Mishell-Dutton assay

Antibody-production, e.g., Mishell-Dutton assay [47-49] Gold standard in animals ex vivo but highly variable in vitro, originally mouse splenocytes later human cells... 

Measurement of a TDAR in vitro is another variation of the PFC assay. This approach was originally developed by Mishell and Dutton (1967). Sple-nocytes are treated in vitro with test compound or vehicle and are sensitized with sRBC for approximately 4-5 days. Following the incubation period, the cultures are then resuspended in agar with sRBC and complement. As in the PFC assay, the number of antibody-producing cells is determined by counting the number of plaques. The Mishell-Dutton assay requires less test substance compared to in vivo approaches and thus lends to the possibility of testing multiple compounds concurrently. 

Other in vitro assays have been developed, which use animal cells, for example, an assay developed by Fischer et al.(2011), which uses rodent blood cells and splenocytes exposed to sheep red blood cells, and the Mishell-Dutton assay, which is an in vitro immunization culture system measuring an primary antibody responses of mouse splenocytes (Michell and Dutton, 1967). Although the HuLA assay of Collinge et al. (2010) only measures the secondary response, it has advantages over these in vitro methods in that it uses human cells and a human disease-relevant antigen. 

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