These intervening sequences are all H-chain V-region sequences an

These intervening sequences are all H-chain V-region sequences and are unlikely to cause a strong bias in the PCR amplification; therefore, we have assumed that the relative number of clones represents the relative number of recombined genes in the stimulated B-cell population. In addition, we assumed that transgene-induced allelic exclusion does not bias against Src inhibitor intrachromosomal switching. Previously reported studies of ARS5 mice, which are quite similar to VV29 mice but have much higher transgene copy number, have shown that about 25% of B cells expressed the transgene μa allotype,

whereas 75% of the B cells either expressed endogenous μb allotype or both μb and μa allotypes (25% μb, 50% both μb and μa) 22. Furthermore, in ARS5 B cells, reduction in transgene copy number is correlated with reduced transgene μa expression 22, suggesting that even more inefficient allelic exclusion would be likely in lower copy mice like VV29. It should be noted, however, that it is possible that allelic exclusion in the VV29 mice is not similar to the previous

published similar strains and that we may be overestimating the translocation frequency in this study. Nevertheless, even if we overestimate the translocation frequency by a couple of orders of magnitude, our translocation frequency is still at least five orders of magnitude higher than the 2×10−8 in vitro translocation frequency observed between the Igh/c-myc loci GSI-IX mouse 17. In addition, our calculations may underestimate the translocation frequency because it is unlikely that all of the 110 endogenous V genes are expressed. The higher translocation frequency in the VV29 mouse could be due to the presence of certain Ig cis elements which may increase targeting of the CSR machinery to the VV29 transgene.

For example, assembly of protein complexes that promote long-range CSR may be recruited more easily to the VV29 transgene due to the presence of the Sμ regions or the intronic Eμ enhancer. The Sμ region and the Eμ enhancer, however, may not be the only cis elements required to recruit recombination factors to the transgene. Indeed, previous studies have shown that transgenes lacking the Sμ region or Eμ enhancer Dapagliflozin can also undergo recombination with the endogenous Igh loci 26. Alternatively, it is possible that the lack of certain cis elements, such as the 3′RR enhancers located 28 kb downstream of the Cα gene, may promote increased interchromosomal translocation in VV29 mice. A recent report has shown that interchromosomal translocations between an Igh transgene and the endogenous Igh locus can be detected if the transgene (designated as Δ3′RR) is lacking Igh 3′RR enhancer regions, specifically the DNase I hypersensitive sites HS3a, HS1,2, HS3b, and HS4 27. Based on this finding, the authors hypothesize that interaction between the 3′RR enhancer and the intronic Eμ enhancer may function as a protective mechanism against translocations.

Background: Listeria monocytogenes is a rare cause of peritonitis

Background: Listeria monocytogenes is a rare cause of peritonitis, usually occurring in the setting of cirrhosis or immunosuppression. LY2157299 cell line There are 12 published cases of Listeria monocytogenes peritoneal

dialysis peritonitis in the literature. The 10 patients on continuous ambulatory peritoneal dialysis and 2 with unknown method of peritoneal dialysis were all treated with intravenous or intraperitoneal antibiotics. We report a case occurring in an automated peritoneal dialysis patient, successfully treated with oral antibiotics. Methods: An 87 year old, non-immunosuppressed end-stage renal failure patient on automated peritoneal dialysis, presented with abdominal pain, bloating and diarrhoea after consuming a meal of sushi. She was systemically well and commenced

on empiric outpatient antimicrobial therapy with intraperitoneal vancomycin 2 g and gentamicin 80 mg. Peritoneal dialysate gram stain demonstrated gram positive rods, subsequently culture positive for Listeria monocytogenes. Her antibiotic therapy was changed to amoxicillin 1 g every eight hours orally and she completed total of 22 days of therapy. Her abdominal discomfort resolved and her peritoneal dialysate Vismodegib mw cleared. Results: Repeat dialysate culture one week following completion of antibiotic therapy confirmed resolution of peritonitis. Conclusions: Oral antimicrobial therapy may be effective in treatment of Listeria monocytogenes peritoneal dialysis peritonitis in the systemically well patient. 292 UNUSUAL BLEEDING

