A RAB family antigen of Angiostrongylus cantonensis induces a Th1-type immune response in vitro and in vivo.

A cDNA library was constructed from an Angiostrongylus cantonensis young adult and the encoded proteins were expressed in Escherichia coli. One reactive antigen, a RAB-2 protein, was selected using an immunoscreening technique. The expression of the Th1-type cytokine IFN-γ was elicited in mouse splenic cells that were co-cultured with the recombinant RAB-2 protein and in the sera of mice that were immunised with this protein and adjuvant (50 µg at 2-week intervals). In the A. cantonensis-infected groups, the mice were orally infected with 35 infective larvae, and a subset of the infected mice were immunised with the recombinant RAB-2 protein in adjuvant. Serum samples were collected every week for ELISA, and the pathological examinations were performed at 14 days post infection (dpi). An increase in IFN-γ expression was noted in the blood, and the brain sections revealed moderate eosinophilic meningitis in the immunised mice. The RAB-2 antigen of A. cantonensis induced a Th1-type immune response both in vitro and in vivo.

RAB2A: a major subacrosomal protein of bovine spermatozoa implicated in acrosomal biogenesis.

The perinuclear theca (PT) of mammalian sperm is a unique subcellular structure encapsulating the nucleus. Compositionally, the PT is made up of at least six prominent polypeptides (60, 36, 31, 28, 24, and 15 kDa), of which only two have been sequence identified, as well as many less prominent ones. As an ongoing process in unveiling the protein composition of the PT, we have uncovered the sequence identity of the prominent 24-kDa polypeptide (PT24). Initial N-terminal sequence analysis obtained by Edman degradation suggested that PT24 is a RAB2 protein. This was corroborated by mass spectrometric analyses of trypsin-digested fragments of PT24, identifying RAB2A of the RAB2 subfamily as the best sequence match. Quadrapole/time-of-flight analysis identified 72%% sequence coverage between PT24 and bull, human, mouse, or rabbit RAB2A. Since a genome search only identified two RAB2 subfamily members, RAB2A and RAB2B, the 72%% sequence coverage of PT24 provides assurance that this protein is RAB2A and not a new RAB2 subfamily member. Furthermore, commercial RAB2A antibodies, raised against oligopeptide fragments in the unique C-terminal region of RAB2A, specifically labeled PT24 on Western blot analysis of PT extracts. These anti-RAB2A antibodies, along with immune serum that we raised and affinity purified against isolated PT24, demonstrated at both light and electron microscope levels that RAB2 is associated with the periphery of the growing proacrosomic and acrosomic vesicles in the Golgi and cap phases of spermiogenesis and consequently assembled as part of the PT. This pattern of subacrosomal assembly is reminiscent of the pathway used by SubH2Bv (PT15), another prominent and exclusive subacrosomal protein, indicating a common route for subacrosomal-PT assembly. Traditionally somatic RAB2 proteins are involved in vesicular transport between the endoplasmic reticulum and the cis-side of the Golgi apparatus. Our study suggests an unprecedented direction of RAB2A-mediated vesicular transport in spermatids during acrosomal biogenesis, from the trans-side of the Golgi apparatus to the nuclear envelope.