Three different variants of the recombinant hybrid outer membrane protein OprF (aa 190-342)-OprI (aa 21-83) could be obtained in high yield after expression in Escherichia coli. The hybrid protein was modified N terminally, either with a minimal histidine tag or with a homologous sequence of OprF. Both recombinant proteins were purified by nickel chelate affinity chromatography under native and denaturing conditions, and this produced three suitable candidates for a vaccination trial, protein His-F-I, which was purified in its native as well as in its refolded form; and the native purified N terminally extended protein, ex-F-I. In mice, significantly higher antibody titers and survival rates after challenge with Pseudomonas aeruginosa were observed following immunization with protein His-F-I, purified under native conditions.

Chronic granulomatous disease (CGD) is caused by a failure of granulocytes and macrophages to kill phagocytized microorganisms by reactive oxygen radicals. Prophylactic and therapeutic administration of cell-penetrating antibiotics to CGD-patients has clearly improved the unfortunate prognosis of this disease. To this day infections by Aspergillus species remain the major life-threatening complication of CGD. But the entailing high mortality can be decreased by improving prophylaxis, early recognition and early treatment. We evaluated four own and 57 published cases to develop procedural recommendations Results: 1. Boys and girls are equally at risk. 2. The frequency of Aspergillus-infections increases with increasing age. 3. Recurrent infections are not rare, 4. In many cases, initial symptoms, laboratorial and Xray findings are nonspecific, do not appear to be severe and are easily overlooked. 5. Preceding or concomitant extrapulmonary symptoms are not rare. 6. Bacterial superinfections do occur. 7. Even invasive diagnostic procedures can fail to reveal an invading fungal infection, 8. The risk of a fatal outcome increases if the patient was exposed to a massive pollution of fungal spores or if the fungal infection was diagnosed belatedly. Conclusions and recommendations: 1. Patients must be advised how to avoid the inhalation of great quantities of fungal spores. 2. All patients should be supervised regularly and checked for inflammation, or-Aspergillus-titers in the serum, lung function etc. in a specialised medical center, 3. If a patient presents with any complaints an Aspergillus infection must always be considered (beside other CGD-specific opportunistic pathogens). 4. Even in case of only slight suspicion (semi-)invasive diagnostic procedures are indicated (like: broncho-alveolar-lavage, punctures, biopses etc.). 5. Patients who are doing well, but present any suspicious sign should be treated with itraconazole (for three months at least) even if the fungal infection remains doubtful. 6. Patients suffering from an Aspergillus infection should be treated with high doses of Amphotericin B (eventually enclosed into liposomes) for 6 weeks at least, followed by a three months period of itraconazole administration. Attemps for a continous prophylactic treatment with IFN gamma or itraconazole are promising, but the effectiveness and safety are still under discussion.

Averting initial colonization of the respiratory tract with P. aeruginosa would be of great benefit for patients with cystic fibrosis (CF). Our approach to this problem is mucosal immunization with a vaccine prepared from the OMP fraction of a PAO-1 strain of P. aeruginosa. Sprague-Dawley rats were given 5 intragastric doses of the vaccine on 5 consecutive days and an intranasal booster dose 21 days later. Immunized animals developed high titers of OMP-specific IgG antibodies in serum and a specific IgA response in bronchioalveolar and small intestinal lavage samples, all determined by ELISA. When challenged 7 days after the booster (day 28) by intratracheal injection of live bacteria of a heterologous strain of P. aeruginosa the immunized rats showed enhanced bronchopulmonary bacterial clearance compared to nonimmunized controls, as indicated by bacterial counts from homogenized lung tissue taken 4 hrs after challenge. Thus, mucosal immunization with OMP vaccines might hinder initial colonisation of the lungs with P. aeruginosa.

We report on a five-year-old patient with a chronic pneumonia of the upper left lobe of the lung, in whom a broncho-enteric sequestration with ectopic pancreas was found in the mediastinum. Reviewing the literature, different theories on the embryogenesis of this extremely rare anomaly are discussed.

Averting initial colonization of the respiratory tract with P.aeruginosa would be of great benefit for patients with cystic fibrosis (CF). Our approach to this problem is mucosal immunization with a vaccine prepared from the OMP fraction of a PAO-1 strain of P.aeruginosa. Sprague-Dawley rats were given 5 intragastric doses of the vaccine on 5 consecutive days and an intranasal booster dose 21 days later. Immunized animals developed high titers of OMP-specific IgG antibodies in serum and a specific IgA response in bronchoalveolar and small intesinal lavage samples, all determined by ELISA. When challenged 7 days after the booster (day 28) by intratracheal injection of live bacteria of a heterologous strain of P.aeruginosa the immunized rats showed enhanced bronchopulmonary bacterial clearance compared to nonimmunized controls, as indicated by bacterial counts from homogenized lung tissue taken 4 hrs after challenge. Thus, mucosal immunization with OMP vaccines might hinder initial colonisation of the lungs with P.aeruginosa.