Skip to main page content
U.S. flag

An official website of the United States government

Dot gov

The .gov means it’s official.
Federal government websites often end in .gov or .mil. Before sharing sensitive information, make sure you’re on a federal government site.

Https

The site is secure.
The https:// ensures that you are connecting to the official website and that any information you provide is encrypted and transmitted securely.

Access keys NCBI Homepage MyNCBI Homepage Main Content Main Navigation
. 2020 Aug 19;6(8):e04661.
doi: 10.1016/j.heliyon.2020.e04661. eCollection 2020 Aug.

Elevated levels of autoantibodies against DNAJC2 in sera of patients with atherosclerotic diseases

Yoichi Yoshida  1   2   3 , Xiao-Meng Zhang  2 , Hao Wang  2   4 , Toshio Machida  5   6 , Seiichiro Mine  1   5   7 , Eiichi Kobayashi  1   3 , Akihiko Adachi  1   3 , Tomoo Matsutani  1   3 , Ikuo Kamitsukasa  8   9 , Takeshi Wada  10 , Akiyo Aotsuka  10 , Katsuro Iwase  2 , Go Tomiyoshi  2   11 , Rika Nakamura  2   11 , Natsuko Shinmen  2   11 , Hideyuki Kuroda  11 , Hirotaka Takizawa  12 , Koichi Kashiwado  13 , Hideo Shin  14 , Yuichi Akaogi  15 , Junichiro Shimada  15 , Eiichiro Nishi  16   17 , Mikiko Ohno  16   17 , Minoru Takemoto  18   19 , Koutaro Yokote  18 , Kenichiro Kitamura  20 , Yasuo Iwadate  1   3 , Takaki Hiwasa  1   2   3
Affiliations

Elevated levels of autoantibodies against DNAJC2 in sera of patients with atherosclerotic diseases

Yoichi Yoshida et al. Heliyon. .

Abstract

Background: Serum antibody markers have been increasingly identified not only for cancer and autoimmune diseases but also for atherosclerosis-related diseases such as acute ischemic stroke (AIS), acute myocardial infarction (AMI), diabetes mellitus (DM), and chronic kidney disease (CKD). Biomarkers for transient ischemic attack (TIA) and non-ST segment elevation acute coronary syndrome (NSTEACS) are potentially useful for detection of early phase of atherosclerotic changes against AIS and AMI, respectively.

Methods: We utilized serological identification of antigens by recombinant cDNA expression cloning (SEREX) using a human aortic endothelial cell cDNA phage library and sera from patients with TIA or NSTEACS. Serum antibody levels were measured by amplified luminescent proximity homogeneous assay-linked immunosorbent assay (AlphaLISA) using purified recombinant antigens.

Results: Screening of sera from patients with TIA identified DnaJ heat shock protein family (Hsp40) member C2 (DNAJC2) as a candidate antigen, which was also isolated by SEREX screening using sera of patients with NSTEACS. The validation cohort revealed significantly higher DNAJC2 antibody (DNAJC2-Ab) levels in the sera of patients with TIA or AIS than those in healthy donors (HDs). Multivariate logistic regression analysis indicated that the predictive odds ratios (OR) of DNAJC2-Ab levels for TIA and AIS were 2.54 (95% confidence interval [CI]: 1.36-4.74, p = 0.0034) and 2.14 (95% CI: 1.39-3.30, p = 0.0005), respectively. Serum DNAJC2-Ab levels were also higher in patients with AMI, DM, and CKD than those in HDs.

Conclusion: Serum DNAJC2-Ab level may be useful for early detection of atherosclerotic lesions, which lead to AIS and AMI.

Keywords: Acute ischemic stroke; Acute myocardial infarction; Atherosclerosis; Autoantibody biomarker; Biomarkers; Cardiology; Cardiovascular system; Clinical research; DNAJC2; Diagnostics; Hematological system; Neurology; Neurosurgery.

