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The effects of actinomycin D on the primary and secondary immune responses induced in vitro

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Title The effects of actinomycin D on the primary and secondary immune responses induced in vitro
Names Thompson, William Carl (creator)
Newburgh, Robert W. (advisor)
Date Issued 1970-02-13 (iso8601)
Note Graduation date: 1970
Abstract In order to assess the importance of RNA synthesis in the induction
and antibody-producing phases of immune responses, actinomycin
D was used to inhibit the primary and secondary in vitro immune
responses of mouse spleen cells against heterologous erythrocytes.
The presence of actinomycin D (0.1 or 0.125 μg /ml) throughout the
entire four day culture period inhibited the development of plaque-forming
cells (PFCs) in both primary and secondary responses by
more than 99%. The same drug concentration, if added after 1, 2, or
3 days of culture, inhibited the number of PFCs on the fourth day by
81-99%.
Utilizing cultures of spleen cells from mice already undergoing
a secondary immune response, no stimulation of the PFC number
upon a 24 hour treatment with actinomycin D (1.25 μg /ml) was found,
unlike the observation of Harris (J. Experimental Medicine 127:675,
1968), with a similar system of cultured rabbit spleen cells. The treatment of these mouse spleen suspensions for 24 hours with actinomycin
D (1.25 μg /ml) usually brought about a 90-95% decrease in the
number of PFCs.
For both the primary and secondary in vitro immune responses
the sensitivity to actinomycin D treatment throughout the culture period
was determined, A drug concentration of 10⁻³ μg/ml caused no
inhibition in either type of response, but rather caused some stimulation
of PFC numbers by the end of the fourth day of culture, The secondary
response was inhibited by 43% and 98% by actinomycin D concentrations
of 3x10⁻³ and 10⁻² μg /ml respectively, The primary response
was inhibited by 95% and 99% by actinomycin D concentrations
of 3x10⁻³ and 10⁻² μg /ml respectively, This sensitivity of the entire
response is much greater than that reported by Uyeki and Llacer
(Biochemical Pharmacology 18:948, 1969) for actinomycin D additions
at a later stage of the response,
The spleen cell suspension was separated into a macrophagerich
fraction and a lymphocyte-rich fraction. Treatment of the
macrophage population for one hour with actinomycin D (0.1 μg /ml),
either during or before the addition of antigen, inhibited by 95% the
immune response which resulted following removal of the drug and
addition of the lymphocyte-rich population to reconstitute the original
cell suspension.
The results are discussed with respect to the possible events
occurring during the response which would account for the observed sensitivity to actinomycin D. A suggestion is made that the extremely
high sensitivity of the early phases of the immune response may be
due to the need for synthesis of some very large RNA molecule.
Genre Thesis/Dissertation
Topic Immunity
Identifier http://hdl.handle.net/1957/45640

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