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PBMCs were next allowed to adhere onto autologous serum-treated petri dishes in order to separate monocytes from the lymphocyte population.
First, CD3+/-cells were gated from the lymphocyte gate.
RNA was manually extracted from the lymphocyte suspension.
Cells from the lymphocyte gate do express neither DENV-Ag (0.80 ± 0.77%) nor iNOS (3.7 ± 2.57%).
PB-MSC can be obtained from the lymphocyte separation fluid fraction of mononuclear cells after a density gradient centrifugation [ 3].
Total RNA stored in TRIzol (Invitrogen) at -80°C, was extracted from the lymphocyte samples at Cogenics, Inc. (Morrisville, NC) by standard procedures.
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In order to study human T-cell reactivity to the most conserved structural protein of Lassa virus, the glycoprotein 2 (GP2), seven GP2-specific CD4+ T-cell clones (TCCs) were generated from the lymphocytes of a Lassa antibody positive individual.
Figure 3 shows IL-2 secretion from the lymphocytes against the mESCs in alginate, alginate TMC and alginate TMC mPEG SVA microcapsules.
Genomic DNA was extracted from the lymphocytes present in 200 mL peripheral blood using the TIANamp Blood DNA Kit (Tian-gen Biotech, Beijing, China), according to standard manufacturer's protocols.
Absolute DC numbers were calculated from the lymphocyte-monocyte counts.
Genomic DNA (gDNA) that contained the FIV proviral DNA was isolated from the lymphocytes using a mammalian genomic DNA Isolation miniprep kit (Sigma, St . Louis MO).
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