Thein vitro activity of the microsomal fraction isolated from the spleen and the lymph nodes after immunization of the donor |
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Authors: | Miroslava Hrube?ová |
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Institution: | (1) Department of Immunology, Institute of Microbiology, Czechoslovak Academy of Sciences, Prague 4;(2) Present address: Isotope Laboratory of the Biological Institutes, Czechoslovak Academy of Sciences, Prague 4 |
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Abstract: | The microsomal fraction from the spleen (after perfusion) of immunized rabbits incubated for 20 min at 37° C under usual conditions
in the presence of energy sources incorporates14C-labelled amino acids both into the solubilized (by adding deoxycholate), and into the nonsolubilized part (15%). The cell
supernatant incorporates under these conditions the14C-labelled amino acids into total proteins in the absence of microsomes but in a lower degree. The cell supernatant contains
gamma globulin detectable by immunoelectrophoresis. Gamma globulin obtained by specific precipitation of the solubilized microsomal
fraction with antigamma-globulin serum had an measurable radioactivity. The precipitate of gamma globulin obtained from the
supernatant of the incubation medium in the same manner (after removing the microsomes) had a specific activity twice as high.
On separating the microsomal fraction extract and the incubation medium supernatant on DEAE cellulose most fractions show
on extinction maximum at 260 nm in the first case and at 280 nm in the second case.
The microsomal fraction isolated from the spleen and lymph nodes of immunized pigs-48 and 72 h after revaccination, when incubatedin vitro, incorporated14C-labelled amino acids into total protein. After ultrasonic disintegration in 0.14m NaCl and filtration through a Sephadex
G 25 column it is specifically precipitated with the antigammaglobulin serum. Gamma globulin isolated after incubation of
the microsomal fraction had a measurable radioactivity. AntiHSA antibodies determined by adsorption on immunosorbent did not
possess significant radioactivity. Only the concentrated supernatant of the incubation medium showed minute radioactivity
of 75–94 counts/min /ml. The problem of investigating the formation of nascent specific antibodies on a subcellular levelin vitro during the early period of secondary response to the antigen is discussed, in particular the problem of their detection.
An erratum to this article is available at . |
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