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

Hae-Bong Jang; A-Jin Lee; Min-Ji Kim; Chang-Ho Jeon; Hun-Suk Suh; Dae-Sung Hyun; Sang-Gyung Kim

doi:10.3343/lmo.2015.5.1.20

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Jang, Lee, Kim, Jeon, Suh, Hyun, and Kim: Evaluation of CD3+CD4-CD8- (Double-negative) T Cells in Bronchoalveolar Lavage Fluid: an Effective Tool for Pulmonary Disease Diagnosis

Original Article

Diagnostic Immunology

Laboratory Medicine Online 2015; 5(1): 20-26.

Published online: 1 January 2015

DOI: https://doi.org/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

Hae-Bong Jang, M.D.¹, A-Jin Lee, M.D.¹, Min-Ji Kim, M.D.¹, Chang-Ho Jeon, M.D.¹, Hun-Suk Suh, M.D.¹, Dae-Sung Hyun, M.D.², Sang-Gyung Kim, M.D.¹

¹Department of Laboratory Medicine, Catholic University of Daegu, School of Medicine, Daegu, Korea.

²Department of Internal Medicine, Catholic University of Daegu, School of Medicine, Daegu, Korea.

Corresponding author: Sang-Gyung Kim. Department of Laboratory Medicine, Daegu Catholic University, School of Medicine, 33 Duryugongwon-ro 17-gil, Nam-gu, Daegu 705-718, Korea. Tel: +82-53-650-4145; Fax: +82-53-653-8672, sgkim@cu.ac.kr

Received 24 January 2014 Revised 21 April 2014 Accepted 21 April 2014

(open-access):

This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/3.0/) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.

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.

Keywords: Bronchoalveolar lavage fluid, T-lymphocyte subsets, Pulmonary diseases, Autoimmune interstitial lung diseases

Figures and Tables

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).

Table 1

Diagnosis and the results of BALF immunophenotyping of total 122 patients

Diagnosis	Number of patients
Diagnosis	T cell>15% DNT≤5%	T cell>15% DNT>5%	T cell≤15% DNT≤5%	T cell≤15% DNT>5%	Total
Pneumonia	27	4	7	6	44
Autoimmune-related ILDs	8	12	2	0	22
Organizing pneumonia	10	3	0	0	13
EP	3	4	4	1	12
HP	10	3	0	0	13
Lung cancer	0	3	0	0	3
Tuberculosis	2	1	1	2	6
Sarcoidosis	5	0	0	0	5
Miscellaneous	3	0	1	0	4
Total	68	30	15	9	122

Abbreviations: BALF, bronchoalveolar lavage fluid; ILDs, interstitial lung diseases; DNT, double-negative T cell; EP, eosinophilic pneumonitis; HP, hypersensitivity pneumonitis.

Notes

This article is available from http://www.labmedonline.org

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