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Infection and Immunity, July 1999, p. 3383-3389, Vol. 67, No. 7
Biochemisches Institut,
Justus-Liebig-Universität Giessen, 35392 Giessen,
Germany1; Department of Biology,
University of York, York YO10 5YW, United
Kingdom2; and Laboratory of Parasitic
Diseases, National Institute of Allergy and Infectious Diseases,
National Institutes of Health, Bethesda, Maryland
208923
Received 20 November 1998/Returned for modification 5 January
1999/Accepted 15 April 1999
We present here a novel approach to identify T-cell antigens from
any infectious agent by use of a library of purified recombinant proteins. Essential features of this strategy include (i) a highly efficient cDNA cloning system which negatively selects against nonrecombinant transformants by making use of the bacterial
EcoK restriction system, (ii) affinity staining of cDNA
clones expressing recombinant proteins, and (iii) a procedure of
simultaneous purification of recombinant proteins from large numbers of
isolated clones (representing the protein library) in a single step
from pools consisting of up to 24 individual clones. The feasibility of
the screening system was confirmed by constructing a protein library of
the human parasite Schistosoma mansoni. The recombinant
antigens of this library were used to stimulate CD4+ T
cells derived from the axillary lymph nodes of mice vaccinated with
irradiated cercariae. In initial screening experiments, we detected
parasite-specific proliferation and gamma interferon (IFN-
0019-9567/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.
Isolation of T-Cell Antigens by Using a Recombinant
Protein Library and Its Application to the Identification of Novel
Vaccine Candidates against Schistosomiasis
)
secretion in response to several pools of cDNA clones. Further analysis
of one particular pool revealed that only one of its constituents
stimulated considerable IFN- secretion by CD4+ T cells
and that the expressed antigen is identical to a small fragment of
myosin heavy chain.
*
Corresponding author. Present address: Department of
Biology, University of York, York YO10 5YW, United Kingdom. Phone: (44) 1904 434387. Fax: (44) 1904 432884. E-mail:
me10{at}york.ac.uk.
Infection and Immunity, July 1999, p. 3383-3389, Vol. 67, No. 7
0019-9567/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.
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