Use of Humanised Rat Basophilic Leukaemia Cell Line RS-ATL8 for the Assessment of Allergenicity of Schistosoma mansoni Proteins

Background

Parasite-specific IgE is thought to correlate with protection against Schistosoma mansoni infection or re-infection. Only a few molecular targets of the IgE response in S. mansoni infection have been characterised. A better insight into the basic mechanisms of anti-parasite immunity could be gained from a genome-wide characterisation of such S. mansoni allergens. This would have repercussions on our understanding of allergy and the development of safe and efficacious vaccinations against helminthic parasites.

Methodology/Principal Findings

A complete medium- to high-throughput amenable workflow, including important quality controls, is described, which enables the rapid translation of S. mansoni proteins using wheat germ lysate and subsequent assessment of potential allergenicity with a humanised Rat Basophilic Leukemia (RBL) reporter cell line. Cell-free translation is completed within 90 minutes, generating sufficient amounts of parasitic protein for rapid screening of allergenicity without any need for purification. Antigenic integrity is demonstrated using Western Blotting. After overnight incubation with infected individuals'' serum, the RS-ATL8 reporter cell line is challenged with the complete wheat germ translation mixture and Luciferase activity measured, reporting cellular activation by the suspected allergen. The suitability of this system for characterization of novel S. mansoni allergens is demonstrated using well characterised plant and parasitic allergens such as Par j 2, SmTAL-1 and the IgE binding factor IPSE/alpha-1, expressed in wheat germ lysates and/or E. coli. SmTAL-1, but not SmTAL2 (used as a negative control), was able to activate the basophil reporter cell line.

Conclusion/Significance

This method offers an accessible way for assessment of potential allergenicity of anti-helminthic vaccine candidates and is suitable for medium- to high-throughput studies using infected individual sera. It is also suitable for the study of the basis of allergenicity of helminthic proteins.     

Phytoplankton dynamics in a deep, tropical, hyposaline lake

The annual variation of the phytoplankton assemblage of deep (64.6 m), hyposaline (8.5 g l^–1) Lake Alchichica, central Mexico (19 ° N, 97° W), was analyzed in relation to thermal regime, and nutrients concentrations. Lake Alchichica is warm monomictic with a 3-month circulation period during the dry, cold season. During the stratified period in the warm, wet season, the hypolimnion became anoxic. N–NH₃ ranged between non detectable (n.d.) and 0.98 mg l^–1, N–NO₂ between n.d. and 0.007 mg l^–1, N–NO₃ from 0.1 to 1.0 mg l^–1 and P–PO₄ from n.d. to 0.54 mg l^–1. Highest nutrient concentrations were found in the circulation period. Chlorophyll a varied from <1 to 19.8 g l^–1 but most values were <5 g l^–1. The euphotic zone (>1% PAR) usually comprised the top 15–20 m. Nineteen algae species were identified, most of them are typical inhabitants of salt lakes. Diatoms showed the highest species number (10) but the small chlorophyte Monoraphidium minutum, the single-cell cyanobacteria, Synechocystis aquatilis, and the colonial chlorophyte, Oocystis parva, were the numerical dominant species over the annual cycle. Chlorophytes, small cyanobacteria and diatoms dominated in the circulation period producing a bloom comparable to the spring bloom in temperate lakes. At the end of the circulation and at the beginning of stratification periods, the presence of a bloom of the nitrogen-fixing cyanobacteria, N. spumigena, indicated nitrogen-deficit conditions. The well-stratified season was characterized by low epilimnetic nutrients levels and the dominance of small single-cell cyanobacteria and colonial chlorophytes. Phytoplankton dynamics in tropical Lake Alchichica is similar to the pattern observed in some deep, hyposaline, North American temperate lakes.     

Lake Patzcuaro (Mexico): a controversy about the ecosystem water regime approached by field references, climatic variables, and GIS

In this paper, a basic question is asked about a well-documented case study in Mexico: how well do we know Lake Patzcuaro? We address water balance as fundamental to the question. Past studies provide ambiguous explanations about the role of either underground infiltration and/or runoff, relative to the lake-level fluctuation. Thus, our suspicions over the database reliability led us to inspect historic records on water levels and climatic variables; check out the altitude of ground references, and analyze traces of runoff watercourses over the terrestrial basin by means of GIS. By making data re-arrangement and corrections, it became evident that the lake is subject to long-term cycles with ca. 40-year peaks, including short-term seasonal cycles within. Sensitivity to climatic conditions was determined, as well as the active influence of runoff as an important hydrologic component that contributes to cause serious damage to the land surface by erosion. Rearrangement of raw data highlights the occurrence of past misinterpretations founded on biased information.     

Benthic macroinvertebrates of tropical streams: functional and trophic diversity of the Lacantún River,Mexico

Limnology - The differences between temperate and tropical seasonality suggest that ecological models developed for temperate streams do not necessarily apply to tropical streams. While detritus is...     

The “mosco” (Hemiptera: Corixidae and Notonectidae) of Lake Cuitzeo,Mexico: an unusual inland water fishery

Limnology - Lake Cuitzeo is Mexico’s second-largest lake (~ 425&nbsp;km2). The West Basin is shallow, seasonally astatic and alkaline, and it contains saltwater; it is in this...