Isolation of the equine granulocytic ehrlichiosis agent, Ehrlichia equi, in tick cell culture

Ulrike G. Munderloh, John E. Madigan, J. Stephen Dumler, Jesse L. Goodman, Stanley F. Hayes, Jeffrey E. Barlough, Curtis M. Nelson, Timothy J. Kurtti

Research output: Contribution to journalArticle

Abstract

The equine granulocytic ehrlichiosis agent, Ehrlichia equi, is closely related or identical to the human granulocytic ehrlichiosis (HGE) agent. Both are suspected of being transmitted by ticks. We have successfully isolated E. equi in a cell line, IDE8, derived from a putative vector, the tick Ixodes scapularis. Peripheral blood leukocytes from an experimentally infected horse were inoculated onto IDE8 monolayers. Cultures were incubated in a candle jar at 34°C in tick cell culture medium with NaHCO, and an organic buffer 13- (N-morpholino)-propanesulfonic acid] (MOPS). Within 2 weeks, infected cells were detected in Giemsa-stained culture samples, and the organisms subsequently spread to uninfected cells in the cultures. E. equi was passaged serially by transferring a portion of an infected culture to new cell layers every 2 to 3 weeks. The identity of the organisms was confirmed by PCR using oligonucleotide primers specific for E. equi and the HGE agent and by immunocytology. Homologous equine antibodies and human anti-HGE convalescent serum recognized E. equi grown in tick cell culture. Electron microscopy revealed electron-lucent and -dense ehrlichia-like forms developing within host cell endosomes. E. equi passaged twice in tick cell culture retained infectivity and pathogenicity for the equine host, as demonstrated by intravenous inoculation of a suspension of infected tick cells and subsequent reisolation from peripheral blood, in fulfillment of Koch's postulates. The horse developed severe clinical signs, i.e., fever, inappetence, thrombocytopenia, icterus, and limb edema, typical of granulocytic equine ehrlichiosis, within 1 week.

Original languageEnglish (US)
Pages (from-to)664-670
Number of pages7
JournalJournal of Clinical Microbiology
Volume34
Issue number3
StatePublished - 1996
Externally publishedYes

Fingerprint

Anaplasma phagocytophilum
Ehrlichiosis
Ticks
Horses
Cell Culture Techniques
Ehrlichia
Morpholinos
Ixodes
DNA Primers
Endosomes
Jaundice
Thrombocytopenia
Virulence
Culture Media
Anti-Idiotypic Antibodies
Edema
Suspensions
Electron Microscopy
Buffers
Leukocytes

ASJC Scopus subject areas

  • Microbiology (medical)
  • Microbiology

Cite this

Munderloh, U. G., Madigan, J. E., Stephen Dumler, J., Goodman, J. L., Hayes, S. F., Barlough, J. E., ... Kurtti, T. J. (1996). Isolation of the equine granulocytic ehrlichiosis agent, Ehrlichia equi, in tick cell culture. Journal of Clinical Microbiology, 34(3), 664-670.

Isolation of the equine granulocytic ehrlichiosis agent, Ehrlichia equi, in tick cell culture. / Munderloh, Ulrike G.; Madigan, John E.; Stephen Dumler, J.; Goodman, Jesse L.; Hayes, Stanley F.; Barlough, Jeffrey E.; Nelson, Curtis M.; Kurtti, Timothy J.

In: Journal of Clinical Microbiology, Vol. 34, No. 3, 1996, p. 664-670.

Research output: Contribution to journalArticle

Munderloh, UG, Madigan, JE, Stephen Dumler, J, Goodman, JL, Hayes, SF, Barlough, JE, Nelson, CM & Kurtti, TJ 1996, 'Isolation of the equine granulocytic ehrlichiosis agent, Ehrlichia equi, in tick cell culture', Journal of Clinical Microbiology, vol. 34, no. 3, pp. 664-670.
Munderloh UG, Madigan JE, Stephen Dumler J, Goodman JL, Hayes SF, Barlough JE et al. Isolation of the equine granulocytic ehrlichiosis agent, Ehrlichia equi, in tick cell culture. Journal of Clinical Microbiology. 1996;34(3):664-670.
Munderloh, Ulrike G. ; Madigan, John E. ; Stephen Dumler, J. ; Goodman, Jesse L. ; Hayes, Stanley F. ; Barlough, Jeffrey E. ; Nelson, Curtis M. ; Kurtti, Timothy J. / Isolation of the equine granulocytic ehrlichiosis agent, Ehrlichia equi, in tick cell culture. In: Journal of Clinical Microbiology. 1996 ; Vol. 34, No. 3. pp. 664-670.
@article{78c3962a4c4d4657bbbd46018870f05a,
title = "Isolation of the equine granulocytic ehrlichiosis agent, Ehrlichia equi, in tick cell culture",
abstract = "The equine granulocytic ehrlichiosis agent, Ehrlichia equi, is closely related or identical to the human granulocytic ehrlichiosis (HGE) agent. Both are suspected of being transmitted by ticks. We have successfully isolated E. equi in a cell line, IDE8, derived from a putative vector, the tick Ixodes scapularis. Peripheral blood leukocytes from an experimentally infected horse were inoculated onto IDE8 monolayers. Cultures were incubated in a candle jar at 34°C in tick cell culture medium with NaHCO, and an organic buffer 13- (N-morpholino)-propanesulfonic acid] (MOPS). Within 2 weeks, infected cells were detected in Giemsa-stained culture samples, and the organisms subsequently spread to uninfected cells in the cultures. E. equi was passaged serially by transferring a portion of an infected culture to new cell layers every 2 to 3 weeks. The identity of the organisms was confirmed by PCR using oligonucleotide primers specific for E. equi and the HGE agent and by immunocytology. Homologous equine antibodies and human anti-HGE convalescent serum recognized E. equi grown in tick cell culture. Electron microscopy revealed electron-lucent and -dense ehrlichia-like forms developing within host cell endosomes. E. equi passaged twice in tick cell culture retained infectivity and pathogenicity for the equine host, as demonstrated by intravenous inoculation of a suspension of infected tick cells and subsequent reisolation from peripheral blood, in fulfillment of Koch's postulates. The horse developed severe clinical signs, i.e., fever, inappetence, thrombocytopenia, icterus, and limb edema, typical of granulocytic equine ehrlichiosis, within 1 week.",
author = "Munderloh, {Ulrike G.} and Madigan, {John E.} and {Stephen Dumler}, J. and Goodman, {Jesse L.} and Hayes, {Stanley F.} and Barlough, {Jeffrey E.} and Nelson, {Curtis M.} and Kurtti, {Timothy J.}",
year = "1996",
language = "English (US)",
volume = "34",
pages = "664--670",
journal = "Journal of Clinical Microbiology",
issn = "0095-1137",
publisher = "American Society for Microbiology",
number = "3",

