Elevated Adiponectin Antibody Levels in Sera of Patients with Atherosclerosis-Related Coronary Artery Disease, Cerebral Infarction and Diabetes Mellitus

Abstract

Adiponectin secreted from the adipocytes plays pleiotropic, anti-atherosclerotic roles, such as enhancement of insulin secretion and an increase in energy expenditure. The measurement of levels of circulating adiponectin is useful to evaluate the progression of atherosclerosis-related diseases, such as coronary artery disease (CAD), cerebral infarction (CI) and diabetes mellitus (DM). We examined the serum antibody levels against recombinant adiponectin protein via the amplified luminescent proximity homogeneous assay-linked immunosorbent assay (AlphaLISA) method. The results revealed that the antibody levels were significantly higher in patients with CAD, CI and type 2 DM, than in healthy donors. Receiver operating curve analysis showed that the sensitivity was in a range of 41–48% for CAD, CI and DM. Thus, the serum anti-adiponectin antibody levels could be a common marker for atherosclerosis-related diseases.

Serum SH3BP5-specific Antibody Level is a Biomarker of Atherosclerosis

“…We next examined the levels of antibodies against the GST-fusion SH3BP5 protein and bSH3BP5-161 peptide in sera of HDs and patients with DM, using the highly sensitive and stable AlphaLISA method [26][27][28][29]. Serum samples of HDs and patients with DM were obtained from Chiba University Hospital (Figures 2-5).…”

“…The reaction mixture was incubated at room temperature for 6-8 h, following which anti-human IgG-conjugated acceptor beads (2.5 µL at 40 µg/mL) and glutathione-or streptavidin-conjugated donor beads (2.5 µL at 40 µg/ mL) were added and incubated prior to another incubation at room temperature in the dark for 1-14 days. Chemical emissions were read on an EnSpire Alpha microplate reader (PerkinElmer) as described previously [26][27][28][29]. Specific reactions were calculated by subtracting the alpha counts of the GST control and of the buffer control without antigenic peptides, from the counts of GST-fusion proteins and biotinylated peptides, respectively.…”

“…Similarly, our group has searched for antibody markers using the serological identification of antigens by recombinant cDNA expression cloning (SEREX) method, and has previously reported the discovery of antibodies against Trop2, TRIM21, makorin 1, and ECSA for esophageal squamous cell carcinoma (SCC) [18][19][20][21]; SH3GL1 and filamin C for glioma [22,23]; talin-1 for multiple sclerosis [24]; RPA2 and SOSTDC1 for ischemic stroke [25,26]; ATP2B4 and BMP-1 for atherosclerosis [27]; and TUBB2C and adiponectin for DM [28,29]. Here, we describe our recent discovery of a SH3 domain-binding protein 5 (SH3BP5)-specific antibody as a common marker of atherosclerosis-related diseases.…”

Identification of Serum Anti-GADD34 Antibody as a Common Marker of Diabetes Mellitus and Parkinson Disease

“…We examined the levels of antibodies against GADD34 protein in the sera of HDs and patients with DM or CVD, using the highly sensitive and stable AlphaLISA method [19][20][21][22][23][24]. Levels of serum antibodies against GADD34 protein (s-GADD34-Abs) were significantly higher in samples from patients with DM than in those from HDs (P<0.0001; Mann-Whitney U test) (Figure 1).…”

“…The reaction mixture was incubated at room temperature for 7-10 h. Anti-human IgG-conjugated acceptor beads (2.5 µL at 40 µg/mL) and glutathione-conjugated donor beads (2.5 µL at 40 µg/mL) were then added and incubated at room temperature in the dark for 7 days. Chemical emissions were read on an EnSpire Alpha microplate reader (PerkinElmer), as previously described [19][20][21][22][23][24]. Specific reactions were calculated by subtracting the Alpha counts of the GST control from the counts of GST-GADD34 fusion proteins.…”