The developmental cycle of Ehrlichia chaffeensis in

The developmental cycle of Ehrlichia chaffeensis in
The developmental cycle of Ehrlichia chaffeensis in

The developmental cycle of Ehrlichia chaffeensis in vertebrate cells

Jian-zhi Zhang,Vsevolod L.Popov,Si Gao,David H.Walker and Xue-jie Yu*

Department of Pathology,University of Texas Medical Branch,Galveston,TX 77555-0609,USA.Summary

Ehrlichia chaffeensis ,an obligatory intracellular bac-terium,has two forms in mammalian cells:small dense-cored cells (DC)with dense nucleoid and larger reticulate cells (RC)with uniformly dispersed nucleoid.We have determined by electron micros-copy that DC but not RC attaches to and enters into the host cells and RC but not DC multiples inside the host cells.Analysis of outer membrane protein expression by confocal microscopy showed that RC expressed the 28kDa outer membrane protein (p28),the intermediate form,which were transforming from RC to DC,expressed both gp120and p28,and the mature DC expressed gp120only.The TCID 50of DC is 6log 10higher than RC.We conclude that E.chaffeensis has a developmental cycle,in which the DC attaches to and enters into the host cells,and transforms into RC and the RC multiplies by binary ?ssion for 48h and then matures into DC at 72h.Introduction

Human monocytic ehrlichiosis (HME)caused by Ehrlichia chaffeensis is an emerging life-threatening infectious disease.HME is a moderate to severe illness with approximately 40–60%of patients requiring hospital-ization.The estimated case-fatality rate for HME is approximately 3%.Death is generally attributed to multi-system organ failure,respiratory failure,catastrophic haemorrhage,or secondary bacterial or fungal infections (Paddock and Childs,2003).

Ehrlichia chaffeensis is an obligatory intracellular bac-terium that resides in early endosomes of mononuclear phagocytes (Barnewall et al .,1997).The life cycle of E.chaffeensis involves a tick vector and a mammalian host. E.chaffeensis is a Gram-negative bacterium but lacks components such as lipopolysaccharide and pepti-doglycan (Lin and Rikihisa,2003).The bacterial cell wall of Ehrlichia and Anaplasma may be stabilized by cross-linking outer membrane proteins through disul?de bonds and non-covalent bonds (Vidotto et al .,1994).

Two well-characterized major outer membrane proteins of E.chaffeensis are the 120kDa protein (gp120)and the 28kDa outer membrane proteins (p28Omp)(Yu et al .,1993;1997;2000;Ohashi et al .,1998).The gp120is a glycoprotein containing glucose,galactose and xylose (McBride et al.,2000).In a previous study we identi?ed gp120as adhesin using E.coli expressing the repeat region of gp120to adhere to HeLa cells.A few E.coli expressing the gp120were identi?ed inside the host cells,suggesting that the gp120may play a role in bacterial adhesion or invasion of the host cell (Popov et al.,2000).The p28Omp are encoded by a polymorphic multigene family,which consists of 22homologues (Yu et al .,2000;Ohashi et al .,2001).Amino acid sequences among the p28Omp range from 20%to 83%identity (Yu et al .,2000).

Electron microscopy showed that Ehrlichia have two forms:the dense-cored cells (DC)(0.4–0.6m m)and reticulate cells (RC)(0.4–0.6m m by 0.7–1.9)residing within the intracellular parasitophorous vacuoles (morulae)in mammalian host cells.DC are smaller than RC and have a dense nucleoid.In contrast,RC have uniformly dispersed nucleoid ?laments and ribosomes,sometimes forming long projections of the cell wall,pro-trusions of the cytoplasmic membrane into the periplas-mic space,or budding protoplast fragments into the periplasmic space (Popov et al .,1995).Both RC and DC were believed to divide through binary ?ssion,which con-trasts to Chlamydia ,in which only the reticulate body replicates.There is no study that has investigated the differences in molecular biology and pathogenesis of the two forms of E.chaffeensis because the developmental cycle of Ehrlichia has not been determined.

Based on the striking features of the outer membrane proteins of E.chaffeensis ,we reasoned that the gp120and p28Omp are important for intracellular survival of E.chaffeensis and/or attachment to host cells.Thus,we determined the kinetics of the expression of gp120and p28-19during E.chaffeensis infection using confocal microscopy.The results indicated that the gp120and p28are differentially expressed on the DC and the RC of E.chaffeensis .This discovery led us to investigate

Received 7June,2006;revised 2August,2006;accepted 7August,2006.*For correspondence.E-mail xuyu@https://www.360docs.net/doc/7110931473.html,;Tel.(+1)4097471786;Fax (+1)4097472455.

