basal Medium with 0.1% normal human serum in a humidified incubator at 37°C, with enough medium to just cover the explants.

Neuroblastoma cells expressing mSOD1 had increased cytoplasmic calcium levels and a significant decrease in mitochondrial membrane potential [85]. Studies of brain,

Osimertinib molecular weight spinal cord and liver mitochondria isolated from mSOD1 transgenic mice demonstrated an early decrease in the calcium buffering capacity of the mitochondria from the brain and spinal cord, leading to reduced membrane potential and dysfunctional mitochondria [60]. After challenge with calcium, mitochondria underwent less efficient repolarization, consistent with defective calcium buffering in the presence of mSOD1, which could sensitize motor neurones to excitotoxic stress and eventual death [60]. G93A mice crossed with mice genetically modified to have a decreased calcium permeability of alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid (AMPA) receptors in the spinal motor neurones showed a significant delay in the onset of the ALS phenotype [86]. The trigger for this early increase in calcium levels in Selleck Midostaurin motor neurones requires resolution. In SALS, it could potentially be attributed to decreased expression of the glutamate transporter, Excitatory Amino Acid Transporter 2 (EAAT2) [87,88]. Additionally, motor neurones normally have a low expression of GluR2 and thus a higher percentage of calcium permeable

AMPA receptors compared to other neuronal groups, and reduction in the normal editing of the GluR2 subunit may further increase AMPA receptor calcium permeability in motor neurones in ALS [89]. Thus, excessive glutamate stimulation of the calcium-permeable AMPA receptor occurs, emphasizing the need for efficient calcium buffering in motor neurones. In FALS, studies in mice have revealed that mSOD1 interacts with AMPA receptors, altering both their expression patterns and function, rendering them more permeable to calcium [90]. Furthermore, the presence of mSOD1 leads to selective loss of EAAT2 expression, specifically in areas of neurodegeneration [91]. In mSOD1 mice, excessive glutamate application was found to be toxic to Resveratrol the neurones, consistent with decreased calcium buffering in motor neurones [74,78,92]. Motor neurones also have reduced expression of cytosolic calcium

buffers, such as parvalbumin and calbindin; thus, motor neurone mitochondria may play a more pivotal role in the buffering of cytosolic calcium [5,44,93]. Although not sufficient in itself to induce excitotoxic cell death, in the presence of mSOD1, any physiological calcium influx will serve to exacerbate mitochondrial dysfunction in the cell, resulting in the eventual degeneration of the motor neurone [5]. Furthermore, at the neuromuscular junction, mitochondria in the synapse of motor neurones show greater membrane potential depolarization in G85R and G93A mice compared to controls [94]. This is linked to a reduced capacity of the ETC to limit depolarization and correlates with onset and progression of ALS symptoms at the motor neurone terminals.

glabra, respectively, did have anti-HCV activity, their IC50 being 2.5 and 6.2 μg/mL, respectively. Another chalcone, isoliquiritigenin, also showed anti-HCV activity, with an IC50 of 3.7 μg/mL. Time-of-addition analysis revealed that all Glycyrrhiza-derived anti-HCV compounds tested in this study act at the post-entry step. In conclusion, the present results suggest that glycycoumarin, glycyrin, glycyrol and liquiritigenin isolated from G. uralensis, as well as isoliquiritigenin, licochalcone

A and glabridin, would be good AP24534 molecular weight candidates for seed compounds to develop antivirals against HCV. “
“OTHER THEMES PUBLISHED IN THIS IMMUNOLOGY IN THE CLINIC REVIEW SERIES Metabolic Diseases, Host Responses, Allergies, Autoinflammatory Diseases, Type 1 diabetes and viruses. Despite complex genomic and epigenetic abnormalities,

many cancers are irrevocably dependent on an initiating oncogenic lesion whose restoration to a normal physiological activation can elicit a dramatic and sudden reversal of their neoplastic properties. This phenomenon of the reversal of tumorigenesis has been described as oncogene addiction. Oncogene addiction had been thought to occur largely through tumour cell-autonomous mechanisms such as proliferative arrest, apoptosis, differentiation and cellular senescence. However, the immune system plays an integral role in PD0332991 solubility dmso almost every aspect of tumorigenesis, including tumour initiation, prevention and progression as well as the response to therapeutics. Here we highlight more Tryptophan synthase recent evidence suggesting that oncogene addiction may be integrally dependent upon host immune-mediated mechanisms, including specific immune effectors and cytokines that regulate

