Limits...
A crucial role for B and T lymphocyte attenuator in preventing the development of CD4+ T cell-mediated herpetic stromal keratitis.

Xia L, Zhang S, Zhou J, Li Y - Mol. Vis. (2010)

Bottom Line: Systemic administration of pBTLA resulted in a diminished incidence and severity of corneal lesions compared to controls.The results suggest that recombinant pBTLA plays a crucial role in preventing HSV-1 specific responses in CD4(+) Th1 cells in the infected corneas.Thus, BTLA, with immunosuppressive effects, may be a good candidate for treatment of HSK.

View Article: PubMed Central - PubMed

Affiliation: Department of Ophthalmology, Shengjing Hospital, China Medical University, Shenyang, Liaoning Province, People's Republic of China. xialk@sjhospital.org

ABSTRACT

Purpose: To investigate the effect of the B and T lymphocyte attenuator (BTLA; CD272) on cluster of differentiation (CD)4(+) T cell-mediated corneal immunopathology during murine herpetic stromal keratitis (HSK).

Methods: BALB/c mice were infected with the herpes simplex virus type 1 (HSV-1) KOS strain by corneal scarification. The levels of BTLA expression in CD4(+) and CD8(+) T cells in murine peripheral blood were determined by flow cytometry on days 0, 3, 7, 10, 14, and 21 after HSV-1 infection. BTLA expression in the infected cornea was detected by immunohistochemistry. BALB/c mice were injected intraperitoneally with recombinant plasmid DNA encoding BTLA (pBTLA), pcDNA3.1, or PBS on 0 and 7 days before infection and 7 days postinfection. The incidence and severity of stromal disease, tear film virus titers, and the delayed-type hypersensitivity (DTH) reaction were then compared among treated and control groups. The effects of pBTLA on CD4(+) T cells that infiltrated into infected corneas and on type 1 helper T-cell (Th1) cytokines (interferon-gamma [IFN-γ]) were evaluated. The levels of glycoprotein D (gD) mRNA in corneas were tested by real-time PCR. The eyes were examined histologically.

Results: BTLA expression increased both in the corneas of HSV-1 infected mice and in CD4(+) T cells in the murine peripheral blood. Systemic administration of pBTLA resulted in a diminished incidence and severity of corneal lesions compared to controls. Treatment with pBTLA led to a decreased infiltration of CD4(+) T cells into infected corneas, and diminished Th1 responses in murine corneas, draining lymph nodes, and splenocytes. The pBTLA treated mice showed an impaired DTH response two weeks after HSV-1 infection compared to control mice. No differences were noted in tear film virus titers or gD mRNA levels in corneas among the experimental groups.

Conclusions: The results suggest that recombinant pBTLA plays a crucial role in preventing HSV-1 specific responses in CD4(+) Th1 cells in the infected corneas. Thus, BTLA, with immunosuppressive effects, may be a good candidate for treatment of HSK.

Show MeSH

Related in: MedlinePlus

pBTLA treatment does not affect virus titers in tear film. Tear film virus titers were compared in the different mice groups following HSV-1 infection. Mice treated with pBTLA and pcDNA3.1 received 100 μg of the respective plasmid on days, −7, 0, 7 relative to HSV-1 corneal inoculation, and the PBS group received 100 µl PBS intraperitoneally. On days 1, 3, and 5, swabs of the corneal surface were collected and transferred to serum-free DMEM, and viral titers were determined by a plaque assay performed on VERO cells as described in methods. The virus titers was calculated as log10PFU/ml. Results are expressed as means±SD for individual eye swabs (n=6). No statistically significant differences were observed among the different groups (p>0.05).
© Copyright Policy - open-access
Related In: Results  -  Collection

License
getmorefigures.php?uid=PMC2965573&req=5

f8: pBTLA treatment does not affect virus titers in tear film. Tear film virus titers were compared in the different mice groups following HSV-1 infection. Mice treated with pBTLA and pcDNA3.1 received 100 μg of the respective plasmid on days, −7, 0, 7 relative to HSV-1 corneal inoculation, and the PBS group received 100 µl PBS intraperitoneally. On days 1, 3, and 5, swabs of the corneal surface were collected and transferred to serum-free DMEM, and viral titers were determined by a plaque assay performed on VERO cells as described in methods. The virus titers was calculated as log10PFU/ml. Results are expressed as means±SD for individual eye swabs (n=6). No statistically significant differences were observed among the different groups (p>0.05).

Mentions: Tear film virus titers on days 1, 3, and 5 following HSV-1 infection are shown in Figure 8. Mice systemically administered pBTLA showed no difference in virus titer from pcDNA3.1 treated or PBS control mice (p>0.05). Systemic administration of recombinant pBTLA therefore did not appear to affect virus titers in tear film.


