Effect of 3-aminobenzamide on antigenic variation of Trypanosoma brucei

• Published in: Biochemical pharmacology Vol. 34; no. 23; p. 4151
• Main Authors: Cornelissen, A W, Michels, P A, Borst, P, Spanjer, W, Versluijs-Broers, J A, Van der Meer, C, Farzaneh, F, Shall, S
• Format: Journal Article
• Language: English
• Published: England 01.12.1985
• Subjects: Animals; Antigens, Protozoan; Benzamides - blood; Benzamides - pharmacology; DNA Repair; Female; Glycoproteins - genetics; Guinea Pigs; Mice; Nucleotidyltransferases - physiology; Poly(ADP-ribose) Polymerases; Rats; Trypanosoma brucei brucei - drug effects; Trypanosoma brucei brucei - immunology; Variant Surface Glycoproteins, Trypanosoma
• ISSN: 0006-2952
• DOI: 10.1016/0006-2952(85)90208-4

African trypanosomes, like Trypanosoma brucei, depend on antigenic variation to evade the immune response of the vertebrate host. An antigenic switch corresponds to the activation of a variable surface glycoprotein (VSG) gene from a large silent repertoire. Most switches require the duplicative transposition of a VSG gene, which involves strand breaks in DNA and subsequent repair. The nuclear enzyme adenosine-diphosphoribosyl transferase (ADPRT), which is dependent on the presence of DNA strand breaks for its activity, might be involved in this process because it has a regulatory role in DNA repair in all eukaryotic cells studied so far. In previous work, the presence of ADPRT activity was demonstrated in T. brucei. Moreover, it was also shown in isolated trypanosomes the ADPRT activity, which is stimulated by the induction of DNA strand breaks, could be blocked by the competitive inhibitor 3-aminobenzamide. Here we report experiments using rats which were infected with small numbers of T. brucei expressing VSG gene 118. After two days, the rats were coupled to a continuous intraperitoneal infusion system administrating 3-aminobenzamide in 0.9% NaCl (81.4 mM) at a rate of 0.65 ml/hr/rat for a period of up to five days. Control rats received only a 0.9% NaCl infusion. At days 1, 3 and 5, 250 microliters blood was obtained from a tail artery. Plasma 3-aminobenzamide was determined using a new high performance liquid chromatography method, developed for these experiments. In most rats the plasma concentrations were maintained between 0.8 and 1.2 mM. The rate of antigenic switching was determined by quantitating the fraction of trypanosomes that had lost their VSG 118 coat, using antibody against VSG 118 and a limiting dilution in mice. The average switching rate found was 2.0 X 10(-6) in controls and 1.3 X 10(-7) in drug-treated rats (15-fold reduction). This suggests that ADPRT is required for completing most antigenic switching events. We discuss the possibility that drug-resistant switching only involves non-duplicative VSG gene activation.