In the present study, the number of the four T-cell immunogenic peptides and glutamine residues occurring in the two polyglutamine domains of the 22 cloned genes were analyzed, along with their similarity to the other 95 genes originating in the three diploid species representative of the A and D genomes or the putative ancestral B genome of common wheat. In agreement with previous findings [13], [15], [21] and [23], our study confirmed that the set of epitopes, as well as the clusters formed in the phylogenetic tree, were indeed distinct for each genome. Thus, according to the distinct genomic characteristics, 8, 6 and 8 genes were assigned respectively

to chromosomes 6A, 6B and 6D, and a total of 16, 0 and 23 epitopes (including a highly immunogenic 33-mer

peptide present in Z4A-5) were detected. Alpha-gliadins from the A and especially Selleckchem Romidepsin the D genomes are more Cabozantinib datasheet deleterious for CD patients, and Zhengmai 004 had the potential to cause the development of CD. However, everything has advantages and disadvantages: a study using Chinese Spring Gli-2 deletion lines showed that removing the α-gliadin locus from the short arm of chromosome 6D resulted in a distinct loss of technological properties, although the T-cell immunogenic epitopes decreased [41]. We also found that four of the five genes in this study that have an odd number of cysteine residues, as well as the majority of the genes in GenBank that share this characteristic, were assigned to chromosome 6D on the basis of the occurrence of the epitopes and fell into a cluster Pyruvate dehydrogenase lipoamide kinase isozyme 1 in the phylogenetic tree (data not shown). Thus, just as it has been demonstrated that the D genome contributes to many characteristics (including the effects on baking quality of HMW-GS on chromosome 1D) of common wheat [13], the Gli-2 locus on chromosome 6D also appears to make specific contributions to baking quality, most likely increasing loaf volume, in addition to being mainly responsible for most of the T-cell stimulatory peptides in α-gliadins. Fortunately, however, there is evidence

[42] in the literature that the amount of gluten exposure has a marked influence on the likelihood of CD development: the higher the exposure to the complex of immunogenic peptides, the higher the incidence of CD. Theoretical comparative analysis also supports this opinion [13] and [17]. A diet based on wheat cultivars low in T-cell stimulatory sequences may thus have high potential for CD prevention. Furthermore, given the heterogeneity of T-cell epitopes in gluten, it is possible to generate wheat varieties with few or even no toxic peptides via conventional breeding strategies [15] and [17]. In the phylogenetic tree we constructed, 11 exceptional α-gliadin genes originating from T. monococcum and Ae. tauschii encode few or even none immunogenic T-cell peptides. These findings further confirmed that the wild genetic resources of T. monococcum and Ae.

This catalytic preference might be explained by the presence of amino acids that promote a non-polar environment in the catalytic site. The sequence of the first forty residues from the N-terminal of LmLAAO determined by Edman degradation was ADDRNPLGECFRETDYEEFLEIAKNGLRATSNPKHVVIGA,

showing that it is a new enzyme from L. muta venom. The complete Selleckchem MDV3100 amino acid sequence of LmLAAO (Figs. S1 and S2) was deduced by Expressed Sequence Tags (ESTs) sequencing (Fig. S1). The obtained ESTs were subsequently aligned with the LAAOs from other snakes, leading to the identification of these transcripts. Among the identified transcripts, twenty ESTs showed high similarity with other snake LAAOs. The complete sequence of the cDNA of L. muta LAAO was resolved by the superposition of these twenty ESTs and confirmed manually. The complete deduced cDNA was named LMUT0069C. The overall proteomic profile of L. muta venom reported by Sanz et al. (2008) showed that L. muta venom contains a single LAAO molecule. This information, along with the N-terminal (ADDRNPLGECFRETDYEEFL) and internal sequences reported by them (SAGQLYEESLGK and KFWEDDGIR,

