Cytokine pattern in cystic fibrosis patients during antibiotic therapy and gene therapy using adenoviral vector.

Cystic fibrosis (CF) is a genetic disease caused by mutations in the cystic fibrosis transmembrane conductance regulator (CFTR) gene. Despite improvements in treatment, pulmonary disease still remains the primary cause of death among these patients. In order to introduce a normal CFTR gene copy into airway epithelial cells, adenoviral vectors (AV) have been developed. AV are known to induce an inflammatory reaction that limits transgene expression, and can be potentially harmful. No human study has clearly monitored simultaneously, systemic and local inflammatory reaction, during AV administration. We report here the levels of C-reactive protein (CRP), interleukin (IL)-6, IL-8, tumor necrosis factor-alpha (TNF-alpha), and interleukin-1 receptor antagonist (IL-1Ra) in plasma and bronchoalveolar lavage fluid (BALF) from six cystic fibrosis patients receiving AV encoding CFTR (AdCFTR). AdCFTR was administered to three cohorts of two patients into the nose on day 0, at doses ranging from 105 to 4 x 108 plaque-forming units (pfu), followed, on day 1, by aerosolization of 107 to 5.4 x 108 pfu. In order to ensure that patients were in the best clinical condition, and to further attenuate the broncho-pulmonary inflammation secondary to bacterial infection, they received antibiotic therapy, two weeks prior to AdCFTR administration, until 9 to 11 days after. We found that antibiotics markedly decreased CRP, TNF-alpha, IL-6, IL-1Ra levels in blood. In BALF, antibiotics slightly decreased TNF-alpha levels but had no effect on IL-8 and IL-1Ra, while IL-6 levels increased. AdCFTR administration did not induce any systemic or local cytokine release. In both blood and BALF, CRP, IL-8, IL-1Ra, TNF-alpha decreased, while IL-6 levels increased between day -7 and day 3. One patient presented an asymptomatic increase of all parameters in the BALF on day 7. Twenty one days later, he displayed a clinical deterioration suggestive of an exacerbation. In conclusion, this study demonstrates that antibiotic administration tends to attenuate systemic but not local broncho-pulmonary inflammation in CF patients. In the setting of our study, AdCFTR administration did not induce cytokine release. Further studies are necessary to investigate other inflammatory markers and the mechanisms involved during AV-mediated gene transfer for a better understanding of the immune reaction, which continues to hamper the development of gene therapy for CF patients.

Obliterative airway disease progresses in heterotopic airway allografts without persistent alloimmune stimulus.

BACKGROUND: Up to 50% of human lung allografts develop chronic rejection manifested as obliterative bronchiolitis (OB). This complication frequently progresses despite maximal immunosuppression, suggesting that, once initiated, factors other than alloimmunity play a role in its progression. In animals, heterotopically transplanted allograft airways develop obliterative airway disease (OAD), an immunologically mediated lesion that is used as a preclinical model of OB. We sought to determine whether OAD would progress even after removal from the alloimmune environment. METHODS: Tracheas from Lewis rats were transplanted subcutaneously into Brown Norway recipients to create allografts. After 7 or 14 days of alloimmune stimulus, these allografts were removed and retransplanted into an isogeneic environment for an additional 21 days. Histology was assessed at each time point, with quantitation of the airway epithelium and intraluminal fibroproliferation. RESULTS: Allografts exposed to 14 days of alloimmune stimulus had a significant loss of airway epithelium compared with grafts exposed to only 7 days ( <0.001). There was little fibroproliferation seen in either of these groups. After retransplantation, the grafts initially exposed to 7 days of alloimmune stimulus had few abnormalities. In contrast, the group exposed initially to 14 days of alloimmunity and retransplanted had near complete obliteration of the lumen with fibroproliferation (96.9% occlusion, =0.001) and absent airway epithelium. CONCLUSIONS: OAD progresses despite removal of alloimmunity if the initial period of alloimmune injury is sufficient. Airway epithelial loss correlated with progression to fibroproliferation, suggesting that the epithelium plays a significant role in the pathogenesis of OB.

Chronic parvovirus B19 infection in a pediatric lung transplanted patient.

In immunocompromised patients, clinical manifestations of human parvovirus B19 (PVB19) infection are mostly reported as acute or chronic hematological disorders. Recently, PVB19 infection has been associated with nonhematological symptoms. Four years after lung transplantation, a 9-year-old girl developed a severe anemia with reticulocytopenia requiring blood transfusion. PVB19 DNA was found by polymerase chain reaction in blood. Blood marrow aspiration revealed typical features of PVB19 infection. She was successfully treated with high dose of i.v. Ig. Then, she exhibited recurrent nonregenerative anemia requiring another course of i.v. Ig. PVB19 DNA has been persisted in blood with no specific immune response. At the same time, she suffered from several lung infection syndromes with no microorganism found except PVB19 DNA. Recurrent mild renal dysfunction was noticed with no other explanation than PVB19 infection. This report indicates that pediatric transplanted patients are at risk of chronic PVB19 infection, which can be associated with lung and/or renal disorders.