Cellular Microbiology (2007)9(3),610–618

doi:10.1111/j.1462-5822.2006.00812.x First published online 20September 2006

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further and revealed the developmental cycle of E.chaffeensis .Results

Electron microscopy

Electron microscopy showed that at 0h post inoculation DC adhered to the host cell surface (Fig.1A).At 1h post inoculation DC were observed closely attached to the host cell surface (Fig.1B),were in the process of being engulfed (Fig.1C),or were located in a vacuole in the host cell cytoplasm (Fig.1D).At 1h post inoculation some organisms had a dispersed nucleoid-like RC but con-tained a cluster of dense ?laments and ribosomes (Fig.1E).These organisms may represent an intermedi-ate cell transforming from DC to RC.RC were never observed attached to the host cells or in a vacuole at 0h after inoculation.At 24h post infection E.chaffeensis were represented predominantly by large RC,each residing as single bac-terium within a vacuole.Some of the RC appeared to be dividing by binary ?ssion (Fig.2A).At 48h,many morulae contained multiple RC,suggesting that they had multi-plied (Fig.2B).At 72h,most morulae contained DC,only few morulae contained RC (Fig.2C).Continuing cultiva-tion of the infected cells from day 4to day 9post inocu-lation showed loss of synchronization with most DH82cells containing both RC and DC of E.chaffeensis (data not shown).The mixed population of DC and RC from days 4to 9of culture apparently resulted from the sec-ondary infection by the DC released from other DH82cells.A DC undergoing binary ?ssion was never observed in these experiments.

Usually an early endosome harbouring one ehrlichial cell became a morula as DC converted to RC and the latter started dividing thus expanding the morula.If a

host

Fig.1.Electron micrographs of E.chaffeensis interaction with DH82cells.

A.At 0h post inoculation a DC is attached to a DH82cell and surrounded by microvilli.

B.A DC closely adherent to the membrane of the host cell at 1h post inoculation.

C.A DC being engulfed by the host cell at 1h post inoculation

D.A DC is in a vacuole at 1h post inoculation.

E.An ehrlichia in a vacuole presumably transforming from DC to RC (intermediate phase)(arrowhead).Dense-cored ehrlichiae are indicated by arrows.

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cell was infected with several DC,each of them went through its cycle in its own morula,though rarely fusion of the morulae could be observed (Fig.3).

As we described previously (Popov et al .,1995;2000)at later times of infection ehrlichial morulae were character-istically surrounded by mitochondria and granular endo-plasmic reticulum cisterns.First contacts of endoplasmic reticulum with the morula membrane were observed at 1h post inoculation but there were still no mitochondria close to a morula (Fig.1D),though some mitochondria were noted in their proximity.At 24h post inoculation these contacts were usually well developed (Fig.2A).

Immuno-electron microscopy showed that the gp120antibody reacted with dense-cored E.chaffeensis attached to the host cells (Fig.4).Confocal microscopy

At 24h post inoculation,confocal microscopy showed that the morulae reacted with anti-p28MAb and a few of them reacted with gp120antibody.The morulae were tiny,which is consistent with the size of single organisms.At 48h post inoculation,the morulae were easily observed with the majority (86%)of them reacting with

anti-p28

Fig.2.Electron micrographs of E.chaffeensis interaction with DH82cells.

A.A single RC dividing by binary ?ssion at 24h post inoculation (black arrowhead).

B.Two morulae (black arrowheads)contain RC,and two bacteria in the morulae in the upper part of the ?eld are dividing at 48h post inoculation.

C.At 72h post inoculation most Ehrlichia have matured into DC.Three morulae contain DC (black arrows),and one morula contains RC (black arrowhead).

D.High power of a morula containing DC at 72h post inoculation.Mitochondria surrounding ehrlichia morulae were indicated by white arrows.

