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A penetrating ocular injury can affect the induction of anterior chamber-associated immune deviation.

Lei F, Zhang J, Zhang J, He H, Du Y, Yang P - Mol. Vis. (2008)

Bottom Line: However, the levels of IFN-gamma produced by splenocytes of the AC-injection group and the 72-h and 120-h group were significantly lower than those in the 24-h and 48-h group.An increased frequency of CD4(+)CD25(+)Foxp3(+)T cells was found in the AC-injection group and the 72-h and 120-h group.Penetrating ocular injury preformed shortly (24 h-48 h) after an AC injection of an antigen was able to abrogate ACAID and was associated with a decreased production of TGF-beta1 and IL-10, an increased production of IFN-gamma, and a decreased expression of CD4(+)CD25(+)Foxp3(+)T cells.

View Article: PubMed Central - PubMed

Affiliation: Department of Ophthalmology, First Affiliated Hospital, Department of Microbiology and Immunology, Zheng Zhou University, Zhengzhou, Henan Province, China.

ABSTRACT

Purpose: To determine the effect of penetrating ocular injury on the induction of anterior chamber-associated immune deviation (ACAID).

Methods: An injection of 5 mul ovalbumin (OVA, 20 mg/ml) into the anterior chamber (AC) of female BALB/c mice was performed to induce ACAID. A penetrating ocular injury was induced via the limbus on OVA-inoculated eyes at 24 h, 48 h, 72 h, and 120 h following AC injection. The mice receiving an OVA inoculation without the ocular injury served as the AC-injection group. Delayed type hypersensitivity (DTH) was examined to evaluate the induction of ACAID. The levels of transforming growth factor (TGF)-beta1, interleukin (IL)-10, and interferon (IFN)-gamma produced by splenocytes were detected by enzyme-linked immunosorbent assays (ELISA). The frequency of CD4(+)CD25(+)Foxp3(+)T cells in the splenocytes was detected by flow cytometry.

Results: A significantly decreased DTH response was observed in the AC-injection group as well as in mice that received a penetrating injury at 72 h and 120 h following AC-injection of OVA. The levels of TGF-beta1 and IL-10 produced by splenocytes of mice in the AC-injection group and in the 72-h and 120-h group were significant higher than those in the 24-h and 48-h group. However, the levels of IFN-gamma produced by splenocytes of the AC-injection group and the 72-h and 120-h group were significantly lower than those in the 24-h and 48-h group. An increased frequency of CD4(+)CD25(+)Foxp3(+)T cells was found in the AC-injection group and the 72-h and 120-h group.

Conclusions: Penetrating ocular injury preformed shortly (24 h-48 h) after an AC injection of an antigen was able to abrogate ACAID and was associated with a decreased production of TGF-beta1 and IL-10, an increased production of IFN-gamma, and a decreased expression of CD4(+)CD25(+)Foxp3(+)T cells.

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The method for preparation of a 1 mm-depth penetrating ocular injury at six o’clock of the murine limbus. A modified Standard Angle Blade is shown in the picture (A). A copper sheet was stuck 1 mm posterior to the blade tip. The diagram (B) showed that a 1 mm-depth penetrating ocular injury was made perpendicularly to the limbus using the modified standard Angle Blade.
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f1: The method for preparation of a 1 mm-depth penetrating ocular injury at six o’clock of the murine limbus. A modified Standard Angle Blade is shown in the picture (A). A copper sheet was stuck 1 mm posterior to the blade tip. The diagram (B) showed that a 1 mm-depth penetrating ocular injury was made perpendicularly to the limbus using the modified standard Angle Blade.

Mentions: ACAID was induced as described previously [14-16]. Mice were anesthetized with inhalation anesthesia consisting of oxygen and 1.7% isoflurane. Five microliters of ovalbumin (OVA; 20mg/ml; Sigma-Aldrich, Steinheim, Germany) dissolved in phosphate buffered saline (PBS) were injected into the AC of mice using a glass micropipette (about 80 μm in diameter) with a sterile infant feeding tube mounted onto a 0.1 ml Hamilton syringe (Hamilton, Reno, NV). Seven days later, the mice were immunized by subcutaneous (s.c.) injection of 250 μg OVA (2.5mg/ml, dissolved in PBS) emulsified 1:1 in complete Freund’s adjuvant (CFA; Sigma-Aldrich). This group of mice acted as the AC-injection group. Mice receiving an s.c. injection of OVA in CFA alone acted as positive controls. Untreated mice acted as normal controls. Mice receiving an injection of 5 μl sterile PBS into the anterior chamber and an s.c. injection of OVA in CFA seven days after AC injection acted as PBS controls. A penetrating ocular injury on OVA-inoculated eyes was performed 24 h (24-h group), 48 h (48-h group), 72 h (72-h group), and 120 h (120-h group) after AC injection. A 1 mm-depth penetrating ocular injury was made using a modified Standard Angle Blade (Alcan Laboratory, Fort Worth, Texas) as described below. A copper sheet was attached 1 mm posterior to the blade tip using metal gel to prevent a deeper injury (Figure 1A). Under a microscope, a perpendicular injury was carefully performed at the six o’clock position of the limbus using this modified blade (Figure 1B). Iris incarceration was observed in the eye immediately after this procedure had been performed. All mice were challenged with an s.c. injection of OVA in CFA seven days after the AC injection of OVA.


