Apical organelle discharge by Cryptosporidium parvum is temperature, cytoskeleton, and intracellular calcium dependent and required for host cell invasion

Xian Ming Chen, Steven P. O'Hara, Bing Q. Huang, Jeremy B. Nelson, Jim Jung Ching Lin, Guan Zhu, Honorine D. Ward, Nicholas F La Russo

Research output: Contribution to journalArticle

54 Citations (Scopus)

Abstract

The apical organelles in apicomplexan parasites are characteristic secretory vesicles containing complex mixtures of molecules. While apical organelle discharge has been demonstrated to be involved in the cellular invasion of some apicomplexan parasites, including Toxoplasma gondii and Plasmodium spp., the mechanisms of apical organelle discharge by Cryptosporidium parvum sporozoites and its role in host cell invasion are unclear. Here we show that the discharge of C. parvum apical organelles occurs in a temperature-dependent fashion. The inhibition of parasite actin and tubulin polymerization by cytochalasin D and colchicines, respectively, inhibited parasite apical organelle discharge. Chelation of the parasite's intracellular calcium also inhibited apical organelle discharge, and this process was partially reversed by raising the intracellular calcium concentration by use of the ionophore A23187. The inhibition of parasite cytoskeleton polymerization by cytochalasin D and colchicine and the depletion of intracellular calcium also decreased the gliding motility of C. parvum sporozoites. Importantly, the inhibition of apical organelle discharge by C. parvum sporozoites blocked parasite invasion of, but not attachment to, host cells (i.e., cultured human cholangiocytes). Moreover, the translocation of a parasite protein, CP2, to the host cell membrane at the region of the host cell-parasite interface was detected; an antibody to CP2 decreased the C. parvum invasion of cholangiocytes. These data demonstrate that the discharge of C. parvum sporozoite apical organelle contents occurs and that it is temperature, intracellular calcium, and cytoskeleton dependent and required for host cell invasion, confirming that apical organelles play a central role in C. parvum entry into host cells.

Original languageEnglish (US)
Pages (from-to)6806-6816
Number of pages11
JournalInfection and Immunity
Volume72
Issue number12
DOIs
StatePublished - Dec 2004

Fingerprint

Cryptosporidium parvum
Cytoskeleton
Organelles
Parasites
Calcium
Temperature
Sporozoites
Cytochalasin D
Colchicine
Polymerization
Plasmodium
Ionophores
Toxoplasma
Calcimycin
Secretory Vesicles
Tubulin
Complex Mixtures
Actins
Cultured Cells
Cell Membrane

ASJC Scopus subject areas

  • Immunology

Cite this

Apical organelle discharge by Cryptosporidium parvum is temperature, cytoskeleton, and intracellular calcium dependent and required for host cell invasion. / Chen, Xian Ming; O'Hara, Steven P.; Huang, Bing Q.; Nelson, Jeremy B.; Lin, Jim Jung Ching; Zhu, Guan; Ward, Honorine D.; La Russo, Nicholas F.

In: Infection and Immunity, Vol. 72, No. 12, 12.2004, p. 6806-6816.

Research output: Contribution to journalArticle

Chen, Xian Ming ; O'Hara, Steven P. ; Huang, Bing Q. ; Nelson, Jeremy B. ; Lin, Jim Jung Ching ; Zhu, Guan ; Ward, Honorine D. ; La Russo, Nicholas F. / Apical organelle discharge by Cryptosporidium parvum is temperature, cytoskeleton, and intracellular calcium dependent and required for host cell invasion. In: Infection and Immunity. 2004 ; Vol. 72, No. 12. pp. 6806-6816.
@article{7995850fce514234a2b9c97efdfa6ff5,
title = "Apical organelle discharge by Cryptosporidium parvum is temperature, cytoskeleton, and intracellular calcium dependent and required for host cell invasion",
abstract = "The apical organelles in apicomplexan parasites are characteristic secretory vesicles containing complex mixtures of molecules. While apical organelle discharge has been demonstrated to be involved in the cellular invasion of some apicomplexan parasites, including Toxoplasma gondii and Plasmodium spp., the mechanisms of apical organelle discharge by Cryptosporidium parvum sporozoites and its role in host cell invasion are unclear. Here we show that the discharge of C. parvum apical organelles occurs in a temperature-dependent fashion. The inhibition of parasite actin and tubulin polymerization by cytochalasin D and colchicines, respectively, inhibited parasite apical organelle discharge. Chelation of the parasite's intracellular calcium also inhibited apical organelle discharge, and this process was partially reversed by raising the intracellular calcium concentration by use of the ionophore A23187. The inhibition of parasite cytoskeleton polymerization by cytochalasin D and colchicine and the depletion of intracellular calcium also decreased the gliding motility of C. parvum sporozoites. Importantly, the inhibition of apical organelle discharge by C. parvum sporozoites blocked parasite invasion of, but not attachment to, host cells (i.e., cultured human cholangiocytes). Moreover, the translocation of a parasite protein, CP2, to the host cell membrane at the region of the host cell-parasite interface was detected; an antibody to CP2 decreased the C. parvum invasion of cholangiocytes. These data demonstrate that the discharge of C. parvum sporozoite apical organelle contents occurs and that it is temperature, intracellular calcium, and cytoskeleton dependent and required for host cell invasion, confirming that apical organelles play a central role in C. parvum entry into host cells.",
author = "Chen, {Xian Ming} and O'Hara, {Steven P.} and Huang, {Bing Q.} and Nelson, {Jeremy B.} and Lin, {Jim Jung Ching} and Guan Zhu and Ward, {Honorine D.} and {La Russo}, {Nicholas F}",
year = "2004",
month = "12",
doi = "10.1128/IAI.72.12.6806-6816.2004",
language = "English (US)",
volume = "72",
pages = "6806--6816",
journal = "Infection and Immunity",
issn = "0019-9567",
publisher = "American Society for Microbiology",
number = "12",

