Limits...
Anaplasma phagocytophilum and Anaplasma marginale elicit different gene expression responses in cultured tick cells.

Zivkovic Z, Blouin EF, Manzano-Roman R, Almazán C, Naranjo V, Massung RF, Jongejan F, Kocan KM, de la Fuente J - Comp. Funct. Genomics (2009)

Bottom Line: The genus Anaplasma (Rickettsiales: Anaplasmataceae) includes obligate tick-transmitted intracellular organisms, Anaplasma phagocytophilum and Anaplasma marginale that multiply in both vertebrate and tick host cells.The results of these studies demonstrated modulation of tick gene expression by A. phagocytophilum and provided evidence of different gene expression responses in tick cells infected with A. phagocytophilum and A. marginale.These differences in Anaplasma-tick interactions may reflect differences in pathogen life cycle in the tick cells.

View Article: PubMed Central - PubMed

Affiliation: Utrecht Centre for Tick-Borne Diseases (UCTD), Department of Infectious Diseases and Immunology, Faculty of Veterinary Medicine, Utrecht University, Yalelaan 1, 3584 CL Utrecht, The Netherlands.

ABSTRACT
The genus Anaplasma (Rickettsiales: Anaplasmataceae) includes obligate tick-transmitted intracellular organisms, Anaplasma phagocytophilum and Anaplasma marginale that multiply in both vertebrate and tick host cells. Recently, we showed that A. marginale affects the expression of tick genes that are involved in tick survival and pathogen infection and multiplication. However, the gene expression profile in A. phagocytophilum-infected tick cells is currently poorly characterized. The objectives of this study were to characterize tick gene expression profile in Ixodes scapularis ticks and cultured ISE6 cells in response to infection with A. phagocypthilum and to compare tick gene expression responses in A. phagocytophilum- and A. marginale-infected tick cells by microarray and real-time RT-PCR analyses. The results of these studies demonstrated modulation of tick gene expression by A. phagocytophilum and provided evidence of different gene expression responses in tick cells infected with A. phagocytophilum and A. marginale. These differences in Anaplasma-tick interactions may reflect differences in pathogen life cycle in the tick cells.

No MeSH data available.


Related in: MedlinePlus

Comparison between differential gene expression in A. phagocytophilum- and A. marginale-infected tick ISE6 cells at different time points after infection. Studies were done on A. phagocytophilum- (A.p.-) and A. marginale- (A.m.-) infected tick ISE6 cells (two independent cultures each) at 2, 5, and 8 days postinfection (dpi) with approximately 30%–40%, 60%–70%, and >90% infected cells, respectively. (a) The A.p. and A.m. infection levels were evaluated by real-time PCR of msp4 and normalized against tick 16S rDNA. Known amounts of the full length A.p. and A.m. msp4 PCR product were used to construct a standard curve for quantitation of pathogens per cell. Data represent average  ± SD. (b) The mRNA levels of selected genes were evaluated by real-time RT-PCR and normalized against tick 16S rRNA. Bars represent the ratio between average Ct values in A.p.-infected cells/average Ct values in A.m.-infected cells. The mRNA levels were compared between A.p.- and A.m.-infected tick cells by Student's t-test (*P ≤ .05). Identical mRNA levels in A. phagocytophilum and A. marginale infected cells equal one.
© Copyright Policy - open-access
Related In: Results  -  Collection


getmorefigures.php?uid=PMC2712686&req=5

fig4: Comparison between differential gene expression in A. phagocytophilum- and A. marginale-infected tick ISE6 cells at different time points after infection. Studies were done on A. phagocytophilum- (A.p.-) and A. marginale- (A.m.-) infected tick ISE6 cells (two independent cultures each) at 2, 5, and 8 days postinfection (dpi) with approximately 30%–40%, 60%–70%, and >90% infected cells, respectively. (a) The A.p. and A.m. infection levels were evaluated by real-time PCR of msp4 and normalized against tick 16S rDNA. Known amounts of the full length A.p. and A.m. msp4 PCR product were used to construct a standard curve for quantitation of pathogens per cell. Data represent average ± SD. (b) The mRNA levels of selected genes were evaluated by real-time RT-PCR and normalized against tick 16S rRNA. Bars represent the ratio between average Ct values in A.p.-infected cells/average Ct values in A.m.-infected cells. The mRNA levels were compared between A.p.- and A.m.-infected tick cells by Student's t-test (*P ≤ .05). Identical mRNA levels in A. phagocytophilum and A. marginale infected cells equal one.

Mentions: Because the kinetics of differentially expressed genes may vary with Anaplasma infection levels, the expression of selected genes was compared by real-time RT-PCR in A. phagocytophilum- and A. marginale-infected ISE6 cells collected at 2, 5, and 8 dpi (Figure 4(a)). The results showed time-dependent variation in the mRNA ratios of studied genes between A. phagocytophilum- and A. marginale-infected cells (Figure 4(b)). However, significant differences were observed between A. phagocytophilum and A. marginale infection at all time points, thus suggesting that differences in gene expression profiles elicited by these pathogens are present throughout the infection cycle in tick ISE6 cells.