PCI-32765 cell line corresponding to LmLAAO amino acid residues 152–163 and 334–342, respectively), are also evidences that the cDNA-deduced protein sequence reported now may actually correspond to the venom expressed protein. LmLAAO showed high sequence identity with LAAOs from other snake venoms, such as Sistrurus catenatus edwardsii (91%), Crotalus atrox (91%), A. halys pallas (90%), Crotalus adamanteus (90.6%), Trimeresurus stejnegeri (89%) and Calloselasma rhodostoma (88%) ( Fig. S2). In fact, the high sequence identity shared by L. muta and A. halys pallas LAAOs ( Fig. S2) allowed us to predict through the tertiary structure of the monomeric form of LmLAAO ( Fig. 5). The final model consists of a 486 amino acid polypeptide chain and one FAD molecule. The fourteen

last residues are missing in the protein model due to the lack of information on template structure. Analysis of Ramachandran plot revealed that 95.9% residues are in most favored, 3.1% in additionally allowed, and 1.0% in disallowed regions. The overall fold of snake venom LAAOs consists of three domains: a FAD-binding domain, the substrate binding domain and the α-helical domain ( Fig. 5). The FAD cofactor is found inside a cavity formed between cofactor binding and the substrate binding domains. In terms of overall structure, no major structural differences have been found when comparing the simulated LmLAAO structure with the template model (PDB entry: 1REO). In fact, structural comparison of all LAAO crystal structures available at the protein data bank (PDB entries: 1REO, 3KVE, 2IID, 1TDN) suggests a high degree of sequence identity and structural similarity amongst snake venoms LAAOs (Fig. S2).

, 1993) has an MHD of 0.2 μg, B-JussuMP-I from Bothrops jararacussu has an MHD of 4 μg ( Mazzi et al., 2006) and BaH4 from Bothrops asper has an MHD of 2 μg ( Franceschi et al., 2000). Based on these results, we consider Batroxase to be a weakly hemorrhagic metalloproteinase.

To determine the mechanism underlying the induction of hemorrhage Selleckchem R428 by Batroxase, its capacity to digest extracellular matrix components was assessed. Batroxase was able to hydrolyze type IV collagen and fibronectin molecules, and it also degraded the α 1, α and γ chains of laminin in Matrigel, but it was not able to digest isolated laminin. No nidogen proteolysis was detected. According to Bou-Gharios et al. (2004), the basement membranes of blood vessels consist mainly of laminin, collagen and fibronectin. Therefore, the ability of Batroxase to hydrolyze these components is consistent with its ability to induce hemorrhage by degrading extracellular matrix components of the blood vessel basement

membranes. Batroxase was able to digest fibrinogen by cleaving the α and β chains. Furthermore, the fibrinogen hydrolysis occurred in a concentration-dependent manner and was inhibited by EDTA and EGTA, which indicates that its metalloproteinase character was important for inducing proteolysis. According to Mosesson (2005), under physiological conditions, fibrin is formed by the cleavage of the fibrinogen α chain by thrombin. However, the results obtained showed that α and β chain cleavage by Batroxase suggests that the fibrin formed

might not be able to polymerize. Thus, the activity of Batroxase on the fibrinogen molecule likely indicates a consumption of this substrate PR-171 nmr and an inhibition of clot and thrombus formation. Several PI SVMPs are able to preferentially digest the α chain of the fibrinogen molecule, e.g., BnPI from Bothrops neuwiedi ( Baldo et al., 2008), BlaH1 from Bothrops lanceolatus ( Stroka et al., 2005), Atroxlysin-I from Bothrops atrox ( Sanchez et al., 2010), BmooMPα-I from Bothrops moojeni ( Bernardes et al., 2008) and Neuwiedase from Bothrops neuwiedi ( Rodrigues et al., 2001). Fibrinolytic activity has been reported for several PI-class SVMPs, such as Neuwiedase www.selleck.co.jp/products/Paclitaxel(Taxol).html (Rodrigues et al., 2001) and BnP1 from Bothrops neuwiedi ( Baldo et al., 2008), Bothrojaractivase from Bothrops jararaca ( Berguer et al., 2008), Berythractivase from Bothrops erythromelas ( Silva et al., 2003), BthMP from Bothrops moojeni ( Lopes et al., 2009) and Atroxlysin-1 from Bothrops asper ( Sanchez et al., 2010). Batroxase was able to induce fibrin digestion in a concentration-dependent manner up to 8 μg. The lack of further digestion at higher concentrations was probably the result of the total consumption of the fibrin in the gel. To confirm that the fibrinolytic hydrolysis mediated by Batroxase was not the result of the activation of plasminogen to generate plasmin, Batroxase was incubated with plasminogen, and the resulting fragments were analyzed.

