The DR+2hL group

was re-exposed to light for 2 hr before euthanization. The primary visual cortex was extracted and homogenized as described previously ( Philpot et al., 2001). We thank Susana da Silva for technical assistance and data analysis. We thank Susana da Silva, Ian Davison, Cyril Hanus, Juliet Hernandez, Hyun-Soo Je, and Thomas Newpher for review of the manuscript. We thank Irina Lebedeva, Marguerita Klein, Sarah Lancaster, and Jaya Miriyala for excellent technical assistance. We thank Nils Brose for providing NLG1-KO mouse brains. Work in the laboratory of M.D.E. was supported by HHMI and grants from NIMH-NIH, NINDS-NIH, and the Simons Foundation. Work in the laboratory of B.D.P. is supported by grants from NEI-NIH and the Simons Foundation. R.T.P. was supported by the Portuguese FCT grant SFRH/BD/15217/2004. Cobimetinib price P.A.K. was supported by NICHD training Grant T32HD040127. “
“Formation and maintenance of the synaptic structure is a dynamic process that requires bidirectional interactions between pre- and postsynaptic components. A diverse assortment of cell adhesion molecules is present at

the synapse and organizes the synaptic specializations of both selleck chemical excitatory and inhibitory central synapses (Dalva et al., 2007; Siddiqui and Craig, 2011). Neuroligin (NLG) is one of the potent synaptogenic adhesion proteins located at the postsynapse, which transsynaptically binds to a presynaptic ligand, neurexin (NRX) (Ichtchenko et al., 1995; Irie et al., 1997; Scheiffele et al., 2000; Südhof, 2008; Bottos et al., 2011). Mammals express four NLG genes (i.e., NLG1 to NLG4). NLG polypeptides are type 1 transmembrane proteins with a large extracellular domain with homology to acetylcholinesterases but lack critical residues in the active site and interact with NRXs at the synaptic membrane surface (Südhof, 2008). Notably, NLG1 is localized at glutamatergic postsynapse, and overexpression of NLG1 induces the accumulation of glutamatergic presynapse and postsynapse molecules in vitro (Song et al.,

1999; Scheiffele et al., 2000; Budreck and Scheiffele, 2007). In contrast, NLG2 triggers the maturation of GABAergic synapses, implicating specific functions of different unless NLGs in the formation and maturation of different chemical types of synapses in vitro and in vivo (Graf et al., 2004; Varoqueaux et al., 2004, 2006). Recent studies revealed that copy number variation or point mutation in NLG genes are linked to autism spectrum disorder (ASD), schizophrenia, or mental retardation (reviewed in Südhof, 2008). Notably, ASD-linked mutations in NLG genes have been shown to affect the expression, folding, or dimerization of NLG proteins to compromise their surface expression and binding to NRXs (Comoletti et al., 2004; Levinson and El-Husseini, 2007; Zhang et al., 2009). Moreover, copy number variations that are associated with an increased risk of ASD were identified in NLG1 locus (Glessner et al.

Because PCDH17 was mostly expressed along the medial prefrontal cortex-anterior striatal pathways in a topographic manner, in both rodents and primates, it is likely that PCDH17 is anatomically and functionally conserved in the corticobasal

ganglia circuits of higher primates as well. Therefore, the PCDH17-expressing neuronal pathway could correspond to the prefrontal cortical loops in primates used for processing http://www.selleckchem.com/products/Bortezomib.html some aspects of motivational and executive functions. As the complementary expression patterns of PCDH17 and PCDH10 appear at E14.5 and gradually develop in the embryonic mouse striatum until birth (our unpublished data), we assumed that the expression of these protocadherins was reciprocally regulated by positional information in the embryonic striatum. Nevertheless, PCDH17 is dispensable Androgen Receptor antagonist in this topographic map formation (Figure S4), although it has crucial roles in the synaptic development of this pathway. In addition to these protocadherins, some axon guidance molecules that are involved in topographic map

formation in the visual and olfactory systems (Luo and Flanagan, 2007; Sakano, 2010), are also expressed in the embryonic striatum in a zone-specific manner. Like PCDH17, Netrin-1 exhibits an expression pattern with a high-anterior to low-posterior gradient in embryonic striatal regions (Powell et al., 2008). In contrast, similar to PCDH10, Ephrin-A5 and Semaphorin-3A exhibit expression patterns with low-anterior to high-posterior gradients

