rologie i - European Congress of Virology

5 th European Congress of Virology26. VIRAL INFECTIONS INTRANSPLANT ANDIMMUNOCOMPROMISED PATIENTSPosters: REF 539 to REF 562REF 539Determination of anti HCMV T Cell response of potential recipientsand donors in cellular immunotherapy of leukemic patients aftertransplantation of hematopoetic stem cells (SCT)Sarka NEMECKOVA 1 , Katarina BABIAROVA 1 , Jitka KRYSTOFOVA 1 ,Petr HAINZ 1 , Kamila ZURKOVA 1 , Marketa STASTNA MARKOVA 21 Institute of Hematology and Blood Transfusion, Department of experimentalvirology, Prague, CZECH REPUBLIC; 2 Institute of Hematologyand Blood Transfusion, Transplantation Ward, Prague, CZECH REPU-BLICCytomegalovirus (CMV) reactivation in immunosuppressed patients isassociated with morbidity and mortality. The testing CMV specific T cellresponse can help to find the patients which are in a high risk of CMVdisease. Such patients would profit from adoptive transfer of CMV specificT cells. Cellular immune response against CMV of 14 patients after allogeneicSCT was monitored for 180 days by ELISPOT interferon gammawith the aim to find patients with insufficient control of CMV reactivation.The second aim was to compare various forms of CMV antigen forin vitro stimulation. The patients after SCT were classified according tothe anti CMV T cell response, the presence of CMV DNA in the blood andthe response of T cells after stimulation with anti CD3 antibody into fourgroups: 1) CMV DNA (neg.), anti CMV T cells + (pos.); 2) CMV DNA,anti CMV T cells; 3) CMV DNA +, anti CMV T cells, CD3+; 4) CMVDNA +, anti HCMV T cells, CD3. Patients of groups 3,4 would benefitfrom adoptive transfer. By testing the recall response of T cells of donorsagainst pp65, IE 1, US3, UL36, US29, UL55 peptide pools, pp65, IE 1epitopes or CMV lysate we found that inclusion of US3, UL36, US29,UL55 as testing antigens can increase significantly the overall responseof anti CMV T cells. We observed that stimulation with the long 30merpeptides from pp65 or IE 1 sequence induced activation of CD8 and CD4antiviral T cells. The long peptides could be used for T cell activationof multiple epitopes if the MHCI binding minimal CTL epitopes are notknown.Supported by NT/13898/2012 grant.donor/recipient status (D R). Virus load (VL) was measured by RochePCR. In all 10 D+R patients primary HCMV infection led to a developmentof total and IgG3 HCMV ABs, but 2 patients showed no IgG1 response.Total HCMV IgG, and IgG3 AB RLUs remained stable post TX within 1log during follow up in all 9DR+andD+R+ patients without detectableVL and in 86% and 96% of 28 patients with detectable VL, respectively.In contrast, IgG1 subclass ABs were variable in 46% of the patients (RLUvariation >1log) who developed HCMV VL. In 7 cases IgG1 declinedprior to viremia and in 6 patients IgG1 increased during virus replication.HCMV total IgG and IgG3 AB responses are stable over time, while IgG1ABs exhibit substantial variability in relation to virus replication. Theclinical implications of these data will be further elucidated.REF 541Detection of TS polyomavirus from tonsillar tissues of children andadultsMohammadreza SADEGHI 1 , Lea HEDMAN 1,2 , Leena MaijaAALTONEN 3 , Maria SÖDERLUND VENERMO 1 , Klaus HEDMAN 1,21 Department of Virology, Haartman Institute, University of Helsinki, Helsinki,FINLAND; 2 Department of Virology and Immunology, HelsinkiUniversity Central Hospital Laboratory Division, Helsinki, FINLAND;3 Department of Otorhinolaryngology Head and Neck Surgery, HelsinkiUniversity Central Hospital, Helsinki, FINLANDBackground: in the past few years several new polyomaviruses of humanshave been discovered. Trichodysplasia spinulosa associated polyomavirus(TSPyV) was identified in a patient with trichodysplasia spinulosa (TS), arare follicular skin disease of immunocompromised patients. Seroepidemiologicalstudies indicate that TSPyV is ubiquitous and infects ∼ 70% ofhealthy individuals. We have shown that the tonsil may be a site of WUPyVinitial infection and of MCPyV persistence. Whether TSPyV persistenceoccurs in the lymphoid system is unknown.Methods: to determine whether tonsils harbor TSPyV, we tested a total of229 matched pairs of tonsillar tissues and sera from asymptomatic childrenor adults (overall mean age 21.0 years). The samples were also screenedfor the single copy human RNase P gene by real time quantitative PCR(qPCR). TSPyV was detected by qPCRs with Taq Man probes and primerpairs targeting the VP1 or LT genes. Results: TSPyV DNA was detectedin 8 of 229 (3.5%) tonsillar tissues, from both cohorts, and in none ofthe corresponding sera. Sequence analysis of the 8 virus positive samplesconfirmed the identity of TSPyV.Conclusions: these data provide the first evidence of TSPyV in tonsillartissue and suggest that (i) tonsils may serve as an initial site of TS polyomavirusinfection and that (ii) lymphoid tissue may be a persistence sitefor this virus.REF 540Human Cytomegalovirus specific IgG subclass antibody kinetics inthe follow up after lung transplantationBenedikt SIMON, Irene GÖRZER, Elisabeth PUCHHAMMERSTÖCKLDepartment of Virology, Med Uni Wien, Vienna, AUSTRIAHuman cytomegalovirus (HCMV) may cause severe infections in lungtransplant recipients (LTRs). HCMV specific AB response limits virusreplication, but the kinetics of subclass AB response after transplantation(TX) are not clear.We analyzed the HCMV specific IgG AB kinetics of total IgG and of thedominant IgG AB subclasses 1 and 3 after TX and assessed whether theseare associated with HCMV replication in the follow up. HCMV AB titers of53 LTRs were measured repeatedly post TX using the MEDAC CMV IgGELISA, modified for subclass 1 and 3 ABs. ODs were expressed in relativelight units (RLUs). Cut offs were established in patients with HCMVREF 542The Reactivation of Immunomodulating Beta Herpesviruses Infectionin Patients Undergoing Microvascular Free Flap SurgeriesArnis VILKS 1,2 , Romans DZALBS 4 , Santa RASA 3 , Zaiga NoraKRUKLE 3 , Viesturs BOKA 1 , Biruta MAMAJA 2,1 , ModraMUROVSKA 31 Riga Eastern Clinical University Hospital Gailezers, Riga, LATVIA;2 Riga Stradins University, Department of Anaesthesiology and Reanimatology,Riga, LATVIA; 3 Riga Stradins University, A. Kirchenstein Instituteof Microbiology and Virology, Riga, LATVIA; 4 Riga Stradins University,Faculty of Medicine, Riga, LATVIABeta herpesviruses HHV 6 and HHV 7 are immunomodulating viruses.They have the ability to impair host defence system seriously and maycontribute to other infectious pathogens due to immune suppression. Wehave investigated the effect of long lasting microvascular free flap surgeryS270 Virologie, Vol 17, supplément 2, septembre 2013