Lab Med Online.  2015 Jan;5(1):20-26. 10.3343/lmo.2015.5.1.20.

Evaluation of CD3+CD4-CD8- (Double-negative) T Cells in Bronchoalveolar Lavage Fluid: an Effective Tool for Pulmonary Disease Diagnosis

Affiliations
  • 1Department of Laboratory Medicine, Catholic University of Daegu, School of Medicine, Daegu, Korea. sgkim@cu.ac.kr
  • 2Department of Internal Medicine, Catholic University of Daegu, School of Medicine, Daegu, Korea.

Abstract

BACKGROUND
Cellular analysis of bronchoalveolar lavage fluid (BALF) is a useful diagnostic tool for interstitial lung diseases (ILDs). The lymphocytes in BALF consist of CD3+CD4+ T cells (T4), CD3+CD8+ T cells (T8), and a few B cells. However, sometimes, an increased number of CD3+CD4-CD8- T cells (double-negative T cells, DNTs) are noted in BALF. It is known that DNTs in the blood are associated with immunoregulation and autoimmune diseases. However, there are only few studies on DNTs in BALF. We evaluated the DNTs in BALF in patients with pulmonary diseases.
METHODS
Immunophenotyping results of the BALF obtained from 122 pulmonary disease patients over an 8-yr period were reviewed. T-lymphocyte subsets (T4, T8, and DNT) and inflammatory markers were analyzed for each group of clinical diagnosis. T-lymphocyte percentage of more than 15% of the total cells was defined as BALF lymphocytosis, and DNT percentage of more than 5% of T lymphocytes was defined as high DNT.
RESULTS
The most frequent diseases found in the patients were pneumonia (31.6%), autoimmune-related ILDs (18.0%), hypersensitivity pneumonitis (10.7%), and organizing pneumonia (10.7%). However, the occurrence of autoimmune-related ILDs was significantly high (40%) in patients with lymphocytosis and high DNT (P=0.002). All lung cancer patients showed lymphocytosis with high DNT. In addition, CD3-signal intensities of DNTs were significantly higher than those of other T-lymphocyte subtypes (P=0.003).
CONCLUSION
The number of DNTs in BALF was increased in patients with autoimmune-related ILDs and lung cancer. High DNTs in BALF are useful as supportive diagnostic tools for autoimmune-related ILDs.

Keyword

Bronchoalveolar lavage fluid; T-lymphocyte subsets; Pulmonary diseases; Autoimmune interstitial lung diseases

MeSH Terms

Alveolitis, Extrinsic Allergic
Autoimmune Diseases
B-Lymphocytes
Bronchoalveolar Lavage Fluid*
Diagnosis*
Humans
Immunophenotyping
Lung Diseases*
Lung Diseases, Interstitial
Lung Neoplasms
Lymphocytes
Lymphocytosis
Pneumonia
T-Lymphocyte Subsets
T-Lymphocytes*

Figure

  • Fig. 1 Representation of nucleated cells in cytocentrifuged bronchoalveolar lavage fluid (Wright stain, ×600). (A) Lymphocytes (black arrow) and alveolar macrophages (white arrow); (B) Neutrophils (black arrow); (C) Eosinophils (black arrow); (D) Ciliated epithelial cells (black arrow).

  • Fig. 2 CD3 mean fluorescence intensity of double-negative T (DNT, green) was higher than that of T8 (blue, 10.54 vs. 8.03, P=0.001) and T4 subtypes (purple, 10.54 vs. 8.19, P=0.003).


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