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MAb and a small portion (14%)reactive with gp120antibody.At 72h post inoculation,the ratio of morulae reacting with gp120antibody and with p28antibody was reversed with the majority (88%)of morulae reacting with anti-gp120antibody and a minor population (12%)react-ing with anti-p28antibody.A few morulae contained ehr-lichiae that reacted with both gp120and p28antibodies simultaneously at 48and 72h post infection (Figs 5and 6).The morulae reacting with gp120were usually larger than the morulae reacting with only the anti-p28antibody.Continued cultivation of the infected cells resulted in more morulae reacting with p28antibody and fewer morulae reacting with gp120antibody (Figs 5and 6).From day 4to day 9,both the size and the number of the morulae increased in infected DH82cells.These obser-

vations suggested that new dissynchronous cycle(s)of E.chaffeensis infection started after release of the mature DC of E.chaffeensis from the previous cycle(s).

To ensure that the apparent differential detection of the proteins was not due to the different sensitivity of the ?uorophores (FITC or Alexa ?uor 647),the ?uorophores were switched for detection of gp120antibody and p28MAb 1A9in different experiments.There was no differ-ence in the apparent protein expression pattern of the particular protein with different ?uorophores (data not shown).Because electron microscopy showed that E.chaffeensis are predominantly RC at 24and 48h post inoculation and DC at 72h and immuno-electron micros-copy showed that gp120is expressed exclusively by the DC (Popov et al .,2000),we conclude that the organisms expressing p28detected by confocal microscopy at the early time points were RC;the cells expressing both p28and gp120are apparently in an intermediate phase trans-forming from RC to DC;and the cells expressing gp120are mature DC.

TCID 50of E.chaffeensis

The genome copy numbers of E.chaffeensis samples obtained at 24,48and 72h post inoculation were 1.45¥108, 4.03¥108and 9.0¥108respectively (Table 1).Based on the increment of the genome copy number,the number of E.chaffeensis doubled approxi-mately every 8.5h from 24h to 48h post inoculation and every 12h from 48h to 72h post inoculation.Thus,the generation time of E.chaffeensis is between 8.5and 12h.The TCID 50were 1.45¥108,1868and 42genomes for E.chaffeensis samples prepared at 24,48and 72h post inoculation respectively.The E.chaffeensis from 72h post inoculation was 3.5million times and 44times more infectious than the samples prepared at 24h and 48h post inoculation respectively.

Discussion

The developmental cycle of E.chaffeensis

In this study we have determined by electron microscopy that the DC,but not the RC,attaches to and enters into the host cells and the RC,but not the DC,divides by binary ?ssion.The DC-rich population is 3.5million times more infectious than the RC-rich population.Thus,the DC is the mature infectious form,and the RC is the multipli-cation form of E.chaffeensis .Based on the results of these experiments,we propose a developmental cycle for E.chaffeensis in vertebrate cells.The developmental cycle starts with the DC,which attaches to and enters into the host cell.Inside a vacuole in the host cell,the

DC

Fig.3.Electron micrographs of E.chaffeensis -infected DH82cells showed fusion between two E.chaffeensis

morulae.

Fig.4.Immuno-electron microscopy of gp120antibody reacting with E.chaffeensis DC.A DC attached to the membrane of the host cell was labelled with immunogold particles.

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rapidly transforms into a RC,which multiplies by binary ?ssion for approximately 48h and then matures into DC at 72h after infection.The mature DC are released and start a new cycle of infection (Fig.7).The reason that the developmental cycle of Ehrlichia was not observed previously in spite of the description of DC and RC is most likely because the previous electron microscopic observations were based on heavily

infected

Fig.5.Confocal microscopic analysis of E.chaffeensis outer membrane protein expression.In each panel,the top left shows DAPI stained nuclei in blue;the top right shows E.chaffeensis morulae in green that reacted with p28MAb 1A9;the bottom left shows E.chaffeensis morulae in red that reacted with gp120antibodies;and the bottom right shows the merged image,in which yellow structures were morulae reactive with both anti-gp120and p28MAb 1A9.D1to D9are days post inoculation.