148(Rrs(490)/Rrs(555))−2.18.POC=0.148Rrs490/Rrs555−2.18. This particular formula may be compared with the formula presented in the previously cited paper by Stramski et al. (2008) on relationships between POC and optical properties in the eastern South Pacific and eastern Atlantic Oceans. The authors of that work gave two very similar variants of the POC vs. Rrs(490)/Rrs(555) relationships, one of which (relating to all the data in Stramski et al., i.e. including the Chilean

upwelling stations) took the following form: POC = 0.3083(Rrs(490)/Rrs (555))− 1.639. The latter formula is plotted in Figure 9 together with formula  (13). Such a comparison shows clearly that the formula describing LBH589 mw the average oceanic relationship has a less steep slope (compare the constants C2: − 2.18 with − 1.639). As a consequence of that within the range of minimal blue-to-green reflectance values in the analysed Baltic Sea dataset (values of about 0.4) both formulas would predict similar POC concentrations, but within the range of maximum blue-to-green values (here ca. 0.9) the POC concentrations predicted according to the oceanic formula would be about twice as high as those estimated with formula  (13).

However, while performing such a comparison it has to be borne in mind that formula  (13) does not offer very attractive values of statistical parameters: among other PFT�� things, the standard error factor X is equal to 1.74, which is much higher than the value of X of 1.56 obtained with formula  (11), which makes use of the blue-to-red ratio. With regard to formulas for estimating Chl a, the fact that no single band formula was found to be acceptable for estimating that pigment concentration for the Baltic Sea data analysed here (no such formula is presented in Table 3) is in agreement with one of the

conclusions suggested by Bukata et al. (1995), namely, that a reliable estimate of chlorophyll concentration in waters other than Case 1 (other than open ocean Clomifene regions) most likely cannot result from a single wavelength reflectance relationship. The other important fact is that among the reflectance ratio formulas found here to be acceptable for estimating the Chl a concentration in the southern Baltic Sea (see the last six lines in Table 4) there is also no formula using the classic blue-to-green ratio that would resemble any of the standard remote sensing algorithms commonly used for Case 1 waters. This is in agreement with earlier studies documenting the generally poor performance of standard Chl a satellite algorithms when they were applied to the Baltic Sea environment (see e.g. Darecki & Stramski (2004)). But it has to be pointed out that the few positive observations/arguments presented above are only qualitative in their nature.

Australia) that converts the digitized images to grayscale images (black and white) after color selection ( Solomon, 2009). Fig. 1A and B illustrates the color segmentation which see more allows the selective capture of the immunoreactive sites against the desired antibody and measures their pixel densities. Quantitative analysis was accomplished by calculating the percentage of pixels of the anti-AQP4 and anti-GFAP in the white matter and granular, Purkinje and molecular layers of the cerebellar cortex separately. All numerical results were analyzed using the GraphPad Prism software package (San Diego, CA, USA) and expressed

as the mean ± standard error (S.E.). Differences between data means of saline-treated and PNV-treated groups were determined by the unpaired Student t-test with a p value ≤ 0.05 indicating statistical significance. Two-way analysis Fulvestrant research buy of variance

was used when appropriate to test age/temporal differences in the response to venom effect. The AQP4 and GFAP immunoreactivity of astrocytes was co-localized among the neuron bodies of the granular and Purkinje layers and widespread throughout the width of the molecular layer with the difference that the glial processes appeared well-defined in the anti-GFAP reaction. The anti-AQP4 reaction, although strong, was more diffuse. In animals injected with PNV, there was gradual time-dependent increase in the intensity of the immunolabeling in the white matter and layers of the cerebellar cortex for both P14 and adults. Fig. 2, Fig. 3 and Fig. 4 were chosen to illustrate the reaction pattern at two time intervals

(2 h and 24 h) for either P14 rats or adults; the figures also display the calculation of the density of pixels relative to the immunoreaction intensity throughout the period of observation. There is no significant difference in the physiological expression of AQP4 and GFAP in the white matter of adult and P14 rats at the different time-points after saline solution injection (Fig. 2C and F). However, rats administered PNV showed a 103.8% increase of AQP4 expression in adult animals (*p ≤ 0.05) and a 77.5% (**p ≤ 0.01) in neonate animals after 24 h ( Fig. 2C). The venom also caused a 57.3% increase in the GFAP expression after 24 h only in the astrocytes MycoClean Mycoplasma Removal Kit of P14 animals (*p ≤ 0.05). Although not significantly, AQP4 expression was 11%–20% higher in P14 PNV-treated animals (ranging from 16.48 ± 1.06 at 2 h to 27.73 ± 2.57 at 24 h, respectively) than in adult PNV-treated ones (where it ranged from 13.68 ± 2.03 at 2 h to 24.94 ± 3.55 at 24 h, respectively). In contrast, the values for GFAP were in general slightly higher for adults than for P14 animals. The two-way analysis of variance showed that the time elapsed between envenomation and animal euthanasia interfered with the expression of AQP4 and GFAP in the white matter of neonates and adults (*p ≤ 0.05). Also, there was interference of the age variable in the expression of AQP4 and GFAP at 24 h only.

