Ehrlichia species are emerging zoonotic pathogens that cause acute systemic diseases (ehrlichiosis) in humans. The human granulocytic ehrlichiosis (HGE) agent infects granulocytes. E. chaffeenis and E. sennetsu infect monocytes/macrophages. Work in the P.I.'s laboratory indicates that the ehrlichiae bind by a protein ligand to a host surface protein receptor, induce protein tyrosine kinase-dependent internalization and multiply in the unique early endosome which selectively accumulates transferrin receptors (TfR). The mechanisms involved in the accumulation of TfR by the ehrlichial inclusions and how lysosomal fusion is prevented is unknown. The P.I. has developed three hypotheses to be tested. Hypothesis 1 is that the three ehrlichial species have a specific outer membrane protein which binds to the specific host cell receptor that would induce clathrin-dependent uptake and not activate the cells. Hypothesis 2 is that the interaction of the OMP and the host cell receptor induces the selective fusion of ehrlichial inclusions with TfR-containing endosomes. TfR is required to provide an essential iron to ehrlichia and prevent lysosomal fusion. Hypothesis 3 is that immunization of the host with the OMP protein can protect the host from the ehrlichiosis. To address these hypotheses, the P.I. proposes to accomplish five specific aims.
Aim 1 will test hypothesis 1 by preparing the outer membrane fraction from purified HGE agent, E. chaffeensis and E. sennetsu, and genetically and antigenically characterizing selected OMP s that are likely to be involved in ehrlichial binding and internalization.
Aim 2 will also test hypothesis 1 by examining whether the three Ehrlichia species share the host cell receptor and cytoplasmic compartment and identifying the ehrlichial ligand and host cell receptor which are involved in ehrlichial internalization into the unique early endosome.
Aim 3 also addresses hypothesis 1 by evaluating the inhibitory effects of the Fab fragment of anti-ehrlichial adhesin (recombinant or natural) IgG or anti-ehrlichial receptor IgG, the recombinant OMP or recombinant receptor protein on ehrlichial binding and internalization.
Aim 4 tests hypothesis 2 by examining the role of ehrlichial receptor in recruiting Tf-TfR endosomes and the role of TfR in the ehrlichial inclusion avoidance of lysosomal fusion.
Aim 5 addresses hypothesis 3 by examining the effects of immunization with the ehrlichial OMP in a mouse model of human ehrlichiosis.

Agency
National Institute of Health (NIH)
Institute
National Institute of Allergy and Infectious Diseases (NIAID)
Type
Research Project (R01)
Project #
5R01AI040934-05
Application #
6373605
Study Section
Bacteriology and Mycology Subcommittee 2 (BM)
Program Officer
Baker, Phillip J
Project Start
1997-04-01
Project End
2004-03-31
Budget Start
2001-04-01
Budget End
2004-03-31
Support Year
5
Fiscal Year
2001
Total Cost
$205,920
Indirect Cost
Name
Ohio State University
Department
Veterinary Sciences
Type
Schools of Veterinary Medicine
DUNS #
098987217
City
Columbus
State
OH
Country
United States
Zip Code
43210
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Unver, Ahmet; Rikihisa, Yasuko; Stich, Roger W et al. (2002) The omp-1 major outer membrane multigene family of Ehrlichia chaffeensis is differentially expressed in canine and tick hosts. Infect Immun 70:4701-4
Zhi, Ning; Ohashi, Norio; Tajima, Tomoko et al. (2002) Transcript heterogeneity of the p44 multigene family in a human granulocytic ehrlichiosis agent transmitted by ticks. Infect Immun 70:1175-84
Kim, Hyung-Yong; Mott, Jason; Zhi, Ning et al. (2002) Cytokine gene expression by peripheral blood leukocytes in horses experimentally infected with Anaplasma phagocytophila. Clin Diagn Lab Immunol 9:1079-84
Kim, Hyung-Yong; Rikihisa, Yasuko (2002) Roles of p38 mitogen-activated protein kinase, NF-kappaB, and protein kinase C in proinflammatory cytokine mRNA expression by human peripheral blood leukocytes, monocytes, and neutrophils in response to Anaplasma phagocytophila. Infect Immun 70:4132-41
Liddell, Allison M; Sumner, John W; Paddock, Christopher D et al. (2002) Reinfection with Ehrlichia chaffeensis in a liver transplant recipient. Clin Infect Dis 34:1644-7
Zhi, Ning; Ohashi, Norio; Rikihisa, Yasuko (2002) Activation of a p44 pseudogene in Anaplasma phagocytophila by bacterial RNA splicing: a novel mechanism for post-transcriptional regulation of a multigene family encoding immunodominant major outer membrane proteins. Mol Microbiol 46:135-45
Felek, S; Unver, A; Stich, R W et al. (2001) Sensitive detection of Ehrlichia chaffeensis in cell culture, blood, and tick specimens by reverse transcription-PCR. J Clin Microbiol 39:460-3
Unver, A; Ohashi, N; Tajima, T et al. (2001) Transcriptional analysis of p30 major outer membrane multigene family of Ehrlichia canis in dogs, ticks, and cell culture at different temperatures. Infect Immun 69:6172-8
Unver, A; Felek, S; Paddock, C D et al. (2001) Western blot analysis of sera reactive to human monocytic ehrlichiosis and human granulocytic ehrlichiosis agents. J Clin Microbiol 39:3982-6

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