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Observations on Early and Late Post-Sporozoite Tissue Stages in Primate Malaria

II. The hypnozoite of Plasmodium cynomolgi bastianellii from 3 to 105 Days After Infection, and Detection of 36- to 40-Hour Pre-Erythrocytic Forms^*

W. A. Krotoski, R. S. Bray^,,, P. C. C. Garnham, R. W. Gwadz, R. Killick-Kendrick^,,, C. C. Draper^||, G. A. T. Targett^||, D. M. Krotoski^,*, M. W. Guy^||, L. C. Koontz AND F. B. Cogswell^,**,

Confirmation of the existence of a persistent, uninucleate, dormant pre-erythrocytic stage, the hypnozoite, of the relapsing simian malaria parasite, Plasmodium cynomolgi bastianellii, has been obtained by means of experiments involving the intravenous injection into susceptible monkeys of 48 to 85 x 10⁶ sporozoites derived from mosquitoes of a different species and source than employed previously. The development of these hypnozoites was traced from 3 days until 105 days after sporozoite inoculation, employing a sensitive immunofluorescence technique followed by restaining with Giemsa. From an average mean diameter of 4 µm at 3 and 5 days, uninucleate hypnozoites grow to 5 µm at 7 days, then persist with little change until at least 105 days after infection. Strong evidence for the viability of these persistent forms was obtained by treatment of a host monkey with primaquine, which eliminated all trace of hypnozoites present 2 weeks before.

Examination of hepatic tissue from a monkey injected with sporozoites 36 and 40 hours earlier revealed rare uninucleate pre-erythrocytic forms of 2.5-µm diameter. These early forms were present in hepatocytes in a density only approximately 1/30th of that expected on the basis of numbers of pre-erythrocytic stages found in the same animal's liver 7 days after infection. Nevertheless, subinoculation experiments appeared to rule out the circulation as a vehicle for dissemination of any putative early intermediate hepatotropic forms from another site.

Accepted for publication June 30, 1981.

^* Portions of this work were presented at the 29th Annual Meeting of the American Society of Tropical Medicine and Hygiene, Atlanta, Georgia, 4–7 November 1980, and at the 10th International Congress of Tropical Medicine and Malaria, Manila, The Philippines, 9–15 November 1980.

Supported, in part, by Bureau of Medical Services Grant ORL 80-27-76, Health Services Administration, U.S. Public Health Service, Department of Health and Human Services, and in part by the Wellcome Foundation.

Tropical Infectious Disease Research Program, Clinical Research Department, U.S. Public Health Service Hospital, New Orleans, Louisiana 70118. Reprint address: Laboratory Research Department, U.S. Public Health Service Hospital, Carville, Louisiana 70721.

Department of Pure and Applied Biology, Imperial College of Science and Technology, London SW7, England.

Member, External Staff, Medical Research Council, England.

Laboratory of Parasitic Diseases, NIAID, National Institutes of Health, Bethesda, Maryland 20205.

^|| Departments of Tropical Hygiene and Medical Protozoology, London School of Hygiene and Tropical Medicine, London WC1, England.

^** Departments of Biology (D.M.K.) and Parasitology (F.B.C.), Tulane University, New Orleans, Louisiana 70112.