加护医学部.pptVIP

Differential effects of ethanol on IFN-γ and TNF-α producing splenic T lymphocytes in a murine model of gram-negative pneumonia 加護醫學部 陳志雄 Introduction Prolonged and excessive consumption of alcohol has been shown to predispose the host to severe infectious complications Alcohol consumption has been shown to impair cytokine production. Introduction T cellmediated immune response is critical for the clearance of K. pneumoniae The pro-inflammatory cytokines, IFN-γ and TNF-α have been individually shown to be crucial for the clearance of K. pneumoniae from the lung as seen in different animal models In order to examine the influence of alcohol on the immune response to infection The study group investigated the frequency of TNF-α and IFN-γ produced by splenic T-lymphocytes in a murine model of gram-negative pneumonia initiated after 8 days of alcohol treatment. Method Method A total 32 experimental animals (Balb/c female mice) were used K. pneumoniae [104 CFUs; strain 43816] intranasaly for induction of infection Mice were repeatedly pretreated over 8 days standardized at 09:00 a.m. each day with ethanol intraperitoneally at a dose of 3 mg/g body weight Motor activity, mucous membranes, piloerection and body weight were assessed by one observer on day 1 and day 8 of experiment. Method 2 On day 7, the mice were anesthetized to guarantee aspiration 104 CFU of KP in a total volume of 50μl saline was administered intranasaly using a pipette On day 8, 24 hours after inoculation, mice were anesthesized to receive median abdominal laparotomy. Lungs, spleen and liver were removed. Lungs and livers were evaluated morphologically using routine histological techniques. Splenic lymphocytes were obtained by preparing single-cell suspensions from spleens. Method – Flow cytometric analysis Cells were stimulated with PMA(phorbol 12-myristate 13-acetate)/ionomycin and incubated for 2 hours. Brefeldin A containing Golgi Plug was added then cells were incubated again. Cultured cells wer