}

TY - JOUR

T1 - Isolation of the equine granulocytic ehrlichiosis agent, Ehrlichia equi, in tick cell culture

AU - Munderloh, Ulrike G.

AU - Madigan, John E.

AU - Stephen Dumler, J.

AU - Goodman, Jesse L.

AU - Hayes, Stanley F.

AU - Barlough, Jeffrey E.

AU - Nelson, Curtis M.

AU - Kurtti, Timothy J.

PY - 1996

Y1 - 1996

N2 - The equine granulocytic ehrlichiosis agent, Ehrlichia equi, is closely related or identical to the human granulocytic ehrlichiosis (HGE) agent. Both are suspected of being transmitted by ticks. We have successfully isolated E. equi in a cell line, IDE8, derived from a putative vector, the tick Ixodes scapularis. Peripheral blood leukocytes from an experimentally infected horse were inoculated onto IDE8 monolayers. Cultures were incubated in a candle jar at 34°C in tick cell culture medium with NaHCO, and an organic buffer 13- (N-morpholino)-propanesulfonic acid] (MOPS). Within 2 weeks, infected cells were detected in Giemsa-stained culture samples, and the organisms subsequently spread to uninfected cells in the cultures. E. equi was passaged serially by transferring a portion of an infected culture to new cell layers every 2 to 3 weeks. The identity of the organisms was confirmed by PCR using oligonucleotide primers specific for E. equi and the HGE agent and by immunocytology. Homologous equine antibodies and human anti-HGE convalescent serum recognized E. equi grown in tick cell culture. Electron microscopy revealed electron-lucent and -dense ehrlichia-like forms developing within host cell endosomes. E. equi passaged twice in tick cell culture retained infectivity and pathogenicity for the equine host, as demonstrated by intravenous inoculation of a suspension of infected tick cells and subsequent reisolation from peripheral blood, in fulfillment of Koch's postulates. The horse developed severe clinical signs, i.e., fever, inappetence, thrombocytopenia, icterus, and limb edema, typical of granulocytic equine ehrlichiosis, within 1 week.

AB - The equine granulocytic ehrlichiosis agent, Ehrlichia equi, is closely related or identical to the human granulocytic ehrlichiosis (HGE) agent. Both are suspected of being transmitted by ticks. We have successfully isolated E. equi in a cell line, IDE8, derived from a putative vector, the tick Ixodes scapularis. Peripheral blood leukocytes from an experimentally infected horse were inoculated onto IDE8 monolayers. Cultures were incubated in a candle jar at 34°C in tick cell culture medium with NaHCO, and an organic buffer 13- (N-morpholino)-propanesulfonic acid] (MOPS). Within 2 weeks, infected cells were detected in Giemsa-stained culture samples, and the organisms subsequently spread to uninfected cells in the cultures. E. equi was passaged serially by transferring a portion of an infected culture to new cell layers every 2 to 3 weeks. The identity of the organisms was confirmed by PCR using oligonucleotide primers specific for E. equi and the HGE agent and by immunocytology. Homologous equine antibodies and human anti-HGE convalescent serum recognized E. equi grown in tick cell culture. Electron microscopy revealed electron-lucent and -dense ehrlichia-like forms developing within host cell endosomes. E. equi passaged twice in tick cell culture retained infectivity and pathogenicity for the equine host, as demonstrated by intravenous inoculation of a suspension of infected tick cells and subsequent reisolation from peripheral blood, in fulfillment of Koch's postulates. The horse developed severe clinical signs, i.e., fever, inappetence, thrombocytopenia, icterus, and limb edema, typical of granulocytic equine ehrlichiosis, within 1 week.

UR - http://www.scopus.com/inward/record.url?scp=0030023811&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0030023811&partnerID=8YFLogxK

M3 - Article

C2 - 8904434

AN - SCOPUS:0030023811

VL - 34

SP - 664

EP - 670

JO - Journal of Clinical Microbiology

JF - Journal of Clinical Microbiology

SN - 0095-1137

IS - 3

ER -