(Dufour et al., 2003; Wright et al., 2007). Therefore, these axon guidance molecules might organize the topographic map delineated by PCDH17 and PCDH10 expression in the embryonic basal ganglia. PCDH10−/− mice exhibit axonal growth defects in the striatum and die within the first several weeks after birth ( Uemura et al., tuclazepam 2007), whereas PCDH17−/− mice do not die prematurely and are not characterized by abnormal striatal axonal growth. These phenotypic differences may be at least partially attributable to different protein distributions in embryonic striatal fibers; PCDH10 ( Uemura et al., 2007), but not PCDH17 (our unpublished data), is distributed around striatal fibers at E14.5. It should be noted that similar to that of PCDH17, the expression of PCDH10 peaks during early synaptogenesis. A recent paper showed that PCDH10 is required for activity-dependent synapse elimination in cultured neurons ( Tsai et al., 2012). Therefore, PCDH10 may function not only in axonal growth, but also in synaptic development of corticobasal ganglia circuits.

51, p < 0.001, pη2 = 0.69). The increased correlation between visual and language this website nodes was stronger in θ with respect to α and δ (all p values < 0.001) and γ (p < 0.02), and in β with respect to α (p < 0.01), δ (p < 0.001) (Figure S6D). The comparison between β and θ and between β and γ did not reach significance (all p values > 0.05). Same results were

obtained using nodes based on the maps of Figure 3 (Figure S5A). In contrast, correlation decreased more strongly for α BLP during movie with respect to fixation between visual and auditory nodes (all p values < 0.001) (Figure S6E) and between visual and dorsal attention nodes (Figure S6F) (all p values < 0.001). In summary, regions in the visual network decrease their α BLP interaction with regions in the dorsal attention network but increase their BLP interaction in β, θ, and γ with regions in the language network. Next, we consider modulation in the default-mode network with respect to sensory and attention networks (Figure 6). Within-DMN BLP correlation dropped specifically in the α band during

the movie condition as compared to fixation. With respect to cross-network interaction, α BLP correlation decreased between DMN and auditory and dorsal attention, while β BLP correlation GW786034 in vitro decreased between DMN and auditory networks. However, the DMN increased correlation with the language network in the γ band. In summary, the default-mode network showed a pattern of BLP correlation modulation similar to other sensory and attention networks including decrements of within- and across-network interaction in the α band and increased interaction with the language network at higher (γ) frequencies. Control analyses were also carried out to examine the robustness of the results in relation to possible decreases in signal-to-noise due to α power decreases produced by visual stimulation in the movie condition (Berger, 1929; Supplemental Information and Figures S5B and S5C) and at the level of single subjects (Figures S5D and S5E).

One of the most influential observations shaping the debate on the function of RSN is that their topography is similar to that Oxymatrine of task networks (Biswal et al., 1995, Greicius et al., 2003 and Smith et al., 2009). This has suggested that RSN may be a prior for task-driven patterns of activity (Raichle, 2011). Inspection of the covariance matrix for fMRI connectivity vis-à-vis MEG BLP correlation in different bands (Figure 5) and by network (Figure 6), shows significant decreases of fMRI connectivity during movie within visual (t test: p < 0.005) and dorsal attention networks as well as between networks (visual-auditory [p = 0.003]; visual-dorsal attention RSN [p < 0.001]). Functional MRI connectivity decreases qualitatively parallel α and β BLP correlation decreases within and across networks earlier described.

Activating mutations in the human Trpv4 gene were recently shown to be present in individuals with a surprising range of genetic disorders including skeletal dysplasias, but also Charcot-Marie-Tooth disease and sensory and motor neuropathies ( Auer-Grumbach et al., 2010, Deng et al., 2010, Krakow et al., 2009, Landouré et al., 2010, Nilius and Owsianik, LY2157299 chemical structure 2010 and Rock et al., 2008). It is entirely possible that the small number of individuals identified with activating mutations in the Trpv4 gene have metabolic or cardiovascular deficits due to malfunction of the afferent pathway