}

TY - JOUR

T1 - Apical organelle discharge by Cryptosporidium parvum is temperature, cytoskeleton, and intracellular calcium dependent and required for host cell invasion

AU - Chen, Xian Ming

AU - O'Hara, Steven P.

AU - Huang, Bing Q.

AU - Nelson, Jeremy B.

AU - Lin, Jim Jung Ching

AU - Zhu, Guan

AU - Ward, Honorine D.

AU - La Russo, Nicholas F

PY - 2004/12

Y1 - 2004/12

N2 - The apical organelles in apicomplexan parasites are characteristic secretory vesicles containing complex mixtures of molecules. While apical organelle discharge has been demonstrated to be involved in the cellular invasion of some apicomplexan parasites, including Toxoplasma gondii and Plasmodium spp., the mechanisms of apical organelle discharge by Cryptosporidium parvum sporozoites and its role in host cell invasion are unclear. Here we show that the discharge of C. parvum apical organelles occurs in a temperature-dependent fashion. The inhibition of parasite actin and tubulin polymerization by cytochalasin D and colchicines, respectively, inhibited parasite apical organelle discharge. Chelation of the parasite's intracellular calcium also inhibited apical organelle discharge, and this process was partially reversed by raising the intracellular calcium concentration by use of the ionophore A23187. The inhibition of parasite cytoskeleton polymerization by cytochalasin D and colchicine and the depletion of intracellular calcium also decreased the gliding motility of C. parvum sporozoites. Importantly, the inhibition of apical organelle discharge by C. parvum sporozoites blocked parasite invasion of, but not attachment to, host cells (i.e., cultured human cholangiocytes). Moreover, the translocation of a parasite protein, CP2, to the host cell membrane at the region of the host cell-parasite interface was detected; an antibody to CP2 decreased the C. parvum invasion of cholangiocytes. These data demonstrate that the discharge of C. parvum sporozoite apical organelle contents occurs and that it is temperature, intracellular calcium, and cytoskeleton dependent and required for host cell invasion, confirming that apical organelles play a central role in C. parvum entry into host cells.

AB - The apical organelles in apicomplexan parasites are characteristic secretory vesicles containing complex mixtures of molecules. While apical organelle discharge has been demonstrated to be involved in the cellular invasion of some apicomplexan parasites, including Toxoplasma gondii and Plasmodium spp., the mechanisms of apical organelle discharge by Cryptosporidium parvum sporozoites and its role in host cell invasion are unclear. Here we show that the discharge of C. parvum apical organelles occurs in a temperature-dependent fashion. The inhibition of parasite actin and tubulin polymerization by cytochalasin D and colchicines, respectively, inhibited parasite apical organelle discharge. Chelation of the parasite's intracellular calcium also inhibited apical organelle discharge, and this process was partially reversed by raising the intracellular calcium concentration by use of the ionophore A23187. The inhibition of parasite cytoskeleton polymerization by cytochalasin D and colchicine and the depletion of intracellular calcium also decreased the gliding motility of C. parvum sporozoites. Importantly, the inhibition of apical organelle discharge by C. parvum sporozoites blocked parasite invasion of, but not attachment to, host cells (i.e., cultured human cholangiocytes). Moreover, the translocation of a parasite protein, CP2, to the host cell membrane at the region of the host cell-parasite interface was detected; an antibody to CP2 decreased the C. parvum invasion of cholangiocytes. These data demonstrate that the discharge of C. parvum sporozoite apical organelle contents occurs and that it is temperature, intracellular calcium, and cytoskeleton dependent and required for host cell invasion, confirming that apical organelles play a central role in C. parvum entry into host cells.

UR - http://www.scopus.com/inward/record.url?scp=9244256786&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=9244256786&partnerID=8YFLogxK

U2 - 10.1128/IAI.72.12.6806-6816.2004

DO - 10.1128/IAI.72.12.6806-6816.2004

M3 - Article

VL - 72

SP - 6806

EP - 6816

JO - Infection and Immunity

JF - Infection and Immunity

SN - 0019-9567

IS - 12

ER -