With the increase in ‘source’ depth, the transport of phosphorus was reduced to 2.5 tons m−1 at 45 m depth for the upwelling off the northern Seliciclib ic50 coast and at 65 m depth off the southern coast. In the case of nitrogen the behaviour was slightly different. The greatest transport was from the depth interval of 40–65 m off the southern coast ( Figure 5d) and 43–49 m in the case of the opposite coast ( Figure 5b). The regional upwelling response pattern differs more than 2.5 times – during the southern coast upwelling more than 10 tons m−1 of nitrogen was brought to the surface layer from depths of 45– 55 m, while off the northern coast the highest values were no more than 4 tons m−1 from depths of 40–45 m. The deeper

layers were quite inefficient as nutrient sources for the euphotic layer during short-term upwelling events. Less than 1 ton m−1 of nitrogen was brought to the surface layer from depths of over 53 m and 73 m during the upwelling events along the northern and the southern coasts respectively. The results of a similar nutrient transport

simulation with a 50% smaller wind stress (τ = 0.5 τ0) are shown in Figure 6. The reduction in wind stress results in the overall decrease of amounts of upwelled nutrients. In particular, the largest transport of phosphorus remained in the upper 15–25 m layer off both coasts, whereas nitrogen transport from deeper layers was vanishingly small for the upwelling along the northern coast (< 0.75 tons m−1 from depths greater than 35 m). As regards the southern coast, N-acetylglucosamine-1-phosphate transferase the largest transport of nitrogen remained Trichostatin A purchase in the depth range of 40–55 m with the maximum at 45 m. Nutrients are considered to be conservative passive tracers, and it is therefore possible to transform the cumulative amount of nutrients per metre Δm10/Δz to a volume of water V10, which is cumulatively transported to the upper 10-m layer from a 1 m thick layer at a certain depth z: equation(1) V10=1C(z)Δm10Δz,where C(z) is the initial

nutrient concentration at depth z ( Figure 3). The cumulative volume transports per unit source layer thickness to the upper 10-m layer during the upwelling along the northern and the southern coasts with different wind stresses are shown in Figure 7, and the snapshot of upwelled volumes during the maxima of nutrient amounts on the 6th simulation day in Figure 8. It is seen in both Figure 7 and Figure 8 that the total volume of water transported to the upper 10-m layer from the top depth interval of 15–19 m was almost the same for the upwelling events off the northern and the southern coasts of the Gulf, with the maximum of 6.7 × 109 m2 ( Figure 8). Such equality of upwelled volumes is achieved as a result of the predominance of vertical turbulent diffusion (vertical mixing) over vertical advection, as the intensity of turbulent mixing in the upper sea is governed by wind force rather than wind direction.

(Category 3) Once the diagnosis of TSC is established and initial

diagnostic http://www.selleckchem.com/products/AZD6244.html evaluations completed, continued surveillance is necessary to monitor progression of known problems or lesions and emergence of new ones (Table 3).20 Some manifestations begin in childhood and are less likely to be present or cause new problems in adulthood, such as cardiac rhabdomyomas or subependymal giant cell astrocytomas. In contrast, problems with LAM are typically limited to adults, and renal manifestations require significantly more monitoring and intervention in adulthood compared with childhood because of the cumulative nature of angiomyolipomata and other renal lesions. Finally, other aspects of TSC may be present throughout the entire lifespan of the individual, such as epilepsy and TAND, but specific manifestations click here and impact on overall health and quality of life can vary. Thus, ongoing periodic surveillance is needed after initial diagnosis for optimal care and prevention of secondary complications associated with TSC.

