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Renal tubular acidosis, serum anti- VH ~-ATP enzyme subunit antibodies B1/B2

Author: LiYanPing
Tutor: ZhaoJiaJun
School: Shandong University
Course: Internal Medicine
Keywords: Renal tubular acidosis Sjogren's syndrome v-H ~-ATP enzyme subunit B1 v-H ~-ATP enzyme subunit B2
CLC: R446.6
Type: Master's thesis
Year: 2008
Downloads: 29
Quote: 0
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Abstract

Objective immune factors in the pathogenesis of renal tubular acidosis, the role has attracted increasing attention. It has been reported in patients with renal tubular acidosis, renal tubular epithelial cells in the presence of anti-self antibodies, but these antibodies specific types not yet clear. This study was designed to detect renal tubular acidosis in the presence of serum anti-v-hydrogen ion adenosine triphosphatase (vH ~-ATP enzyme) B1 and B2 subunit subunit autoantibodies. Methods Study subjects: renal tubular acidosis in 11 patients, including 10 patients with Sjogren's syndrome: a simple 8 patients with Sjogren's syndrome; healthy controls eight cases. All patients were untreated new-onset or recurrence. Take the study of whole blood using flow technology for immunological tests, some serum biochemical indicators of detection, some indirect immunofluorescence assay for serum autoantibodies. First determine whether the presence of anti-serum antibodies tubular epithelial cells, as well as observe vH ~-ATP enzyme subunit B1 and B2 subunit expression in normal human kidney tissue characteristics. In normal kidney tissue paraffin sections, the increase in patients with renal tubular acidosis, serum dope, dope pure Sjogren's syndrome patients and healthy control sera liquid, anti-vH ~-ATP enzyme B1 subunit antibody or anti-vH ~-ATP enzyme B2 subunit antibody as an anti-incubated overnight at 4 ℃ to fluorescein isothiocyanate (Fluoresceinisothiocynate, FITC, green fluorescence) - goat anti-human immunoglobulin (Immunoglobulin, IgG) were incubated in serum markers of renal tubular epithelial cells themselves anti- antibodies, or tetramethyl rhodamine isothiocyanate (tetramethylrhodamineisothiocyanate, TRITC, red fluorescence) - rabbit anti-goat IgG incubated mark vH ~-ATP enzyme subunit B1 or B2 subunit, 4,6 - United amidine -2 - phenylindole (4,6-diamidino-2-phenyl-indolediacetare, DAPI, blue fluorescence) labeled nuclei and mounted. Positive results, further diluting the appropriate primary antibody, repeat the experiment until the result was negative. And for the following indirect immunofluorescence double labeling experiments to select the appropriate dilution ratio. For the detection of renal tubular acidosis in the presence of serum anti-vH ~-ATP enzyme subunit B1 and B2 subunit autoantibodies, first with the serum of patients with renal tubular acidosis, dope, dope pure Sjogren's syndrome or healthy control sera sera liquid paraffin sections with normal kidney tissue was incubated overnight at 4 ℃, then add different dilutions of goat anti-human vH ~-ATP enzyme B1 subunit antibody incubation overnight at 4 ℃, and then successively adding TRITC-rabbit anti-goat IgG and FITC-goat anti- human IgG fluorescent secondary antibody Incubate, DAPI labeled nuclei and mounted. Positive results, the serum was diluted in different proportions, repeat the experiment until the result is negative. Serum anti vH ~-ATP enzyme subunit B2 autoantibodies with anti vH ~-ATP enzyme subunit B1 autoantibodies. Located negative control for each experiment. General clinical outcome indicators findings: Patients with Sjogren's syndrome alone and health for photographic comparison of patients with renal tubular acidosis, serum chloride levels were significantly elevated serum carbon dioxide decreased, significantly increased urinary pH, flow studies, immunoglobulin and complement findings are suggestive of immunological changes. Serum anti-tubular epithelial cell autoantibodies Results: 11 cases of renal tubular acidosis patients, 6 patients with renal tubular epithelial cells showed bright green fluorescence, suggesting that anti-tubular epithelial cell antibodies; When autoantibodies gradual dilution of serum, its green fluorescence intensity was gradually weakened to varying degrees, suggesting that as the dilution of serum autoantibodies concentrations decreased; while 8 cases of pure Sjogren's syndrome patients and healthy controls eight cases of renal tubular epithelial cells is no obvious green fluorescence, suggesting that serum anti-renal tubular epithelial cell autoantibodies were negative. vH ~-ATP enzyme subunit B1 and B2 subunit expression in normal kidney tissue results: Mark vH ~-ATP enzyme subunit B1 red fluorescence was intercalated cells in the kidney specificity scattered, vH ~-ATP enzyme labeled B2 red fluorescent subunits in renal tubular epithelial cells along the lumen appear before a continuous linear distribution. Anti-vH ~-ATP enzyme subunit B1 and B2 subunit autoantibodies Results: 11 cases of renal tubular acidosis patients, 6 patients labeled vH ~-ATP enzyme B1 subunit of the red fluorescence intensity was significantly reduced, suggesting that sera for the anti-vH ~-ATP enzyme subunit B1 antibody blocking effect has obvious; when the serum autoantibodies gradually diluted vH ~-ATP enzyme labeled subunit B1 gradually increased intensity of red fluorescence, indicating with serum dilution, which for the anti-vH ~-ATP enzyme blocking effect B1 subunit antibodies showed different degrees of weakening, suggesting different dilutions of serum antibody titers varying intensity; while 8 cases of pure Sjogren's syndrome patients and 8 healthy controls The serum had no blocking effect. Anti-vH ~-ATP enzyme subunit B2 autoantibodies results with the anti-vH ~-ATP enzyme subunit B1 autoantibody test results. These results suggest that renal tubular acidosis, serum anti vH ~-ATP enzyme subunit B1 and B2 subunit autoantibodies. Conclusion Patients with renal tubular acidosis, serum anti tubular epithelial cell autoantibodies, the positive rate was 6/11; renal tubular acidosis presence of serum anti-vH ~-ATP enzyme subunit B1 and B2 subunit autoantibody positive rates were 6/11: renal tubular acidosis serum anti-vH ~-ATP enzyme subunit B1 and B2 subunit autoantibodies are anti-tubular epithelial cell autoantibodies in two.

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