[Selection and identification of DNA aptamers against DC-SIGN].
OBJECTIVE
To develop the high affinity DNA aptamers which can selectively recognize DC-SIGN protein as putative preventive reagents and drugs against infectious diseases.


METHODS
A technique of cell surface-SELEX (TECS-SELEX) was employed to screen the DNA aptamers recognizing DC-SIGN displayed on the surface of NIH3T3 cells. Some aptamers from the aptamer pools with the highest affinity were cloned. The binding affinities between the aptamers and target cells were measured by flow cytometry and ELISA. The values of IC(50); were determined to detect the toxicity of aptamers for cells by MTT method.


RESULTS
The 14th aptamer pool had the highest affinity and the binding rates between single aptamer (ZD8) and DC-SIGN protein were in a dose dependent manner. DC-SIGN antibody can competitively inhibit the binding reactions between the aptamers and their target cells. The selected aptamers have very low or no toxicity for hepatic cells.


CONCLUSIONS
The selected DNA aptamers bound to DC-SIGN protein with high affinity and specificity have been successfully selected, and they can be used both as preventive reagents and drugs against infectious diseases, such as AIDS, hepatitis C and tuberculosis.
Chen
Fang
F
Medical College, Wuhan University, Wuhan 430071, China. chf188@163.com
Zeng
Jie
J
Sun
Ping
P
Pan
Qin
Q
Zhang
Xiao-Lian
XL
D000818:Animals; D052157:Aptamers, Nucleotide / Q000378:metabolism*; D015815:Cell Adhesion Molecules / Q000378:metabolism*; D037181:Lectins, C-Type / Q000378:metabolism*; D051379:Mice; D041681:NIH 3T3 Cells; D011485:Protein Binding; D011956:Receptors, Cell Surface / Q000378:metabolism*; D052156:SELEX Aptamer Technique / Q000379:methods*; D000099215:DC-Specific ICAM-3 Grabbing Nonintegrin
D052157:Aptamers, Nucleotide; D015815:Cell Adhesion Molecules; D037181:Lectins, C-Type; D011956:Receptors, Cell Surface; D000099215:DC-Specific ICAM-3 Grabbing Nonintegrin