Management of specific complications of TSC will often require input from a multidisciplinary team. Genetic testing and counseling should be offered to individuals with TSC when they reach reproductive age, and first-degree relatives of affected individuals should be offered clinical assessment and, where a mutation has been identified in the index case, genetic

testing. (Category 1) Symptomatic SEGA or SEGA Adenosine triphosphate associated with increasing ventricular enlargement, or with unexplained changes in neurological status or TAND symptoms, require intervention or more frequent clinical monitoring and reimaging. For acutely symptomatic individuals, surgical resection is the recommended intervention, and cerebrospinal fluid diversion may also be necessary. For growing but otherwise asymptomatic SEGA, either surgical resection or medical therapy with mTOR inhibitors can be effective.31 and 32 Shared decision-making with the patients or their parents in selecting the best treatment option should take the following considerations into account: risk of complications or adverse effects, cost of treatment, expected length of treatment, and potential impact on TSC comorbidities. Patients with unilateral, single, gross total resectable SEGA without individual risk factors or other comorbidities preferentially may benefit from surgery, whereas patients with multisystem disease or multiple or infiltrating SEGA lesions that are not amenable to gross total resection may favor mTOR inhibitor treatment.

The experiments were designed in such a way that the number of animals used and their Fulvestrant in vivo suffering was minimized. The chemically synthesized NOD1 agonist FK565 was provided by Astellas Pharma Inc. (Ibaraki, Japan) (Watanabe et al., 1985). MDP (N-acetylmuramyl-l-alanyl-d-isoglutamine hydrate, catalogue number A9519, Sigma–Aldrich, Vienna, Austria) was used as synthetic NOD2 agonist and LPS extracted from Escherichiacoli 0127:B8 (purified by gel-filtration chromatography,

catalogue number L3137, Sigma–Aldrich, Vienna, Austria) was used as a TLR4 agonist. The experiments were started after the animals had become accustomed to the institutional animal house over the course of at least 2 weeks. Prior to the behavioral tests, the mice were allowed to adapt to the test room (lights on at 6:00 h, lights off at 18:00 h, set points 22 °C and 50% relative air humidity, maximal light intensity 100 lux) for at least one day. The pattern of locomotion, exploration, feeding as well as sucrose preference (SP) were assessed with the LabMaster system (TSE Systems, Bad Homburg, Germany), allowing

continuous recording of the animals without intervention by any investigator, as described previously ABT-737 mouse (Painsipp et al., 2013). The LabMaster system consisted of test cages (type III, 42.0 × 26.5 × 15.0 cm, length × width × height), surrounded by two external infrared frames and a cage lid equipped with three weight transducers. For recording locomotion and exploration, the two external infrared frames were positioned in a horizontal manner above one another at a distance of 4.3 cm, with the lower frame being fixed 2.0 cm above the bedding floor. The bottom frame was used Mannose-binding protein-associated serine protease to record horizontal locomotion of the mice, whereas the top frame served to record vertical movements (rearing, exploration). The measures of activity (locomotion, exploration) were derived from the light beam interruptions (counts) of the corresponding

infrared frames (Painsipp et al., 2013). The three weight transducers were employed to quantify ingestive behavior. To this end, a feeding bin was filled with standard rodent chow (altromin 1324 FORTI, Altromin, Lage, Germany). In order to assess SP, one drinking bottle was filled with tap water and one with a 1% sucrose solution and the bottles were each attached to a transducer on the cage lid for the total duration of the experiment. SP was calculated using the formula: sucrose intake/(sucrose intake + water intake). In a few cases in which the fluid bottles got obstructed, the data were excluded from analysis. Each test parameter was collected over a 24 h interval and activity scores and food intake recorded during the day before injection were set as 100%, and the daily scores measured post-injection expressed as a percentage of the pre-injection score.

Female BALB/c wild-type (wt) mice (6–8 weeks) were purchased from Harlan Laboratories, Zeist, The Netherlands. Six to eight weeks old C57BL6/J (wt) and B6.129-Tlr2tm1Kir/J mice (TLR2KO) were purchased from Jackson Laboratories, France. All mice were kept under standard housing conditions at the University of Groningen, The Netherlands. Animal experiments were evaluated and approved by the Committee for

