Quantification of measles virus by a virus receptor-dependent and haemagglutinin-specific T cell stimulation assay.
The human measles virus receptor CD46 plays a major role in the uptake of measles virus (MV) for antigen presentation by major histocompatibility complex class II molecules to T cells. On this basis, a new bioassay has been set up to quantify measles virus in a cell free tissue culture supernatant. A stable mouse B cell transfectant expressing CD46 was used as the antigen presenting cell for presentation of measles virus to a haemagglutinin-specific and class II-restricted mouse T cell hybridoma. The measles virus haemagglutinin was quantified by its ability to stimulate IL-2 secretion by the T cells. A good correlation was found between the amount of haemagglutinin measured in supernatants from infected cells using the CD46-dependent T cell stimulation assay and the number of infectious viral particles as determined in a plaque assay. When MV was purified on a discontinuous sucrose gradient, most of the infectious virus and the haemagglutinin antigen were recovered in the same fraction. These data indicate that the CD46-dependent haemagglutinin-specific T cell assay could be used to measure the production of measles virus in the supernatant of infected cells. The assay required only 48 h, was sensitive, highly specific, and did not rely on the replication of the virus. This new bioassay would be applicable for the detection of any other virus provided that antigen presenting cells expressing the corresponding virus receptor and virus envelope glycoprotein-specific T cells are available. Moreover, it would be an interesting tool to monitor the receptor binding properties of attenuated vaccine virus and envelope glycoprotein subunit vaccines.
D000818:Animals; D000938:Antigen-Presenting Cells / Q000276:immunology; D015703:Antigens, CD / Q000378:metabolism*; D000956:Antigens, Viral / Q000032:analysis; D001681:Biological Assay; D002478:Cells, Cultured; D006389:Hemagglutinins, Viral / Q000276:immunology; D000949:Histocompatibility Antigens Class II / Q000276:immunology; D006801:Humans; D008213:Lymphocyte Activation; D008459:Measles virus / Q000276:immunology*; D051916:Membrane Cofactor Protein; D008562:Membrane Glycoproteins / Q000378:metabolism*; D051379:Mice; D011991:Receptors, Virus / Q000276:immunology*; D013601:T-Lymphocytes / Q000276:immunology; D010948:Viral Plaque Assay
D015703:Antigens, CD; D000956:Antigens, Viral; C497857:CD46 protein, human; D006389:Hemagglutinins, Viral; D000949:Histocompatibility Antigens Class II; C497859:Mcp protein, mouse; D051916:Membrane Cofactor Protein; D008562:Membrane Glycoproteins; D011991:Receptors, Virus