Difference between CD25+Tregs and Helios+Tregs in a murine model of allergic rhinitis.

INTRODUCTION: Regulatory T or Treg cells, balance the peripheral immune response to allergens in allergic rhinitis. Traditionally, Treg (CD25+ Treg) is identified by the coexpression of Foxp3 and CD25, but this strategy does not represent the true inhibitory function of Treg cells. Helios has been thought of as novel marker of activated Tregs, with an important inhibitory function. Consequently, Helios was proposed as a marker of Treg. Recent articles have shown that Foxp3 and Helios co-expression (Helios+Tregs) is an important functional stage of Treg. OBJECTIVE: To compare the prevalence of CD25+Tregs and Helios+Tregs using a mouse model of allergic rhinitis. METHODS: Twenty mice were randomized into two groups. The test group comprised 10 allergic rhinitis model mice exposed to ovalbumin; the control group was exposed to saline. The fractions of CD25+Tregs, Helios+Tregs, Helios+CD25+, and Helios+Foxp3+CD25+Tregs present in the two groups were determined using flow cytometry. RESULTS: CD25+Tregs and Helios+Tregs were less abundant in the spleen and nasal mucosa cells of the allergic rhinitis model compared with the control. We also observed fewer Helios+Tregs than CD25+Tregs in nasal mucosa and splenic cells of both control and test groups. Moreover, we observed fewer Helios+Foxp3+, Helios+CD25+, and Helios+Foxp3+CD25+ Tregs in the nasal mucosa in the allergic rhinitis model. Helios was expressed the most in CD4+ CD25+Foxp3+ T-cells, followed by CD4+ CD25-Foxp3- T-cells. Approximately 75% of CD25+Tregs were Helios+ in spleens of allergic rhinitis and control mice. CONCLUSION: This is the first report of the proportions of Helios+Tregs in nasal mucosa and spleens of allergic rhinitis mice. Gating true inhibitory Tregs with the coexpression of Foxp3 and Helios might be more useful than relying on the expression of CD25. This study provides a new insight for Treg studies of allergic rhinitis, and the potential utility of the marker as a therapeutic target.

Flt3 ligand enhanced the immunogenicity of classical swine fever vectored vaccine based on human adenovirus by fusing to the E2 glycoprotein

Background: Classical swine fever (CSF) is a highly contagious disease of domestic pigs and wild boar. CSFV E2 protein is the major target for neutralizing antibodies and the main component of marker vaccine. CSF vaccine based on human adenovirus type 5 (HAdV-5) had achieved an efficient protection in swine. However, the efficacy of rAdV-E2 only expressed the E2 glycoprotein was unstable. As a vaccine adjuvant, FL has been proven to enhance the immune effect, and its extracellular domain retained full biological activity. The adjuvant activity of the porcine Flt3L (the extracellular domain) in CSF vectored vaccine based on HAdV-5 was investigated.Materials, Methods & Results: The FL (the extracellular domain) was linked to the CSFV E2 (N-terminus) by a 10GS (Gly4/Ser) linker. The rAdV-FL-E2 based on HAdV-5 was constructed and the FL-E2 protein expressed in 293 cells infected with rAdV-FL-E2 was confirmed by Western blot with the use of anti-CSFV serum. Twelve CSFV-free cross-bred piglets, which were 6 to 7 weeks old and double confirmed to be free to CSFV specific serum antibodies and antigens using the IDEXX HerdChek* CSFV Antibody Test Kit and real-time RT-PCR, respectively, and were randomly divided into four groups (A, B, C, and D) with three animals in each group. Groups A and B were vaccinated with a dose of 2×106 TCID50 of rAdV-FL-E2, and rAdV-E2 by intramuscular (i.m.) inoculation respectively. All pigs were given a booster immunization at 14 d intervals. Group C was immunized with one-dose CSFV C-strain vaccine and served as the positive control. Group D was injected with DMEM medium. Finally, all animals were challenged with 1×103 TCID50 of CSFV Shimen strain by i.m. injection at 21 d after the second vaccination. CSFV-specific neutralizing antibodies in sera were tested by using the IDEXX HerdChek* CSFV Antibody Test Kit and NPLA.[…](AU)

Flt3 ligand enhanced the immunogenicity of classical swine fever vectored vaccine based on human adenovirus by fusing to the E2 glycoprotein