tumour cell senescence and tumour-associated angiogenesis. Hence, the host immune system is essential to oncogene addiction. Oncogene addiction is the phenomenon by which even highly complex tumour cells that are a consequence of multiple genetic and epigenetic changes become exquisitely dependent upon a single oncogene for their continued growth and survival [1,2]. Early studies illustrated that, in tumour cells, the in vitro suppression of an oncogene or the restoration of expression of a tumour suppressor could be sufficient to induce the sustained loss of their neoplastic features [3]. More recently, conditional transgenic mouse models have been used to explore the tumour-specific consequences of the suppression of oncogenes including MYC, RAS, BRAF and BCR-ABL[4–10]. The specific consequences of oncogene inactivation in a tumour are dependent upon cellular and genetic context and can include proliferative arrest, apoptosis [4], differentiation [5,6] and senescence [11] as well as the inhibition of angiogenesis [12,13].

63 A major component in the generation of systemic inflammatory stress is the activation of the nuclear transcription factor-κB (NF-κB). There is an interaction between the VDR and NF-κB,64 with stimulation of the VDR downregulating NF-κB signalling,65 and results in a reduction of activated Pexidartinib T-cell and Antigen

Presenting Cell activity. Various studies have further demonstrated 1,25-OHD’s ability to decrease expression of pro-inflammatory cytokines (including CRP, IL-6, TNFα) both in vitro and in vivo.28,66 In a mouse model of renal inflammation Tan et al. demonstrated that administration of paricalcitol (an analogue of 1,25-OHD) resulted MI-503 molecular weight in a reduced expression of the NF-κB-dependent RANTES and TNFα, with less recruitment of activated T-cells and macrophages.67 Looking at this in vitro (human proximal tubule cells), while paricalcitol did not affect NF-κB nuclear translocation, it did increase VDR expression and nuclear localization, and promoted intra-nuclear association of VDR with the NFκB p65 subunit, thereby reducing RANTES gene transcription.67 Intervention trials in CKD addressing inflammation have again been limited, performed predominantly in the haemodialysis

(HD) population and yielded mixed results.68–77 There is much heterogeneity PD184352 (CI-1040) between the available published work, and it is difficult to compare studies. However, it would appear that prolonged use (>3 months) of substantial doses of 1,25-OHD (6.14 ± 1.25 µg/week) may reduce circulating inflammatory burden (as determined by IL-1β, IL-6, TNFα or hsCRP), by up to 60%.69,73,74 Whether this observation translates into clinically meaningful outcomes and is applicable to earlier stages of CKD or administration of other forms of vitamin D has yet to be elucidated. Vitamin D influences the RAS; a link first

highlighted by the inverse association between vitamin D status and high-renin hypertension,78–80 and more recently by analysis of the LURIC cohort, where Tomaschitz and colleagues demonstrated that both 25- and 1,25-OHD were independently negatively correlated with both plasma renin and circulating angiotensin II.81 However, manipulation of the RAS with vitamin D has implications beyond just hypertension and glucose homeostasis in terms of cardiac risk. In VDR knockout mice a sevenfold increase in the expression of renin and angiotensin II was demonstrated,82 and using this together with a CYP27B1 knockout model, researchers have shown that this results in blood pressure-independent increased left ventricular (LV) mass, systolic dysfunction and myocyte hypertrophy and fibrosis.

However, it is necessary to realize that the number of Tregs alone ICG-001 in vivo is not decisive for effective suppression function [43]. Functional analyses of Tregs are probably more informative. Further, it is necessary to keep in mind that not all lymphocytes exerting suppressor function express FoxP3 [44]. Another obstacle can be caused by cell isolation. Many studies analyse Tregs in peripheral blood after Ficoll-Paque separation. We compared the detection of Tregs in whole blood and in the population of isolated cord blood mononuclear cells (CBMC) – the results were similar, but the analyses

obtained with the whole blood were more convincing and consistent and less time-consuming (data not shown). We acknowledge some limitations of our study, namely the heterogeneity of mothers’ allergies, but differentiation

