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The role of Qa-2, the functional homolog of HLA-G, in a Behcet's disease-like mouse model induced by the herpes virus simplex.

Lee M, Choi B, Kwon HJ, Shim JA, Park KS, Lee ES, Sohn S - J Inflamm (Lond) (2010)

Bottom Line: In this study, we evaluated the level of Qa-2, a murine nonclassical class I MHC molecule and possible functional homolog of HLA-G, to determine if it was associated with various symptoms of BD-like mice.The silencing of Qa-2 by intravenous injection of siRNA (500 nmol/mouse, 4 times at 3-day intervals) specifically reduced the Qa-2 levels and worsened the BD-like symptoms.Silencing Qa-2 by injecting siRNA into mice resulted in deterioration of symptoms in BD-like mice.

View Article: PubMed Central - HTML - PubMed

Affiliation: Laboratory of Cell Biology, Ajou University Institute for Medical Sciences, Suwon, Korea. sohnsh@ajou.ac.kr.

ABSTRACT

Background: It has been suggested that the HLA-G molecule is a genetic risk factor for Behcet's disease (BD). In this study, we evaluated the level of Qa-2, a murine nonclassical class I MHC molecule and possible functional homolog of HLA-G, to determine if it was associated with various symptoms of BD-like mice. In addition, we investigated siRNA (small interfering RNA) treatment to determine if it inhibited Qa-2 expression, thereby changing the symptoms of mice.

Methods: RNA interference (RNAi) and vector transfection were employed to manipulate gene expression in vivo in mice. siRNA (small interfering RNA) or Qa-2 expression vector was applied to inhibit or up-regulate Qa-2 expression, respectively.

Results: The Qa-2 levels in granulocytes were lower in BD-like mice than in normal controls. The silencing of Qa-2 by intravenous injection of siRNA (500 nmol/mouse, 4 times at 3-day intervals) specifically reduced the Qa-2 levels and worsened the BD-like symptoms.

Conclusions: Silencing Qa-2 by injecting siRNA into mice resulted in deterioration of symptoms in BD-like mice.

No MeSH data available.


Related in: MedlinePlus

Down-regulation of Qa-2 after intravenous injection of siRNA into BD mice. A. BD mice were injected once with control siRNA or Qa-2 siRNA (500 nmol/mouse), which was composed of the α3 domain, transmembrane domain, and cytoplasmic domain. PBMC collected from the orbital sinus before and 1 day after injection were analyzed by flow cytometry. Qa-2 siRNA effectively reduced the Qa-2 levels in the PBMC of BD mice. B. Two days after injection, the frequency of Qa-2 positive lymphocytes and granulocytes was analyzed in Qa-2 siRNA injected BD mice. In lymphocytes and granulocytes, Qa-2 siRNA significantly reduced the number of Qa-2 positive cells when compared to glucose-injected control mice. C. The frequency of Qa-2 positive cells in mice that were injected with Qa-2 siRNA four times. Specifically, 500 nmol control siRNA or Qa-2 siRNA in 200 μl of 5% glucose solution was intraperitoneally injected four times with a three day interval between injections, and the PBMC were analyzed by FACS two days after the last injection. In lymphocytes and granulocytes, Qa-2 siRNA significantly reduced the Qa-2 positive cells when compared to glucose injected control mice.
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Figure 3: Down-regulation of Qa-2 after intravenous injection of siRNA into BD mice. A. BD mice were injected once with control siRNA or Qa-2 siRNA (500 nmol/mouse), which was composed of the α3 domain, transmembrane domain, and cytoplasmic domain. PBMC collected from the orbital sinus before and 1 day after injection were analyzed by flow cytometry. Qa-2 siRNA effectively reduced the Qa-2 levels in the PBMC of BD mice. B. Two days after injection, the frequency of Qa-2 positive lymphocytes and granulocytes was analyzed in Qa-2 siRNA injected BD mice. In lymphocytes and granulocytes, Qa-2 siRNA significantly reduced the number of Qa-2 positive cells when compared to glucose-injected control mice. C. The frequency of Qa-2 positive cells in mice that were injected with Qa-2 siRNA four times. Specifically, 500 nmol control siRNA or Qa-2 siRNA in 200 μl of 5% glucose solution was intraperitoneally injected four times with a three day interval between injections, and the PBMC were analyzed by FACS two days after the last injection. In lymphocytes and granulocytes, Qa-2 siRNA significantly reduced the Qa-2 positive cells when compared to glucose injected control mice.