Anaplasma phagocytophilum and Anaplasma marginale elicit different gene expression responses in cultured tick cells.

Zivkovic Z, Blouin EF, Manzano-Roman R, Almazán C, Naranjo V, Massung RF, Jongejan F, Kocan KM, de la Fuente J - Comp. Funct. Genomics (2009)

Comparison between differential gene expression in A. phagocytophilum- and A. marginale-infected tick ISE6 cells at different time points after infection. Studies were done on A. phagocytophilum- (A.p.-) and A. marginale- (A.m.-) infected tick ISE6 cells (two independent cultures each) at 2, 5, and 8 days postinfection (dpi) with approximately 30%–40%, 60%–70%, and >90% infected cells, respectively. (a) The A.p. and A.m. infection levels were evaluated by real-time PCR of msp4 and normalized against tick 16S rDNA. Known amounts of the full length A.p. and A.m. msp4 PCR product were used to construct a standard curve for quantitation of pathogens per cell. Data represent average  ± SD. (b) The mRNA levels of selected genes were evaluated by real-time RT-PCR and normalized against tick 16S rRNA. Bars represent the ratio between average Ct values in A.p.-infected cells/average Ct values in A.m.-infected cells. The mRNA levels were compared between A.p.- and A.m.-infected tick cells by Student's t-test (*P ≤ .05). Identical mRNA levels in A. phagocytophilum and A. marginale infected cells equal one.
© Copyright Policy - open-access
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC2712686&req=5

fig4: Comparison between differential gene expression in A. phagocytophilum- and A. marginale-infected tick ISE6 cells at different time points after infection. Studies were done on A. phagocytophilum- (A.p.-) and A. marginale- (A.m.-) infected tick ISE6 cells (two independent cultures each) at 2, 5, and 8 days postinfection (dpi) with approximately 30%–40%, 60%–70%, and >90% infected cells, respectively. (a) The A.p. and A.m. infection levels were evaluated by real-time PCR of msp4 and normalized against tick 16S rDNA. Known amounts of the full length A.p. and A.m. msp4 PCR product were used to construct a standard curve for quantitation of pathogens per cell. Data represent average ± SD. (b) The mRNA levels of selected genes were evaluated by real-time RT-PCR and normalized against tick 16S rRNA. Bars represent the ratio between average Ct values in A.p.-infected cells/average Ct values in A.m.-infected cells. The mRNA levels were compared between A.p.- and A.m.-infected tick cells by Student's t-test (*P ≤ .05). Identical mRNA levels in A. phagocytophilum and A. marginale infected cells equal one.
Mentions: Because the kinetics of differentially expressed genes may vary with Anaplasma infection levels, the expression of selected genes was compared by real-time RT-PCR in A. phagocytophilum- and A. marginale-infected ISE6 cells collected at 2, 5, and 8 dpi (Figure 4(a)). The results showed time-dependent variation in the mRNA ratios of studied genes between A. phagocytophilum- and A. marginale-infected cells (Figure 4(b)). However, significant differences were observed between A. phagocytophilum and A. marginale infection at all time points, thus suggesting that differences in gene expression profiles elicited by these pathogens are present throughout the infection cycle in tick ISE6 cells.

Bottom Line: The genus Anaplasma (Rickettsiales: Anaplasmataceae) includes obligate tick-transmitted intracellular organisms, Anaplasma phagocytophilum and Anaplasma marginale that multiply in both vertebrate and tick host cells.The results of these studies demonstrated modulation of tick gene expression by A. phagocytophilum and provided evidence of different gene expression responses in tick cells infected with A. phagocytophilum and A. marginale.These differences in Anaplasma-tick interactions may reflect differences in pathogen life cycle in the tick cells.

View Article: PubMed Central - PubMed

Affiliation: Utrecht Centre for Tick-Borne Diseases (UCTD), Department of Infectious Diseases and Immunology, Faculty of Veterinary Medicine, Utrecht University, Yalelaan 1, 3584 CL Utrecht, The Netherlands.

ABSTRACT
The genus Anaplasma (Rickettsiales: Anaplasmataceae) includes obligate tick-transmitted intracellular organisms, Anaplasma phagocytophilum and Anaplasma marginale that multiply in both vertebrate and tick host cells. Recently, we showed that A. marginale affects the expression of tick genes that are involved in tick survival and pathogen infection and multiplication. However, the gene expression profile in A. phagocytophilum-infected tick cells is currently poorly characterized. The objectives of this study were to characterize tick gene expression profile in Ixodes scapularis ticks and cultured ISE6 cells in response to infection with A. phagocypthilum and to compare tick gene expression responses in A. phagocytophilum- and A. marginale-infected tick cells by microarray and real-time RT-PCR analyses. The results of these studies demonstrated modulation of tick gene expression by A. phagocytophilum and provided evidence of different gene expression responses in tick cells infected with A. phagocytophilum and A. marginale. These differences in Anaplasma-tick interactions may reflect differences in pathogen life cycle in the tick cells.

No MeSH data available.


Related in: MedlinePlus