Report 2008-2010

For that purpose 93 serum samples were tested by indirect and capture
assays. The level of IgG and IgM antibodies was measured. The
serum specimens tests results were compared with commercially available
Progen Hantavirus IgG and IgM detection kit (Heidelberg, Germany).
By comparison serum results with the Progen kit the sensitivity
of IgG and IgM indirect ELISA assays was 100%, the specificity 97.3%
for IgM test and 100% for IgG test. Comparing the capture assay with
Progen kit results the sensitivity was 100% for IgM assay and 80% for
IgG assay. The specificity of IgM capture assay was 97.3% and 95% of
IgG capture assay. In summary, the results of the tests using recombinant
protein were concordant with Progen test and the yeast expressed
N protein is suitable for hantaan diagnostics.
Characterization of monoclonal antibodies against hantavirus nucleocapsid
protein and their use for immunohistochemistry on rodent
and human samples
Monoclonal antibodies are important tools for various applications
in hantavirus diagnostics. Recently we have generated Puumala virus
(PUUV)-reactive monoclonal antibodies (mAbs) by immunization of
mice with chimeric polyomavirus-derived virus-like particles (VLPs)
harboring the 120 amino acids-long amino-terminal region of PUUV
nucleocapsid (N) protein. Here we describe the generation of two
mAbs by co-immunisation of mice with hexahistidine-tagged fulllength
N proteins of Sin Nombre virus (SNV) and Andes virus (ANDV),
their characterization by different immunoassays and comparison with
the previously generated mAbs raised against a segment of PUUV N
protein inserted into VLPs. All mAbs reacted strongly in the ELISA and
Western blot test with the antigens used for immunization and crossreacted
to varying extents with N proteins of other hantaviruses. All
mAbs raised against a segment of PUUV N protein presented on
chimeric VLPs and both mAbs raised against the full-length AND/SNV
N protein reacted with Vero cells infected with different hantaviruses.
The reactivity of mAbs with native viral nucleocapsids was also confirmed
by their reactivity in immunohistochemistry assays with kidney
tissue specimens from experimentally SNV-infected rodents and
human heart tissue specimens from hantavirus cardiopulmonary syndrome
patients. Therefore the described mAbs represent useful tools
for the immunodetection of hantavirus infection.
Biotechnological approaches to improve plant cold tolerance
This project was supported by the Lithuanian State Science and Studies
Foundation grant N-07014. Project participants: Institute of
Biotechnology, Institute of Horticulture and University of Agriculture.
An economical value and competitiveness of the important and industrial
energy crops rape (Brassica napus), Miscanthus and horticultural
plants as a source of colorants, medical and food compounds
and their rootstocks is reduced by limited adaptation – winter hardiness
and cold adaptation in Lithuania. The aim of this project was
combine an experience of the participants in research on industrially
important plants, gene engineering, and development of cold resistant
plant selection technologies. The main goal of this integrated project
was to develop transgenic plants with improved cold hardening
and resistance properties. In addition, genetic transformation of the
plants require prior investigation of traits implicated in cold hardiness
that could be used to assess effects of interaction of transgenes with
the endogenous mechanisms controlling acclimation processes.
The best known and probably the most cold-specific CBF/DREB cold
resistance pathway was studied in Brassica napus. By homology-based
PCR, analogs of cold inducible transcription factors CBF, ICE and MYB
from B. napus were cloned. Transgenic lines of model plant A. thaliana
have been constructed and were used in function analysis of BnCBF,
BnICE and BnMYB in Arabidopsis. Proposed function of the BnICE
protein as transcription factor was supported by the finding that it is
localised in nucleus.
Figure 4. Expression of Bn-ICE-GFP in roots of transgenic A.
thaliana 35S:BnICE-GFP plants. Green fluorescence - GFP, red fluorescence
- propidium jodide stained cell walls.
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