Limits...
Whole blood gene expression in infants with respiratory syncytial virus bronchiolitis.

Fjaerli HO, Bukholm G, Krog A, Skjaeret C, Holden M, Nakstad B - BMC Infect. Dis. (2006)

Bottom Line: Among the 30 genes most differentially expressed by microarray nearly 50% were involved in immunological processes.We found the highly upregulated interferon, alpha-inducible protein 27 (IFI27) and the highly downregulated gene Charcot-Leyden crystal protein (CLC) to be the two most differentially expressed genes in the microarray study.When performing QRT-PCR on these genes IFI27 was upregulated in all but one infant, and CLC was downregulated in all 18 infants, and similar to that given by microarray.

View Article: PubMed Central - HTML - PubMed

Affiliation: University of Oslo, Faculty Division Akershus University Hospital, Department of Paediatrics, Akershus University Hospital, Norway. h.o.fjarli@medisin.uio.no

ABSTRACT

Background: Respiratory syncytial virus (RSV) is a major cause of viral bronchiolitis in infants worldwide, and environmental, viral and host factors are all of importance for disease susceptibility and severity. To study the systemic host response to this disease we used the microarray technology to measure mRNA gene expression levels in whole blood of five male infants hospitalised with acute RSV, subtype B, bronchiolitis versus five one year old male controls exposed to RSV during infancy without bronchiolitis. The gene expression levels were further evaluated in a new experiment using quantitative real-time polymerase chain reaction (QRT-PCR) both in the five infants selected for microarray and in 13 other infants hospitalised with the same disease.

Results: Among the 30 genes most differentially expressed by microarray nearly 50% were involved in immunological processes. We found the highly upregulated interferon, alpha-inducible protein 27 (IFI27) and the highly downregulated gene Charcot-Leyden crystal protein (CLC) to be the two most differentially expressed genes in the microarray study. When performing QRT-PCR on these genes IFI27 was upregulated in all but one infant, and CLC was downregulated in all 18 infants, and similar to that given by microarray.

Conclusion: The gene IFI27 is upregulated and the gene CLC is downregulated in whole blood of infants hospitalised with RSV, subtype B, bronchiolitis and is not reported before. More studies are needed to elucidate the specificity of these gene expressions in association with host response to this virus in bronchiolitis of moderate severity.

Show MeSH

Related in: MedlinePlus

QRT-PCR study of six whole blood immune response genes as given by microarray. A quantitative real-time polymerase chain reaction (QRT-PCR) study using TaqMan Low Density Array (TLDA) cards was performed with gene expressions given as mean relative quantification (RQ) from triplets of each gene. Analyzed in 18 infants hospitalised with respiratory syncytial virus, subtype B, bronchiolitis versus a pooled sample from four one year old male children exposed to RSV during infancy but not treated and/or hospitalised for bronchiolitis during infancy as exogenous control and with beta-glucuronidase (GUSB) as endogenous control. The following genes (gene symbol and name) were studied; CLC: Charcot-Leyden crystal protein, G1P2: Interferon, alpha-inducible protein (clone IFI-15K), IFI27: Interferon, alpha-inducible protein 27, IFI44: Interferon-induced protein 44, IFI44L: Interferon-induced protein 44-like, MARCO:Macrophage receptor with collagenous structure
© Copyright Policy - open-access
Related In: Results  -  Collection

License
getmorefigures.php?uid=PMC1713240&req=5

Figure 2: QRT-PCR study of six whole blood immune response genes as given by microarray. A quantitative real-time polymerase chain reaction (QRT-PCR) study using TaqMan Low Density Array (TLDA) cards was performed with gene expressions given as mean relative quantification (RQ) from triplets of each gene. Analyzed in 18 infants hospitalised with respiratory syncytial virus, subtype B, bronchiolitis versus a pooled sample from four one year old male children exposed to RSV during infancy but not treated and/or hospitalised for bronchiolitis during infancy as exogenous control and with beta-glucuronidase (GUSB) as endogenous control. The following genes (gene symbol and name) were studied; CLC: Charcot-Leyden crystal protein, G1P2: Interferon, alpha-inducible protein (clone IFI-15K), IFI27: Interferon, alpha-inducible protein 27, IFI44: Interferon-induced protein 44, IFI44L: Interferon-induced protein 44-like, MARCO:Macrophage receptor with collagenous structure

Mentions: To evaluate the impact of the 17 genes expressed by microarray a new cohort was tested using QRT-PCR. In the new cohort of 13 infants hospitalised with the same disease 11 of the 17 genes were significantly differentially expressed in accordance with the results from the microarray study (p < 0.01; independent samples t-test). In order to see if some of the genes associated with immunological processes could also be differentially expressed in the new cohort, as well as in the five infants selected for microarray, we evaluated the expressions of the six immune response genes available for studies with TLDA cards. The gene IFI27 was upregulated in 17 infants and slightly downregulated in one infant while the gene IFI44 was downregulated in 15 infants and upregulated in three infants. The genes IFI44L and MARCO were downregulated in 14 infants and upregulated in four infants, and the gene G1P2 was downregulated and upregulated in nine infants, respectively. Finally, the gene CLC was downregulated in all 18 infants (fig 2).


