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274: Molecular Typing of Pseudomonas aeruginosa Isolated from CF Lung Transplant Recipients: Implications for BOS Risk
Abstracts environment, making the lung more susceptible for exposure to microbial specimens, toxic agents and other hazardous specimens, may offer a plausible explanation. Particulate matter (PM) is known to induce (neutrophilic) inflammation in the airways, which is the halmark of BOS. Methods and Materials: In this epidemiological single center study we retrospectively investigated the effect of air pollution on the development of BOS and mortality in our lung transplant cohort (n⫽217). Results: The incidence of chronic rejection was higher in those living close to major roads (⬍157 m), with an incidence of 14.7 per 100 person-years versus 9.0 per 100 person-years, respectively (p⫽0.011; figure). The corresponding data for mortality during follow-up were 10.7 deaths per 100 person-years in those living close to major roads and 3.1 per 100 personyears in those living further than 157 meters from a major traffic road (p⫽0.017, figure).[figure1]
Conclusions: These results indicate a strong association between rejection/ mortality in lung transplant patients and the role of road traffic fumes generated from motor vehicle exhaust as calculated by proximity to roads with high traffic density. This is independent of all other known risk factors for the development of BOS. 274 Molecular Typing of Pseudomonas aeruginosa Isolated from CF Lung Transplant Recipients: Implications for BOS Risk A. Nicholson,1 J.D. Perry,1 S. Peart,1 A. Fisher,2 K.F. Gould.1 1 Freeman Hospital, Newcastle upon Tyne, Tyne and Wear, United Kingdom; 2Freeman Hospital, Newcastle upon Tyne, Tyne and Wear, United Kingdom. Purpose: An association between colonisation of the lung allograft with Pseudomonas aeruginosa (Pa) and an increased risk of developing Bronchiolitis Obliterans Syndrome (BOS) has recently been established. Whether this association holds true in Cystic Fibrosis (CF) lung recipients, who are very likely to have been colonised with Pa pre-transplant, remains unclear. As Pa can produce bacteriocins which inhibit colonisation with different Pa strains, we wished to determine if Pa in CF lung recipients represented only re-colonisation of the allograft with pre-transplant stains or the acquisition of new Pa strains which may induce a more florid pro-inflammatory response. Methods and Materials: Eleven CF lung recipients provided 74 isolates of Pa from sputum or BAL samples. Isolates from at least 3 different samples were examined with at least one sample taken pre and post lung transplant. The genotypic relatedness of each recipient’s Pa strains pre and posttransplant was determined using random amplification of polymorphic DNA (RAPD) typing to amplify specific regions of genomic DNA. Results: In vitro culture and susceptibility methods gave the impression that each recipient were colonised with more than one strain of Pa. However, using molecular genotyping, only two recipients (19%) were colonised with more than one RAPD type, one being only in post transplant samples. Nine patients (81%) were colonised with a single RAPD type Pa. in both pre and post transplant samples. Two patients (19%) harboured the same RAPD type, the remaining nine (81%) were colonised with different RAPD types. Conclusions: This pilot study demonstrates that in our centre CF recipients remain colonised with the same, usually a single strain of Pa post transplant as they had pre-transplant. This technology provides a valuable tool to determine if re-colonisation as opposed to new acquisition of Pa is associated with differential risks of subsequent BOS.
