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Published on: April 19, 2017
[Establishment of a simple fluorescent microscopy method for measurement of the CD4/CD8 ratio in BALF]
Y Konoike1, K Nishikawa, T Imai
1Second Department of Internal Medicine, Nara Medical University, Japan.
Insights
A new fluorescent microscopy method accurately measures the CD4/CD8 ratio in bronchoalveolar lavage fluid (BALF) for diagnosing diffuse pulmonary diseases. This simple technique aids in disease monitoring and diagnosis.
Area of Science:
- Pulmonary Medicine
- Immunology
- Microscopy Techniques
Background:
- The CD4/CD8 ratio in bronchoalveolar lavage fluid (BALF) is a key indicator in diffuse pulmonary diseases.
- Accurate measurement of BALF CD4/CD8 ratio is crucial for diagnosis and monitoring.
Purpose of the Study:
- To establish a simple and reliable fluorescent microscopy method for measuring the CD4/CD8 ratio in BALF.
- To evaluate the utility of this method in diagnosing and monitoring diffuse pulmonary diseases.
Main Methods:
- Development of a fluorescent microscopy technique to quantify CD4+ and CD8+ T lymphocytes in BALF.
- Correlation analysis of the novel method with flow cytometry for CD4/CD8 ratio determination.
- Measurement of BALF CD4/CD8 ratios in patients with hypersensitive pneumonitis, sarcoidosis, collagen vascular disease, idiopathic interstitial pneumonitis, and measles pneumonia.
Main Results:
- The fluorescent microscopy method showed strong correlation with flow cytometry for CD4/CD8 ratio in both peripheral blood and BALF.
- BALF CD4/CD8 ratios varied significantly among different diffuse pulmonary diseases (e.g., low in HP and MP, high in SAR).
- A decrease in BALF CD4/CD8 ratio in summer-type HP correlated with disease activity and improved upon environmental changes.
Conclusions:
- The developed fluorescent microscopy method is simple, accurate, and effective for measuring BALF CD4/CD8 ratios.
- This method is valuable for the diagnosis and monitoring of various diffuse pulmonary diseases.
- BALF CD4/CD8 ratio serves as a useful biomarker for assessing disease status and therapeutic response in pulmonary conditions.
Abstract:
The authors established an easy fluorescent microscopy method to measure the ratio of CD4 (helper/inducer T lymphocytes) to CD8 (suppressor/cytotoxic T lymphocytes) (CD4/CD8 ratio) in bronchoalveolar lavage fluid (BALF) in patients with diffuse pulmonary diseases and measles pneumonia. The CD4/CD8 ratio determined by this method strongly correlated with the CD4/CD8 ratio obtained by flow cytometry in both peripheral blood lymphocytes and BALF lymphocytes. When the proportion of lymphocytes in BALF nuclear cells is more than 6%, the CD4/CD8 ratio in BALF was measurable. The CD4/CD8 ratio in peripheral blood was not significantly different among hypersensitive pneumonitis (HP), sarcoidosis (SAR), collagen vascular disease (CVD), idiopathic interstitial pneumonitis (IIP), and measles pneumonia (MP). However, the BALF CD4/CD8 ratio was 0.61 +/- 0.37 in HP, 6.83 +/- 2.73 in SAR, 0.77 +/- 0.40 in CVD, 1.49 +/- 0.29 in IIP. These results were consistent with previously published data. The CD4/CD8 ratio in patients with MP was as low as 0.39 +/- 0.34. In patients with summer type HP, this ratio was low in the acute phase, but became within normal range after moving or reconstructing their homes, accompanied with the improvement of subjective symptoms and the disappearance of inflammation. These data indicate that our method is very simple and useful in the diagnosis and the monitoring of diffuse pulmonary diseases.
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