PubMed Disclaimer

Figures

Figure 1
Figure 1
Western blot analysis. Glutathione-S-transferase (GST) (lane 1) and affinity-purified GST-tagged DnaJ heat shock protein family (Hsp40) member C2 (DNAJC2) (lane 2) proteins were separated on sodium dodecyl sulfate-polyacrylamide gels and blotted using an anti-GST antibody (a); the sera of healthy donors (HDs) (b); or the sera of patients with transient ischemic attack (TIA) (c, d), acute ischemic stroke (AIS) (e), non-ST segment elevation acute coronary syndrome (NSTEACS). (f), and acute myocardial infarction (AMI) (g). Arrows at 58 kDa and 28 kDa indicate GST-DNAJC2 and GST proteins, respectively. Asterisks indicate partially degraded proteins. Molecular weights are shown on the left. The full, non-adjusted image of Figure 1 is shown in the supplementary figure S1.
Figure 2
Figure 2
Serum levels of DNAJC2 antibodies (DNAJC2-Abs) in stroke patients examined by amplified luminescent proximity homogeneous assay-linked immunosorbent assay (AlphaLISA) in the validation cohort. The DNAJC2-Ab levels measured as Alpha photon counts were compared between the HDs and the patients with TIA, AIS, or chronic-phase cerebral infarction (cCI) in box-whisker plots displaying the 10th, 20th, 50th, 80th, and 90th percentiles (a). ∗∗∗p < 0.001 by the Mann–Whitney U test with type I error adjustment using the Bonferroni procedure, not significant (n.s.), p = 1 by the Kruskal–Wallis test with type I error adjustment using the Bonferroni procedure. Receiver operating characteristic curve (ROC) analysis was performed to assess the ability of DNAJC2-Abs to detect TIA (b), AIS (c), and chronic cerebral infarction (cCI) (d). Table 3 summarizes areas under the curves (AUCs), 95% confidence intervals (CIs), cutoff values, sensitivity, specificity, and p values calculated using ROC analysis.
Figure 3
Figure 3
Association between DNAJC2-Ab levels and other clinical parameters in stroke patients. Correlations between DNAJC2-Ab levels and age (a), sex (b), hypertension (c), diabetes mellitus (DM) (d), hyperlipidemia (e), cardiovascular disease (CVD) (f), obesity (body mass index ≥25) (g), and smoking (h) were examined using Spearman's correlation analysis (a) or the Mann–Whitney U test (b–h).
Figure 4
Figure 4
Association between DNAJC2-Ab levels and other atherosclerotic diseases including AMI and DM. (a) Serum DNAJC2-Ab levels of the HDs and the patients with AMI or DM determined by AlphaLISA are shown as box-whisker plots, as described in the legend of Figure 2. The mean ages (±standard deviation) of the HDs and the patients with AMI and DM were 58.29 ± 5.63, 58.28 ± 8.5, and 58.37 ± 9.11 years, respectively. Results of the ROC analysis of DNAJC2-Abs to detect AMI (b), and DM (c) are also shown. ∗∗∗p < 0.001 by the Mann–Whitney U test with type I error adjustment using the Bonferroni procedure.
Figure 5
Figure 5
Association between DNAJC2-Ab levels and chronic kidney disease (CKD). (a) Serum DNAJC2-Ab levels of the HDs and the patients with CKD are shown as box-whisker plots, as described in the legend of Figure 2. CKD patients were divided into three groups: type 1, diabetic kidney disease; type 2, nephrosclerosis; and type 3, glomerulonephritis. The mean age (±standard deviation) of the HDs and the patients with type 1, 2, and 3 CKD were 45.82 ± 11.66, 65.78 ± 10.28, 75.97 ± 9.94, and 66.05 ± 14.60 years, respectively. Results of the ROC analysis of DNAJC2-Abs to detect type 1, 2, and 3 CKD (b–d) are also shown. ∗∗∗p < 0.001 by the Mann–Whitney U test with type I error adjustment using the Bonferroni procedure.
See this image and copyright information in PMC