A crucial role for B and T lymphocyte attenuator in preventing the development of CD4+ T cell-mediated herpetic stromal keratitis.

Xia L, Zhang S, Zhou J, Li Y - Mol. Vis. (2010)

pBTLA treatment does not affect virus titers in tear film. Tear film virus titers were compared in the different mice groups following HSV-1 infection. Mice treated with pBTLA and pcDNA3.1 received 100 μg of the respective plasmid on days, −7, 0, 7 relative to HSV-1 corneal inoculation, and the PBS group received 100 µl PBS intraperitoneally. On days 1, 3, and 5, swabs of the corneal surface were collected and transferred to serum-free DMEM, and viral titers were determined by a plaque assay performed on VERO cells as described in methods. The virus titers was calculated as log10PFU/ml. Results are expressed as means±SD for individual eye swabs (n=6). No statistically significant differences were observed among the different groups (p>0.05).
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC2965573&req=5

f8: pBTLA treatment does not affect virus titers in tear film. Tear film virus titers were compared in the different mice groups following HSV-1 infection. Mice treated with pBTLA and pcDNA3.1 received 100 μg of the respective plasmid on days, −7, 0, 7 relative to HSV-1 corneal inoculation, and the PBS group received 100 µl PBS intraperitoneally. On days 1, 3, and 5, swabs of the corneal surface were collected and transferred to serum-free DMEM, and viral titers were determined by a plaque assay performed on VERO cells as described in methods. The virus titers was calculated as log10PFU/ml. Results are expressed as means±SD for individual eye swabs (n=6). No statistically significant differences were observed among the different groups (p>0.05).
Mentions: Tear film virus titers on days 1, 3, and 5 following HSV-1 infection are shown in Figure 8. Mice systemically administered pBTLA showed no difference in virus titer from pcDNA3.1 treated or PBS control mice (p>0.05). Systemic administration of recombinant pBTLA therefore did not appear to affect virus titers in tear film.

Bottom Line: Systemic administration of pBTLA resulted in a diminished incidence and severity of corneal lesions compared to controls.The results suggest that recombinant pBTLA plays a crucial role in preventing HSV-1 specific responses in CD4(+) Th1 cells in the infected corneas.Thus, BTLA, with immunosuppressive effects, may be a good candidate for treatment of HSK.

View Article: PubMed Central - PubMed

Affiliation: Department of Ophthalmology, Shengjing Hospital, China Medical University, Shenyang, Liaoning Province, People's Republic of China. xialk@sjhospital.org

ABSTRACT

Purpose: To investigate the effect of the B and T lymphocyte attenuator (BTLA; CD272) on cluster of differentiation (CD)4(+) T cell-mediated corneal immunopathology during murine herpetic stromal keratitis (HSK).

Methods: BALB/c mice were infected with the herpes simplex virus type 1 (HSV-1) KOS strain by corneal scarification. The levels of BTLA expression in CD4(+) and CD8(+) T cells in murine peripheral blood were determined by flow cytometry on days 0, 3, 7, 10, 14, and 21 after HSV-1 infection. BTLA expression in the infected cornea was detected by immunohistochemistry. BALB/c mice were injected intraperitoneally with recombinant plasmid DNA encoding BTLA (pBTLA), pcDNA3.1, or PBS on 0 and 7 days before infection and 7 days postinfection. The incidence and severity of stromal disease, tear film virus titers, and the delayed-type hypersensitivity (DTH) reaction were then compared among treated and control groups. The effects of pBTLA on CD4(+) T cells that infiltrated into infected corneas and on type 1 helper T-cell (Th1) cytokines (interferon-gamma [IFN-γ]) were evaluated. The levels of glycoprotein D (gD) mRNA in corneas were tested by real-time PCR. The eyes were examined histologically.

Results: BTLA expression increased both in the corneas of HSV-1 infected mice and in CD4(+) T cells in the murine peripheral blood. Systemic administration of pBTLA resulted in a diminished incidence and severity of corneal lesions compared to controls. Treatment with pBTLA led to a decreased infiltration of CD4(+) T cells into infected corneas, and diminished Th1 responses in murine corneas, draining lymph nodes, and splenocytes. The pBTLA treated mice showed an impaired DTH response two weeks after HSV-1 infection compared to control mice. No differences were noted in tear film virus titers or gD mRNA levels in corneas among the experimental groups.

Conclusions: The results suggest that recombinant pBTLA plays a crucial role in preventing HSV-1 specific responses in CD4(+) Th1 cells in the infected corneas. Thus, BTLA, with immunosuppressive effects, may be a good candidate for treatment of HSK.

Show MeSH
Related in: MedlinePlus