Whole blood gene expression in infants with respiratory syncytial virus bronchiolitis.

Fjaerli HO, Bukholm G, Krog A, Skjaeret C, Holden M, Nakstad B - BMC Infect. Dis. (2006)

QRT-PCR study of six whole blood immune response genes as given by microarray. A quantitative real-time polymerase chain reaction (QRT-PCR) study using TaqMan Low Density Array (TLDA) cards was performed with gene expressions given as mean relative quantification (RQ) from triplets of each gene. Analyzed in 18 infants hospitalised with respiratory syncytial virus, subtype B, bronchiolitis versus a pooled sample from four one year old male children exposed to RSV during infancy but not treated and/or hospitalised for bronchiolitis during infancy as exogenous control and with beta-glucuronidase (GUSB) as endogenous control. The following genes (gene symbol and name) were studied; CLC: Charcot-Leyden crystal protein, G1P2: Interferon, alpha-inducible protein (clone IFI-15K), IFI27: Interferon, alpha-inducible protein 27, IFI44: Interferon-induced protein 44, IFI44L: Interferon-induced protein 44-like, MARCO:Macrophage receptor with collagenous structure
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC1713240&req=5

Figure 2: QRT-PCR study of six whole blood immune response genes as given by microarray. A quantitative real-time polymerase chain reaction (QRT-PCR) study using TaqMan Low Density Array (TLDA) cards was performed with gene expressions given as mean relative quantification (RQ) from triplets of each gene. Analyzed in 18 infants hospitalised with respiratory syncytial virus, subtype B, bronchiolitis versus a pooled sample from four one year old male children exposed to RSV during infancy but not treated and/or hospitalised for bronchiolitis during infancy as exogenous control and with beta-glucuronidase (GUSB) as endogenous control. The following genes (gene symbol and name) were studied; CLC: Charcot-Leyden crystal protein, G1P2: Interferon, alpha-inducible protein (clone IFI-15K), IFI27: Interferon, alpha-inducible protein 27, IFI44: Interferon-induced protein 44, IFI44L: Interferon-induced protein 44-like, MARCO:Macrophage receptor with collagenous structure
Mentions: To evaluate the impact of the 17 genes expressed by microarray a new cohort was tested using QRT-PCR. In the new cohort of 13 infants hospitalised with the same disease 11 of the 17 genes were significantly differentially expressed in accordance with the results from the microarray study (p < 0.01; independent samples t-test). In order to see if some of the genes associated with immunological processes could also be differentially expressed in the new cohort, as well as in the five infants selected for microarray, we evaluated the expressions of the six immune response genes available for studies with TLDA cards. The gene IFI27 was upregulated in 17 infants and slightly downregulated in one infant while the gene IFI44 was downregulated in 15 infants and upregulated in three infants. The genes IFI44L and MARCO were downregulated in 14 infants and upregulated in four infants, and the gene G1P2 was downregulated and upregulated in nine infants, respectively. Finally, the gene CLC was downregulated in all 18 infants (fig 2).

Bottom Line: Among the 30 genes most differentially expressed by microarray nearly 50% were involved in immunological processes.We found the highly upregulated interferon, alpha-inducible protein 27 (IFI27) and the highly downregulated gene Charcot-Leyden crystal protein (CLC) to be the two most differentially expressed genes in the microarray study.When performing QRT-PCR on these genes IFI27 was upregulated in all but one infant, and CLC was downregulated in all 18 infants, and similar to that given by microarray.

View Article: PubMed Central - HTML - PubMed

Affiliation: University of Oslo, Faculty Division Akershus University Hospital, Department of Paediatrics, Akershus University Hospital, Norway. h.o.fjarli@medisin.uio.no

ABSTRACT

Background: Respiratory syncytial virus (RSV) is a major cause of viral bronchiolitis in infants worldwide, and environmental, viral and host factors are all of importance for disease susceptibility and severity. To study the systemic host response to this disease we used the microarray technology to measure mRNA gene expression levels in whole blood of five male infants hospitalised with acute RSV, subtype B, bronchiolitis versus five one year old male controls exposed to RSV during infancy without bronchiolitis. The gene expression levels were further evaluated in a new experiment using quantitative real-time polymerase chain reaction (QRT-PCR) both in the five infants selected for microarray and in 13 other infants hospitalised with the same disease.

Results: Among the 30 genes most differentially expressed by microarray nearly 50% were involved in immunological processes. We found the highly upregulated interferon, alpha-inducible protein 27 (IFI27) and the highly downregulated gene Charcot-Leyden crystal protein (CLC) to be the two most differentially expressed genes in the microarray study. When performing QRT-PCR on these genes IFI27 was upregulated in all but one infant, and CLC was downregulated in all 18 infants, and similar to that given by microarray.

Conclusion: The gene IFI27 is upregulated and the gene CLC is downregulated in whole blood of infants hospitalised with RSV, subtype B, bronchiolitis and is not reported before. More studies are needed to elucidate the specificity of these gene expressions in association with host response to this virus in bronchiolitis of moderate severity.

Show MeSH
Related in: MedlinePlus