IN THIN GLOMERULAR BASEMENT MEMBRANE DISEASE A LEE, J SEVASTOS St Vincent’s Hospital, Sydney, NSW, Australia Aim: We present a case of thin glomerular basement membrane (GBM) disease with unusual manifestations of haematuria, haemoptysis and peritoneal bleeding. Background: Thin GBM disease is caused by a defect of collagen, occasionally Glutamate dehydrogenase associated with loin pain haematuria syndrome. It is considered a disease affecting only the renal tract. There are only few case reports of haemoptysis associated with this condition but there is no literature suggesting bleeding elsewhere. Methods: A young patient presented age 16 with recurrent severe abdominal pain over many months. Laparoscopy for appendicectomy demonstrated no appendicitis but a small amount of blood was found in the pelvis. She subsequently developed intermittent macroscopic haematuria. Cystoscopy showed mild to moderate mucosal bladder erythema and trabeculation, possibly interstitial cystitis. Repeat laparoscopy again noted the presence of free blood in the pelvis. There was no endometriosis and sexually transmitted infection screen was negative. Endoscopy revealed moderate chronic fundal gastritis and colonoscopy to investigate rectal bleeding found a rectal hyperplastic polyp.

11) This difference was statistically significant In addition,

11). This difference was statistically significant. In addition, the areas of the NeuN- and Olig2-positive nuclei exhibited some notable overlap. All six of the cases studied were 1p loss-negative (figures not shown). Previous studies have shown that small numbers of OLCs exhibit Histone Methyltransferase inhibitor neuronal differentiation.[15] However, the exact morphological differences between OLCs and neurocytes remain controversial. OLCs exhibit non-specific ultrastructural features and round, heterochromatic nuclei. Intracytoplasmic organelles

are poor. Microtubules but not intermediate filaments are seen.[15] Oligodendrogliomas with the chromosome 1p/19q codeletion exhibit identical features.[16] The nucleus is heterochromatic and the cytoplasm contains mitochondria, a small rough endoplasmic reticulum (ER) and ribosomes, as well as a few microtubules. The neurocytes contain a small rough ER and are rich in mitochondria; however, direct synaptic attachments on the cell surface are rarely seen. In general, ganglion cells are regarded as being part of the tumor when they exhibit atypia. Daumas-Duport listed two reasons why floating neurons that lack atypia are not entrapped pre-existing neurons.[8] First,

no cytological RXDX-106 solubility dmso variations are seen within normal cortical neurons. Second, these neurons are always present in the subcortical white matter. Since the nuclear size generally correlates with the cytoplasmic size, our morphometric study indicated that the neurons in the specific glioneuronal element possessed cytological variations that are also seen in normal cortical neuron and that they were same in size but rounder compared to normal neurons. In addition, floating neurons were absent or extremely those rare in DNT lesions involving

the subcortical white matter in our study. Moreover, Miyanaga reported a case of DNT that extended into the subarachnoid space.[17] In that case, no floating neurons were identified in the specific glioneuronal element within the subarachnoid space. These observations strongly suggest that Daumas-Duport’s theory might indeed not be a valid assumption. Based on the above results, particularly the fact that Olig2 and NeuN are mutually exclusive, we naturally came to the conclusion that the NeuN-positive small and large cells observed within the element are in fact entrapped granular and pyramidal cells within the cortex. We also concluded that OLCs are essentially glial and not neuronal in nature. If our assumption is correct, then DNT might very well be pure glial tumors as opposed to glioneuronal tumors. Although OLCs lack both 1p/19 loss[18] and PDGFRα overexpression[19] which are characteristic features in oligodendrogliomas, OLCs otherwise share a common phenotype with oligodendrogliomas. In conclusion, our results suggest that DNTs are more akin to oligodendroglioma than glioneuronal tumors, although their biological and genetic nature is clearly distinguishing form oligodendroglioma.

Epithelial

cells influence adaptive immunity by affecting

Epithelial

cells influence adaptive immunity by affecting the function of antigen presenting cells (APCs). Before the adaptive immune system can respond to inhaled allergens, the allergens have to be presented to them by professional selleckchem APCs such as macrophages, B cells, dendritic cells (DCs) or even by less professional APCs such as basophils and eosinophils [32, 33]. We have recently created TCR transgenic mice reactive to an immunogenic peptide of Der p 1, one of the major allergens of the HDM Dermatophagoides pteronyssinus, to address which APCs present inhaled allergens to naive CD4+ T cells in the draining mediastinal LNs of the lung [34]. Using this novel tool, only mucosal lining DCs were able to present HDM-derived antigens to T cells in the mediastinal nodes, whereas B cells or macrophages were unable to do so. These results are consistent with other reports demonstrating that only DCs, but not basophils, are able to induce Th2 immunity to HDM upon adoptive transfer

