Previous Article | Next Article 
Infection and Immunity, September 2003, p. 5324-5331, Vol. 71, No. 9
0019-9567/03/$08.00+0 DOI: 10.1128/IAI.71.9.5324-5331.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.
Ehrlichia chaffeensis and Anaplasma phagocytophilum Lack Genes for Lipid A Biosynthesis and Incorporate Cholesterol for Their Survival
Mingqun Lin and Yasuko Rikihisa*
Department of Veterinary Biosciences, The Ohio State University, Columbus, Ohio 43210
Received 12 March 2003/
Returned for modification 14 May 2003/
Accepted 23 June 2003
Ehrlichia chaffeensis and Anaplasma phagocytophilum are agents of human monocytic and granulocytic ehrlichioses, respectively. They are extremely sensitive to mechanical stress and are pleomorphic gram-negative bacteria. Membrane incorporation of cholesterol from the eukaryotic host is known to be essential for other fragile and pleomorphic bacteria and mycoplasmas that lack a cell wall. Thus, we tested whether cholesterol is required for E. chaffeensis and A. phagocytophilum. Using a freeze fracture technique and biochemical analysis, these bacteria were found to contain significant levels of membrane cholesterol. These bacteria lack genes for cholesterol biosynthesis or modification. However, host cell-free bacteria had the ability to take up directly exogenous cholesterol or NBD-cholesterol, a fluorescent cholesterol derivative. Treatment of the bacteria with cholesterol extraction reagent methyl-ß-cyclodextrin caused their ultrastructural changes. Furthermore, pretreatment of the bacteria with methyl-ß-cyclodextrin or NBD-cholesterol deprived these bacteria of the ability to infect leukocytes, thus killing these obligate intracellular bacteria. Analysis of E. chaffeensis and A. phagocytophilum genome sequences revealed that these bacteria lack all genes for the biosynthesis of lipid A and most genes for the biosynthesis of peptidoglycan, which confer structural strength to gram-negative bacteria. Taken together, these results suggest that human ehrlichiosis agents became cholesterol dependent due to the loss of these genes. As the first report of gram-negative bacteria incorporating cholesterol for survival, these findings offer insight into the unique nature of their parasitism and imply that cholesterol is important in the control of human ehrlichioses.
* Corresponding author. Mailing address: Department of Veterinary Biosciences, The Ohio State University, 1925 Coffey Rd., Columbus, OH 43210. Phone: (614) 292-9677. Fax: (614) 292-6473. E-mail:
Rikihisa.1{at}osu.edu.
Editor: B. B. Finlay
Infection and Immunity, September 2003, p. 5324-5331, Vol. 71, No. 9
0019-9567/03/$08.00+0 DOI: 10.1128/IAI.71.9.5324-5331.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.
This article has been cited by other articles:
-
Luce-Fedrow, A., Von Ohlen, T., Chapes, S. K.
(2009). Ehrlichia chaffeensis Infections in Drosophila melanogaster. Infect. Immun.
77: 4815-4826
[Abstract]
[Full Text]
-
Zhu, B., Nethery, K. A., Kuriakose, J. A., Wakeel, A., Zhang, X., McBride, J. W.
(2009). Nuclear Translocated Ehrlichia chaffeensis Ankyrin Protein Interacts with a Specific Adenine-Rich Motif of Host Promoter and Intronic Alu Elements. Infect. Immun.
77: 4243-4255
[Abstract]
[Full Text]
-
Lin, M., Zhang, C., Gibson, K., Rikihisa, Y.
(2009). Analysis of complete genome sequence of Neorickettsia risticii: causative agent of Potomac horse fever. Nucleic Acids Res
37: 6076-6091
[Abstract]
[Full Text]
-
MacNamara, K. C., Racine, R., Chatterjee, M., Borjesson, D., Winslow, G. M.
(2009). Diminished Hematopoietic Activity Associated with Alterations in Innate and Adaptive Immunity in a Mouse Model of Human Monocytic Ehrlichiosis. Infect. Immun.
77: 4061-4069
[Abstract]
[Full Text]
-
Huang, H., Lin, M., Wang, X., Kikuchi, T., Mottaz, H., Norbeck, A., Rikihisa, Y.
(2008). Proteomic Analysis of and Immune Responses to Ehrlichia chaffeensis Lipoproteins. Infect. Immun.
76: 3405-3414
[Abstract]
[Full Text]
-
Racine, R., Chatterjee, M., Winslow, G. M.
(2008). CD11c Expression Identifies a Population of Extrafollicular Antigen-Specific Splenic Plasmablasts Responsible for CD4 T-Independent Antibody Responses during Intracellular Bacterial Infection. J. Immunol.
181: 1375-1385
[Abstract]
[Full Text]
-
Noh, S. M., Brayton, K. A., Brown, W. C., Norimine, J., Munske, G. R., Davitt, C. M., Palmer, G. H.
(2008). Composition of the Surface Proteome of Anaplasma marginale and Its Role in Protective Immunity Induced by Outer Membrane Immunization. Infect. Immun.
