Immune Netw.  2017 Apr;17(2):103-109. 10.4110/in.2017.17.2.103.

Detection of Autoantibodies against Aquaporin-1 in the Sera of Patients with Primary Sjögren's Syndrome

• Alam J
• Choi YS
• Koh JH
• Kwok SK
• Park SH
• Song YW
• Park K
• Choi Y

• KMID: 2376867
• DOI: http://doi.org/10.4110/in.2017.17.2.103

Abstract

The pathophysiology of glandular dysfunction in Sjögren's syndrome (SS) has not been fully elucidated. Previously, we reported the presence of autoantibodies to AQP-5 in patients with SS, which was associated with a low resting salivary flow. The purpose of this study was to investigate the presence of anti-AQP1 autoantibodies. To detect anti-AQP1 autoantibodies, cell-based indirect immunofluorescence assay was developed using MDCK cells that overexpressed human AQP1. By screening 112 SS and 52 control sera, anti-AQP1 autoantibodies were detected in 27.7% of the SS but in none of the control sera. Interestingly, the sera that were positive for anti-AQP1 autoantibodies also contained anti-AQP5 autoantibodies in the previous study. Different from anti-AQP5 autoantibodies, the presence of anti-AQP1 autoantibodies was not associated with the salivary flow rate. Although anti-AQP1 autoantibodies are not useful as a diagnostic marker, the presence of autoantibodies to AQP1 may be an obstacle to AQP1 gene therapy for SS.

Keyword

Sjögren's syndrome; Aquaporin 1; Autoantibodies; Fluorescent antibody technique

MeSH Terms

Aquaporin 1
Autoantibodies*
Fluorescent Antibody Technique
Fluorescent Antibody Technique, Indirect
Genetic Therapy
Humans
Madin Darby Canine Kidney Cells
Mass Screening
Aquaporin 1
Autoantibodies

Figure

• 

  Figure 1 Detection of anti-AQP1 IgG and IgA in the sera of SS patients. (A) MDCK cells over-expressing AQP1 were stained with anti-AQP1 antibodies and either control or SS sera (1:200 dilution), followed by Alexa Fluor 488-conjugated anti-goat IgG (green) and CF™ 594-conjugated anti-human IgG (red). The colocalization of green and red signals in the three images was calculated using Mander's coefficient. *p=0.0001 (B) The intensities of the red signals for anti-AQP1 IgG were expressed by the magnitude of brightness that was reduced until the AQP1 staining disappeared. (C) MDCK cells over-expressing AQP1 were stained with anti-AQP1 antibodies and either control or SS sera (1:20 dilution), followed by Alexa Fluor 488-conjugated anti-goat IgG (green) and Alexa Fluor 555-conjugated anti-human IgA (red). The colocalization of green and red signals in the three images was calculated using Mander's coefficient. *p=0.02 (D) The intensities of the red signals for anti-AQP5 IgA were expressed by the magnitude of brightness that was reduced until the AQP1 staining disappeared. (E) The intensities of anti-AQP1 IgG or IgA and of anti-AQP5 IgG or IgA that were identified in each individual were plotted.

• 

  Figure 2 Sequence alignment of AQP5 and AQP1. Identical and conserved amino acids are highlighted with dark gray and light gray, respectively. The extracellular and cytosolic domains are boxed with solid and dotted lines, respectively.

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  Figure 3 Sequence alignment of AQP1 and bacterial AQPs. Identical and conserved amino acids are highlighted with dark gray and light gray, respectively. The extracellular and cytosolic domains are boxed with solid and dotted lines, respectively.

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