to naive mice, and that CD11chi cells (depleted via the CD11cDTR system) are necessary for the development of Th2 immunity to HDM allergens [8]. It is well established that DCs play a role both in the initiation and maintenance of allergic airway inflammation and asthma, and control many aspects of the disease, including BHR and GCM. DCs do so by controlling the recruitment and activation of Th2 cells, Fer-1 mouse by producing chemokines that attract eosinophils and Th2 cells, and by expressing co-stimulatory molecules for terminal Teff-cell generation (reviewed in [35]). The exact subtype of DCs exerting all these functions is a matter of intense study [36, 37]. In our hands, Th2 priming

was mainly performed by CD11b+ conventional (c)DCs, and not by CD103+ Meloxicam cDCs [34]. The restimulation of Th2 effector cells and recruitment of inflammatory cells was the function of CD11b+CD64+ FceRI+ monocyte-derived DCs [34]. In our previous work, we have found that plasmacytoid DCs induced anti-inflammatory effects and prevented asthma development, possibly by activating Treg cells [38, 39]. As epithelial cells represent the first line of defense to inhaled allergens and also express TLRs, they have the ability to sense the same stimuli as innate immune cells. Triggering of these epithelial cell pattern recognition receptors (PRRs) by PAMPs initiates NF-κB activation and leads to the release of pro-Th2 cytokines such as TSLP, granulocyte-macrophage colony stimulating factor (GM-CSF), IL-1α, IL-25, and IL-33 in mice [40-42]. These cytokines all share the capacity to activate DCs, which then coordinate the subsequent Th2-type immune response. DCs can, however, also be directly activated by stimulation of their PRRs. Additionally, PRR-dependent epithelial cell activation also results in the production of endogenous danger signals such as uric acid, adenosine triphosphate, and lysophosphatidic acid [43].

HIV tetramer (Sanquin, Amsterdam, the Netherlands) served as nega

HIV tetramer (Sanquin, Amsterdam, the Netherlands) served as negative control (< 0·05% positive). We measured CD1d tetramer binding to T cells that were negative for a mixture of FITC-conjugated anti-CD13 (Beckman Coulter), anti-CD14, anti-CD16 and anti-CD19 (B&D Biosciences, San Jose, CA USA) instead of positive for CD3 antibody to avoid blocking or hindering of tetramer binding. NK T cells

in tissues were examined by triple immunofluorescence staining by anti-CD3 antibody combined with anti-TCR Vα24 and Vβ11 antibodies and analysis by confocal laser scanning BMS-907351 solubility dmso microscopy, as described previously [25,26]. In brief, 4-µm cryostat sections from primary tumour and lymph nodes from patients B2 and B7 were air-dried overnight, fixed in acetone for 10 min at room temperature, preincubated in 5% (vol/vol) normal goat serum (Sanquin) and incubated successively with mouse anti-CD3 antibody (Dako A/S, Glostrup, Denmark), biotinylated goat anti-mouse antibody (Dako), normal mouse serum (Sanquin), VX-770 cost mouse anti-human TCR Vα24-FITC, mouse anti-human TCR Vβ11-PE (Beckman Coulter) and rabbit anti-PE antibody (Biogenesis, Poole, UK), followed

by Cy3-conjugated goat anti-rabbit antibody and Cy5-conjugated streptavidin (Jackson Immunoresearch Laboratories, Inc., Palo Alto, CA, USA). Between incubations, sections were rinsed extensively in PBS. For each fluorochrome label, isotype-matched control antibodies were included and found negative. For counting of NK T cells, 2000 CD3+ T cells in two separate tissue sections were examined. Confocal fluorescence images were obtained on a Leica TCS SP (Leica Microsystems, Heidelberg, Germany) confocal Resveratrol system, equipped with an Argon/Krypton/HeliumNeon laser combination. Images were taken using a 40× 1·25 NA objective. Possible spectral leak-through between FITC, Cy3 and Alexa 647, which could give rise to false-positive co-localization

of different signals, was avoided by careful selection of the imaging conditions. Colour photomicrographs were taken from electronic overlays. Statistical significance was determined using the Student’s t-test. Immunomonitoring of RCC patients in the IFN-α trial revealed an exceptionally high percentage of circulating CD3+CD56+ T cells in patient B2 (Table 1). Further analysis indicated that this patient and patient B7 showed significantly elevated levels of NK T cells expressing TCR Vα24/Vβ11 in their peripheral blood compared to a panel of healthy donors (Table 1). There were no large differences between NK T cell numbers pre-, during and post-treatment in each patient, as is reflected in the relatively low standard deviation (s.d.) values for the mean (Table 1).

