Serum samples and immunoglobulin fractions of eight mammalian species were applied to a Sepharose--protein A column. As with the human immunoglobulin subclasses IgG1, IgG2 and IgG4, all examined IgG classes and subclasses were bound to a greater or lesser extent to protein A. However, the binding of IgG1 of ruminants was very poor. Polyclonal IgM and IgA of the pig, the dog and the cat may be separated in protein A reactive and protein A non-reactive fractions. In addition, monoclonal canine IgM and IgA partially reacted with protein A. In combination with methods such as ammonium sulphate precipitation, ion exchange chromatography and gel-filtration, affinity chromatography with protein A is recommended for the rapid purification of certain Ig (sub)classes of a number of mammalian species.
Hartman, E.G., van Houten, M., Frik, J.F. and van der Donk, J.A., 1984. Humoral immune response of dogs after vaccination against leptospirosis measured by an IgM-and IgG-specific ELISA. Vet. Immunol. Immunopathol., 7: 245-254.An IgM-and IgG-specific ELISA was used to measure the antibody response stimulated in dogs by vaccination with a leptospiral bacterin containing chemically inactivated Leptospira interrogans serotype icterohaemorrhagiae and serotype canicola leptospires. All dogs produced anti-leptospiral IgM and IgG. The IgM production was of the primary response type after each vaccination (primary vaccination, booster vaccination and annual revaccination). A substantial antileptospiral IgG response could be demonstrated only after the first booster vaccination and the annual revaccination. Annual revaccination resulted in a higher and much longer persisting IgG response than did the first booster vaccination. A revision of the vaccination scheme is suggested.
Quiescent gonocytes were isolated from fetal testes of rat 18-day post coitum and cultured alone or on monolayers of somatic cells from different origins. The gonocytes specifically adhered to Sertoli cells, isolated from 21 to 23-day-old rat testes; this adherence was necessary for their survival in vitro. Addition of follicle-stimulating hormone and testosterone to these cultures did not increase the viability of the gonocytes. Serum was found to be deleterious to the germ cells. Electron-microscopic examination of Sertoli-cell-gonocyte co-cultures revealed the presence of numerous adhesion plaques between these cells, indicating that Sertoli cells and gonocytes are able to communicate in vitro. Gonocytes, in co-culture with Sertoli cells, were viable for at least 9 days. The gonocytes did not spontaneously resume proliferation. The simple culture system described in the present paper should be useful in studying the nature of the factors that are responsible for sending the quiescent gonocytes into the cell cycle and for stimulating the formation of A spermatogonia, a process characterizing the start of spermatogenesis.
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