A penetrating ocular injury can affect the induction of anterior chamber-associated immune deviation.

Lei F, Zhang J, Zhang J, He H, Du Y, Yang P - Mol. Vis. (2008)

The method for preparation of a 1 mm-depth penetrating ocular injury at six o’clock of the murine limbus. A modified Standard Angle Blade is shown in the picture (A). A copper sheet was stuck 1 mm posterior to the blade tip. The diagram (B) showed that a 1 mm-depth penetrating ocular injury was made perpendicularly to the limbus using the modified standard Angle Blade.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC2255028&req=5

f1: The method for preparation of a 1 mm-depth penetrating ocular injury at six o’clock of the murine limbus. A modified Standard Angle Blade is shown in the picture (A). A copper sheet was stuck 1 mm posterior to the blade tip. The diagram (B) showed that a 1 mm-depth penetrating ocular injury was made perpendicularly to the limbus using the modified standard Angle Blade.
Mentions: ACAID was induced as described previously [14-16]. Mice were anesthetized with inhalation anesthesia consisting of oxygen and 1.7% isoflurane. Five microliters of ovalbumin (OVA; 20mg/ml; Sigma-Aldrich, Steinheim, Germany) dissolved in phosphate buffered saline (PBS) were injected into the AC of mice using a glass micropipette (about 80 μm in diameter) with a sterile infant feeding tube mounted onto a 0.1 ml Hamilton syringe (Hamilton, Reno, NV). Seven days later, the mice were immunized by subcutaneous (s.c.) injection of 250 μg OVA (2.5mg/ml, dissolved in PBS) emulsified 1:1 in complete Freund’s adjuvant (CFA; Sigma-Aldrich). This group of mice acted as the AC-injection group. Mice receiving an s.c. injection of OVA in CFA alone acted as positive controls. Untreated mice acted as normal controls. Mice receiving an injection of 5 μl sterile PBS into the anterior chamber and an s.c. injection of OVA in CFA seven days after AC injection acted as PBS controls. A penetrating ocular injury on OVA-inoculated eyes was performed 24 h (24-h group), 48 h (48-h group), 72 h (72-h group), and 120 h (120-h group) after AC injection. A 1 mm-depth penetrating ocular injury was made using a modified Standard Angle Blade (Alcan Laboratory, Fort Worth, Texas) as described below. A copper sheet was attached 1 mm posterior to the blade tip using metal gel to prevent a deeper injury (Figure 1A). Under a microscope, a perpendicular injury was carefully performed at the six o’clock position of the limbus using this modified blade (Figure 1B). Iris incarceration was observed in the eye immediately after this procedure had been performed. All mice were challenged with an s.c. injection of OVA in CFA seven days after the AC injection of OVA.

Bottom Line: However, the levels of IFN-gamma produced by splenocytes of the AC-injection group and the 72-h and 120-h group were significantly lower than those in the 24-h and 48-h group.An increased frequency of CD4(+)CD25(+)Foxp3(+)T cells was found in the AC-injection group and the 72-h and 120-h group.Penetrating ocular injury preformed shortly (24 h-48 h) after an AC injection of an antigen was able to abrogate ACAID and was associated with a decreased production of TGF-beta1 and IL-10, an increased production of IFN-gamma, and a decreased expression of CD4(+)CD25(+)Foxp3(+)T cells.

View Article: PubMed Central - PubMed

Affiliation: Department of Ophthalmology, First Affiliated Hospital, Department of Microbiology and Immunology, Zheng Zhou University, Zhengzhou, Henan Province, China.

ABSTRACT

Purpose: To determine the effect of penetrating ocular injury on the induction of anterior chamber-associated immune deviation (ACAID).

Methods: An injection of 5 mul ovalbumin (OVA, 20 mg/ml) into the anterior chamber (AC) of female BALB/c mice was performed to induce ACAID. A penetrating ocular injury was induced via the limbus on OVA-inoculated eyes at 24 h, 48 h, 72 h, and 120 h following AC injection. The mice receiving an OVA inoculation without the ocular injury served as the AC-injection group. Delayed type hypersensitivity (DTH) was examined to evaluate the induction of ACAID. The levels of transforming growth factor (TGF)-beta1, interleukin (IL)-10, and interferon (IFN)-gamma produced by splenocytes were detected by enzyme-linked immunosorbent assays (ELISA). The frequency of CD4(+)CD25(+)Foxp3(+)T cells in the splenocytes was detected by flow cytometry.

Results: A significantly decreased DTH response was observed in the AC-injection group as well as in mice that received a penetrating injury at 72 h and 120 h following AC-injection of OVA. The levels of TGF-beta1 and IL-10 produced by splenocytes of mice in the AC-injection group and in the 72-h and 120-h group were significant higher than those in the 24-h and 48-h group. However, the levels of IFN-gamma produced by splenocytes of the AC-injection group and the 72-h and 120-h group were significantly lower than those in the 24-h and 48-h group. An increased frequency of CD4(+)CD25(+)Foxp3(+)T cells was found in the AC-injection group and the 72-h and 120-h group.

Conclusions: Penetrating ocular injury preformed shortly (24 h-48 h) after an AC injection of an antigen was able to abrogate ACAID and was associated with a decreased production of TGF-beta1 and IL-10, an increased production of IFN-gamma, and a decreased expression of CD4(+)CD25(+)Foxp3(+)T cells.

Show MeSH
Related in: MedlinePlus