RESUMO
To investigate the effect of recombinant human granulocyte colony-stimulating factor (rhG-CSF) on sepsis, chronically catheterized conscious pigs were challenged with Pseudomonas aeruginosa (8 x 10(7) colony-forming units kg-1 h-1) for 84 h (Group A, n = 8). Group B (n = 7) also received rhG-CSF at 5 micrograms kg-1 d-1, the first dose being given 30 min before starting bacterial infusion. Two of the animals in Group A died from pulmonary failure, whereas all those treated with rh-GCSF survived. Fever, severe pulmonary hypertension and systemic hypotension--the latter accompanied at first by a transient hypodynamic, and later a hyperdynamic response--were observed in all of the animals. In Group B, however, the rise in temperature, mean pulmonary arterial pressure (at a later stage of the observation), plasma levels of tumor necrosis factor, and endotoxin were significantly less than in Group A. In the rhG-CSF-treated pigs, an initial leukopenia completely recovered within 24 h (p < .05 vs. Group A). These data suggest that rhG-CSF might be beneficial in the treatment of sepsis.
Assuntos
Fator Estimulador de Colônias de Granulócitos/uso terapêutico , Hemodinâmica/efeitos dos fármacos , Pulmão/fisiopatologia , Infecções por Pseudomonas/tratamento farmacológico , Sepse/tratamento farmacológico , Fator de Necrose Tumoral alfa/efeitos dos fármacos , Animais , Modelos Animais de Doenças , Feminino , Pulmão/efeitos dos fármacos , Masculino , Distribuição Aleatória , Proteínas Recombinantes/uso terapêutico , Testes de Função Respiratória , Sepse/sangue , Sepse/fisiopatologia , SuínosRESUMO
Recombinant outer membrane proteins (Oprs) of Pseudomonas aeruginosa were expressed in Escherichia coli as glutathione S-transferase (GST)-linked fusion proteins. GST-linked Oprs F and I (GST-OprF190-350 [GST linked to OprF spanning amino acids 190 to 350] and GST-OprI21-83, respectively) and recombinant hybrid Oprs (GST-OprF190-342-OprI21-83 and GST-OprI21-83-OprF190-350) were isolated and tested for their efficacy as vaccines in immunodeficient mice. GST-OprF-OprI protected the mice against a 975-fold 50% lethal dose of P. aeruginosa. Expression of GST-unfused OprF-OprI failed in E. coli, although this hybrid protein has been expressed without a fusion part in Saccharomyces cerevisiae and used for immunizing rabbits. The immune rabbit sera protected severe combined deficient (SCID) mice against a 1,000-fold 50% lethal dose of P. aeruginosa. Evidence is provided to show that the most C-terminal part of OprF (i.e., amino acids 332 to 350) carries an important protective epitope. Opr-based hybrid proteins may have implications for a clinical vaccine against P. aeruginosa.
Assuntos
Anticorpos Antibacterianos/uso terapêutico , Proteínas de Bactérias/uso terapêutico , Imunoterapia , Lipoproteínas/uso terapêutico , Porinas/uso terapêutico , Infecções por Pseudomonas/prevenção & controle , Animais , Anticorpos Monoclonais/uso terapêutico , Proteínas de Bactérias/genética , Proteínas de Bactérias/imunologia , Sequência de Bases , Mapeamento de Epitopos , Escherichia coli/genética , Feminino , Terapia de Imunossupressão , Imunoterapia Ativa , Imunoterapia Adotiva , Lipoproteínas/genética , Lipoproteínas/imunologia , Camundongos , Camundongos Endogâmicos BALB C , Camundongos SCID , Dados de Sequência Molecular , Porinas/genética , Porinas/imunologia , Infecções por Pseudomonas/mortalidade , Proteínas Recombinantes de Fusão/uso terapêutico , Saccharomyces cerevisiae/genética , Análise de SobrevidaRESUMO
AroA Salmonella dublin was used as recipient for a plasmid coding for the outer membrane protein I (OprI) of Pseudomonas aeruginosa. Oral immunization of Balb/c mice with recombinant S. dublin induced serum IgG and IgA antibodies against P. aeruginosa. In spleen and Peyer's patches anti-P. aeruginosa IgG- and IgA-secreting cells could be measured by the ELISPOT technique. In an oral challenge of immunosuppressed mice with P. aeruginosa the orally immunized animals had a 58-fold higher LD50 than control animals.