The benefit of such deactivation is to decrease the instances of

The benefit of such deactivation is to decrease the instances of aberrant immune responses, such as allergic and autoimmune disorders. Pathogenic microorganisms may also have evolved to express antigens that cross-react with gut flora antigens. In infections, the removal or modification of the gut flora is associated with a modification of the phenotype of the host responses. Therefore, some microorganisms may hijack Tregs that are induced or activated

in the gut to limit pathogenic check details responses against gut flora to ensure their own survival. Over time, established GI infections may create a new homeostatic set point, in which reactivity to the chronic pathogen is minimized, with wider implications for responsiveness selleck antibody to self-antigens and allergens which may not be altogether detrimental. At this point, it remains unclear to what extent any recalibration of host immunity is induced purely by the pathogen, or by perturbation of the commensal population, or is a result

of endogenous controls within the immune system itself. On the basis of both human and experimental studies discussed above, it seems likely that all three components play an essential role in reaching a stable and nonpathogenic steady state for the longer term. None. “
“Pregnancy challenges immune cells and immunomodulatory circuits of the mother and the developing fetus to dynamically adapt to each other in an homeostatic and tolerant environment Org 27569 for fetal growth. This entails the coordination of multiple cellular processes all devoted to accommodate and nourish the fetus while protecting the mother from endogenous and exogenous threatens. From the earliest stages of pregnancy, several strategies to efficiently

communicate immune and trophoblast cells within the interface or at a distance were identified and chemokines might act at on different targets through direct or indirect mechanisms. Here, we briefly review some mechanisms of T regulatory cell recruitment to the early maternal–placental interfaces to accomplish immunotolerance and homeostatic control and we discuss evidence on two locally released polypeptides, RANTES (regulated on activation, normal, T-cell expressed, and secreted) and vasoactive intestinal peptide (VIP), as novel contributors to the multiplicity of immune tolerant responses and uterine quiescence requirements.

albicans from non-C albicans species directly in clinical sample

albicans from non-C. albicans species directly in clinical samples. “
“Regulatory T (Treg) cells may play an important role in the pathogenesis of paracoccidioidomycosis (PCM), but data on the role of Treg cells in the context of oral PCM are still scarce. The objectives of this study were to investigate the density of FoxP3+ T regulatory

cells in oral PCM and to correlate the results with the density of Paracoccidioides brasiliensis in the lesions. Cases of chronic oral PCM seen between 2000 and 2008 were included in this study. The diagnosis of all lesions was confirmed with histopathological examination and Grocott-Gomori staining. The quantitative analysis of the viable fungi was conducted in all cases with Grocott-stained slides. Treg cells were identified using antibodies against FoxP3. Pearson correlation coefficient was used check details to test the correlation between the density of fungi and Treg cells. Results were considered significant when P < 0.05. A total of 11 cases of oral PCM were obtained. AG 14699 There was a positive correlation between fungal density and FoxP3+ Treg cells density in oral lesions, however, without statistical significance. A positive relation between Treg cells and fungal density was seen in oral PCM. Further studies are required to

further elucidate the role of these cells in the pathogenesis of oral PCM, as well the clinical significance of these findings. “
“The objective of this study was to investigate the management of suspected fungal nail infections by general practitioners (GPs) and determine whether guidance is sought when submitting specimens for investigation or treating cases. Questionnaires were sent to all GPs (n = 2420) served by five Health Protection Agency (HPA) collaborating laboratories in the South West of England. A total of 769 GPs responded – topical and oral antifungals were never used by 29% and 16% of GPs respectively. When antifungals were prescribed, topicals were normally given because of the severity of infection (32%); Amorolofine (53%) was the preferred choice. Oral Methane monooxygenase antifungals were most often

prescribed after receipt of a laboratory report (77%); Terbinafine was the preferred choice (86%). Seventy percent of GPs would only treat a suspected nail infection with oral antifungals after sending a sample for investigation, yet 27% never waited for a microscopy report before prescribing oral antifungal treatment. GPs routinely send specimens from suspected fungal nail infections for microbiological investigation, yet treatment is often prescribed before a result is received. With clinical signs of fungal infections often non-specific, GPs should rely on laboratory results before prescribing expensive and lengthy antifungal treatments. Laboratories could further reduce antifungal use by including guidance on microscopy and culture reports.