Background: Classical swine fever (CSF) is a highly contagious disease of domestic pigs and wild boar. CSFV E2 protein is the major target for neutralizing antibodies and the main component of marker vaccine. CSF vaccine based on human adenovirus type 5 (HAdV-5) had achieved an efficient protection in swine. However, the efficacy of rAdV-E2 only expressed the E2 glycoprotein was unstable. As a vaccine adjuvant, FL has been proven to enhance the immune effect, and its extracellular domain retained full biological activity. The adjuvant activity of the porcine Flt3L (the extracellular domain) in CSF vectored vaccine based on HAdV-5 was investigated.Materials, Methods & Results: The FL (the extracellular domain) was linked to the CSFV E2 (N-terminus) by a 10GS (Gly4/Ser) linker. The rAdV-FL-E2 based on HAdV-5 was constructed and the FL-E2 protein expressed in 293 cells infected with rAdV-FL-E2 was confirmed by Western blot with the use of anti-CSFV serum. Twelve CSFV-free cross-bred piglets, which were 6 to 7 weeks old and double confirmed to be free to CSFV specific serum antibodies and antigens using the IDEXX HerdChek* CSFV Antibody Test Kit and real-time RT-PCR, respectively, and were randomly divided into four groups (A, B, C, and D) with three animals in each group. Groups A and B were vaccinated with a dose of 2×106 TCID50 of rAdV-FL-E2, and rAdV-E2 by intramuscular (i.m.) inoculation respectively. All pigs were given a booster immunization at 14 d intervals. Group C was immunized with one-dose CSFV C-strain vaccine and served as the positive control. Group D was injected with DMEM medium. Finally, all animals were challenged with 1×103 TCID50 of CSFV Shimen strain by i.m. injection at 21 d after the second vaccination. CSFV-specific neutralizing antibodies in sera were tested by using the IDEXX HerdChek* CSFV Antibody Test Kit and NPLA.[…]

Effect of ischemia preconditioning on renal ischemia/reperfusion injury in rats

To study the effect of ischemia preconditioning (IP) on renal ischemia/reperfusion (I/R)-associated functional injury and expression of renal adhesion molecules in rats.

The ischemia preconditioning plan adopted in this experiment involved renal warm ischemia for 6 min. and blood flow for 4 min., repeated four times. The Wistar rat kidneys used for warm ischemia preconditioning were subjected to 60 min of renal warm ischemia followed by reperfusion. The rat kidneys with ischemia/reperfusion were compared with the ischemia preconditioning group to observe rat renal function and changes in the expression of renal adhesion molecules ICAM-1, P--Selectin, and E-Selectin.

The expression of rat renal adhesion molecules (ICAM-1, P-Selectin, and E-Selectin) with ischemia preconditioning was significantly lower than that of the ischemia/reperfusion group. Serum creatinine was significantly lower than that in the ischemia/reperfusion group after 48 hours.

Ischemia preconditioning has a protective effect on renal function. Reduced expression of renal adhesion molecules is likely a mechanism involved in the observed protection. .

Effect of ischemia preconditioning on renal ischemia/reperfusion injury in rats.

OBJECTIVE: To study the effect of ischemia preconditioning (IP) on renal ischemia/reperfusion (I/R)-associated functional injury and expression of renal adhesion molecules in rats. MATERIALS AND METHODS: The ischemia preconditioning plan adopted in this experiment involved renal warm ischemia for 6 min. and blood flow for 4 min., repeated four times. The Wistar rat kidneys used for warm ischemia preconditioning were subjected to 60 min of renal warm ischemia followed by reperfusion. The rat kidneys with ischemia/reperfusion were compared with the ischemia preconditioning group to observe rat renal function and changes in the expression of renal adhesion molecules ICAM-1, P-Selectin, and E-Selectin. RESULTS: The expression of rat renal adhesion molecules (ICAM-1, P-Selectin, and E-Selectin) with ischemia preconditioning was significantly lower than that of the ischemia/reperfusion group. Serum creatinine was significantly lower than that in the ischemia/reperfusion group after 48 hours. CONCLUSIONS: Ischemia preconditioning has a protective effect on renal function. Reduced expression of renal adhesion molecules is likely a mechanism involved in the observed protection.