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Association of the HLA‑DRB1*04 allele and auto‑antibodies with disease severity in smokers and non‑smokers with rheumatoid arthritis

  • Authors:
    • Kawthar Eidan
    • Dhuha S. Saleh
  • View Affiliations / Copyright

    Affiliations: Department of Biology, College of Science, University of Baghdad, Baghdad 10015, Iraq
    Copyright: © Eidan et al. This is an open access article distributed under the terms of Creative Commons Attribution License [CC BY 4.0].
  • Article Number: 64
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    Published online on: May 18, 2026
       https://doi.org/10.3892/wasj.2026.479
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Abstract

The present study aimed to investigate the association between the HLA‑DRB1*04 allele and the severity of rheumatoid arthritis (RA), as well as the presence of autoantibodies, in patients who were smokers and non‑smokers. For this purpose, a case‑control study was conducted involving 100 patients with RA (50 smokers and 50 non‑smokers) and 50 healthy controls (25 smokers and 25 non‑smokers). HLA‑DRB1 gene expression was quantified using reverse transcription‑quantitative PCR and HLA‑DRB1*04 allele genotyping was performed with sequence‑specific primers. The levels of serum rheumatoid factor (RF) and anti‑cyclic citrullinated peptide (ACCP) antibodies were measured using ELISA. The Disease Activity Score‑28 (DAS‑28) was assessed by a specialist physician. Positive HLA‑DRB1 gene expression was detected in 64% of the patients and 16% of the controls. The HLA‑DRB1*04 allele was identified in 32% of the patients and 6% of the controls. Smokers carrying the HLA‑DRB1*04‑positive allele (n=19) exhibited higher HLA‑DRB1 gene expression levels (4.46±4.2‑fold), ACCP antibody levels (77.54±24.6 U/ml) and RF levels (138.48±140.1 U/ml) compared with HLA‑DRB1*04‑positive patients who were non‑smokers (n=13). DAS‑28 values were significantly higher in HLA‑DRB1*04‑positive smokers (4.52±0.9) than in HLA‑DRB1*04‑negative non‑smokers (3.8±1.02; P≤0.05). No significant difference was observed between HLA‑DRB1*04‑positive non‑smokers and HLA‑DRB1*04‑negative smokers. On the whole, the present study demonstrates that cigarette smoking has an adverse effect on the severity of RA, comparable to that of the HLA‑DRB1*04 allele alone. The interaction between smoking and HLA‑DRB1*04 positivity links to higher autoantibody levels and disease activity, highlighting the importance of both genetic and environmental risk factors in the pathogenesis of RA.
View Figures

Figure 1

Gel electrophoresis of PCR products on
a 2% agarose gel. The 434-bp band represents the control PCR
product, while the 259-bp band corresponds to the allele-specific
primer product. A DNA 100-bp ladder (lane L) was used for size
estimation.

Figure 2

HLA-DRB1*04 allele in patients who
were smokers and controls in relation to the mean levels of (A)
HLA-DRB1gene expression, (B) ACCP, and (C) RF. ACCP, anti-cyclic
citrullinated peptide; RF, rheumatoid factor; SE, shared epitope.
NS, no significant difference. **P<0.001, highly
significant difference (comparison between patients and controls
who were smokers).

Figure 3

HLA-DRB1*04 allele in patients who
were non-smokers and controls in relation to the mean levels of (A)
HLA-DRB1 gene expression, (B) ACCP, and (C) RF. ACCP, anti-cyclic
citrullinated peptide; RF, rheumatoid factor; SE, shared epitope.
**P<0.001, highly significant difference (comparison
between patients and controls who were non-smokers).

Figure 4

Positive HLA-DRB1*04 allele in
patients who were smokers and non-smokers in relation to the mean
levels ofHLA-DRB1 gene expression, ACCP and RF. ACCP, anti-cyclic
citrullinated peptide; RF, rheumatoid factor; SE, shared epitope.
*P<0.05, significant difference (comparison between
smokers and non-smokers who positive for the HLA-DRB1*04 allele in
the patient group).

Figure 5

Negative HLA-DRB1*04 allele in
patients who were smokers and non-smokers in relation to the mean
levels of HLA-DRB1 gene expression, ACCP, and RF. ACCP and RF.
ACCP, anti-cyclic citrullinated peptide; RF, rheumatoid factor; SE,
shared epitope. *P<0.05, significant difference
(comparison between smokers and non-smokers who were negative for
the HLA-DRB1*04 allele in the patient group).

Figure 6

Disease activity (DAS-28) in relation
to HLA-DRB1*04 (SE) and smoking. ACCP and RF. ACCP, anti-cyclic
citrullinated peptide; RF, rheumatoid factor; SE, shared epitope;
NS, not significant. *P<0.05, significant difference
between smokers who were SE-positive and non-smokers who were
SE-negative.
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Copy and paste a formatted citation
Spandidos Publications style
Eidan K and Saleh DS: Association of the HLA‑DRB1*04 allele and auto‑antibodies with disease severity in smokers and non‑smokers with rheumatoid arthritis. World Acad Sci J 8: 64, 2026.
APA
Eidan, K., & Saleh, D.S. (2026). Association of the HLA‑DRB1*04 allele and auto‑antibodies with disease severity in smokers and non‑smokers with rheumatoid arthritis. World Academy of Sciences Journal, 8, 64. https://doi.org/10.3892/wasj.2026.479
MLA
Eidan, K., Saleh, D. S."Association of the HLA‑DRB1*04 allele and auto‑antibodies with disease severity in smokers and non‑smokers with rheumatoid arthritis". World Academy of Sciences Journal 8.4 (2026): 64.
Chicago
Eidan, K., Saleh, D. S."Association of the HLA‑DRB1*04 allele and auto‑antibodies with disease severity in smokers and non‑smokers with rheumatoid arthritis". World Academy of Sciences Journal 8, no. 4 (2026): 64. https://doi.org/10.3892/wasj.2026.479
Copy and paste a formatted citation
x
Spandidos Publications style
Eidan K and Saleh DS: Association of the HLA‑DRB1*04 allele and auto‑antibodies with disease severity in smokers and non‑smokers with rheumatoid arthritis. World Acad Sci J 8: 64, 2026.
APA
Eidan, K., & Saleh, D.S. (2026). Association of the HLA‑DRB1*04 allele and auto‑antibodies with disease severity in smokers and non‑smokers with rheumatoid arthritis. World Academy of Sciences Journal, 8, 64. https://doi.org/10.3892/wasj.2026.479
MLA
Eidan, K., Saleh, D. S."Association of the HLA‑DRB1*04 allele and auto‑antibodies with disease severity in smokers and non‑smokers with rheumatoid arthritis". World Academy of Sciences Journal 8.4 (2026): 64.
Chicago
Eidan, K., Saleh, D. S."Association of the HLA‑DRB1*04 allele and auto‑antibodies with disease severity in smokers and non‑smokers with rheumatoid arthritis". World Academy of Sciences Journal 8, no. 4 (2026): 64. https://doi.org/10.3892/wasj.2026.479
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