Proton pump modulator of the children into subgroups according to different kinds of maternal allergy decreased the power of statistical analyses. Individual types of maternal allergies are listed in Table 1. Tregs are thought to play an important role in immune regulations even during intrauterine life [7]. Increased numbers of Tregs in this period can be partially responsible for decreased neonatal immune responses. The function of Tregs is critical in the early postnatal period, when the tuning of the immature immune system takes place. The impairment of Tregs could be the underlying mechanism contributing to heightened allergy development

in predisposed children. Our proof of decreased functionality of Tregs in cord blood of children of allergic mothers is in full agreement with the work of Prescott [22], who tested the immune function of neonatal CD4+CD25+CD127 low/– Tregs. However, both Prescott [22] and Schaub [30] did not find significant differences in transcription factor FoxP3 between high- and low-risk infants, whereas other studies pointed to decreased function of Tregs based on the lower presence of FoxP3 tetracosactide (MFI) [23]. This could be explained either by low numbers of individuals included [22] or by different methods used for the quantification of FoxP3. Quantitiative PCR (qPCR) was often used for the detection of FoxP3 gene expression [22,30]. Conversely, we exploited flow cytometry for FoxP3 protein detection. Schaub [30] suggests that the mRNA level of FoxP3 in Tregs is not regulated differently in dependence on maternal atopy. Nevertheless, the same group observed quantitatively and qualitatively increased Tregs in the cord blood of children of farming mothers whose children were postulated to be low-risk individuals for allergy development [7]. It is believed that lower exposure to non-pathogenic microbes together with reduced regulatory T function early in life could lead to Th1/Th2 imbalance, increasing the risk of allergy development [3]. The relationship between immune function of cord blood Tregs and allergy development requires further detailed studies.

The percentages of suppression were determined based on the proliferation index for RAD001 cell line effector cells cultured alone (100% proliferation, 0% suppression) compared with the proliferation

index of effector cells co-cultured with Treg cells. Statistical analysis was performed using SPSS version 19 and the normality of the data was assessed by the Shapiro–Wilk test. Differences between independent data sets, with normal distribution, were analysed using the Student’s unpaired t-test with the assumption of equal variance assessed by Levene’s test and for data sets without normal distribution the Mann–Whitney U-test was used. Differences between related data sets were analysed using the Student’s paired t-test and the Wilcoxon Signed Rank test for data sets normally or not normally distributed, respectively. Values were considered significant when P < 0·05. The frequency of 5-Fluoracil CD4+ CD25inter CD127low/− (termed CD25inter) and CD4+ CD25high CD127low/− (termed CD25high) Treg cells in the peripheral circulation of newly presenting HNSCC patients as a whole cohort (8·59 ± 0·41% and 6·67 ± 0·45%) was similar to that of healthy controls (8·77 ± 0·85% and 5·81 ± 0·66%). However, the expression of Foxp3 by both

CD25inter and CD25high Treg cells was significantly greater in HNSCC patients (n = 9; 30·08 ± 3·47% and 81·67 ± 2·21%, respectively) compared with healthy controls (n = 6; 15·83 ± 2·26% and 70·63 ± 3·17%), P ≤ 0·01. Additionally, the expression of Foxp3 in the CD25high Treg cell population was significantly greater compared with the CD25inter Treg cells in both HNSCC patients and healthy controls, P ≤ 0·01. Dividing the HNSCC patient cohort by tumour subsite demonstrated that patients with cancer of the larynx and oropharynx had similar percentages of circulating Treg cells irrespective of whether the level of expression of CD25 was intermediate or high (Fig. 2a). However, on analysis of tumour stage, patients with advanced stage tumours had a significantly elevated level of CD25high cells

compared with early stage patients, a trend mirrored, although not significantly, in both the the laryngeal and oropharyngeal subgroups (Fig. 2b). It was also observed that patients with tumours that had metastasized to the lymph nodes had significantly elevated levels of CD25high Treg cells compared with patients without nodal involvement, a trend shared by CD25inter Treg cells but not reaching significance (Fig. 2c). The functional activity of CD25inter and CD25high Treg cells from HNSCC patients (n = 28) and healthy controls (n = 9) was assessed by their ability to suppress the proliferation of two distinct autologous effector T-cell populations (CD4+ CD25− CD127−/+ and CD4+ CD25+ CD127+).

16 ml or 7.7 ml flow chambers. The flow rate (F) was adjusted to a very low rate of 1.3 ml h−1 resulting in an exchange rate of up to 180 and 6.25 times chamber volumes per 24 hours in the small and large chambers, respectively. The results of culture at a very low flow rate were markedly different from cultures in micro well plates. Low flow rates may better mimic the in vivo situation and thus may be of higher relevance for the clinical setting.

