Immune Netw.  2015 Dec;15(6):304-312. 10.4110/in.2015.15.6.304.

Anti-inflammatory Effect of Alloferon on Ovalbumin-induced Asthma

• Jeon J
• Kim Y
• Kim H
• Kang JS
• Lee WJ

• KMID: 2132704
• DOI: http://doi.org/10.4110/in.2015.15.6.304

Abstract

Asthma is a well-known inflammatory lung disease; however, the specific underlying mechanism is largely unknown. We previously demonstrated that alloferon effectively downregulates pulmonary inflammation. In this study, we examined whether alloferon has a therapeutic effect on asthma. Alloferon remarkably decreased the number of eosinophils, macrophages, and neutrophils in the bronchoalveolar lavage fluid (BALF) from ovalbumin (OVA)-induced asthma mice. It was synergistically decreased with 2.5 mg/kg prednisolone (PDA). Inflammatory cell infiltration around the bronchioles and in the alveolus of OVA-induced asthma mice was effectively prevented by alloferon alone and combined treatment with alloferon and PDS. The production of IL-5 and IL-17 was decreased by alloferon alone and combined treatment with alloferon and PDS. There was no change the level of total immunoglobulin (Ig) following alloferon administration; however, total Ig was decreased by PDS. IgG2a levels were not changed by either alloferon alone or alloferon in combination with PDS. However, the levels of OVA-specific IgG1 and IgE were decreased by alloferon and PDS. In conclusion, our results suggest that a combination of alloferon and prednisolone is effective for the treatment of asthma, as it prevents inflammatory cell infiltration via the down-regulation of IL-5 and IL-17 production and decreases IgG1 and IgE production via the suppression of T helper type 2 immune response.

Keyword

Alloferon; Asthma; Interleukin-17

MeSH Terms

Animals
Asthma*
Bronchioles
Bronchoalveolar Lavage Fluid
Down-Regulation
Eosinophils
Immunoglobulin E
Immunoglobulin G
Immunoglobulins
Interleukin-17
Interleukin-5
Lung Diseases
Macrophages
Mice
Neutrophils
Ovalbumin
Pneumonia
Prednisolone
Immunoglobulin E
Immunoglobulin G
Immunoglobulins
Interleukin-17
Interleukin-5
Ovalbumin
Prednisolone

Figure

• 

  Figure 1 Alloferon does not affect airway resistance in OVA-induced asthma mice. (A) Plethysmography of experimental mice (n=6 per group) was measured after inhalation of an aerosol containing methacholine (6.25, 12.5, 25, and 25 mg/mL). Breathing was recorded for 150 sec as described in the Materials and Methods. Data are presented as the mean±SD. (B) Weight of the mice was measured on days 1 and 26 to monitor for possible adverse effects of alloferon and prednisolone administration. Data are presented as the mean±SD.

• 

  Figure 2 Alloferon reduced eosinophil infiltration into the lungs of mice with OVA-induced asthma. Bronchoalveolar lavage fluid (BALF) was obtained, and the number of cells in BALF was counted to determine the effect of alloferon on inflammatory cell infiltration as described in the Materials and Methods. (A) The total number of cells in BALF was measured using a Trypan blue dye exclusion assay. Data are presented as the mean±SD. ***p< 0.0001 (B) The cell composition in BALF with and without alloferon treatment was determined after Wright's Giemsa staining. Cells were collected from BALF by centrifugation and resuspended in 300 µL of PBS for cytospin preparation. Then, the composition of the cell fraction was determined after Wright's Giemsa staining. Data are presented as the mean±SD. *p<0.05, **p<0.001, ***p<0.0001.

• 

  Figure 3 Alloferon prevented airway remodeling in OVA-induced asthma mice. Lung and tracheal tissues were stained with hematoxylin and eosin as described in the Materials and Methods. The stained tissues were examined under a light microscope at 200×magnification. (A) Without OVA, (B) With OVA, (C) OVA+Alloferon (2.5 mg/kg), (D) OVA+Prednisolone (2.5 mg/kg), (E) OVA+Prednisolone (5 mg/kg), (F) OVA+Alloferon (2.5 mg/kg)+Prednisolone (2.5 mg/kg), (G) OVA+Alloferon (2.5 mg/kg)+Prednisolone (5 mg/kg).

• 

  Figure 4 Alloferon suppressed IL-5 and IL-13 production OVA-induced asthma mice. BALF was obtained, centrifuged, incubated with fluorescence-tagged antibody-conjugated beads using the CBA assay kit, and analyzed by flow cytometry. Data are presented as the mean±SD. The concentration of Th1/Th2 cytokines in BALF was determined according to the manufacturer's protocol. *p<0.05, **p<0.001, ***p<0.0001.

• 

  Figure 5 Alloferon did not alter OVA-specific antibody production. Blood was collected from the intra-orbital plexus with a heparinized capillary. After centrifugation at 1,500 rpm for 5 min at 4℃, the serum was collected. The concentration of OVA-specific antibodies was measured by ELISA as described in the Materials and Methods. Titers are shown relative to that in standard serum. (A) Total Igs, (B) OVA-specific IgG1, (C) OVA-specific IgG2a, (D) OVA-specific IgE. **p<0.001, ***p<0.0001.

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