Mentions: Next, an siRNA mixture composed of the siRNA of the α3 domain, transmembrane domain and the cytoplasmic domain was injected into BD mice. Five to six individual BD mice in each group were intravenously injected once with 5% glucose or 500 nmol of Qa-2 siRNA or control siRNA, and their PBMCs were analyzed one day and two days later by flow cytometry. One day after Qa-2 siRNA injection, the number of Qa-2 positive granulocytes was 32.18 ± 14.64%, which was significantly lower (p = 0.049) than that of mice treated with 5% glucose (54.21 ± 1.89%) or leader peptide (61.32 ± 12.27%) (Figure 3A). In lymphocytes, the Qa-2 positive cell counts did not differ significantly among groups. Two days later, the frequency of Qa-2 positive cells was 84.12 ± 10.34% in Qa-2 siRNA injected mice, while it was 94.23 ± 3.86% of glucose injected control mice in lymphocytes (p = 0.029). In granulocytes, the frequency of Qa-2 positive cells was 42.18 ± 28.40% in Qa-2 siRNA injected mice, while it was 75.65 ± 23.59% in glucose injected control mice (p = 0.008). These findings demonstrated that Qa-2 siRNA effectively reduced the frequencies of Qa-2 positive cells in lymphocytes and granulocytes in BD mice (Figure 3B). To determine if repeated administration can reduce the Qa-2 level more efficiently, the frequency of Qa-2 positive cells in BD mice that were injected with Qa-2 siRNA four times was analyzed. To accomplish this, 500 nmol control siRNA or Qa-2 siRNA in 200 μl of 5% glucose solution was intraperitoneally injected four times with a three day interval in between injections. Two days after the last injection, the PBMCs were analyzed by FACS. In lymphocytes and granulocytes, Qa-2 siRNA led to a significant reduction in Qa-2 positive cells when compared to glucose injected control mice (p = 0.05, p = 0.02 each) (Figure 3C). However, the reduction in Qa-2 level observed in response to one and four injections did not differ significantly.


The role of Qa-2, the functional homolog of HLA-G, in a Behcet's disease-like mouse model induced by the herpes virus simplex.

Lee M, Choi B, Kwon HJ, Shim JA, Park KS, Lee ES, Sohn S - J Inflamm (Lond) (2010)

Down-regulation of Qa-2 after intravenous injection of siRNA into BD mice. A. BD mice were injected once with control siRNA or Qa-2 siRNA (500 nmol/mouse), which was composed of the α3 domain, transmembrane domain, and cytoplasmic domain. PBMC collected from the orbital sinus before and 1 day after injection were analyzed by flow cytometry. Qa-2 siRNA effectively reduced the Qa-2 levels in the PBMC of BD mice. B. Two days after injection, the frequency of Qa-2 positive lymphocytes and granulocytes was analyzed in Qa-2 siRNA injected BD mice. In lymphocytes and granulocytes, Qa-2 siRNA significantly reduced the number of Qa-2 positive cells when compared to glucose-injected control mice. C. The frequency of Qa-2 positive cells in mice that were injected with Qa-2 siRNA four times. Specifically, 500 nmol control siRNA or Qa-2 siRNA in 200 μl of 5% glucose solution was intraperitoneally injected four times with a three day interval between injections, and the PBMC were analyzed by FACS two days after the last injection. In lymphocytes and granulocytes, Qa-2 siRNA significantly reduced the Qa-2 positive cells when compared to glucose injected control mice.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC2902457&req=5