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tissue culture samples that were usually harvested 5days or later after Ehrlichia infection.At late times of the infec-tion cycle,all ehrlichial morulae contain many RC or many DC or an intermediate form that is transforming from RC to DC.Some of the ehrlichial intermediate forms that may have not yet completed dividing might have been mistaken previously as dividing https://www.360docs.net/doc/7110931473.html,ck of study of the molecular biology and pathogenesis of the RC and DC is most likely explained by the inability to separate DC from RC using puri?cation procedures including sizing column chroma-tography and renogra?n or percoll gradient centrifugation and by the assumption that both DC and RC are infectious.Regulation of the developmental cycle

The nucleoid of DC must undergo decondensation during the transformation to RC,and the nucleoid of the RC must undergo condensation when it matures to DC.In Chlamy-dia trachomatis ,the transformation of reticulate body to the condensed nucleoid of elementary bodies is regulated by the activities of two proteins (Hc1and Hc2)with primary amino acid sequence homology to eukaryotic histone H1(Hackstadt et al .,1991;Tao et al .,1991;Perara et al .,1992;Barry,III et al .,1993).Decondensation of the chlamydial nucleoid is believed to be mediated by the interaction of the histone with an intermediary metabo-lite of an enzyme encoded by ispE in the non-mevalonate

methylerythritol phosphate pathway of isoprenoid biosyn-thesis (Grieshaber et al .,2004).We have searched for ispE ,Hc1and Hc2genes by BLAST in the genomes of Ehrlichia canis (NC_007354)and E.chaffeensis (NC_007799).We identi?ed only the predicted ispE gene,but not the genes for Hc1or Hc2in E.canis and E.chaffeensis .Thus,the regulation of the developmental cycle of Ehrlichia may be quite different from Chlamydia ,and the knowledge of the Chlamydia developmental cycle and expression of outer membrane proteins in the elementary body and reticulate body may not apply to Ehrlichia because the two organisms neither share a common ecologic niche nor are phylogenetically related.The regulation of the developmental cycle of E.chaffeensis requires further investigation.

E. chaffensis morulae ex pressing

gp120 and p28

204060

80100

1201

2

3

4

5

6

7

Days post-inoculation

% o f m o r u l a e

gp120p28-19

Fig.6.Percentage of E.chaffeensis morulae reacting with antibodies to gp120or anti-p28MAb 1A9.

Table 1.TCID 50of E.chaffeensis.E.chaffeensis

24h

48h

72h

Genome copy number in 200m l inoculum 1.45¥108

4.03¥1089.0¥108Dilution of one TCID 50100(non-diluted)

10-5.33

10-7.33

TCID 50

1.45¥108genome copies

1868genome copies

42genome copies

IM1RC

Multiplying

IM2

DC

DC

Adhering

Engulfing

Nucleus

Fig.7.Illustration of the developmental cycle of E.chaffeensis .A dense-cored (DC)cell attaches to the host cells,and it enters into the host through phagocytosis.In the phagosome the DC

transforms into the intermediate phase (IM)-1and subsequently RC.RC multiplies by binary ?ssion for 48h and transforms into intermediate phase (IM)-2cell and eventually matures into DC at 72h,which is released by lysis of the host cell.

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Differential expression of E.chaffeensis proteins in the developmental cycle

With this new de?nition of the developmental cycle of E.chaffeensis,we hypothesize that many proteins asso-ciated with the pathogenesis of Ehrlichia such as adhesin and invasin should be expressed only on the surface of the DC and the proteins speci?c to RC may play roles in the metabolism of E.chaffeensis.The gp120is a cellular marker for DC.Another DC-speci?c protein is the47kDa glycoprotein(Doyle et al.,2005a).The p28is expressed by RC and intermediate phase cells which are transform-ing from RC to DC.Proteomic analysis of the DC and RC will reveal the DC-speci?c proteins and the virulence determinants of E.chaffeensis.

Host cell reaction to E.chaffeensis infection

It is characteristic of E.chaffeensis infection that the morulae in the host cell cytoplasm are surrounded by mitochondria and cisterns of granular endoplasmic reticu-lum,sometimes having visible contacts with the morula membrane.This study demonstrated that these contacts are formed during?rst2h of infection.The function of these contacts is unknown as are the mechanisms of recruitment of mitochondria and endoplasmic reticulum to the morula membrane though it might be hypothesized that endoplasmic reticulum encircling the morula repre-sents an unsuccessful attempt of a host cell to autoph-agocytoze ehrlichiae which is prevented by them.

It is also unclear how the morula membrane expands as RC continue to multiply and thus enlarge the intramorular volume.It is not known if the morula membrane consists only of the host cell proteins or,as in chlamydiae,the parasite proteins are also involved.These questions deserve special investigation.

