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. 1999 Jul;6(4):471-8.

Cytokine mRNA Expression in Lesions in Cats With Chronic Gingivostomatitis

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Free PMC article

Cytokine mRNA Expression in Lesions in Cats With Chronic Gingivostomatitis

R Harley et al. Clin Diagn Lab Immunol. .
Free PMC article

Abstract

Semiquantitative reverse transcription-PCR assays were developed to measure feline interleukin-2 (IL-2), IL-4, IL-5, IL-6, IL-10, and IL-12 (p35 & p40); gamma interferon (IFN-gamma); and glyceraldehyde-3-phosphate dehydrogenase mRNA concentrations in biopsies of feline oral mucosa. Biopsies were collected from 30 cats with chronic gingivostomatitis (diseased) prior to each cat receiving one of four treatments. In 23 cases replicate biopsies were collected 3 months after treatment commenced. Biopsies were also analyzed from 11 cats without clinical disease (nondiseased). Expression of IL-2, IL-10, IL-12 (p35 and p40), and IFN-gamma was detected in most nondiseased biopsies, while IL-6 was detected in a minority, and IL-4 and IL-5 were both undetectable. Compared to nondiseased cats, the diseased population showed a significant increase in the relative mRNA expression of IL-2, IL-4, IL-6, IL-10, IL-12 (p35 and p40), and IFN-gamma. In contrast, IL-5 mRNA expression was unchanged and was only detected in one case. No significant relationship was demonstrable between the change in relative expression of specific cytokine mRNA and the change in clinical severity of the local mucosal lesions over the treatment period. The results demonstrate that the normal feline oral mucosa is biased towards a predominantly (Th) type 1 profile of cytokine expression and that during the development of lesions seen in feline chronic gingivostomatitis there is a shift in the cytokine profile from a type 1 to a mixed type 1 and type 2 response.

