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Infection and Immunity, November 1998, p. 5406-5413, Vol. 66, No. 11
Department of Veterinary Microbiology and
Pathology, Washington State University, Pullman, Washington
99164,1 and
Department of
Parasitology, Kimron Veterinary Institute, Bet-Dagan,
Israel2
Received 26 May 1998/Returned for modification 28 July
1998/Accepted 4 September 1998
Protective immunity against the ehrlichial pathogen
Anaplasma marginale has been hypothesized to
require induction of immunoglobulin G2 (IgG2) antibody against outer
membrane protein epitopes and coordinated activation of macrophages for
phagocytosis and killing. In the present study, cell-mediated immune
responses, including induction of IgG isotype switching, were
characterized in calves immunized with purified outer membranes of the
Florida strain of A. marginale. Importantly, these
calves were subsequently shown to be protected upon experimental
challenge with the Florida strain, and calves which developed the
highest IgG2 titers were completely protected against infection.
Peripheral blood mononuclear cells (PBMC) obtained after immunization
proliferated strongly in response to both whole A. marginale homogenates and purified outer membranes, and this
responsiveness persisted until the time of challenge. Responding cells
were shown to be CD4+ T cells, and CD4+ T-cell
lines cultured for 2 to 4 weeks also proliferated specifically in
response to A. marginale and produced high titers of
gamma interferon. The helper T-cell response included recognition of conserved epitopes, as PBMC proliferation was stimulated by the homologous Florida strain, four genetically distinct A. marginale strains, and Anaplasma ovis. The
outer membrane proteins stimulating the PBMC responses in protected
calves included major surface proteins (MSPs) MSP-1, MSP-2, and
MSP-3, which were previously shown to induce partial protection
against infection. These studies demonstrate, for the first time,
potent helper T-cell responses in cattle protectively immunized with
outer membranes against A. marginale challenge and
identify three MSPs that are recognized by immune T cells. These
experiments provide the basis for subsequent identification of the
helper T-cell epitopes on MSP-1, MSP-2, and MSP-3 that are
needed to evoke anamnestic antibody and effector T-cell responses
elicited by protein or nucleic acid immunization.
0019-9567/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.
CD4+ T-Lymphocyte and Immunoglobulin G2 Responses in
Calves Immunized with Anaplasma marginale Outer Membranes
and Protected against Homologous Challenge
*
Corresponding author. Mailing address: Department of
Veterinary Microbiology and Pathology, Washington State University,
Pullman, WA 99164-7040. Phone: (509) 335-6067. Fax: (509) 335-8529. E-mail: wbrown{at}vetmed.wsu.edu.
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