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Journal of Dental Research, Vol. 81, No. 9,
633-636 (2002)
DOI: 10.1177/154405910208100911
HEMA Bound to Self-protein Promotes Auto-antibody Production in Mice
E. Sandberg1,*,
G. Bergenholtz1,
C. Eklund1 and
U.I. Dahlgren1,2
1 Department of Endodontology/Oral Diagnosis, Faculty of Odontology, Box 450, SE 405 30 Göteborg; and
2 Department of Rheumatology, Faculty of Medicine, The Sahlgrenska Academy at Göteborg University, Sweden;
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Figure 1. Number of 2-hydroxyethylmethacrylate (HEMA) molecules bound to bovine serum albumin (BSA) after incubation at a molecular ratio (HEMA to BSA) of 500:1. The number of conjugated HEMA molecules was calculated from the subsequent inhibition of trinitrophenyl binding to the BSA molecules. (A) Samples were incubated at 37°C, pH 7.4, for 45 hrs at increasing protein concentrations. Means and standard deviations are from 3 samples. (B) Influence of incubation time and pH at a BSA concentration of 40 mg/mL. Means and standard deviations of 2 samples.
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Figure 2. IgG anti-MSA (A) and anti H-MSA (B) antibody activity in serum of mice immunized with mouse serum albumin carrying HEMA (H-MSA) (n = 9), mouse serum albumin (MSA) (n = 9), or adjuvant (n = 8) only. The mice were immunized in Freund’s incomplete adjuvant 4x with intervals of 2 to 3 wks. Serum was taken 6 days after the last immunization. Each dot represents 1 mouse, and the median is indicated. Statistical comparison was done between the MSA- and H-MSA-treated groups. The IgG antibody activity was determined with ELISA and is expressed as absorbance units at 405 nm, serum dilution 1/100.
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Figure 3. The IgG1 anti-MSA antibody activity in relation to the corresponding IgG anti-MSA antibody activity in serum of mice immunized with mouse serum albumin carrying HEMA (H-MSA) (n = 9), mouse serum albumin (MSA) (n = 9), or adjuvant (n = 8) only. The mice were immunized 4x 2 to 3 wks apart, and the serum was taken 6 days after the last immunization. Each dot represents 1 mouse, and the median is indicated. Statistical comparison was done between the MSA- and H-MSA-treated groups.
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Figure 4. IL-2 secretion of spleen cell cultures stimulated with HEMA-conjugated mouse serum albumin (H-MSA). Mice were immunized subcutaneously with H-MSA (n = 8) without adjuvant. Controls (n = 8) were given phosphate-buffered saline only. The mice were immunized 3x and killed 7 days after the last immunization. Supernatants were analyzed for IL-2 after 4 days’ stimulation with H-MSA. Each dot represents 1 mouse, and the median is indicated.
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