76: 2219-2226
[Abstract]
[Full Text]
-
Stevenson, H. L., Crossley, E. C., Thirumalapura, N., Walker, D. H., Ismail, N.
(2008). Regulatory Roles of CD1d-Restricted NKT Cells in the Induction of Toxic Shock-Like Syndrome in an Animal Model of Fatal Ehrlichiosis. Infect. Immun.
76: 1434-1444
[Abstract]
[Full Text]
-
Luce-Fedrow, A., Von Ohlen, T., Boyle, D., Ganta, R. R., Chapes, S. K.
(2008). Use of Drosophila S2 Cells as a Model for Studying Ehrlichia chaffeensis Infections. Appl. Environ. Microbiol.
74: 1886-1891
[Abstract]
[Full Text]
-
Manzano-Roman, R., Almazan, C., Naranjo, V., Bloui, E. F., Kocan, K. M., de la Fuente, J.
(2008). Expression of perilipin in human promyelocytic cells in response to Anaplasma phagocytophilum infection results in modified lipid metabolism. J Med Microbiol
57: 159-163
[Abstract]
[Full Text]
-
Ge, Y., Rikihisa, Y.
(2007). Identification of Novel Surface Proteins of Anaplasma phagocytophilum by Affinity Purification and Proteomics. J. Bacteriol.
189: 7819-7828
[Abstract]
[Full Text]
-
Choi, K.-S., Dumler, J. S.
(2007). Mitogenic Component in Polar Lipid-Enriched Anaplasma phagocytophilum Membranes. CVI
14: 1260-1265
[Abstract]
[Full Text]
-
Ismail, N., Crossley, E. C., Stevenson, H. L., Walker, D. H.
(2007). Relative Importance of T-Cell Subsets in Monocytotropic Ehrlichiosis: a Novel Effector Mechanism Involved in Ehrlichia-Induced Immunopathology in Murine Ehrlichiosis. Infect. Immun.
75: 4608-4620
[Abstract]
[Full Text]
-
Miura, K., Rikihisa, Y.
(2007). Virulence Potential of Ehrlichia chaffeensis Strains of Distinct Genome Sequences. Infect. Immun.
75: 3604-3613
[Abstract]
[Full Text]
-
Mavromatis, K., Doyle, C. K., Lykidis, A., Ivanova, N., Francino, M. P., Chain, P., Shin, M., Malfatti, S., Larimer, F., Copeland, A., Detter, J. C., Land, M., Richardson, P. M., Yu, X. J., Walker, D. H., McBride, J. W., Kyrpides, N. C.
(2006). The Genome of the Obligately Intracellular Bacterium Ehrlichia canis Reveals Themes of Complex Membrane Structure and Immune Evasion Strategies. J. Bacteriol.
188: 4015-4023
[Abstract]
[Full Text]
-
Ismail, N., Stevenson, H. L., Walker, D. H.
(2006). Role of Tumor Necrosis Factor Alpha (TNF-{alpha}) and Interleukin-10 in the Pathogenesis of Severe Murine Monocytotropic Ehrlichiosis: Increased Resistance of TNF Receptor p55- and p75-Deficient Mice to Fatal Ehrlichial Infection. Infect. Immun.
74: 1846-1856
[Abstract]
[Full Text]
-
Wang, X., Kikuchi, T., Rikihisa, Y.
(2006). Two Monoclonal Antibodies with Defined Epitopes of P44 Major Surface Proteins Neutralize Anaplasma phagocytophilum by Distinct Mechanisms. Infect. Immun.
74: 1873-1882
[Abstract]
[Full Text]
-
Collins, N. E., Liebenberg, J., de Villiers, E. P., Brayton, K. A., Louw, E., Pretorius, A., Faber, F. E., van Heerden, H., Josemans, A., van Kleef, M., Steyn, H. C., van Strijp, M. F., Zweygarth, E., Jongejan, F., Maillard, J. C., Berthier, D., Botha, M., Joubert, F., Corton, C. H., Thomson, N. R., Allsopp, M. T., Allsopp, B. A.
(2005). The genome of the heartwater agent Ehrlichia ruminantium contains multiple tandem repeats of actively variable copy number. Proc. Natl. Acad. Sci. USA
102: 838-843
[Abstract]
[Full Text]
-
Brayton, K. A., Kappmeyer, L. S., Herndon, D. R., Dark, M. J., Tibbals, D. L., Palmer, G. H., McGuire, T. C., Knowles, D. P. Jr.
(2005). Complete genome sequencing of Anaplasma marginale reveals that the surface is skewed to two superfamilies of outer membrane proteins. Proc. Natl. Acad. Sci. USA
102: 844-849
[Abstract]
[Full Text]
-
Branger, S., Rolain, J. M., Raoult, D.
(2004). Evaluation of Antibiotic Susceptibilities of Ehrlichia canis, Ehrlichia chaffeensis, and Anaplasma phagocytophilum by Real-Time PCR. Antimicrob. Agents Chemother.
48: 4822-4828
[Abstract]
[Full Text]