S93 275 The Impact of Viral Infections on the Occurrence of Bronchiolitis Obliterans Syndrome and Its Influence on BOS Stage E. Balestro, M. Loy, M. Damin, F. Braccioni, M. Schiavon, F. Lunardi, A. De Filippis, F. Calabrese, F. Rea. University of Medical School of Padova, Padova, Italy. Purpose: Bronchiolitis obliterans syndrome (BOS) remains the major limitation to long-term survival after lung transplantation (LTx). A thorough understanding of the factors that confer high risk of developing BOS is important to help therapeutic strategies. The aim of our study is to investigate the presence of immunological and non-immunological risks factors, in particular viral infections and acute rejection (A1-A3), in the development of BOS. Methods and Materials: We analysed a cohort of patients with a follow-up period from a minimum of twelve months to three years. Sixty-four recipients (F:19/M:45;age 44⫾10) were divided in two groups: 30 patients with BOS and 34 without the occurrence of BOS. Both were followed with scheduled functional test and bronchoscopy for BAL and transbronchial biopsy (TBB). Conventional molecular viral investigation were used to detect common respiratory virus (including CMV). Acute rejection index (ARI: number of acute rejection/number of total TBBsX100) and the infection index (II: number of infection index/number of total BALsX100) were obtained in the two groups. Results: The occurrence of BOS in our population was histologically confirmed by the presence of BO (n⫽30): in 80% of them was diagnosed between 1 and 3 years after LTx and the left 20% after 3 years. Thirteen recipients belong to BOS stage 1, six patients to BOS stage 2 and eleven patients to BOS stage 3. Infection Index was significantly different in the two groups of patients with and without BOS (mean value 37%Vs28%; p⫽0,04), but we didn’t find any correlation between II and BOS stage (p⫽0,09; r⫽0,31). Acute rejection index was similar in the two groups of patients with and without BOS (mean value 31%Vs33% respectively; p⫽0,66), and no correlation was found between ARI and BOS stage (p⫽0,7; r⫽⫺0,05). Conclusions: Our data support the hypothesis that a non-immunological risk factors as recurrent viral infections with predominance of CMV, have a crucial role in the pathogenesis of BOS, even if its influence on BOS stage has not been shown. 276 Epstein Barr Virus Reactivation in the Lung Allograft Is Inversely Associated with Histopathological Acute Rejection in the Current Era of Routine Antiviral Prophylaxis D.T. Keating,1 A. Michaelides,2 G.P. Westall,1,3 G.I. Snell,1,3 T. Kotsimbos.1,3 1The Alfred Hospital, Melbourne, Vic, Australia; 2The Alfred Hospital, Melbourne, Vic, Australia; 3Monash University, Melbourne, Vic, Australia. Purpose: EBV associated post transplant lymphoproliferative disease (PTLD) occurred in up to 30% of EBV Donor seropositive/Recipient seronegative lung transplant recipients (LTR) prior to the use of routine antiviral prophylaxis. The availability of new molecular tools to detect EBV now allows us to fully examine the sub-clinical reactivation dynamics of EBV in the current era of routine antiviral prophylaxis. Methods and Materials: All LTR between May 2002 and July 2003 were included in this study. All patients received initial ganciclovir prophylaxis for 2 weeks followed by either oral valaciclovir or valganciclovir depending on their EBV and CMV donor/recipient serostatus. Bronchoalveolar lavage from surveillance bronchoscopies (week 1, 2, 3, 6, 9, 12) of 53 patients was analysed using EBV PCR to detect EBV reactivation. Additionally, simultaneously obtained lung tissue biopsies were analysed histologically for acute rejection using ISHLT guidelines and all LTR had regular clinical assesssments as part of routine post transplant follow up protocols. Results: Of the 56 LTR, 46 were recipient and/or donor seropositive for EBV IgG, (3/46 EBV primary mismatches).Despite routine antiviral prophylaxis, EBV reactivation was present in the BAL of 22/46 LTR, 18 in the first 6 months.During the first 6 months post transplantation AR was
Conclusions: These results indicate a strong association between rejection/ mortality in lung transplant patients and the role of road traffic fumes generated from motor vehicle exhaust as calculated by proximity to roads with high traffic density. This is independent of all other known risk factors for the development of BOS. 274 Molecular Typing of Pseudomonas aeruginosa Isolated from CF Lung Transplant Recipients: Implications for BOS Risk A. Nicholson,1 J.D. Perry,1 S. Peart,1 A. Fisher,2 K.F. Gould.1 1 Freeman Hospital, Newcastle upon Tyne, Tyne and Wear, United Kingdom; 2Freeman Hospital, Newcastle upon Tyne, Tyne and Wear, United Kingdom. Purpose: An association between colonisation of the lung allograft with Pseudomonas aeruginosa (Pa) and an increased risk of developing Bronchiolitis Obliterans Syndrome (BOS) has recently been established. Whether this association holds true in Cystic Fibrosis (CF) lung recipients, who are very likely to have been colonised with Pa pre-transplant, remains unclear. As Pa can produce bacteriocins which inhibit colonisation with different Pa strains, we wished to determine if Pa in CF lung recipients represented only re-colonisation of the allograft with pre-transplant stains or the acquisition of new Pa strains which may induce a more florid pro-inflammatory response. Methods and Materials: Eleven CF lung recipients provided 74 isolates of Pa from sputum or BAL samples. Isolates from at least 3 different samples were examined with at least one sample taken pre and post lung transplant. The genotypic relatedness of each recipient’s Pa strains pre and posttransplant was determined using random amplification of polymorphic DNA (RAPD) typing to amplify specific regions of genomic DNA. Results: In vitro culture and susceptibility methods gave the impression that each recipient were colonised with more than one strain of Pa. However, using molecular genotyping, only two recipients (19%) were colonised with more than one RAPD type, one being only in post transplant samples. Nine patients (81%) were colonised with a single RAPD type Pa. in both pre and post transplant samples. Two patients (19%) harboured the same RAPD type, the remaining nine (81%) were colonised with different RAPD types. Conclusions: This pilot study demonstrates that in our centre CF recipients remain colonised with the same, usually a single strain of Pa post transplant as they had pre-transplant. This technology provides a valuable tool to determine if re-colonisation as opposed to new acquisition of Pa is associated with differential risks of subsequent BOS.
