الفهرس 
IntroductionOnly 14 pages are availabe for public view
 |  | from | 213 |  |  |
| | | from | 213 | | |
Abstract
In this study we aimed to produce recombinant CL-10 and CL-11 proteins to be then used for immunization of female NZW rabbits to produce specific polyclonal anti-CL-10 and anti-CL-11. In addition, the purpose of the current study was to assess the binding ability of CL-10 and CL-11 to different microorganisms. Moreover, we aimed to identify the lectin pathway recognition molecules that can bind to K. pneumoniae and to assess the effect of CL-11 on susceptibility to K. pneumoniae infection. We finally aimed to assess the effect of administration of anti-CL-11 antibodies on lung inflammation induced by K. pneumoniae. For that, we firstly prepared recombinant CL-10 and CL-11 via recombinant DNA technology in E. coli BL-21 (DE3) pLysS cells using pRSET-B as expression vector. Second, polyclonal antibodies were produced in rabbits, which were immunized subcutaneously with the purified recombinant proteins emulsified with Freund’s adjuvant. The antibodies were purified via protein G Sepharose column and their binding capacity and functional activity were evaluated. The ability of CL-10 and CL-11 to bind to different bacteria was also examined using ELISA technique, as well as using immunostaining for the lungs of infected mice. Finally, the effect of CL-11 on susceptibility to infection with Klebsiella pneumoniae was evaluated in vitro and in vivo, where the results showed that there was no significant difference in the survival of mice treated with anti-CL-11 or isotype-type antibodies. Intranasal administration of Klebsiella pneumoniae caused severe pneumonia accompanied by a marked increase in the levels of TNF-α, IL-1β and IL-6 in the bronchoalveolar lavage fluid. Interestingly, mice that were pre-treated with CL-11 antagonist showed improved lung pathology as well as a significant decrease in cytokine levels.