Humoral immune responses to Theileria parva in cattle as measured by two-dimensional western blotting.

Humoral immune responses to schizont antigens from six stocks of Theileria parva were compared by two-dimensional Western blotting using sera from cattle that had been infected with a T. parva stock or a clone. Isoelectric points of a polymorphic immunodominant molecule (PIM) of schizonts that induces strong antibody responses in cattle ranged from acidic to basic. Molecular masses (Mr) of the PIM of the respective T. parva stocks were as follows: T. parva Muguga, 86 kDa; Mariakani, 83 kDa; Marikebuni, 83 kDa; Uganda, 83 kDa; T. parva Boleni, 83 kDa; and T. parva 7014, 100 kDa. Among nine cattle infected with T. parva Muguga, four produced antibodies to a basic antigen having an Mr of 32 kDa. The PIM of T. parva Muguga, T. parva Boleni, and T. parva 7014 reacted strongly with serum obtained from an animal that had been infected with T. parva Muguga. Two-dimensional Western blotting using antischizont monoclonal antibodies enabled us to differentiate between stocks of T. parva.

Characterization of buffalo-derived theilerial parasites with monoclonal antibodies and DNA probes.

The characteristics of intra-lymphocytic Theileria isolated from African buffalo and from cattle that were infected with buffalo-derived parasites were evaluated using anti-schizont monoclonal antibodies (mAbs) and DNA probes. Antigenic differences were revealed by the reactivities of 27 mAbs with the buffalo-derived parasites isolated from different animals. Antigenic diversity was also seen with Theileria-infected lymphoblastoid cell isolates taken from the lymph nodes and lambda gt11, showed specific hybridization to parasite DNA in Southern blots of restriction enzyme-digested, lymphoblastoid cells infected with buffalo-derived theilerial parasites. Genotypic differences between the buffalo-derived parasites were revealed by the restriction fragment length polymorphisms seen with hybridization of those probes to DNA from cloned and uncloned Theileria-infected cell lines. The evaluation of theilerial parasites derived from buffalo and from cattle which underwent typical T. p. lawrencei reactions, after being infected with buffalo-derived theilerial parasites, did not show any specific phenotypic or genotypic characteristics of these parasites that would distinguish them from T. p. parva and T. p. bovis parasites. The validity of these subspecies distinctions is discussed.

Isolation of Theileria parasites from African buffalo (Syncerus caffer) and characterization with anti-schizont monoclonal antibodies.

Antigenic differences between intra-lymphocytic theilerial parasites isolated from the blood of 18 African buffalo and grown in vitro were assessed with anti-schizont monoclonal antibodies (mAbs). There was marked antigenic diversity both between isolates from different buffalo and between isolates taken at different times from the same buffalo. Many of the isolates from both wild and captive buffalo appeared to consist of mixed parasite populations. Some isolates were found by limiting dilution cloning and mAb testing to contain at least 3 or 4 distinct populations of Theileria. Once cloned, Theileria-infected lymphoblastoid cell lines retained their mAb profiles during prolonged in vitro cultivation and, when recloned, the subclones had the same mAb profile as their parent clone. The implications of these results for further studies on buffalo-derived theilerial parasites are discussed.