[Effect of growth inhibition of the secretory protein SPLUNC1 on Pseudomonas aeruginosa].
OBJECTIVE
To express the recombinant SPLUNC1 protein in HNE1 cells and to study its function of bactericidal and binding to lipopolysaccharide (LPS).


METHODS
Full length of SPLUNC1 gene was cloned into pCMV-tag4A vector and stably transfected into HNE1 cell lines, the supernatant of cell cultures was collected. After being treated with the supernatant, the Pseudomonas aeruginosa was seeded to LB soft agar plate, and the bacteria clones were counted and analyzed. For in vitro LPS binding assay, LPS was coated to 96-well plates. We incubated in the plate with SPLUNC1 protein, and detected the binded SPLUNC1 protein by ELISA. Incubating the FITC-LPS with the SPLUNC1 stably transfected or control cells, the intracellular intensity of fluorescence was observed under the fluorescence microscope.


RESULTS
SPLUNC1 inhibited the bacteria clone formation obviously. Although the binding efficiency of LPS and SPLUNC1 in vitro was very low, more FITC-LPS entered into the SPLUNC1 stably transfected cells.


CONCLUSIONS
SPLUNC1 can inhibit the growth of Pseudomonas aeruginosa and bind LPS, and play an important defensive role in innate immunity of the upper airway.
Zhou
Hou-de
HD
Cancer Research Institute, Central South University, Changsha 410011, China.
Wu
Ming-hua
MH
Shi
Lei
L
Zhou
Ming
M
Yang
Yi-xin
YX
Zhao
Jin
J
Deng
Tan
T
Li
Xiao-ling
XL
Sheng
Shou-rong
SR
Li
Gui-yuan
GY
D045744:Cell Line, Tumor; D006023:Glycoproteins / Q000302:isolation & purification / Q000494:pharmacology*; D006801:Humans; D008565:Membrane Proteins / Q000737:chemistry; D009303:Nasopharyngeal Neoplasms / Q000235:genetics / Q000473:pathology; D010750:Phosphoproteins / Q000302:isolation & purification / Q000494:pharmacology*; D011550:Pseudomonas aeruginosa / Q000187:drug effects*; D020545:Respiratory Mucosa / Q000737:chemistry / Q000276:immunology; D012137:Respiratory System / Q000737:chemistry* / Q000276:immunology; D014162:Transfection
C485599:BPIFA1 protein, human; D006023:Glycoproteins; D008565:Membrane Proteins; D010750:Phosphoproteins