Experimental procedures

Host cell-free E.chaffeensis

Ehrlichia chaffeensis Arkansas strain was cultivated in DH82 cells,a canine histiocytoma cell line,with5%bovine calf serum-supplemented Eagle’s minimum essential medium(MEM)at 37°C.Five days post inoculation,the cells were100%infected as determined by examination of Diff-Quik stained cells,and were harvested.The cells were centrifuged for20min at12100g.The pellet was suspended in SPK buffer(0.2M sucrose and0.05M potassium phosphate buffer,pH7.4)and sonicated twice for10s on ice at40W using an Ultrasonic Processor(Sonic and Mate-rials,Newtown,CT).The suspension was centrifuged at200g for 10min to remove cell debris.The supernatant was centrifuged for20min at12100g.The pellet was resuspended in PBS and stored on ice for infection of DH82cells and determination of the E.chaffeensis infectious content by quantitative real-time PCR as described below.Confocal microscopy

DH82cells(2¥105)were seeded onto each well of24well plates with a glass coverslip in each well.After overnight cultivation,the monolayers were washed three times with PBS and inoculated with host cell-free E.chaffeensis at an average number of50 bacteria per host cell.After incubation at room temperature for 1h,the monolayers were washed with PBS and further treated with pronase E(2mg ml-1)for3min to remove uninternalized E.chaffeensis.One millilitre of5%MEM containing0.5% agarose at a temperature of50°C was added to each well.After cooling to room temperature,the plates were incubated at37°C with5%CO2atmosphere.Four coverslips were removed from the plates daily until day9after E.chaffeensis infection.The coverslips were?xed for10min in a solution of methanol and acetone(1:1,v/v)pre-chilled to-20°C,and the coverslips were frozen at-20°C until use.The coverslips were incubated at37°C for1h with primary antibodies,then conjugated secondary anti-bodies sequentially,and washed with PBS after each incubation. The primary antibodies included rabbit anti-E.chaffeensis gp120recombinant protein and murine monoclonal antibodies to p28-19(Yu et al.,1993;1997;1999).The conjugated antibodies included FITC-or Alexa?uor647-labelled anti-rabbit IgG(H+l) (Molecular Probes Invitrogen Detection Technologies)and FITC-or Alexa?uor647-labelled anti-mouse IgG(H+l)(Molecular Probes Invitrogen Detection Technologies).Each coverslip was mounted face-down onto a glass slide with Vectashield mounting medium containing DAPI(Vector Laboratories)and was exam-ined with a Zeiss LSM510UV META laser scanning confocal microscope at the Infectious Disease and Toxicology Optical Imaging Core at the University of Texas Medical Branch.The images were analysed using the LSM5Image Browser(Carl Zeiss,Germany).

Electron microscopy

T25?asks containing monolayers of DH82cells were inoculated with E.chaffeensis at an average number of50bacteria per host cell and were centrifuged at700g for1h at4°C.The?asks were washed with PBS three times at room temperature.For0h time point sample,one T25?ask was?xed for transmission electron microscopy(TEM)immediately after centrifugation,prior to adding medium.MEM supplemented with5%bovine calf serum was added to the remaining?asks,and they were incubated at 37°C.The monolayer in a T25?ask was?xed for TEM at1h,2h and4h and then every day from day1(24h after infection)to day9post inoculation.The?xative consisted of2.5%formalde-hyde made from paraformaldehyde,and0.1%glutaraldehyde in 0.05M cacodylate buffer pH7.3to which0.03%trinitrophenol and0.03%CaCl2were added.After?xation the cells were washed with0.1M cacodylate buffer,scraped off the?asks and pelleted.The pellets were separated into two parts.For regular TEM the pellets were post-?xed in1%OsO4in0.1M cacodylate buffer,en bloc stained with1%uranyl acetate in0.1M maleate buffer,dehydrated in ethanol and embedded in Poly/Bed812 (Polysciences,Warrington,PA).Ultrathin sections were cut on Reichert-Leica Ultracut S ultramicrotome,stained with uranyl acetate and lead citrate and examined in a Philips201or CM-100 electron microscope at60kV.

For immuno-electron microscopy osmium?xation was omitted, and after uranyl acetate staining the pellets were dehydrated in

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50%and75%ethanol and embedded in LR White resin.Ultrathin sections on nickel Formvar-carbon coated grids were incubated with anti-gp120primary antibodies diluted1:100and then with goat anti-rabbit IgG(H+l)labelled with15nm gold particles (Amersham Biosciences,Buckinghamshire,England)as previ-ously described(Popov et al.,1995).

