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Early Diagnostic Markers of Latent and Acute Episodes of Recurrent Genital Herpes
Abstracts
F.3. Early Diagnostic Markers of Latent and Acute Episodes of Recurrent Genital Herpes Irene Rudenko 2, Michael Rudenko 1, Andrey Zabelev 2, Andrey Shemshura 3. 1Brighton and Sussex University Hospitals, Brighton, United Kingdom; 2Eurodon, Rosotov-on-Don, Russian Federation; 3Zdorovye, Rosotov-on-Don, Russian Federation Genital herpes is the fourth most common cause of sexually transmitted diseases. Herpes genitalis infection increases the risk of acquiring HIV after sexual exposure (Renzi C. et al 2003). Transmission of genital herpes as well as self reinfection occur more frequently during an acute phase. It is well known that antiviral treatment at the early stages of a recurrent acute episode is the most effective. Early detection of latent and acute episodes of herpes virus infection is very important. 135 women with recurrent genital herpes who attended outpatient clinics or participated in one of our clinical studies were monitored during 2000-09. 300 episodes and 171 remission screenings were recorded and HSV1,2 IgM and IgG were analyzed in accordance with clinical data and PCR test findings using statistical software SPSS. The presence of virus in vaginal swabs, confirmed by PCR was defined as an episode. We divided all episodes into 3 groups: latent episodes where clinical examination did not reveal any symptoms during 10 days (25.7%), early phase episodes (15.5%) patients reported discomfort, itching and/or minor objective finding including vague erythema and late phase episodes (22.7%) where on examination multiple papules, vesicles or ulcers with erythema, oedema and crusting. All patients were IgG seropositive. IgM titters varied: group 1 (1.78 ± 0.12), group 2 (1.76 ± 0.14), group 3 (1.65 ± 0.09) and control group (0.73 ± 0.11). There was statistically significant difference between control group and other groups (p b 0.05) and no significant difference in-between groups. We can conclude that early detection of latent and recurrent episodes of genital herpes can be done on the bases of elevated IgM to HSV as well as positive PCR during screening in patients with known genital herpes with mild or without any clinical symptoms. doi:10.1016/j.clim.2010.03.225
F.4. Detecting Disease-related Biological Neighborhoods by Human Mucosal Interface Metaproteome Analysis XiaoXiao Li 1, James LeBlanc 1, David Elashoff 1, James Borneman 2, Lee Goodglick 1, Jonathan Braun 1. 1University of California-Los Angeles, Los Angeles, CA; 2University of California-Riverside, Riverside, CA Aberrant interactions between the host and the intestinal bacteria contribute to IBD pathogenesis. We hypothesize that the mucosal surface is a mosaic of biological neighborhoods, each displaying dynamic interactions between the local bacterial biofilms and the underlying host mucosa. The existence of such neighborhoods was tested by a metaproteomic and microbial phylogenetic analysis of mucosal lavage specimens. 88 human subjects (55 normal, 13 ulcerative
S75 colitis, 21 Crohn's disease) underwent endoscopic saline lavage sampling at 6 sites (cecum, ascending, transverse, descending, sigmoid, rectum). Each sample was analyzed by MALDI-TOF/ MS. High-resolution spectra were preprocessed in the SpecAlign software. Permutation and mixed-effect model were used to examine regional and disease-related features. Biological neighborhoods were constructed using weighted correlation network analysis. Microbial phylotypes were analyzed by highthroughput DNA fingerprinting. Bacterial copy numbers were correlated with quantities of peptides to determine the specific microbes associated with the metaproteomic neighborhoods. Significant differences in the mucosal metaproteome were observed between the distal (rectum, sigmoid) and proximal colon in healthy subjects. This geographic difference was diminished in IBD patients, suggesting the mucosal environment is homogenized under the disease condition. Analysis of the metaproteomic data showed several major neighborhoods, defined by clusters of host and bacterial proteins/peptides sharing similar expression patterns. This study establishes a robust and reliable methodology to study the mucosal surface biology. We further demonstrated the existence of mucosal biologic neighborhoods, which are apparently organized by interfacial microbial communities and can be altered by IBD. doi:10.1016/j.clim.2010.03.226