Under these conditions, a general resistance of fungal biofilms against anidulafungin cannot be confirmed. Strains of C. albicans and C. glabrata showed very uniform results whereas the C. parapsilosis group and C. lusitaniae varied from high susceptibility to resistance. Species differentiation of the C. parapsilosis group KPT-330 concentration selleckchem appears to be appropriate in clinical microbiological diagnostics. For the majority of the tested Candida species, anidualafungin was more effective than voriconazole. For the species C. lusitaniae and C. guilliermondii susceptibility testing should be considered prior to clinical use of echinocandin antifungals. “
“A biofilm composed of various microorganisms including Candida is found on denture surfaces and is likely to be involved in the etiology of denture-induced

stomatitis. The purpose of this study was to examine the role of hydrophobic interactions in candidal adherence to acrylic surfaces, particularly that of the hyphal form of Candida albicans. Candida clinical isolates were used. Acrylic plates coated with carrageenan and hydrocolloid (Hitachi chemical, Tokyo, Japan) were used as a hydrophilic substratum. A microbial suspension was placed on each acrylic plate and incubated. All plates were washed

Tau-protein kinase in phosphate-buffered saline containing CaCl2 and MgCl2 [PBS (+)] and cells still adhering to the acrylic surface were collected by 0.25% trypsin treatment. Cell-surface hydrophobicity was estimated using a modification of the technique used to measure adherence to hydrocarbons. When the acrylic plates were coated with hydrophilic materials, the adherence of hydrophobic clinical isolates of Candida and the hydrophobic hyphal C. albicans decreased, whereas the adherence of non-hydrophobic Candida was not affected or increased. We suggest that hydrophilic coating of denture surfaces could be a potent method for reduction of the adherence of relatively hydrophobic fungal cells, particularly hyphal C. albicans, which causes denture stomatitis and related infections. “
“This study was to develop a real-time florescence quantitative PCR (RT-FQ-PCR) assay to measure virulence-associated DEAD-box RNA helicase (VAD1) mRNA from Cryptococcus neoformans and evaluate its potential use in diagnosis and follow-up treatment of C. neoformans meningitis (CNM). Cryptococcus neoformans was detected using RT-FQ-PCR, ink staining, fungal culturing and C. neoformans antigen detection in CNM compared with a normal control.

First, it must be demonstrated that chronic infections,

in general, are indeed associated with bacteria adopting a biofilm mode of growth. Second, it must be demonstrated that there is a supply or a means to generate a supply of DNA for HGT within the biofilm community. Third, there need to be mechanisms (vide supra) for the transfer of DNA into live organisms. Fourth, and perhaps most importantly, the infecting bacterial IWR-1 in vitro population must be polyclonal in nature, i.e. be made up of multiple independent strains of the same bacterial species that are present simultaneously. The necessity for polyclonality derives from the need to generate diversity. If the infection–colonization is monoclonal, it means that each bacterium in the biofilm contains the same set of genes and the same set of allele forms of each gene; thus, exchanging DNA between any two cells in such an environment would not produce a new strain with new combinations of genes and alleles. In such a case, an extensive energy output would be rewarded with no possible gain in terms of creating a more competitive organism. Finally, it must be demonstrated that gene exchange indeed does occur, in real time, among strains within a polyclonal biofilm population and that some of the recombinant strains persist and expand their presence over time (i.e. prove to have a reproductive advantage under

the prevailing conditions in https://www.selleckchem.com/products/lee011.html the host) and in turn serve as recipients or donors of DNA in further HGT processes. An examination of the conditions present during the bacterial colonization of eukaryotic hosts, and during the subsequent chronic infectious disease processes, demonstrates that all of the criteria exist for fruitful genic reassortments (Hu & Ehrlich, 2008). Bacterial infections

associated with chronic disease states are nearly universally found to have adopted a biofilm phenotype (Hu & Ehrlich, 2008). The bacterially elaborated extracellular matrix of the biofilm, associated Montelukast Sodium with the final irreversible attachment of bacterial cells to a surface, is composed of multiple extracellular polymeric substances (EPS) including exopolysaccharides, eDNA, proteins, and lipids, and provides a protective physical barrier for the bacteria within. The cooperative creation of the matrix on host tissues or implantable devices by a community of bacteria is a population-level virulence trait as it provides for a community of bacteria that are collectively more difficult for the host to eradicate than individual free-swimming or individual attached bacteria would be. Once initiated, a biofilm acts like a single dynamic living organism that can grow, change its physical properties in response to its environment, evolve through mutation to be better adapted to its environment (Boles et al., 2004; Kraigsley & Finkel, 2009), and incorporate other pathogenic species into an integrated polymicrobial community.