Isolation and quantification of HIV from lymph nodes. 2005

Catherine Tamalet
Laboratory of Virology, Timone Hospital, Marseille, France.

This chapter describes a standardized microculture technique adapted from a peripheral blood mononuclear cells (PBMC) microculture assay to isolate and quantify HIV from lymph nodes. This quantitative lymph node microculture estimates the number of infectious cells per million lymph node mononuclear cells. The assay, as described below, is performed in two 24-well tissue culture plates using six fivefold dilutions. Each sample of patient cells is cocultivated with phytohemagglutinin-stimulated normal donor PBMC for 21 d. The supernatant from each well is assayed for HIV p24 antigen production by the standard HIV p24 enzyme-linked immunosorbent assay (ELISA). HIV-1 infectious titers are expressed as tissue culture infective dose (TCID50/10(6) cells) according to Reed and Muench.

UI MeSH Term Description Entries
D008198 Lymph Nodes They are oval or bean shaped bodies (1 - 30 mm in diameter) located along the lymphatic system. Lymph Node,Node, Lymph,Nodes, Lymph
D009500 Neutralization Tests The measurement of infection-blocking titer of ANTISERA by testing a series of dilutions for a given virus-antiserum interaction end-point, which is generally the dilution at which tissue cultures inoculated with the serum-virus mixtures demonstrate cytopathology (CPE) or the dilution at which 50% of test animals injected with serum-virus mixtures show infectivity (ID50) or die (LD50). Neutralization Test,Test, Neutralization,Tests, Neutralization
D002478 Cells, Cultured Cells propagated in vitro in special media conducive to their growth. Cultured cells are used to study developmental, morphologic, metabolic, physiologic, and genetic processes, among others. Cultured Cells,Cell, Cultured,Cultured Cell
D004797 Enzyme-Linked Immunosorbent Assay An immunoassay utilizing an antibody labeled with an enzyme marker such as horseradish peroxidase. While either the enzyme or the antibody is bound to an immunosorbent substrate, they both retain their biologic activity; the change in enzyme activity as a result of the enzyme-antibody-antigen reaction is proportional to the concentration of the antigen and can be measured spectrophotometrically or with the naked eye. Many variations of the method have been developed. ELISA,Assay, Enzyme-Linked Immunosorbent,Assays, Enzyme-Linked Immunosorbent,Enzyme Linked Immunosorbent Assay,Enzyme-Linked Immunosorbent Assays,Immunosorbent Assay, Enzyme-Linked,Immunosorbent Assays, Enzyme-Linked
D006801 Humans Members of the species Homo sapiens. Homo sapiens,Man (Taxonomy),Human,Man, Modern,Modern Man
D015497 HIV-1 The type species of LENTIVIRUS and the etiologic agent of AIDS. It is characterized by its cytopathic effect and affinity for the T4-lymphocyte. Human immunodeficiency virus 1,HIV-I,Human Immunodeficiency Virus Type 1,Immunodeficiency Virus Type 1, Human
D018920 Coculture Techniques A technique of culturing mixed cell types in vitro to allow their synergistic or antagonistic interactions, such as on CELL DIFFERENTIATION or APOPTOSIS. Coculture can be of different types of cells, tissues, or organs from normal or disease states. Cocultivation,Co-culture,Coculture,Co culture,Co-cultures,Cocultivations,Coculture Technique,Cocultures

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