F.5. Altered Synapse Formation by Self-reactive T Cells from MS Patients David Schubert 1, Susana Gordo 1, Santosh Vardhana 2, Jason Pyrdol 1, Nilufer Seth 3, Khadir Raddassi 4, David Hafler 4, Michael Dustin 2, Kai Wucherpfennig 1. 1Dana-Farber Cancer Institute, Boston, MA; 2New York University, New York City, NY; 3Pfizer, Cambridge, MA; 4Brigham and Women's Hospital, Boston, MA Multiple Sclerosis (MS) is an autoimmune disease attacking the central nervous system myelin. The CD4 + T-cell clones Ob.1A12 and Ob.2F3 were isolated from a patient with relapsing remitting MS and recognize an epitope of myelin basic protein (MBP) presented by HLA-DR2. We compared the immunological synapse (IS) formation and signaling between the myelin specific T-cell clones Ob.1A12 and Ob.2F3 and the anti-viral T-cell clones HA1.7 and HA:D7 (specific for an influenza hemagglutinin peptide bound to HLA-DR1 and HLADR4, respectively) using real time imaging of IS formation. We used glass supported planar lipid bilayers that were loaded with fluorescently labeled ICAM-1 and peptide-MHC (pMHC). The anti-viral T-cell clones formed pMHC microclusters within minutes which later turned into central supra-molecular activation cluster (cSMAC) from which ICAM-1 was included. In contrast the self-reactive T-cells formed only small microclusters amd only few cSMACs. Microcluster formation was delayed (19-46 min) and ICAM-1 exclusion was not observed. We used total internal reflection fluorescence microscopy (TIRFM) to investigate the recruitment of molecules into the IS that are involved in signaling and degradation of TCR. Numerous bright microclusters of phosphorylated ZAP-70 were observed in the IS of HA:D7 T-
F.3. Early Diagnostic Markers of Latent and Acute Episodes of Recurrent Genital Herpes Irene Rudenko 2, Michael Rudenko 1, Andrey Zabelev 2, Andrey Shemshura 3. 1Brighton and Sussex University Hospitals, Brighton, United Kingdom; 2Eurodon, Rosotov-on-Don, Russian Federation; 3Zdorovye, Rosotov-on-Don, Russian Federation Genital herpes is the fourth most common cause of sexually transmitted diseases. Herpes genitalis infection increases the risk of acquiring HIV after sexual exposure (Renzi C. et al 2003). Transmission of genital herpes as well as self reinfection occur more frequently during an acute phase. It is well known that antiviral treatment at the early stages of a recurrent acute episode is the most effective. Early detection of latent and acute episodes of herpes virus infection is very important. 135 women with recurrent genital herpes who attended outpatient clinics or participated in one of our clinical studies were monitored during 2000-09. 300 episodes and 171 remission screenings were recorded and HSV1,2 IgM and IgG were analyzed in accordance with clinical data and PCR test findings using statistical software SPSS. The presence of virus in vaginal swabs, confirmed by PCR was defined as an episode. We divided all episodes into 3 groups: latent episodes where clinical examination did not reveal any symptoms during 10 days (25.7%), early phase episodes (15.5%) patients reported discomfort, itching and/or minor objective finding including vague erythema and late phase episodes (22.7%) where on examination multiple papules, vesicles or ulcers with erythema, oedema and crusting. All patients were IgG seropositive. IgM titters varied: group 1 (1.78 ± 0.12), group 2 (1.76 ± 0.14), group 3 (1.65 ± 0.09) and control group (0.73 ± 0.11). There was statistically significant difference between control group and other groups (p b 0.05) and no significant difference in-between groups. We can conclude that early detection of latent and recurrent episodes of genital herpes can be done on the bases of elevated IgM to HSV as well as positive PCR during screening in patients with known genital herpes with mild or without any clinical symptoms. doi:10.1016/j.clim.2010.03.225
