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Loss of infectivity of mycobacterial and protozoal exoplasmic parasites after exposure in vitro to the polyenzymic cocktail 'PIGO'.

A myeloperoxidase-mediated oxygen-dependent antimicrobial system has been discovered by Karnovsky, Klebanoff, Rossi, Sbarra, Schultz and their co-workers as the most powerful among several physiological systems that specifically equip the lysosomes of polymorphonuclear leucocytes and endow them with a considerable capacity to destroy micro-organisms after endocytosis (Klebanoff, 1975). Since then the activity in vitro of an analogous polyenzymic free-radial generator named ‘PIGO’ after its composition (peroxidase, iodide, glucose, type 11-oxidase and oxygen) has been systematically checked on a variety of facultative or obligate intracellular parasites that reside within lysosomes or lysosome-related vacuoles in the infected animal, and which have been called ‘exoplasmic parasites’ (Jacques, 1972; Jacques & Demoulin-Brahy, 1974) for that very reason. On the basis of survival tests in vitro (dye exclusion, motility, non-proliferative growth, multiplication), the cidal activity of ‘PIGO’ has already been claimed in the case of numerous exoplasmic parasites including the three mycobacteria of human tuberculosis and pathogenic yeasts (Demoulin-Brahy et al., 1975), two parasitic protozoa (Jacques et al., 1975), Mycobacterium Iepraemurium (R. J. W. Rees & P. J. Jacques, personal communication) and M. leprue (Delville et al., 1976). Only in the latter case, however, was the survival of the parasite assessed by a method in vivo, namely infectivity to mice after inoculation in the food pads of the ‘PIGO’-treated parasite. In view of recently expressed reservations about the validity of classical criteria for survival in vitro (Medzihradsky & Marks, 1975), and because the lack of chemotherapy of most exoplasmic parasitoses renders acceptable any new drugs depressing infectivity (even though they might prove unable to kill the parasite), we decided to carry out the present study to check the infectivity in susceptible animals, of ‘PIGO’-treated Leishmania braziliensis, Trypanosoma cruzi and M. lepraemurium. The parasites were briefly exposed to ‘PIGO’ in cifro (5min at 37°C for L. braziliensis, 20min at 26°C for T. cruzi, and 15min at 27°C for M. lepraemurium) and then immediately inoculated into susceptible animals (in the rear foot pads of hamster for L. braziliensis; intraperitoneally into mice, for both T. cruzi and M . lepraemurium). The animals were checked daily for mortality, regularly biopsied and finally killed after a suitable time (4, 1 or 7 months respectively for L. braziliensis, T. cruzi and M. fepraemurium). Parasites were counted in various organs (foot pads for L. braziliensis-infected