described here. Our study on the molecular characterization of this novel osmosensitive afferent pathway could potentially lead to the development of new procedures to target this pathway in vivo. It has already been demonstrated that orthostatic hypotension and postprandial hypotension respond to water drinking ( Jordan et al., 2000, Schroeder et al., 2002 and Shannon et al., 2002). Moreover, water drinking in man can prevent neutrally mediated syncope during blood donation or after prolonged standing ( Claydon et al., 2006, Hanson and France, 2004, Lu et al., 2003 and Schroeder et al., 2002). Finally, water drinking is also associated

with weight loss in overweight individuals ( Stookey et al., 2008). The characterization of the Panobinostat mouse afferent arc of the reflex responsible for such effects will allow a better understanding of the physiological role of this reflex and allow the development of tools for its manipulation. Water,

phosphate-buffered saline (PBS), or water doped with 100 μM RR were administered orally using an application cannula with a rounded tip, fluids being dripped into the back of the mouth to evoke a swallowing reflex. Fluid intake was completed within 60 s. Rolziracetam Small blood samples (50 μl) were taken from the hepatic portal vein and osmolality determined using a vapor-pressure osmometer (VAPRO, Wescor). We obtained serum samples from liver transplant recipients and measured osmolality as above. We also determined C-terminal pro-arginine-vasopressin using an immunoluminometric assay (kindly provided by BRAHMS GmbH). All studies using human material were approved by the ethics committee of the Medical School Hannover. Thirty minutes after drinking, animals were anaesthetized with a Ketavet (10 mg/ml)/Rompun (0.04%) mixture (Pfizer, Bayer), and perfused with chilled (4°C) 4% paraformaldehyde (PFA) in PBS. The liver was removed, postfixed in 4% PFA at 4°C for 2 hr, dehydrated with 25% sucrose in PBS for 1–3 days and 20 μm cryosections were prepared and collected on gelatine coated slides. Tissue sections were pre-incubated in 1% serum albumin (BSA) and 0.3% Triton X-100 in TBS (Tris buffered saline) for 2 hr and incubated overnight at RT with primary antibodies (pERK 1:250, Cell Signaling, PGP9.5 1:1000, UltraClone Ltd, TRPV4 1:200 ab39260 Abcam) in TBS with 0.3% Triton X-100 and 5% normal goat serum.

Participants were paid ∼80

euros for their participation. In each of 5 scanning sessions of ∼8 min each, subjects viewed 120 successive, full-contrast Fasudil manufacturer Gabor patches that were oriented at between −90° and 90° relative to the vertical meridian. Each stimulus was visible for 1500 ms, during which period subjects were required to make a categorization judgment by pressing the right or left button on the response pad. Auditory feedback consisted of an ascending (400/800 Hz) or descending (800/400 Hz) tone of 200 ms, and followed stimulus onset by a variable interval in the range of 3–7 s. On 25% of trials, correct or incorrect feedback engendered a small monetary gain or loss, which was totaled up and supplemented subjects’

compensation (range 20–30 euros). An interstimulus interval of 1 s intervened between feedback and the subsequent stimulus. Stimuli were drawn randomly from category A (60 trials) or B (60 trials) with no constraints, and response-category assignments were counterbalanced across subjects. Category means and variances were unstable and independent, and jumped unpredictably every 10 or 20 trials (4 episodes of 10 trials, 4 episodes of 20 trials, randomly www.selleckchem.com/products/Paclitaxel(Taxol).html intermixed) to a new mean drawn from a uniform random distribution with a variance of either 5° or 20°. Values representing the probability of choosing category A over B under the Bayesian model were estimated using a hierarchical Bayesian learner that calculates best-guess estimates of the generative mean and variance of each category in a Markovian fashion. For each category, a generative model of the observations is assumed as follows (see Supplemental Experimental Procedures and Behrens et al. [2007] for a more extensive description of a related model). At each trial i, after see more the true category has been revealed, the probability of observing the orientation i (given any possible mean and variance) may be written: equation(Equation 4) p(Yi|μi,σi)∼N(μi,σi)p(Yi|μi,σi)∼N(μi,σi)Hence,

each new data point contains information about the underlying mean and variance. However, the mean and variance are constant over runs of trials before jumps, or change points occur. Hence, the prior distribution, conditional on the previous trial, may be written as follows: equation(Equation 5) p(μi|μi−1,Ji)={δ(μi−μi−1)U(0,180)Ji=0Ji=1This equation states that the underlying category mean at trial i will be the same as that at trial i-1 if there has not been a jump (J = 0), or could take on any value if there has been a jump (J = 1). A similar equation may be written to describe the dynamics of σ, which varied in a log space. equation(Equation 6) p(σi|σi−1,Ji)={δ(σi−σi−1)U(2,40)Ji=0Ji=1Jumps J occur at random with probability v, termed the volatility.