Quantitative real-time PCR for determining the DNA copy number of E.chaffeensis

A pCR Topo recombinant plasmid containing a376bp dsb gene fragment of E.chaffeensis was constructed previously and used as a standard to determine the genome copy number of E.chaffeensis(McBride et al.,2002;Doyle et al.2005b).The plasmid DNA concentrations were determined by spectrophotom-eter,and the copy numbers of the plasmid standard DNA molecules were calculated using the following formula:[Xg m l-1 DNA/(plasmid length in basepairs¥660)]¥6.022¥1023=Y molecules m l-1.

Quantitative real-time PCR was performed with E.chaffeensis DNA and plasmid standards using dsb primers(forward primer: TTGCAAAATGATGTCTGAAGATATGAAACA and reverse primer:GCTGCTCCACCAATAAATGTATCYCCTA).The PCR conditions were40cycles at94°C for10s,60°C for1min.A standard curve(plot of Ct values/crossing points of different standard dilutions against log of amount of standard)was gen-erated using a dilution series of the standards.Ampli?cation of the standard dilution series and of E.chaffeensis was carried out in separate https://www.360docs.net/doc/7110931473.html,paring the Ct of an unknown amount of E.chaffeensis with the standard curve allowed calculation of the initial amount of E.chaffeensis used by real-time PCR.

TCID50

The median infectious dose of E.chaffeensis was determined by infection of cell culture.Host cell-free ehrlichiae were prepared from two150cm2?asks of E.chaffeensis-infected DH82cells and resuspended in4ml of PBS.E.chaffeensis-infected DH82 cells were harvested on day5post infection when100%of cells were infected.Three T25?asks with con?uent DH82cell mono-layers were inoculated with1ml of host cell-free E.chaffeensis suspension each.After1h rocking at room temperature,the inoculum was removed from the?asks,and monolayers were washed with PBS three times.The?asks were incubated at37°C in a5%CO2atmosphere.At each time point24,48and72h post inoculation,DH82cells from one?ask were washed with PBS three times and prepared for host-cell free E.chaffeensis as described above.The host cell-free E.chaffeensis from each time point was suspended in1ml of PBS,and100m l was diluted in10-fold increments from1:10in PBS.The remaining E.chaffeensis suspension was frozen at-20°C for quantitative real-time PCR to determine the copy number of E.chaffeensis genomes in the inoculum.Two hundred microlitres of E.chaffeensis was added to one well of a24well plate.Each dilution was added to6wells.After1h rocking at room tempera-ture,the monolayers were washed with PBS three times,and 1ml of5%MEM was added.After incubation of the plates at 37°C with5%CO2for5days,a cytospin smear was prepared from each well and stained with Diff-Quik stain.Each slide was observed for ehrlichial morulae under a light microscope using an oil immersion lens.Twenty?elds were randomly observed on each slide for E.chaffeensis morulae.If E.chaffeensis morulae were observed in one visual?eld of the slide,the culture was considered as E.chaffeensis positive,and if no E.chaffeensis morulae were observed in any?eld of the slide,the culture was considered as negative.The Reed-Muench formula was used to determine the TCID50.

Acknowledgements

The authors are grateful to Violet Han for expert assistance in electron microscopy.This study was supported by a grant from the National Institute of Allergy and Infectious Diseases (AI45871).

References

Barnewall,R.E.,Rikihisa,Y.,and Lee,E.H.(1997)Ehrlichia chaffeensis inclusions are early endosomes which selec-tively accumulate transferrin receptor.Infect Immun65: 1455–1461.

Barry,C.E.,Brickman T.J.,III,and Hackstadt,T.(1993)Hc1-mediated effects on DNA structure:a potential regulator of chlamydial development.Mol Microbiol9:273–283. Doyle,C.K.,Cardenas,A.M.,Aguiar,D.M.,Labruna,M.B., and Ndip,L.M.,Yu,X.J.,and McBride,J.W.(2005a) Molecular characterization of E.canis gp36and E.chaffeensis gp47tandem repeats among isolates from different geographic locations.Ann NY Acad Sci1063: 433–435.

Doyle,C.K.,Labruna,M.B.,Breitschwerdt,E.B.,Tang,Y.W., Corstvet,R.E.,Hegarty,B.C.,et al.(2005b)Detection of medically important Ehrlichia by quantitative multicolor TaqMan real-time polymerase chain reaction of the dsb gene.J Mol Diagn7:504–510.