Samples were acquired on a BD LSRFortessa using FACSDiva software

Samples were acquired on a BD LSRFortessa using FACSDiva software (version 6.2, BD Biosciences) and analyzed using FlowJo software (version 9.5.3, Treestar, Ashland, OR, USA). CD8+ cells were enriched by positive selection using magnetic beads (MACS, Miltenyi Biotec). Cells were fluorescence-activated cell sorted (FACS) by BD FACSAriaIII cell sorter using CD39-PE (Biolegend). Purity of all cell sorts was ≥97% as assessed by flow cytometry. Cell lines were tested for their capacity to inhibit proliferation of a Th1

responder clone (Rp15 1–1) and its cognate M. tuberculosis hsp65 p3–13 peptide, presented by HLA-DR3 positive, irradiated (20 Gy) PBMCs as APCs in a coculture assay that has been previously reported [8, 34]. Proliferation was measured

after 3 days of coculture by addition of 0.5 μCi/well and (3H)thymidine incorporation was assessed after 18 h. Values represent means from triplicate HM781-36B wells. For the CFSE-labeling assay, the Rp15 1–1 Th1-responder clone was labeled with 0.005 μM of CFSE and the irrelevant, isogenic T-cell clone (R2F10), with different peptide specificity and HLA-DR2 restriction, with 0.5 μM of CFSE, similar in design to previously described [13]. After 16 h of coculture with 5 × 104 CD8+CD39+ T cells, the p3–13 peptide (50 ng/mL) and HLA-DR3 positive KU-60019 in vivo APCs, cells were harvested and stained for CD3, CD4, and CD8. CFSE intensity was measured on a BD LSRFortessa using FACSDiva software and analyzed using FlowJo software. ARL 67156 trisodium salt hydrate (Sigma-Aldrich) was added to the well in 150 μM and daily during the 3 days of coculture. Anti-CD39 monoclonal antibody BY40/OREG-103 (Orega Biotech, Ecully, France) was added to the well at the first day of coculture at a final concentration of 10 μg/mL, as was the IgG1

isotype control (R&D Systems). Values represent mean ± SE from triplicate wells. Suppressive capacity of CD8+CD39+ Oxymatrine T cells was independent of original proliferation of the Th1 clone, as tested by reducing the cognate peptide concentration in the coculture assays. Reversal of suppression was calculated in proportion to original clone proliferation in the absence of Treg cells, since ARL and anti-CD39 monoclonal antibody interfered directly with Th1 clone proliferation signals in the CD39 pathway, as demonstrated by reduced (3H)thymidine incorporation after 3 days. Percentage blocking was calculated after natural logarithmic transformation, and inhibition of proliferation in the presence and absence of blocking agents was calculated and expressed as percentage [8]. Raw data can be provided per request. Mann–Whitney tests and Wilcoxon signed-ranks tests were performed using GraphPad Prism (version 5, GraphPad Software, San Diego, CA, USA) and SPSS statistical software (version 20, SPSS IBM, Armonk, NY, USA). We acknowledge EC FP6 TBVAC contract no. LSHP-CT-2003–503367, EC FP7 NEWTBVAC contract no. HEALTH-F3–2009—241745, and EC FP7 ADITEC contract no. HEALTH.2011.1.

90 ± 33 00 μmol/L) and the mean serum creatinine in the control g

90 ± 33.00 μmol/L) and the mean serum creatinine in the control group was higher (117.14 ± 44.55 μmol/L), but these differences were not significant (P = 0.69) (Table 3). At the 3-year follow-up, the eGFR was 56.13 ± 12.51 mL/min in the treatment group and 59.39 ± 11.58 mL/min

in the control group (P = 0.40) (Table 3). The rate of change of eGFR was 0.67 ± 2.23 mL/min per year in the treatment group and −0.69 ± 2.15 mL/min per year in the control group (P = 0.068). At baseline and throughout the follow-up, the mean blood pressure was less than 130/80 mmHg in both groups. At the 1-year follow-up, the systolic pressure was 114.79 ± 11.14 mmHg in the treatment group and 116.00 ± 12.74 mmHg in the control group (P = 0.11 and P = 0.02, selleck chemicals llc compared with baseline levels) (Table 3). These changes in blood pressure were comparable. The mean diastolic pressure of each group remained unchanged during the study period. At the 3-year follow-up, the blood pressure was 126.25 ± 8.50/76.67 ± 5.77 mmHg in the treatment group and 127.50 ± 17.08/78.75 ± 6.29 mmHg in the control group (P = 0.90

