The paraflagellar rod proteins (PAR) purified from Trypanosoma cruzi epimastigotes were shown to protect mice against an otherwise lethal challenge inoculum of 10(3) bloodstream-form trypomastigotes. The injection route used for immunization was shown to have a marked impact on the development of protective immunity. Mice receiving subcutaneous (s.c.) injections of PAR proteins had reduced bloodstream parasitemias and showed 100% survival following challenge. In contrast, mice immunized via the intraperitoneal (i.p.) route developed parasitemia levels equivalent to those of unimmunized controls and did not survive infection. Western blotting (immunoblotting) demonstrated that sera from both i.p. and s.c. immunized mice reacted specifically with PAR proteins; however, the antibody titer of the i.p. immunized mice was approximately 64-fold greater than that of the s.c. immunized mice, suggesting that the protective response in the s.c. immunized mice is cell mediated rather than humoral.

Archaeological remains of brachyurans (e.g. crabs) are often overlooked as potential paleoenvironmental and paleoclimatical proxies in contrast to other marine and terrestrial invertebrates such as mollusks and landsnails. The potential for fine-scale regional paleoclimate reconstruction based upon these organisms' behavioral ecology has yet to be examined. We present novel isotopic analyses of the remains of a semiterrestrial freshwater crab (Potamon potamios) endemic to southwest Asia recovered from the archaeological site of Khirbat al-Mudayna al-’Aliya (KMA). KMA lies on a southern tributary of the Wadi al-Mujib in the Hashemite Kingdom of Jordan, approximately 40 km east of the Dead Sea. Excavations here recovered architecture, artifacts, and ecofacts dating to a single-period occupation during the early Iron Age (∼1000 cal BCE). Oxygen and carbon isotopes from these brachyuran remains were analyzed in conjunction with a two-year isotopic and morphometric study of the modern potomonautid population near the archaeological site and at nearby wadi systems to assess whether the isotopic values of modern crab populations correlated with variables such as temperature, including isotopic study of the pools in which crabs were found. In turn, these data could be used to reconstruct paleoclimate or paleoenvironment in the archaeological population. The high correlation in oxygen isotope values between crab carapaces and the water of the pools in which they were sampled suggest that variability in isotopic values of crab specimens recovered from KMA is tracking ancient human capture of these organisms across potentially different pools. Further, intra-year variability in isotopic values in modern crab carapaces as well as the similarity of isotopic values across wadi systems with different environments suggest that P. potamios remains cannot be used as reliable paleoenvironmental indicators. The implications of this research will have significance for archaeologists and other researchers in the study of paleoenvironment and paleoecology, and bears upon human mobility, resource acquisition, and human-nonhuman animal relationships.

Previous studies have shown that immunization of mice with the paraflagellar rod proteins (PAR) of Trypanosoma cruzi induces an immune response capable of protecting mice against an otherwise lethal challenge with this parasite. Herein, we define immunologic responses that do or do not play a critical role in PAR-mediated protection. Firstly, PAR-immunized Ab-deficient (muMT) strain mice survived an otherwise lethal T. cruzi challenge, indicating that a B cell response is not required for PAR-induced immunity. However, beta2m -/- mice, which are severely deficient in MHC class I and TCR alphabeta+ CD8+ CD4- T cells, did not survive challenge infection following PAR immunization, indicating that MHC class I/CD8+ T cell function is necessary for protection induced by PAR immunization. Surprisingly, PAR-immunized mice depleted of CD4+ T cells survived a T. cruzi challenge for >84 days postinfection while maintaining a parasitemia that is generally thought to be lethal (i.e., >10(6) trypomastigotes/ml), thus associating CD4+ T cell function with the process of parasite clearance. Consistent with this association, CD4+ T cells from PAR-immunized mice released INF-gamma and stimulated T. cruzi-infected macrophages to release nitric oxide. The importance of IFN-gamma in PAR-induced protective immunity is further indicated by the observation that PAR-immunized INF-gamma knockout mice developed an extremely high parasitemia and did not survive a challenge infection. Thus, while Ab-mediated immune mechanisms are not required for protection induced by PAR immunization, T cell responses are necessary for both elimination of bloodstream parasites and survival.