We hypothesized that the response to LOT inputs might be suppressed by prior activation of the cortical circuitry because of the recruitment of strong feedback inhibition. This prediction was tested by delivering a short train of LOT stimulation (3 pulses at 40 Hz) to achieve spiking in half the trials (0.56 ± check details 0.042). Indeed,

when a similar train of piriform stimuli (3 pulses at 40 Hz; probability of spiking, 0.36 ± 0.16) preceded the LOT input by 100 ms, we observed an 18% reduction in the probability of spiking (LOT train following PCx train, 0.46 ± 0.049; n = 9 cells; paired t test comparing two LOT trains, p = 0.017; Figure 4D). Two forms of inhibition have been described in the piriform cortex. Feedforward inhibition is mediated by interneurons in layer I that receive direct input from the LOT and synapse onto apical dendrites of pyramidal cells, whereas feedback inhibition is mediated by the layer II/III interneurons that are activated by pyramidal cells and synapse onto pyramidal cell bodies (Luna and Schoppa, 2008, Neville and Haberly, 2004, Stokes and Isaacson, 2010 and Suzuki and Bekkers, 2010). Two experimental approaches were employed to demonstrate that feedback inhibition is significantly stronger than feedforward inhibition. We observed a dramatically greater

selleck chemical effect of gabazine on synaptic responses following subthreshold recurrent stimulation versus LOT stimulation (Figure S4A). We also determined the lowest stimulation intensities of either

the LOT or recurrent inputs that reliably drove spiking when inhibition was blocked. LOT stimulation at this intensity could still generate spiking when inhibition was intact (Figure S4), consistent with a relatively small role for feedforward inhibition. In contrast, piriform stimulation at this intensity always failed to evoke spikes in downstream piriform neurons when inhibition was intact. These data support a dominant role for feedback versus feedforward inhibition in controlling the activation of piriform cortex pyramidal cells. In the piriform cortex, the specificity of aminophylline an odorant is represented by a unique ensemble of neurons that is distributed without discernable spatial order. These cells also make extensive recurrent connections with other excitatory and inhibitory neurons that may shape the odorant representation. We have introduced ChR2 into focal regions of the piriform cortex to study the role of recurrent circuitry in shaping the cortical response to bulbar input. Axons of layer II/III pyramidal cells project across the piriform cortex, where they make excitatory synaptic contacts with other pyramidal cells. The likelihood that any two pyramidal cells are synaptically connected is very small but remains roughly constant over remarkably long distances compared to neocortical sensory areas.

This finding suggests that, besides the phase difference between pyramidal cells and inhibitory interneurons, local groups of neurons (as captured by a MUA recording) are locked to approximately the same phase of the gamma rhythm.

This leaves a cell’s position in the horizontal cortical map or vertical cortical column as the main candidate determinants of its preferred gamma phase. A position in the horizontal cortical map, during visual stimulation, translates to a particular position in the cortical activation map. A given selleck inhibitor stimulus typically generates an ordered spatial pattern of activation in the map, such that a cell’s position in the map translates into a particular activation level. We have shown previously that

the level of V1 activation further translates to the gamma phase (Vinck et al., 2010a). However, this effect accounted INCB024360 mouse for only a relatively small part of the phase variance (see Figure 2 of Vinck et al., 2010a). The activation independent part of the phase variance (that is already visible in that figure and replicated here in Figure 4) likely requires a different explanation. We propose that it is related to the remaining possible source of phase variance, i.e., the position of a neuron in the vertical cortical column. In fact, there is direct evidence in favor of this suggestion: Livingstone (1996) has shown that pairs of gamma-synchronized neurons within the granular and supragranular layers of monkey V1 had the more superficial neuron lagging the deeper neuron by ∼3 ms for a distance of ∼400 μm. The dependence of gamma phase on the vertical position in the cortex might be due to the pattern of synaptic connections within a column and the resulting flow of activation. Gamma

