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Infect. Immun., 03 1995, 811-817, Vol 63, No. 3
B Carcy, E Precigout, A Valentin, A Gorenflot and J Schrevel
The supernatants of in vitro cultures of Babesia divergens Rouen 1987 in
human erythrocytes, obtained by using a semidefined medium based on human
high-density lipoproteins, were fractionated by gel filtration
chromatography into four fractions, F1 to F4. The crude supernatant as well
as each fraction adjuvanted with Quil-A protected gerbils from mortality
due to a homologous infectious challenge. Analysis of the humoral response
of the 10 protected gerbils with fraction F4, containing major proteins
with molecular masses lower than 50 kDa, showed that a few antigens (from
50 to 17 kDa) could be important candidates for an improved vaccine against
B. divergens babesiosis. As an immunodominant response was directed against
the 37-kDa antigen (Bd37) in two different B. divergens strains tested, a
polyclonal antibody directed against Bd37 was produced in a rabbit. In an
immunofluorescence assay, the anti-Bd37 antiserum strongly labelled small
internal vesicles of the merozoites and the cell surface was diffusely
labelled after fixation, whereas on live merozoites, this labelling was not
observed. [3H]glucosamine-radiolabelling experiments demonstrate that Bd37
is a glycoprotein. The Bd37 protein can also be labelled with
[14C]palmitate but not with [3H]myristic acid. In Triton X-114 temperature
phase partitioning of B. divergens-infected erythrocyte extracts, Bd37 was
exclusively found into the detergent phase, indicating that the
palmitoylated Bd37 protein was in the membrane fraction. In the in vitro
supernatant, the glycoprotein Bd37 was found in a nonpalmitoylated form,
indicating excretion and/or release of the glycoprotein from the merozoite.
Copyright © 1995, American Society for Microbiology
A 37-kilodalton glycoprotein of Babesia divergens is a major component of a protective fraction containing low-molecular-mass culture-derived exoantigens
Laboratoire de Biologie Parasitaire et Chimiotherapie, URA CNRS 114, Museum National d'Histoire Naturelle, Paris, France.
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