Aptamer-containing surfaces for selective capture of CD4 expressing cells.

Aptamers have recently emerged as an excellent alternative to antibodies because of their inherent stability and ease of modification. In this paper, we describe the development of an aptamer-based surface for capture of cells expressing CD4 antigen. The glass or silicon surfaces were modified with amine-terminated silanes and then modified with thiolated RNA aptamer against CD4. Modification of the surface was first characterized by ellipsometry to demonstrate assembly of biointerface components and to show specific capture of recombinant CD4 protein. Subsequently, surfaces were challenged with model lymphocytes (cell lines) that were either positive or negative for CD4 antigen. Our experiments show that aptamer-functionalized surfaces have similar capture efficiency to substrates containing anti-CD4 antibody. To mimick capture of specific T-cells from a complex cell mixture, aptamer-modified surfaces were exposed to binary mixtures containing Molt-3 cells (CD4+) spiked into Daudi B cells (CD4-). 94% purity of CD4 cells was observed on aptamer-containing surfaces from an initial fraction of 15% of CD4. Given the importance of CD4 cell enumeration in HIV/AIDS diagnosis and monitoring, aptamer-based devices may offer an opportunity for novel cell detection strategies and may yield more robust and less expensive blood analysis devices in the future.

[1]  R. Siliciano,et al.  Analysis of host-virus interactions in AIDS with anti-gp120 T cell clones: Effect of HIV sequence variation and a mechanism for CD4+ cell depletion , 1988, Cell.

[2]  L. Qun,et al.  Aberration of CCR7+ CD8+ memory T cells from patients with systemic lupus erythematosus: an inducer of T helper type 2 bias of CD4+ T cells , 2004, Immunology.

[3]  Mehmet Toner,et al.  A robust electrical microcytometer with 3-dimensional hydrofocusing. , 2009, Lab on a chip.

[4]  Gerhard Ziemer,et al.  A New Technique for the Isolation and Surface Immobilization of Mesenchymal Stem Cells from Whole Bone Marrow Using High‐Specific DNA Aptamers , 2006, Stem cells.

[5]  A. Ellington,et al.  Aptamer beacons for the direct detection of proteins. , 2001, Analytical biochemistry.

[6]  Kevin W Plaxco,et al.  A reagentless signal-on architecture for electronic, aptamer-based sensors via target-induced strand displacement. , 2005, Journal of the American Chemical Society.

[7]  G. Whitesides,et al.  Soft lithography in biology and biochemistry. , 2001, Annual review of biomedical engineering.

[8]  A. Plückthun,et al.  PIN-bodies: a new class of antibody-like proteins with CD4 specificity derived from the protein inhibitor of neuronal nitric oxide synthase. , 2006, Biochemical and biophysical research communications.

[9]  Weihong Tan,et al.  Enrichment of cancer cells using aptamers immobilized on a microfluidic channel. , 2009, Analytical chemistry.

[10]  Andrew D Ellington,et al.  Nanotextured substrates with immobilized aptamers for cancer cell isolation and cytology , 2012, Cancer.

[11]  Weihong Tan,et al.  Molecular aptamer beacons for real-time protein recognition. , 2002, Biochemical and biophysical research communications.

[12]  A. Heeger,et al.  Label-free electronic detection of thrombin in blood serum by using an aptamer-based sensor. , 2005, Angewandte Chemie.

[13]  R. Tompkins,et al.  Effect of flow and surface conditions on human lymphocyte isolation using microfluidic chambers. , 2004, Langmuir : the ACS journal of surfaces and colloids.

[14]  R. Flavell,et al.  CD4 T-cell differentiation and inflammatory bowel disease. , 2009, Trends in molecular medicine.

[15]  A. Lackner,et al.  The mucosal immune system: primary target for HIV infection and AIDS. , 2001, Trends in immunology.

[16]  D. Umetsu,et al.  TH1 and TH2 CD4+ cells in human allergic diseases. , 1997, The Journal of allergy and clinical immunology.