activity is primarily found in supragranular layers (Buffalo et al., 2011), and within those, the gamma phase of firing increases systematically with distance from the input layer 4 (Livingstone 1996). At the same time, a larger distance from layer 4 corresponds to a longer conduction time. Thus, the precise others connectivity of the cortical column might generate the precise temporal sequence of gamma activation. Therefore, we would like to suggest that a cell’s preferred gamma phase is determined by two activation-independent factors (vertical position and cell class) and one activation-dependent factor (cf. gamma phase shifting). The interplay between these contributions to the gamma phase might explain the firing sequences and their stimulus dependence in anesthetized cat primary visual cortex (Havenith et al., 2011). The potential consequences of the different gamma-phase components are intriguing. First, the delay between pyramidal cell and interneuron spiking allows the gamma rhythm and in fact overall activation to be maintained (Börgers and Kopell, 2005).

Pooled sera from mice immunized with two doses of 1 μg PCV7 served as the quality control. Goat anti-mouse HRP conjugate was purchased from Southern Technologies (Birmingham, AL). To measure total functional antibodies, a standard opsonophagocytic assay (OPA) described by Romero-Steiner et al. [31] and [32] was utilized. Titers were calculated as the reciprocal dilution at which ≥50% bacterial killing occurred KPT330 in comparison

to complement control wells. To assess differences in functional activity due to species specific phagocytic cells, an alternative OPA protocol using Raw 264.7, mouse monocytes (ATCC) and guinea pig complement (MP Biomedicals, Solon, OH) was also evaluated [15], [33] and [34]. A week after administering

the last dose, mice were intranasally challenged with approximately 1 × 106 CFU of log phase S. pneumoniae serotype 4, 14, or 19A suspended in 10 μl PBS. Challenge doses were later confirmed by counting the overnight growth of a 10-fold serial diluted challenge inoculum [18]. Three to five days post-challenge, each mouse was euthanized and its nasopharyngeal (NP) cavity washed as described by Moreno et al. [26] and Wu et al. [35]. As seen in the study by Moreno et al., control mice significantly cleared pneumococci six days post intranasal challenge [26]. In this study, we found three to five days post-challenge to be the optimal time point in detecting a difference between control and immunized mice. NP washes aminophylline (100 μl) were collected, diluted with equal volume of saline, and further serially diluted, RG7204 mw 3-fold, an additional five times in a 96-well plate. Fifty microliters of each dilution was cultured on blood agar plates supplemented with 2.5 mg/L gentamicin. In preliminary studies, mice cleared serotypes 4 and 19A within 4 days and serotype 14 within 5 days post-challenge. Because of these results, NP washes were conducted 3 days post-challenge of serotype 4 or 19A and 4 days post-challenge

with serotype 14. As previously defined, carriage values are the average count of Pnc colony-forming units (cfu) collected in 50 μl of nasal wash [18]. Counts were adjusted for dilution factors prior to averaging. Antibody concentrations were calculated with a 4-parameter logistic equation (ELISA for Windows, CDC). Mean or geometric mean of OPA titers (with log-transformation) and colony counts were calculated. Significant differences, P ≤ 0.05, were determined between two groups using Mann–Whitney rank sum test or t-test, within an experiment using one way analysis of variance on ranks, and for multiple pairwise comparisons using the Student–Newman–Keuls method (SigmaStat software version 2.0; Jandel scientific, Point Richmond, CA). To examine the effect of PCV7 + PsaA co-administration on IgG antibody levels, mouse immune sera were assayed before and after challenge.

, 2003 and Oltedal et al., PD0332991 chemical structure 2007). Furthermore, direct access to presynaptic boutons via the patch pipette should not only allow one to control the presynaptic membrane potential but also to measure and manipulate the presynaptic Ca2+ concentration by direct loading of synthetic Ca2+ dyes and Ca2+-caging compounds. Until now, these types of experiments were only possible in large

synapses such as the calyx of Held. In summary, we anticipate that the combined application of HPICM-assisted patch-clamp recordings, together with previously described electrophysiological and imaging methods to image vesicular release and Ca2+-dynamics in individual synaptic boutons (e.g., Ariel and Ryan, 2010, Ermolyuk et al., 2012, Hoppa et al., 2012, Li et al., 2011 and Li and Tsien, 2012), will provide answers

