Characterization of an immunodominant Onchocerca volvulus antigen with patient sera and a monoclonal antibody.

Adult Onchocerca voluvlus and infective larvae, but not microfilariae contain an immunodominant antigen (33,000 and 21,000 Mr in females, 39,000, 33,000, and 21,000 Mr in males, 133,000 Mr in infective larvae) which is recognized by an Onchocerca-specific mAb. The component is part of the reproductive organs and muscles. 96.2% of onchocerciasis sera contained antibodies detectable by immunoblotting against it. Antigen purified by immunoaffinity chromatography was specifically recognized in immunoblots by onchocerciasis sera, but not by sera from other filarial infections. The high immunogenicity, the specificity, and the occurrence in infective larvae of this antigen indicate an immunodiagnostic potential and a possible role in the immunobiology of the parasite.

Frequency and activity of immune interferon (IFN-gamma)- and colony-stimulating factor-producing human peripheral blood T lymphocytes.

This report describes the first frequency estimate of immune interferon (IFN-gamma)- and colony-stimulating factor (CSF)-producing human peripheral blood T lymphocytes. Human peripheral blood lymphocytes (HPBL) were activated with T cell mitogens [concanavalin A (Con A) or phytohemagglutinin (PHA)] in bulk culture and subsequently plated in limiting dilution (LD) microcultures in the presence of irradiated allogeneic HPBL filler cells and culture medium supplemented with human interleukin 2 and T cell mitogen (Con A or PHA). HPBL growing in these cultures were T cells as determined by E-rosetting (greater than 85%) and were positive for the OKT8 marker. The growth frequency (f) of HPBL in LD was f = 1/20-1/100. Such cells were induced with T cell mitogens (Con A or PHA) to release lymphokines into the supernatant. Statistical analysis of the data from the LD experiments showed that (a) the precursor cell frequency for IFN-gamma- and CSF-producing T cells was f = 1/63-1/151 and 1/89-1/217, respectively, and (b) most T cell clones released IFN-gamma and CSF simultaneously. In addition, individual T cell clones were shown to be capable of releasing different CSF types. In principle, this experimental system can be used to evaluate the precursor frequency, relationship and activity of normal or pathological human T cells performing any in vitro T cell function.

Identification of chitinase as the immunodominant filarial antigen recognized by sera of vaccinated rodents.

Acanthocheilonema viteae is a parasitic nematode of rodents. We identified the chitinase of A. viteae infective stage larvae (L3) as the main target of the humoral immune response of jirds, which were protected against challenge infection after vaccination with irradiation attenuated L3. The cDNA of the L3 chitinase has been sequenced, and the deduced amino acid sequence shows significant homologies to chitinases of Brugia malayi microfilariae, insects, yeast, bacteria, and Streptomyces sp. The protein has been characterized by monoclonal antibodies and substrate activity gels. The chitinase of L3 may contribute to degrading the nematode cuticle during molting and thus represents a target of protective immune responses in a phase where the parasite is highly vulnerable. In addition, it has been shown that a similar enzyme exists in uterine microfilariae, which probably has a role in casting the egg shell.

Induction of IL 2 receptor expression and cytotoxicity of thymocytes by stimulation with TCF1.

We investigated the role of T cell cytotoxicity inducing factor 1 (TCF1) in the induction of a cytotoxic T cell response. We found that help-deficient thymocyte cultures supplied with saturating amounts of purified IL 2 did not develop CTL in a 5-day culture. The expression of cytotoxicity was dependent on the addition of TCF1 derived from the T cell hybridoma K15. TCF1 also induced proliferation of thymocytes in the presence of IL 2. Only the PNA- thymocyte subpopulation responded to TCF1 with proliferation and cytotoxicity in the presence of IL 2. The monokine IL 1 also induced proliferation in this subpopulation but failed to induce cytotoxicity. IL 1 was further distinguished from TCF1 by inhibition of IL 1-induced but not TCF1-induced proliferation by anti-IL 1 antibodies. In addition, using anti-IL 2 receptor antibodies (AMT 13), we showed that TCF1 in the presence of IL 2 substantially increased IL 2 receptor expression in thymocytes. IL 1 had the same effect on induction of IL 2 receptor expression as TCF1. Because some effects of IL 1 and TCF1 are distinct and some overlap, we discuss whether IL 1 and TCF1 induce different subsets of PNA- thymocytes.