Grieshaber,N.A.,Fischer,E.R.,Mead,D.J.,Dooley,C.A., and Hackstadt,T.(2004)Chlamydial histone–DNA interac-tions are disrupted by a metabolite in the methylerythritol phosphate pathway of isoprenoid biosynthesis.Proc Natl Acad Sci USA101:7451–7456.

Hackstadt,T.,Baehr,W.,and Ying,Y.(1991)Chlamydia trachomatis developmentally regulated protein is homolo-gous to eukaryotic histone H1.Proc Natl Acad Sci USA88: 3937–3941.

Lin,M.,and Rikihisa,Y.(2003)Ehrlichia chaffeensis and Anaplasma phagocytophilum lack genes for lipid A biosyn-thesis and incorporate cholesterol for their survival.Infect Immun71:5324–5331.

McBride,J.W.,Yu,X.J.,and Walker,D.H.(2000)Glycosy-lation of homologous immunodominant proteins of Ehrli-chia chaffeensis and Ehrlichia canis.Infect Immun68: 13–18.

McBride,J.W.,Ndip,L.M.,Popov,V.L.,and Walker,D.H. (2002)Identi?cation and functional analysis of an immu-noreactive DsbA-like thio-disul?de oxidoreductase of Ehr-lichia spp.Infect Immun70:2700–2703.

Ohashi,N.,Zhi,N.,Zhang,Y.,and Rikihisa,Y.(1998)Immu-nodominant major outer membrane proteins of Ehrlichia chaffeensis are encoded by a polymorphic multigene family.Infect Immun66:132–139.

Ehrlichia developmental cycle617

?2006The Authors

Journal compilation?2006Blackwell Publishing Ltd,Cellular Microbiology,9,610–618

Ohashi,N.,Rikihisa,Y.,and Unver,A.(2001)Analysis of transcriptionally active gene clusters of major outer mem-brane protein multigene family in Ehrlichia canis and E.chaffeensis.Infect Immun69:2083–2091. Paddock,C.D.,and Childs,J.E.(2003)Ehrlichia chaffeensis: a prototypical emerging pathogen.Clin Microbiol Rev16: 37–64.

Perara,E.,Ganem,D.,and Engel,J.N.(1992)A develop-mentally regulated chlamydial gene with apparent homol-ogy to eukaryotic histone H1.Proc Natl Acad Sci USA89: 2125–2129.

Popov,V.L.,Chen,S.M.,Feng,H.M.,and Walker, D.H. (1995)Ultrastructural variation of cultured Ehrlichia chaffeensis.J Med Microbiol43:411–421.

Popov,V.L.,Yu,X.J.,and Walker,D.H.(2000)The120kDa outer membrane protein of Ehrlichia chaffeensis:preferen-tial expression on dense-core cells and gene expression in Escherichia coli associated with attachment and entry. Microb Pathog28:71–80.

Tao,S.,Kaul,R.,and Wenman,W.M.(1991)Identi?cation and nucleotide sequence of a developmentally regulated gene encoding a eukaryotic histone H1-like protein

from Chlamydia trachomatis.J Bacteriol173:2818–2822.

Vidotto,M.C.,Mcguire,T.C.,McElwain,T.F.,Palmer G.H., and Knowles,D.P.,Jr(1994)Intermolecular relationships of major surface proteins of Anaplasma marginale.Infect Immun62:2940–2946.

Yu,X.J.,Brouqui,P.,Dumler,J.S.,and Raoult,D.(1993) Detection of Ehrlichia chaffeensis in human tissue by using a species-speci?c monoclonal antibody.J Clin Microbiol 31:3284–3288.

Yu,X.J.,Crocquet-Valdes,P.,and Walker, D.H.(1997) Cloning and sequencing of the gene for a120-kDa immu-nodominant protein of Ehrlichia chaffeensis.Gene184: 149–154.

Yu,X.J.,McBride,J.W.,and Walker,D.H.(1999)Genetic diversity of the28-kilodalton outer membrane protein gene in human isolates of Ehrlichia chaffeensis.J Clin Microbiol 37:1137–1143.

Yu,X.J.,McBride,J.W.,Zhang,X.,and Walker,D.H.(2000) Characterization of the complete transcriptionally active Ehrlichia chaffeensis28kDa outer membrane protein mul-tigene family.Gene248:59–68.

618J.-Z.Zhang et al.

?2006The Authors

Journal compilation?2006Blackwell Publishing Ltd,Cellular Microbiology,9,610–618

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