[Detection of intracytoplasmic Th1/Th2 cytokine profiles in patients with sepsis and severe sepsis]

Sepsis is the leading cause of death in critically ill patients throughout the world. The incidence is increasing despite the major advances in the development of antimicrobial agents and other supportive treatments. Based on multiple studies, it has been shown that patient outcome depends on Th1 and Th2 cytokine response. Moreover, whenever the Th2 response is predominant, the sepsis is more severe. The aim of this study was to evaluate the correlation between cytokine levels and the severity of sepsis in patients. A cross-sectional study on the cellular levels of several pro-inflammatory cytokines was carried out in patients with sepsis and severe sepsis. The study included 37 patients [24 men and 13 women], 26 of them had sepsis and 11 had the severe form of sepsis Thirty-seven healthy volunteers served as controls. The average age of the patients was 57 years [ +/- 23.3 years], with a range of 21 to 92 years. From the whole blood of the subjects, we separated the monocytes and leukocytes, which were then cultured. Using an ELISA method, we measured levels and IL-12 [associated with Th1], and IL-4 and IL-10 [associated with gamma of IFN- Th2] in the cultured cells with and without cell stimulation. No gamma production in the cells of patients with sepsis gamma correlation was found for IFN- and severe sepsis, regardless of whether the patients had died or survived. However, IL-12 levels were significantly decreased in severe sepsis compared with those of sepsis patients [P=0.048]. Furthermore, the cells of expired patients also had significantly decreased IL-12 levels compared with those of, IL-4, and surviving patients [P=0.028]. We also found that the levels of IFN- IL-10 were decreased in patients compared with those of controls, which correlated to their production. However, there was no correlation for IL-12 production between the cells of the patients compared with those of the controls. There was also no correlation for cytokine production between men and women with sepsis and in adults compared with that of elderly patients [>55 years old]. We have shown that the predominating T helper cell subset in patients with severe sepsis, as well as expired patients, is Th2. In conclusion, the correlation of Th1 cytokine production and progression of sepsis was demonstrated. Most probably IL-12 levels would be significantly lower in patients with severe sepsis and those who expired

[Monoclonal antibody production against human spermatozoal surface antigens]

As monoclonal antibodies are potential tools for characterization of soluble or cellular surface antigens, use of these proteins has always been considered in infertility and reproduction research. Therefore, in this study, monoclonal antibodies against human sperm surface antigens were produced. To produce specific clones against human sperm surface antigens, proteins were extracted using solubilization methods. Balb/c mice were immunized intraperitoneally with the proteins using complete Freund's adjuvant in the first injection and incomplete Adjuvant in the following booster injections. Hybridoma cells producing ASA were cloned by limiting dilution. Five stable ASA producing hybridoma clones were achieved and their antibody isotypes were determined by ELISA. All the isotypes were of IgG class. Their cross reactivity with rat and mice spermatozoa was examined but they did not have any cross reactivity. The produced antibodies can be used in further studies to characterize and evaluate each of the antigens present on human sperm surface and determining their role in fertilization