Our laboratory is interested in using protein (antibody) microarrays for capture and immunophenotyping of leukocytes from whole blood. In contrast to traditional flow cytometry-based approaches, our cytometry platform employs arrays of leukocyte specific antibodies to capture the desired cell subset (T-cell, monocytes or granulocytes) for “solid-phase” immunophenotyping. These experiments are carried out inside microfluidic devices to allow multiplexing while utilizing a small blood volume. We have successfully employed this device to capture pure CD4 and CD8 T-cells from blood and for detection of cytokine release from these cells. The proportions of CD4 and CD8cells residing on their respective antibody spots closely correlated to flow cytometry results. We have used this platform for quantifying CD4/CD8 ratio in HIV infected blood and envision immediate application for our device for inexpensive, point of care analysis of HIV. Unlike other point of care devices we aim to provide multi-parametric readout as well as analysis of not just leukocyte numbers but also function (cytokine production). In addition to HIV, we are exploring applications for this microdevice for immune profiling of autistic children and for pediatric immunology in general.
T-lymphocytes from human blood isolated on antibody microarrays. (A) Immunofluorescent staining of T-cells captured on an antibody spot shows that cells are CD4+ T-cells. (B) Fluorescently stained antibody spots (green) with captured T-cells (blue and red). (C) Arrays for capturing T-cells and detection of cell-secreted cytokines. Red signal is due to cytokine production by the adjacent T-cells. Cytokine is captured on imprinted anti-cytokine Abs and is detected by fluorescently labeled soluble Abs.
People contributing to this research direction: Gulnaz Stybayeva, Ying Liu, Tam Vu, Tim Kwa, Qing Zhou, Lydia Kwon, Ali Rahimian
Important recent papers on this topic: Stybayeva, G., Mudanyali, O., Seo, S., Silangcruz, J., Macal, M., Ramanculov, E., Dandekar, S., Erlinger, A., Ozcan, A., Revzin, A. “Lensfree Holographic Imaging of Antibody Microarrays for High-Throughput Detection of Leukocyte Numbers and Function” Analytical Chemistry, 2010, 82, 3736-3744
Zhu, H., Stybayeva, G., Silancruz, J., Yan, J., Ramanculov, E., Dandekar, S., George, M.D., Revzin A. “Detecting Cytokine Release from Single T-cells”, Analytical Chemistry, 2009, 81, 8150-8158