F.4. Detecting Disease-related Biological Neighborhoods by Human Mucosal Interface Metaproteome Analysis XiaoXiao Li 1, James LeBlanc 1, David Elashoff 1, James Borneman 2, Lee Goodglick 1, Jonathan Braun 1. 1University of California-Los Angeles, Los Angeles, CA; 2University of California-Riverside, Riverside, CA Aberrant interactions between the host and the intestinal bacteria contribute to IBD pathogenesis. We hypothesize that the mucosal surface is a mosaic of biological neighborhoods, each displaying dynamic interactions between the local bacterial biofilms and the underlying host mucosa. The existence of such neighborhoods was tested by a metaproteomic and microbial phylogenetic analysis of mucosal lavage specimens. 88 human subjects (55 normal, 13 ulcerative
S75 colitis, 21 Crohn's disease) underwent endoscopic saline lavage sampling at 6 sites (cecum, ascending, transverse, descending, sigmoid, rectum). Each sample was analyzed by MALDI-TOF/ MS. High-resolution spectra were preprocessed in the SpecAlign software. Permutation and mixed-effect model were used to examine regional and disease-related features. Biological neighborhoods were constructed using weighted correlation network analysis. Microbial phylotypes were analyzed by highthroughput DNA fingerprinting. Bacterial copy numbers were correlated with quantities of peptides to determine the specific microbes associated with the metaproteomic neighborhoods. Significant differences in the mucosal metaproteome were observed between the distal (rectum, sigmoid) and proximal colon in healthy subjects. This geographic difference was diminished in IBD patients, suggesting the mucosal environment is homogenized under the disease condition. Analysis of the metaproteomic data showed several major neighborhoods, defined by clusters of host and bacterial proteins/peptides sharing similar expression patterns. This study establishes a robust and reliable methodology to study the mucosal surface biology. We further demonstrated the existence of mucosal biologic neighborhoods, which are apparently organized by interfacial microbial communities and can be altered by IBD. doi:10.1016/j.clim.2010.03.226
F.5. Altered Synapse Formation by Self-reactive T Cells from MS Patients David Schubert 1, Susana Gordo 1, Santosh Vardhana 2, Jason Pyrdol 1, Nilufer Seth 3, Khadir Raddassi 4, David Hafler 4, Michael Dustin 2, Kai Wucherpfennig 1. 1Dana-Farber Cancer Institute, Boston, MA; 2New York University, New York City, NY; 3Pfizer, Cambridge, MA; 4Brigham and Women's Hospital, Boston, MA Multiple Sclerosis (MS) is an autoimmune disease attacking the central nervous system myelin. The CD4 + T-cell clones Ob.1A12 and Ob.2F3 were isolated from a patient with relapsing remitting MS and recognize an epitope of myelin basic protein (MBP) presented by HLA-DR2. We compared the immunological synapse (IS) formation and signaling between the myelin specific T-cell clones Ob.1A12 and Ob.2F3 and the anti-viral T-cell clones HA1.7 and HA:D7 (specific for an influenza hemagglutinin peptide bound to HLA-DR1 and HLADR4, respectively) using real time imaging of IS formation. We used glass supported planar lipid bilayers that were loaded with fluorescently labeled ICAM-1 and peptide-MHC (pMHC). The anti-viral T-cell clones formed pMHC microclusters within minutes which later turned into central supra-molecular activation cluster (cSMAC) from which ICAM-1 was included. In contrast the self-reactive T-cells formed only small microclusters amd only few cSMACs. Microcluster formation was delayed (19-46 min) and ICAM-1 exclusion was not observed. We used total internal reflection fluorescence microscopy (TIRFM) to investigate the recruitment of molecules into the IS that are involved in signaling and degradation of TCR. Numerous bright microclusters of phosphorylated ZAP-70 were observed in the IS of HA:D7 T-