S93 275 The Impact of Viral Infections on the Occurrence of Bronchiolitis Obliterans Syndrome and Its Influence on BOS Stage E. Balestro, M. Loy, M. Damin, F. Braccioni, M. Schiavon, F. Lunardi, A. De Filippis, F. Calabrese, F. Rea. University of Medical School of Padova, Padova, Italy. Purpose: Bronchiolitis obliterans syndrome (BOS) remains the major limitation to long-term survival after lung transplantation (LTx). A thorough understanding of the factors that confer high risk of developing BOS is important to help therapeutic strategies. The aim of our study is to investigate the presence of immunological and non-immunological risks factors, in particular viral infections and acute rejection (A1-A3), in the development of BOS. Methods and Materials: We analysed a cohort of patients with a follow-up period from a minimum of twelve months to three years. Sixty-four recipients (F:19/M:45;age 44⫾10) were divided in two groups: 30 patients with BOS and 34 without the occurrence of BOS. Both were followed with scheduled functional test and bronchoscopy for BAL and transbronchial biopsy (TBB). Conventional molecular viral investigation were used to detect common respiratory virus (including CMV). Acute rejection index (ARI: number of acute rejection/number of total TBBsX100) and the infection index (II: number of infection index/number of total BALsX100) were obtained in the two groups. Results: The occurrence of BOS in our population was histologically confirmed by the presence of BO (n⫽30): in 80% of them was diagnosed between 1 and 3 years after LTx and the left 20% after 3 years. Thirteen recipients belong to BOS stage 1, six patients to BOS stage 2 and eleven patients to BOS stage 3. Infection Index was significantly different in the two groups of patients with and without BOS (mean value 37%Vs28%; p⫽0,04), but we didn’t find any correlation between II and BOS stage (p⫽0,09; r⫽0,31). Acute rejection index was similar in the two groups of patients with and without BOS (mean value 31%Vs33% respectively; p⫽0,66), and no correlation was found between ARI and BOS stage (p⫽0,7; r⫽⫺0,05). Conclusions: Our data support the hypothesis that a non-immunological risk factors as recurrent viral infections with predominance of CMV, have a crucial role in the pathogenesis of BOS, even if its influence on BOS stage has not been shown. 276 Epstein Barr Virus Reactivation in the Lung Allograft Is Inversely Associated with Histopathological Acute Rejection in the Current Era of Routine Antiviral Prophylaxis D.T. Keating,1 A. Michaelides,2 G.P. Westall,1,3 G.I. Snell,1,3 T. Kotsimbos.1,3 1The Alfred Hospital, Melbourne, Vic, Australia; 2The Alfred Hospital, Melbourne, Vic, Australia; 3Monash University, Melbourne, Vic, Australia. Purpose: EBV associated post transplant lymphoproliferative disease (PTLD) occurred in up to 30% of EBV Donor seropositive/Recipient seronegative lung transplant recipients (LTR) prior to the use of routine antiviral prophylaxis. The availability of new molecular tools to detect EBV now allows us to fully examine the sub-clinical reactivation dynamics of EBV in the current era of routine antiviral prophylaxis. Methods and Materials: All LTR between May 2002 and July 2003 were included in this study. All patients received initial ganciclovir prophylaxis for 2 weeks followed by either oral valaciclovir or valganciclovir depending on their EBV and CMV donor/recipient serostatus. Bronchoalveolar lavage from surveillance bronchoscopies (week 1, 2, 3, 6, 9, 12) of 53 patients was analysed using EBV PCR to detect EBV reactivation. Additionally, simultaneously obtained lung tissue biopsies were analysed histologically for acute rejection using ISHLT guidelines and all LTR had regular clinical assesssments as part of routine post transplant follow up protocols. Results: Of the 56 LTR, 46 were recipient and/or donor seropositive for EBV IgG, (3/46 EBV primary mismatches).Despite routine antiviral prophylaxis, EBV reactivation was present in the BAL of 22/46 LTR, 18 in the first 6 months.During the first 6 months post transplantation AR was