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[Use of flow cytometry in evaluation of cellular changes in interstitial lung diseases].
Folia Med Cracov. 1997; 38(3-4):69-115.FM

Abstract

Flow cytometry (FC) usefulness for pulmonary reactive cell changes examination in interstitial lung diseases was assessed. Bronchoalveolar lavage (BAL) lymphocyte direct (two-color and three-color) phenotyping was carried out in 63 patients with sarcoidosis (subdivided according to the disease radiological stage and smoking status), 23 patients with systemic sclerosis (SSc) and 12 individuals exposed to silica dusts. 33 healthy volunteers were used as controls. Routine BAL cytology and peripheral blood lymphocyte (PBL) typing was performed in all tested subjects. FC results quality control was performed according to BD Simultest IMK Plus criteria (for PBL) modified by the author. The conventional lymphocytic gate (PBL light scatter based) was excluded, as the source of error in L-BAL subset analysis. Two-color L-BAL phenotyping and alveolar lymphocyte CD45FITC/SSC gating was found to be usually sufficient. However, in poor-lymphocyte materials (e.g. in healthy smokers) three-color method, using anti-CD45 PECy5 monoclonal antibody for staining each patient sample, should be considered. No significant changes in lymphocyte subsets were found in FC as compared with alkaline phosphatase immunocytochemical method. The prevalence of CD3+ cells with only a few lymphocytes expressing NK (CD3-CD16 or 56+), T suppressor (CD8+ 11b+) or B cell phenotype--as compared with PBL, was found in BAL of all examined subjects. CD8+ cell subset was dominated by T cytotoxic cell phenotype (CD8+ CD11b-) in all tested groups. L-BAL T cells expressed sensitized memory cell phenotype--CD4+ (CD8+)CD45RO+ in sarcoidosis, SSc and controls (the staining not performed in silica exposed persons). Characteristic BAL cytological and immunological pattern (lymphocytic alveolitis, increased CD4/CD8, high CD3(CD4)+HLA-DR+ percentage) was observed in pulmonary sarcoidosis. SSc patients showed reduced BAL CD4/CD8 ratio, due to T cytotoxic cell predominance (the changes were more distinct in the group with clinical pulmonary changes). Early activation marker (CD25) was expressed on the elevated percentage of BAL T helper cells in SSc. High CD4+CD25+ cell percentage was the most characteristic sign of lymphocyte activation in individuals exposed to silica. Summing up, the reference values for L-BAL subsets were proposed. CD4/CD8 ratio alterations in interstitial lung disorders should be interpreted as the local imbalance between T helper memory cells and sensitized T cytotoxic lymphocytes. FC characterization of alveolar lymphocytes provides insight into the pathogenesis of pulmonary diseases.

Authors+Show Affiliations

Zakład Patofizjologii Klinicznej Katedra Patofizjologii Collegium Medicum UJ.

Pub Type(s)

Clinical Trial
Controlled Clinical Trial
English Abstract
Journal Article
Research Support, Non-U.S. Gov't

Language

pol

PubMed ID

10481384

Citation

Kopiński, P. "[Use of Flow Cytometry in Evaluation of Cellular Changes in Interstitial Lung Diseases]." Folia Medica Cracoviensia, vol. 38, no. 3-4, 1997, pp. 69-115.
Kopiński P. [Use of flow cytometry in evaluation of cellular changes in interstitial lung diseases]. Folia Med Cracov. 1997;38(3-4):69-115.
Kopiński, P. (1997). [Use of flow cytometry in evaluation of cellular changes in interstitial lung diseases]. Folia Medica Cracoviensia, 38(3-4), 69-115.
Kopiński P. [Use of Flow Cytometry in Evaluation of Cellular Changes in Interstitial Lung Diseases]. Folia Med Cracov. 1997;38(3-4):69-115. PubMed PMID: 10481384.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - [Use of flow cytometry in evaluation of cellular changes in interstitial lung diseases]. A1 - Kopiński,P, PY - 1997/1/1/pubmed PY - 1999/9/11/medline PY - 1997/1/1/entrez SP - 69 EP - 115 JF - Folia medica Cracoviensia JO - Folia Med Cracov VL - 38 IS - 3-4 N2 - Flow cytometry (FC) usefulness for pulmonary reactive cell changes examination in interstitial lung diseases was assessed. Bronchoalveolar lavage (BAL) lymphocyte direct (two-color and three-color) phenotyping was carried out in 63 patients with sarcoidosis (subdivided according to the disease radiological stage and smoking status), 23 patients with systemic sclerosis (SSc) and 12 individuals exposed to silica dusts. 33 healthy volunteers were used as controls. Routine BAL cytology and peripheral blood lymphocyte (PBL) typing was performed in all tested subjects. FC results quality control was performed according to BD Simultest IMK Plus criteria (for PBL) modified by the author. The conventional lymphocytic gate (PBL light scatter based) was excluded, as the source of error in L-BAL subset analysis. Two-color L-BAL phenotyping and alveolar lymphocyte CD45FITC/SSC gating was found to be usually sufficient. However, in poor-lymphocyte materials (e.g. in healthy smokers) three-color method, using anti-CD45 PECy5 monoclonal antibody for staining each patient sample, should be considered. No significant changes in lymphocyte subsets were found in FC as compared with alkaline phosphatase immunocytochemical method. The prevalence of CD3+ cells with only a few lymphocytes expressing NK (CD3-CD16 or 56+), T suppressor (CD8+ 11b+) or B cell phenotype--as compared with PBL, was found in BAL of all examined subjects. CD8+ cell subset was dominated by T cytotoxic cell phenotype (CD8+ CD11b-) in all tested groups. L-BAL T cells expressed sensitized memory cell phenotype--CD4+ (CD8+)CD45RO+ in sarcoidosis, SSc and controls (the staining not performed in silica exposed persons). Characteristic BAL cytological and immunological pattern (lymphocytic alveolitis, increased CD4/CD8, high CD3(CD4)+HLA-DR+ percentage) was observed in pulmonary sarcoidosis. SSc patients showed reduced BAL CD4/CD8 ratio, due to T cytotoxic cell predominance (the changes were more distinct in the group with clinical pulmonary changes). Early activation marker (CD25) was expressed on the elevated percentage of BAL T helper cells in SSc. High CD4+CD25+ cell percentage was the most characteristic sign of lymphocyte activation in individuals exposed to silica. Summing up, the reference values for L-BAL subsets were proposed. CD4/CD8 ratio alterations in interstitial lung disorders should be interpreted as the local imbalance between T helper memory cells and sensitized T cytotoxic lymphocytes. FC characterization of alveolar lymphocytes provides insight into the pathogenesis of pulmonary diseases. SN - 0015-5616 UR - https://www.unboundmedicine.com/medline/citation/10481384/[Use_of_flow_cytometry_in_evaluation_of_cellular_changes_in_interstitial_lung_diseases]_ DB - PRIME DP - Unbound Medicine ER -