and P = 0.67, compared with baseline levels) (Table 3). At the 1-year follow-up, the mean plasma cholesterol was 4.12 ± 1.28 mmol/L in the treatment group and 5.03 ± 1.01 mmol/L in the control group (P = 0.02) (Table 3). At the 3-year follow-up, the plasma cholesterol had declined in both groups (3.90 ± 0.65 mmol/L and 4.75 ± 1.18 mmol/L, respectively) and was comparable to the baseline levels (P = 0.07 and P = 0.67, respectively) (Table 3). Adverse selleck kinase inhibitor events are listed in Table 4. There were no significant differences

in the baseline levels of AST and ALT with the levels at the 3-year follow-up, indicating they did not have evident liver toxicity. In the treatment group, the ECGs of two patients indicated prolonged QT interval. None of the patients in either group had significant changes in serum potassium. In general, probucol and valsartan were well tolerated. To the best of our knowledge, the present multi-centre study is the first clinical trial to assess the effect of an anti-oxidant Resveratrol in combination with an ARB on the progression of IgA nephropathy. Our results showed probucol plus valsartan led to a more rapid decrease of 24-h urinary protein excretion than valsartan alone. In addition, at the 1- and 2-year follow-up, patients given probucol combined with valsartan had significantly reduced 24-h urinary protein relative to baseline levels, but this reduction was not sustained at the 3-year follow-up. Although kidney function remained stable for 3 years in all of our high risk IgA nephropathy patients. All patients in our study were diagnosed with IgA nephropathy and had increased risk for rapid progression, so they can be regarded as a population with high risk for ESRD.

Although peptide-binding algorithms have greatly enhanced rationa

Although peptide-binding algorithms have greatly enhanced rational peptide design, they are far from perfect. Further, despite their orientation away from the T-cell receptor (TCR), anchor residue substitutions can change pMHC conformation to negatively impact TCR recognition. What is needed then is a bit of magic: a general method for increasing peptide affinity while minimizing changes in TCR specificity. In this issue of the European Journal of Immunology, while seeking to improve the CD8+ T-cell response to the melanocyte differentiation Ag

Gp100, Uchtenhagen et al. [18] appear to achieve the impossible, or at least the improbable. Gp100 expression is greatly enhanced in melanoma, making it an attractive therapeutic vaccine target. Human this website Gp10025–33 peptide (KVPRNQDWL (KVP)) presented by the mouse class I Db allomorph elicits self-reactive mouse CD8+ T cells, while the orthologous mouse peptide (EGSRNQDWL (EGS)) does not (Fig. 1) [19, 20]. Both peptides possess canonical p5N and p9L anchor residues for Db (which has a motif of XXXX NXXX[IML], where selleck screening library X represents any aa, N

= asparagine, I = isoleucine, M = methionine, L = leucine) [4]. Despite identical anchors, EGS binds Db with 100-fold lower affinity than KVP [15], evincing the contribution of nonanchor residues to Db binding [21, 22]. Systematic crosswise substitution of p1–3 between KVP and EVS revealed greatly enhanced peptide binding [15]) and pMHC stability when simply replacing p3 of EGS with proline (Pro; EGPRNQDWL second (EGP)) [23]. Immunization with EGP elicited higher numbers of EGS-specific CD8+ T cells than EGS itself, and critically, protected against tumor challenge while the homologous peptide did not [23]. Uchtenhagen et al. [18] scrutinized the structural basis for enhanced EGP peptide affinity with surprising and potentially

generally applicable findings. X-ray crystallography of Db complexed with Gp100 peptides KVP, EGS, or EGP revealed a conserved peptide conformation and similar peptide- Db hydrogen bonding in each complex [23]. Thus, the EGP’s increased affinity was not due to large structural alterations in the complex. Notably, in EGP, the pyrrilodine ring of p3P and the hydroxyphenyl group of Db-Y159 formed CH-π interactions, which affords substantial intermolecular-binding energy [24, 25] (and see http://www.tim.hi-ho.ne.jp/dionisio/page/whatis.html). To examine the contribution of CH-π interactions (which occur with aromatic residues) to EGP/Db stability, Uchtenhagen et al. substituted Y159 with either another aromatic (F) or aliphatic residues with a short (A) or long (L) side chain. Intriguingly, the enhanced pMHC stability of EGP versus EGS was abrogated with Db-Y159A or Db-Y159L. An intermediate effect was observed with Y159F, consistent with reduced energetic stabilization of Phe-Pro CH-π interactions compared with that of Tyr-Pro.