Figures

FIG. 1
FIG. 1
PCR products for G3PDH (400 bp) and IFN-γ (342 bp). Reaction mixes (25 μl) containing 2 μl cDNA sample or 10.5 μl of G3PDH (A) or IFN-γ (B) standard cDNA solution were amplified simultaneously for 35 cycles as described in the text. Then, 15 μl of each reaction product was separated by electrophoresis in a 1% agarose gel containing ∼1 μg of ethidium bromide per ml. Lanes 1, 14, 15, and 28 (a and b), 250-bp ladder molecular weight marker (MW); lanes 2 to 13: G3PDH (A) and IFN-γ (B) PCR products from four aliquots (1 to 4) of a feline tonsil homogenate, each performed in triplicate; lanes 16 to 24, G3PDH (A) and IFN-γ (B) PCR products from sequential 1-in-4 dilutions of the G3PDH or IFN-γ standard solution (106 U/10.5 μl); lane 26 (A and B), negative control (Neg) containing purified water in place of cDNA template solution; lanes 16 and 27 (A and B), empty.
FIG. 2
FIG. 2
Clinical score and cytokine/G3PDH mRNA expression ratios in faucal biopsies from nondiseased (○) and diseased (●) cats. The biopsies from diseased cats were taken after the withdrawal of previous treatment for a minimum time period of 2 to 6 weeks and immediately prior to their entry into the treatment trial (see the text for details). None of the nondiseased cats were receiving medication prior to biopsy collection. Panels: a, IFN-γ; b, IL-2; c, IL-12 p35; d, IL-12 p40; e, IL-4; f, IL-5; g, IL-6; h, IL-10. The median value of each clinically graded group is represented by an adjacent bar (–). The median value of the entire diseased population is represented by a triangle (▵).
FIG. 2
FIG. 2
Clinical score and cytokine/G3PDH mRNA expression ratios in faucal biopsies from nondiseased (○) and diseased (●) cats. The biopsies from diseased cats were taken after the withdrawal of previous treatment for a minimum time period of 2 to 6 weeks and immediately prior to their entry into the treatment trial (see the text for details). None of the nondiseased cats were receiving medication prior to biopsy collection. Panels: a, IFN-γ; b, IL-2; c, IL-12 p35; d, IL-12 p40; e, IL-4; f, IL-5; g, IL-6; h, IL-10. The median value of each clinically graded group is represented by an adjacent bar (–). The median value of the entire diseased population is represented by a triangle (▵).
FIG. 2
FIG. 2
Clinical score and cytokine/G3PDH mRNA expression ratios in faucal biopsies from nondiseased (○) and diseased (●) cats. The biopsies from diseased cats were taken after the withdrawal of previous treatment for a minimum time period of 2 to 6 weeks and immediately prior to their entry into the treatment trial (see the text for details). None of the nondiseased cats were receiving medication prior to biopsy collection. Panels: a, IFN-γ; b, IL-2; c, IL-12 p35; d, IL-12 p40; e, IL-4; f, IL-5; g, IL-6; h, IL-10. The median value of each clinically graded group is represented by an adjacent bar (–). The median value of the entire diseased population is represented by a triangle (▵).
FIG. 2
FIG. 2
Clinical score and cytokine/G3PDH mRNA expression ratios in faucal biopsies from nondiseased (○) and diseased (●) cats. The biopsies from diseased cats were taken after the withdrawal of previous treatment for a minimum time period of 2 to 6 weeks and immediately prior to their entry into the treatment trial (see the text for details). None of the nondiseased cats were receiving medication prior to biopsy collection. Panels: a, IFN-γ; b, IL-2; c, IL-12 p35; d, IL-12 p40; e, IL-4; f, IL-5; g, IL-6; h, IL-10. The median value of each clinically graded group is represented by an adjacent bar (–). The median value of the entire diseased population is represented by a triangle (▵).
FIG. 2
FIG. 2
Clinical score and cytokine/G3PDH mRNA expression ratios in faucal biopsies from nondiseased (○) and diseased (●) cats. The biopsies from diseased cats were taken after the withdrawal of previous treatment for a minimum time period of 2 to 6 weeks and immediately prior to their entry into the treatment trial (see the text for details). None of the nondiseased cats were receiving medication prior to biopsy collection. Panels: a, IFN-γ; b, IL-2; c, IL-12 p35; d, IL-12 p40; e, IL-4; f, IL-5; g, IL-6; h, IL-10. The median value of each clinically graded group is represented by an adjacent bar (–). The median value of the entire diseased population is represented by a triangle (▵).
FIG. 2
FIG. 2
Clinical score and cytokine/G3PDH mRNA expression ratios in faucal biopsies from nondiseased (○) and diseased (●) cats. The biopsies from diseased cats were taken after the withdrawal of previous treatment for a minimum time period of 2 to 6 weeks and immediately prior to their entry into the treatment trial (see the text for details). None of the nondiseased cats were receiving medication prior to biopsy collection. Panels: a, IFN-γ; b, IL-2; c, IL-12 p35; d, IL-12 p40; e, IL-4; f, IL-5; g, IL-6; h, IL-10. The median value of each clinically graded group is represented by an adjacent bar (–). The median value of the entire diseased population is represented by a triangle (▵).
FIG. 2
FIG. 2
Clinical score and cytokine/G3PDH mRNA expression ratios in faucal biopsies from nondiseased (○) and diseased (●) cats. The biopsies from diseased cats were taken after the withdrawal of previous treatment for a minimum time period of 2 to 6 weeks and immediately prior to their entry into the treatment trial (see the text for details). None of the nondiseased cats were receiving medication prior to biopsy collection. Panels: a, IFN-γ; b, IL-2; c, IL-12 p35; d, IL-12 p40; e, IL-4; f, IL-5; g, IL-6; h, IL-10. The median value of each clinically graded group is represented by an adjacent bar (–). The median value of the entire diseased population is represented by a triangle (▵).
FIG. 2
FIG. 2
Clinical score and cytokine/G3PDH mRNA expression ratios in faucal biopsies from nondiseased (○) and diseased (●) cats. The biopsies from diseased cats were taken after the withdrawal of previous treatment for a minimum time period of 2 to 6 weeks and immediately prior to their entry into the treatment trial (see the text for details). None of the nondiseased cats were receiving medication prior to biopsy collection. Panels: a, IFN-γ; b, IL-2; c, IL-12 p35; d, IL-12 p40; e, IL-4; f, IL-5; g, IL-6; h, IL-10. The median value of each clinically graded group is represented by an adjacent bar (–). The median value of the entire diseased population is represented by a triangle (▵).

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