Figure 3: Down-regulation of Qa-2 after intravenous injection of siRNA into BD mice. A. BD mice were injected once with control siRNA or Qa-2 siRNA (500 nmol/mouse), which was composed of the α3 domain, transmembrane domain, and cytoplasmic domain. PBMC collected from the orbital sinus before and 1 day after injection were analyzed by flow cytometry. Qa-2 siRNA effectively reduced the Qa-2 levels in the PBMC of BD mice. B. Two days after injection, the frequency of Qa-2 positive lymphocytes and granulocytes was analyzed in Qa-2 siRNA injected BD mice. In lymphocytes and granulocytes, Qa-2 siRNA significantly reduced the number of Qa-2 positive cells when compared to glucose-injected control mice. C. The frequency of Qa-2 positive cells in mice that were injected with Qa-2 siRNA four times. Specifically, 500 nmol control siRNA or Qa-2 siRNA in 200 μl of 5% glucose solution was intraperitoneally injected four times with a three day interval between injections, and the PBMC were analyzed by FACS two days after the last injection. In lymphocytes and granulocytes, Qa-2 siRNA significantly reduced the Qa-2 positive cells when compared to glucose injected control mice.
Mentions: Next, an siRNA mixture composed of the siRNA of the α3 domain, transmembrane domain and the cytoplasmic domain was injected into BD mice. Five to six individual BD mice in each group were intravenously injected once with 5% glucose or 500 nmol of Qa-2 siRNA or control siRNA, and their PBMCs were analyzed one day and two days later by flow cytometry. One day after Qa-2 siRNA injection, the number of Qa-2 positive granulocytes was 32.18 ± 14.64%, which was significantly lower (p = 0.049) than that of mice treated with 5% glucose (54.21 ± 1.89%) or leader peptide (61.32 ± 12.27%) (Figure 3A). In lymphocytes, the Qa-2 positive cell counts did not differ significantly among groups. Two days later, the frequency of Qa-2 positive cells was 84.12 ± 10.34% in Qa-2 siRNA injected mice, while it was 94.23 ± 3.86% of glucose injected control mice in lymphocytes (p = 0.029). In granulocytes, the frequency of Qa-2 positive cells was 42.18 ± 28.40% in Qa-2 siRNA injected mice, while it was 75.65 ± 23.59% in glucose injected control mice (p = 0.008). These findings demonstrated that Qa-2 siRNA effectively reduced the frequencies of Qa-2 positive cells in lymphocytes and granulocytes in BD mice (Figure 3B). To determine if repeated administration can reduce the Qa-2 level more efficiently, the frequency of Qa-2 positive cells in BD mice that were injected with Qa-2 siRNA four times was analyzed. To accomplish this, 500 nmol control siRNA or Qa-2 siRNA in 200 μl of 5% glucose solution was intraperitoneally injected four times with a three day interval in between injections. Two days after the last injection, the PBMCs were analyzed by FACS. In lymphocytes and granulocytes, Qa-2 siRNA led to a significant reduction in Qa-2 positive cells when compared to glucose injected control mice (p = 0.05, p = 0.02 each) (Figure 3C). However, the reduction in Qa-2 level observed in response to one and four injections did not differ significantly.

Bottom Line: In this study, we evaluated the level of Qa-2, a murine nonclassical class I MHC molecule and possible functional homolog of HLA-G, to determine if it was associated with various symptoms of BD-like mice.The silencing of Qa-2 by intravenous injection of siRNA (500 nmol/mouse, 4 times at 3-day intervals) specifically reduced the Qa-2 levels and worsened the BD-like symptoms.Silencing Qa-2 by injecting siRNA into mice resulted in deterioration of symptoms in BD-like mice.

View Article: PubMed Central - HTML - PubMed

Affiliation: Laboratory of Cell Biology, Ajou University Institute for Medical Sciences, Suwon, Korea. sohnsh@ajou.ac.kr.

ABSTRACT

Background: It has been suggested that the HLA-G molecule is a genetic risk factor for Behcet's disease (BD). In this study, we evaluated the level of Qa-2, a murine nonclassical class I MHC molecule and possible functional homolog of HLA-G, to determine if it was associated with various symptoms of BD-like mice. In addition, we investigated siRNA (small interfering RNA) treatment to determine if it inhibited Qa-2 expression, thereby changing the symptoms of mice.

Methods: RNA interference (RNAi) and vector transfection were employed to manipulate gene expression in vivo in mice. siRNA (small interfering RNA) or Qa-2 expression vector was applied to inhibit or up-regulate Qa-2 expression, respectively.

Results: The Qa-2 levels in granulocytes were lower in BD-like mice than in normal controls. The silencing of Qa-2 by intravenous injection of siRNA (500 nmol/mouse, 4 times at 3-day intervals) specifically reduced the Qa-2 levels and worsened the BD-like symptoms.

Conclusions: Silencing Qa-2 by injecting siRNA into mice resulted in deterioration of symptoms in BD-like mice.

No MeSH data available.


Related in: MedlinePlus