Animal Experimentation of the University of Groningen, The Netherlands, according to the guidelines provided by Dutch Animal Protection Act. Influenza monovalent split vaccines of strain A/Beijing/262/95 (H1N1) and A/Sydney/5/97 (H3N2) were purchased from AdImmune Corp, Taiwan (egg derived, formalin inactivated). The concentration of the selleckchem haemagglutinin (HA) in the vaccine was determined using the single radial immunodiffusion BIBF 1120 mouse assay. The standard BLP-SV vaccines consisted of influenza monovalent SV containing 5 µg HA antigen mixed with BLPs (0.15 mg dry-weight). BLPs were prepared as described before [13] and [14]. BLPs were stored at -80 °C until use. BLPs and SV, were

mixed just prior to i.n. administration. All i.n. vaccine doses were delivered in a final volume of 10 µl of PBS. Mice to be i.n. immunized were lightly anaesthetized with 2.5%, v/v, isoflurane over oxygen (0.8 L/min). Once anaesthetized, the mice were vaccinated i.n. every 10 days with 10 µl of sterile PBS containing BLP-SV (BLPs mixed with the influenza A strain (A/Beijing/262/95 (H1N1)) or SV alone and sacrificed at day

34 of the experiment. Mice were vaccinated i.n. 3 times on day 0, 14 and 28 with 10 µl of sterile PBS containing BLP-SV (BLPs mixed with the influenza A strain (A/Sydney/5/97(H3N2)) or SV alone and sacrificed at day 42 of the experiment. SV without BLPs was administered i.m. in 50 µl of PBS as a positive control for the immunogenicity of the antigenic materials. Blood was collected via puncture of the orbital plexus for antibody measurements and the mice were sacrificed on day 34 or 42 via exsanguination by heart puncture under O2/isoflurane anaesthesia. Subsequently, nasal, lung and vaginal washes were conducted for SIgA antibody measurements. For nasal and lung lavages, 1 ml PBS that contained Roche ADP ribosylation factor “complete” protease inhibitor (according to manufacturer’s description) was used. The tube containing the lavage fluid was placed on ice and centrifuged at 300–400 × g for 5 min at 4 °C and supernatants were collected. Vaginal lavages were conducted by repeated pipetting of 0.2 ml of PBS supplemented with Roche “complete” protease inhibitor. All lavage samples were stored at -20 °C. ELISA was performed as previously described [27]. Briefly, ELISA plates (Greiner, The Netherlands) were coated overnight at 4 °C with influenza monovalent split vaccines of strain A/Sidney/5/97 H3N2 or A/Beijing/262/95 H1N1 (AdImmune). The plates were washed twice and blocked in 200 µl of a 2.5% solution of Protifar Plus (Nutricia™) in coating buffer (0.

The wide variation in local immunoglobulin and antibody levels for any individual animal may have been due to the effects of the menstrual cycle as reported in macaques and women [41], [42] and [38]; however, the present study was not powered to analyse this variable. An effective vaccine will require not only sustained antibody production into mucosal fluids but the antibodies DZNeP chemical structure will need to have potent and broad virus neutralising activity. It is known that monomeric gp120 generally fails to elicit such activity [43], [44], [45] and [46] and for this reason we used a trimeric envelope immunogen, gp140, that has demonstrated remarkable stability in vitro (D. Katinger, personal learn more communication)

and is therefore more likely to mimic the native virion envelope spike [2]. Although cross-clade neutralising activity was restricted to MW965.26 and clade B SF162.LS envelope-bearing pseudoviruses and disappointingly no activity was seen against any of a broad range of clade C envelopes, this study has shown that this narrow specificity is not exclusively due to formulation of the immunogen in Carbopol and/or the vaginal route of administration,

as similar results were obtained after intramuscular immunisation in the presence of AS01 adjuvant. Moreover, as in rabbits [21], serum antibodies did not recognise the highly immunogenic gp41-ED residues 598–597 [47] (data not shown), suggesting that the gp41 region of the molecule may be occluded possibly because of the lack of membrane anchoring. Interestingly macaques have been protected against vaginal challenge with SHIVSF162 following systemic or nasal/systemic immunisation with HIV-1SF162 ΔV2 gp140 and protection was associated with serum neutralising antibody [48]. Although the restricted serum neutralising activity Immune system obtained is of questionable

relevance for a protective HIV-1 vaccine it is interesting that the correlation between anti-gp140 IgG binding antibody titre and neutralising activity seen in animals that were primed intramuscularly did not hold true for animals primed intravaginally. This observation suggests factors other than antibody titre alone may be important, including antibody subclass, avidity and fine specificity. Furthermore, we were unable to measure neutralising activity in mucosal fluids and there is a clear need for the development of micro-neutralisation assays that can be used with small volumes of biological fluid. The results obtained here inform the design of our next clinical trial that will run in parallel with a “paraclinical” macaque study that will include envelope-SHIV challenge. Through this iterative process it will be possible to cross-validate the macaque model – essential for the identification of correlates of protective immunity.