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[Analysis of lymphocyte populations with flow cytometry in routine bronchoalveolar lavage diagnosis: comparison of a 3-color method with the alkaline phosphatase anti-alkaline phosphatase immunohistochemistry].
Pneumologie. 1997 Aug; 51(8):860-4.P

Abstract

BACKGROUND

Identification of lymphocyte phenotypes in bronchoalveolar lavage (BAL) plays a crucial role in the diagnosis of interstitial lung disease. Cells staining positive for specific monoclonal antibodies may be detected by immunocytochemistry or flow cytometry (FCM). The present study compares a three-colour FCM-approach to a standard APAAP protocol for immunocytochemistry.

METHODS

BAL-specimens of 22 patients with various lung diseases were investigated. Inclusion criteria was a lymphocytosis of > 10% of all BAL-cells. After the preparation of cytocentrifuge slides, staining was performed with monoclonal antibodies to CD3, CD4 and CD8 following the APAAP-protocol. FCM-analysis was performed with the following panel of conjugates: CD3-FITC, CD4- or CD8-PE, CD45-perCP. Lymphocytes were gated by their SSC/CD45 characteristics. T-helper and T-suppressor percentages were evaluated by quadrant analysis of CD3/CD4 or CD3/CD8 histograms.

RESULTS

With the exception of CD3, where the range of values was quite narrow (10% variance), the correlation between the two methods was excellent (CD4: r = 0.98; CD8: r = 0.99; CD4/CD8: r = 0.96; p < 0.0001).

CONCLUSION

Flow cytometry reveals similar results compared to immunocytochemistry in the determination of lymphocyte subsets characterised by CD3, CD4 and CD8 antigens.

Authors+Show Affiliations

Klinik für Allgemeine Innere Medizin, Christian-Albrechts-Universität Kiel.No affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info available

Pub Type(s)

Comparative Study
Journal Article

Language

ger

PubMed ID

9380662

Citation

Bergmann, M, et al. "[Analysis of Lymphocyte Populations With Flow Cytometry in Routine Bronchoalveolar Lavage Diagnosis: Comparison of a 3-color Method With the Alkaline Phosphatase Anti-alkaline Phosphatase Immunohistochemistry]." Pneumologie (Stuttgart, Germany), vol. 51, no. 8, 1997, pp. 860-4.
Bergmann M, Bewig B, Tiroke A, et al. [Analysis of lymphocyte populations with flow cytometry in routine bronchoalveolar lavage diagnosis: comparison of a 3-color method with the alkaline phosphatase anti-alkaline phosphatase immunohistochemistry]. Pneumologie. 1997;51(8):860-4.
Bergmann, M., Bewig, B., Tiroke, A., Barth, J., & Fölsch, U. R. (1997). [Analysis of lymphocyte populations with flow cytometry in routine bronchoalveolar lavage diagnosis: comparison of a 3-color method with the alkaline phosphatase anti-alkaline phosphatase immunohistochemistry]. Pneumologie (Stuttgart, Germany), 51(8), 860-4.
Bergmann M, et al. [Analysis of Lymphocyte Populations With Flow Cytometry in Routine Bronchoalveolar Lavage Diagnosis: Comparison of a 3-color Method With the Alkaline Phosphatase Anti-alkaline Phosphatase Immunohistochemistry]. Pneumologie. 1997;51(8):860-4. PubMed PMID: 9380662.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - [Analysis of lymphocyte populations with flow cytometry in routine bronchoalveolar lavage diagnosis: comparison of a 3-color method with the alkaline phosphatase anti-alkaline phosphatase immunohistochemistry]. AU - Bergmann,M, AU - Bewig,B, AU - Tiroke,A, AU - Barth,J, AU - Fölsch,U R, PY - 1997/8/1/pubmed PY - 1998/2/12/medline PY - 1997/8/1/entrez SP - 860 EP - 4 JF - Pneumologie (Stuttgart, Germany) JO - Pneumologie VL - 51 IS - 8 N2 - BACKGROUND: Identification of lymphocyte phenotypes in bronchoalveolar lavage (BAL) plays a crucial role in the diagnosis of interstitial lung disease. Cells staining positive for specific monoclonal antibodies may be detected by immunocytochemistry or flow cytometry (FCM). The present study compares a three-colour FCM-approach to a standard APAAP protocol for immunocytochemistry. METHODS: BAL-specimens of 22 patients with various lung diseases were investigated. Inclusion criteria was a lymphocytosis of > 10% of all BAL-cells. After the preparation of cytocentrifuge slides, staining was performed with monoclonal antibodies to CD3, CD4 and CD8 following the APAAP-protocol. FCM-analysis was performed with the following panel of conjugates: CD3-FITC, CD4- or CD8-PE, CD45-perCP. Lymphocytes were gated by their SSC/CD45 characteristics. T-helper and T-suppressor percentages were evaluated by quadrant analysis of CD3/CD4 or CD3/CD8 histograms. RESULTS: With the exception of CD3, where the range of values was quite narrow (10% variance), the correlation between the two methods was excellent (CD4: r = 0.98; CD8: r = 0.99; CD4/CD8: r = 0.96; p < 0.0001). CONCLUSION: Flow cytometry reveals similar results compared to immunocytochemistry in the determination of lymphocyte subsets characterised by CD3, CD4 and CD8 antigens. SN - 0934-8387 UR - https://www.unboundmedicine.com/medline/citation/9380662/[Analysis_of_lymphocyte_populations_with_flow_cytometry_in_routine_bronchoalveolar_lavage_diagnosis:_comparison_of_a_3_color_method_with_the_alkaline_phosphatase_anti_alkaline_phosphatase_immunohistochemistry]_ L2 - https://medlineplus.gov/pulmonaryfibrosis.html DB - PRIME DP - Unbound Medicine ER -