[17]  A. Ozcan,et al.  Lensfree holographic imaging of antibody microarrays for high-throughput detection of leukocyte numbers and function. , 2010, Analytical chemistry.

[18]  Alexander Revzin,et al.  Development of an aptamer beacon for detection of interferon-gamma. , 2010, Analytical chemistry.

[19]  Ying Liu,et al.  Aptamer-based electrochemical biosensor for interferon gamma detection. , 2010, Analytical chemistry.

[20]  He Zhu,et al.  A microdevice for multiplexed detection of T-cell-secreted cytokines. , 2008, Lab on a chip.

[21]  Mehmet Toner,et al.  Blood-on-a-chip. , 2005, Annual review of biomedical engineering.

[22]  S. Gujar,et al.  Aberrant Lymphocyte Activation Precedes Delayed Virus-Specific T-Cell Response after both Primary Infection and Secondary Exposure to Hepadnavirus in the Woodchuck Model of Hepatitis B Virus Infection , 2008, Journal of Virology.

[23]  W. Tan,et al.  Capturing cancer cells using aptamer-immobilized square capillary channels. , 2011, Molecular bioSystems.

[24]  A. Revzin,et al.  Biosensors for immune cell analysis-A perspective. , 2012, Biomicrofluidics.

[25]  J. Convit,et al.  Differing lymphokine profiles of functional subsets of human CD4 and CD8 T cell clones. , 1991, Science.

[26]  He Zhu,et al.  A miniature cytometry platform for capture and characterization of T-lymphocytes from human blood. , 2008, Analytica chimica acta.

[27]  D. Umetsu,et al.  Th1 and Th2 CD4+ Cells in the Pathogenesis of Allergic Diseases , 1997, Proceedings of the Society for Experimental Biology and Medicine. Society for Experimental Biology and Medicine.

[28]  Mehmet Toner,et al.  Enrichment using antibody-coated microfluidic chambers in shear flow: model mixtures of human lymphocytes. , 2005, Biotechnology and bioengineering.

[29]  S D Jayasena,et al.  Staining of cell surface human CD4 with 2'-F-pyrimidine-containing RNA aptamers for flow cytometry. , 1998, Nucleic acids research.

[30]  Alexander Revzin,et al.  Micropatterning of Aptamer Beacons to Create Cytokine-Sensing Surfaces , 2010, Cellular and molecular bioengineering.

[31]  Jun Liu,et al.  Using aptamers to visualize and capture cancer cells , 2010, Analytical and bioanalytical chemistry.

[32]  Mehmet Toner,et al.  Panning of multiple subsets of leukocytes on antibody-decorated poly(ethylene) glycol-coated glass slides. , 2006, Journal of immunological methods.

[33]  Mehmet Toner,et al.  Enhancing the performance of a point-of-care CD4+ T-cell counting microchip through monocyte depletion for HIV/AIDS diagnostics. , 2009, Lab on a chip.

[34]  Mehmet Toner,et al.  Cell detection and counting through cell lysate impedance spectroscopy in microfluidic devices. , 2007, Lab on a chip.

[35]  Weihong Tan,et al.  Aptamer-based microfluidic device for enrichment, sorting, and detection of multiple cancer cells. , 2009, Analytical chemistry.

[36]  Paul I. Okagbare,et al.  Highly efficient capture and enumeration of low abundance prostate cancer cells using prostate‐specific membrane antigen aptamers immobilized to a polymeric microfluidic device , 2009, Electrophoresis.

[37]  Mehmet Toner,et al.  A microfluidic device for practical label-free CD4(+) T cell counting of HIV-infected subjects. , 2007, Lab on a chip.

[38]  R P Johnson,et al.  Gastrointestinal tract as a major site of CD4+ T cell depletion and viral replication in SIV infection. , 1998, Science.

[39]  J. Szostak,et al.  In vitro selection of RNA molecules that bind specific ligands , 1990, Nature.