to these and other questions relating to the behavior of small central synapses. Hippocampal neurons were isolated from P1–P2 rat pups and cultured in Neurobasal-based medium either on an astrocyte feeder layer or on poly-D-lysine-treated coverslips. All recordings were conducted KU-55933 at ambient temperature (23°C–26°C) 12–19 days after plating. The standard extracellular solution Olopatadine used in all experiments contained 125 mM NaCl, 2.5 mM KCl, 2 mM MgCl2, 2 mM CaCl2, 30 mM glucose, 0.01 mM NBQX, 0.05 mM APV, and 25 mM HEPES (pH 7.4). Active synapses were labeled with 20 μM (bath concentration) FM1-43 (Invitrogen) or 200 μM SynaptoRed C1 (SRC1,

Biotium) by incubation in the extracellular solution, with 90 mM NaCl replaced by 90 mM KCl for 90 s followed by a 10–15 min wash in the original solution. Tetrodotoxin (1 μM) was added to the extracellular solution in some experiments to slow down spontaneous destaining of the FM dyes. HPICM topographic images were obtained using a custom-modified SICM sample scanner ICNano-S (Ionscope) and custom software as described previously (Novak et al., 2009). Briefly, the scan head consisted of a PIHera P-621.2 X-Y Nanopositioning Stage (Physik Instrumente [PI]) with 100 × 100 μm travel range that moved the sample and a LISA piezo actuator P-753.21C (PI) with travel range 25 μm for pipette positioning along the z axis. Coarse sample positioning was achieved with translation stages M-111.2DG (x-y directions) and M-112.1DG (z axis) (PI). The z piezo actuator was driven by a 200 W peak power high-voltage PZT amplifier E-505 (PI), while the x-y nanopositioning stage was driven by 3 × 14 W amplifier E-503 (PI).

For instance, while IFNγ is

required to control infection with SL3261 as shown here and by Vancott et al. [41] it is dispensable for control of infection with a phoP mutant. In summary, we have investigated the role of the F0F1 ATPase in S. Typhimurium infection and shown VX770 that this protein complex makes a significant contribution to bacterial growth in vivo. Furthermore, mutants lacking the atp operon have potential utility as novel live attenuated vaccine strains against Salmonella infection. This work was supported by a BBRSC Project Grant and a BBSRC Industrial Partner Pfizer CASE Studentship BBS/S/N/2006/13095. The work in knock-out mice was supported by the Wellcome Trust Sanger Institute. The technical assistance of C. Willers and D.B. Cone is gratefully acknowledged. “
“Although a successful eradication of certain infectious diseases such as smallpox has been realized, vaccination strategies against human pathogenic parasites remain a fundamental challenge for biomedical research [1]. Long-lasting protective antibody production is one of the hallmarks of effective vaccination and is an important feature of immunological

memory [2]. The clinically silent liver stage of Plasmodium infection epitomizes an attractive target for antimalarial vaccine development [3] and [4]. However, despite decade long endeavors, no antimalarial vaccines have been licensed today. Nevertheless, promising results are emerging despite the fact that the leading pre-erythrocytic subunit vaccine candidate (RTS,S) has proven to be only partially protective in clinical trials [5]. In the previous study, we have http://www.selleckchem.com/products/Bosutinib.html shown that a recombinant (r) BCG expressing the Plasmodium falciparum circumsporozoite protein (BCG-CS) induced activation and priming of CSp-specific immunity in BALB/c mice [6]. A prime-boost regimen consisting of this BCG-CS combined with adenovector 35 (Ad35) expressing the same antigen (Ad35-CS) is utilized in this work. Based on evidences in literature we conclude

that a reasonable strategy to induce broad and prolonged immune response against malaria infection may be realized by priming with recombinant virus and not boosting with rBCG [7], [8] and [9]. Therefore, a rBCG provides an option that can fit within the existing World Health Organization (WHO) expanded program of immunization (EPI) considering that BCG is being given at birth. Since a major concern is, how to induce protective cell-mediated immunity (CMI) particularly IFN-γ-producing CD8+ T cells, which have been shown to provide long-term immunity to malaria [10]. These cells are essential in combating parasitic infections, including malaria. Due to intracellular expression of the CSp insert in the rAd35 genome and the intracellular residence of BCG expressing the same antigen, we propose that BCG-CS is likely an efficient route of antigen delivery.