Similar articles

  • Association of serum levels of antibodies against ALDOA and FH4 with transient ischemic attack and cerebral infarction.
    Wang H, Lu H, Zhang XM, Goto KI, Kobayashi E, Yoshida Y, Adachi A, Matsutani T, Iwadate Y, Mine S, Machida T, Sata M, Yamagishi K, Iso H, Sawada N, Tsugane S, Kamitsukasa I, Wada T, Aotsuka A, Sugimoto K, Takizawa H, Kashiwado K, Shin H, Tomiyoshi G, Nakamura R, Shinmen N, Kuroda H, Xu A, Hiwasa T. Wang H, et al. BMC Neurol. 2021 Jul 9;21(1):274. doi: 10.1186/s12883-021-02301-w. BMC Neurol. 2021. PMID: 34243715 Free PMC article.
  • Elevation of autoantibody level against PDCD11 in patients with transient ischemic attack.
    Yoshida Y, Wang H, Hiwasa T, Machida T, Kobayashi E, Mine S, Tomiyoshi G, Nakamura R, Shinmen N, Kuroda H, Takizawa H, Kashiwado K, Kamitsukasa I, Shin H, Wada T, Aotsuka A, Nishi E, Ohno M, Takemoto M, Yokote K, Takahashi S, Matsushima J, Zhang XM, Takiguchi M, Iwadate Y. Yoshida Y, et al. Oncotarget. 2017 Dec 24;9(10):8836-8848. doi: 10.18632/oncotarget.23653. eCollection 2018 Feb 6. Oncotarget. 2017. PMID: 29507658 Free PMC article.
  • Serum anti-DIDO1, anti-CPSF2, and anti-FOXJ2 antibodies as predictive risk markers for acute ischemic stroke.
    Hiwasa T, Wang H, Goto KI, Mine S, Machida T, Kobayashi E, Yoshida Y, Adachi A, Matsutani T, Sata M, Yamagishi K, Iso H, Sawada N, Tsugane S, Kunimatsu M, Kamitsukasa I, Mori M, Sugimoto K, Uzawa A, Muto M, Kuwabara S, Kobayashi Y, Ohno M, Nishi E, Hattori A, Yamamoto M, Maezawa Y, Kobayashi K, Ishibashi R, Takemoto M, Yokote K, Takizawa H, Kishimoto T, Matsushita K, Kobayashi S, Nomura F, Arasawa T, Kagaya A, Maruyama T, Matsubara H, Tomiita M, Hamanaka S, Imai Y, Nakagawa T, Kato N, Terada J, Matsumura T, Katsumata Y, Naito A, Tanabe N, Sakao S, Tatsumi K, Ito M, Shiratori F, Sumazaki M, Yajima S, Shimada H, Shirouzu M, Yokoyama S, Kudo T, Doi H, Iwase K, Ashino H, Li SY, Kubota M, Tomiyoshi G, Shinmen N, Nakamura R, Kuroda H, Iwadate Y. Hiwasa T, et al. BMC Med. 2021 Jun 9;19(1):131. doi: 10.1186/s12916-021-02001-9. BMC Med. 2021. PMID: 34103026 Free PMC article.
  • Elevation of Autoantibody in Patients with Ischemic Stroke.
    Yoshida Y, Hiwasa T, Machida T, Kobayashi E, Mine S, Matsushima J, Takiguchi M, Iwadate Y. Yoshida Y, et al. Neurol Med Chir (Tokyo). 2018 Jul 15;58(7):303-310. doi: 10.2176/nmc.ra.2018-0022. Epub 2018 May 31. Neurol Med Chir (Tokyo). 2018. PMID: 29848906 Free PMC article. Review.
  • The concept of acute cerebrovascular syndrome.
    Uchiyama S. Uchiyama S. Front Neurol Neurosci. 2014;33:11-8. doi: 10.1159/000351888. Epub 2013 Oct 11. Front Neurol Neurosci. 2014. PMID: 24157553 Review.
See all similar articles

Cited by

See all "Cited by" articles

References

    1. Goldstein L.B., Bushnell C.D., Adams R.J., Appel L.J., Braun L.T., Chaturvedi S. Guideline for the primary prevention of stroke: a guideline for healthcare professionals from the American Heart Association/American Stroke Association. Stroke. 2011;42:517–584. PMID: 21127304. - PubMed
    1. Carandang R., Seshadri S., Beiser A., Kelly-Hayes M., Kase C.S., Kannel W.B. Trends in incidence, lifetime risk, severity, and 30-day mortality of stroke over the past 50 years. J. Am. Med. Assoc. 2006;296:2939–2946. PMID: 17190894. - PubMed
    1. Kubo M., Hata J., Doi Y., Tanizaki Y., Iida M., Kiyohara Y. Secular trends in the incidence of and risk factors for ischemic stroke and its subtypes in Japanese population. Circulation. 2008;118:2672–2678. PMID: 19106389. - PubMed
    1. Libby P., Ridker P.M., Hansson G.K. Progress and challenges in translating the biology of atherosclerosis. Nature. 2011;473:317–325. PMID: 21593864. - PubMed
    1. Rollins K.E., Varadhan K.K., Dhatariya K., Lobo D.N. Systematic review of the impact of HbA1c on outcomes following surgery in patients with diabetes mellitus. Clin. Nutr. 2015;35:308–316. PMID: 25840840. - PubMed

LinkOut - more resources