Anti-HLA antibody binding to HLA class I molecules induces proliferation of airway epithelial cells: A potential mechanism for bronchiolitis obliterans.

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Anti-HLA antibody binding to HLA class I molecules induces proliferation of airway epithelial cells: A potential mechanism for bronchiolitis obliterans syndrome Scott I. Reznik, MDa, Andrés Jaramillo, PhDab, Leiying Zhang, MDa, G.Alexander Patterson, MDa, Joel D. Cooper, MDa, T. Mohanakumar, PhDab The Journal of Thoracic and Cardiovascular Surgery Volume 119, Issue 1, Pages 39-45 (January 2000) DOI: 10.1016/S0022-5223(00)70215-7 Copyright © 2000 Mosby, Inc. Terms and Conditions

Fig. 1 Induction of tyrosine phosphorylation by anti-HLA antibody binding. A549 cells were incubated for 5 minutes in the presence of control mouse IgG (lane 1) , W6/32 monoclonal antibody (lane 2) , NHS (lane 3) , and pooled anti-HLA serum (lane 4) . Cell samples were then analyzed by immunoblotting with the PY20 antiphosphotyrosine monoclonal antibody. The results are representative of 2 different experiments. The Journal of Thoracic and Cardiovascular Surgery 2000 119, 39-45DOI: (10.1016/S0022-5223(00)70215-7) Copyright © 2000 Mosby, Inc. Terms and Conditions

Fig. 2 Induction of AEC tyrosine phosphorylation by anti-HLA antibody binding. A549 cells were incubated for 10 minutes in the presence of control mouse IgG (lane 1) , W6/32 monoclonal antibody (lane 2) , NHS (lane 3) , and pooled anti-HLA serum (lane 4) . Cell samples were then analyzed by immunoblotting with the PY20 antiphosphotyrosine monoclonal antibody. The results are representative of 2 different experiments. The Journal of Thoracic and Cardiovascular Surgery 2000 119, 39-45DOI: (10.1016/S0022-5223(00)70215-7) Copyright © 2000 Mosby, Inc. Terms and Conditions

Fig. 3 Stimulation of AEC proliferation by the W6/32 monoclonal antibody. A549 cells were starved for 48 hours and then incubated in the presence of the W6/32 monoclonal antibody, control mouse IgG (negative control), or complete medium (positive control) for 24, 48, and 72 hours. Cell proliferation was determined by tritiated thymidine incorporation. The results of quadruplicate cultures (n = 4) are expressed as the mean values ± SD and are representative of 3 different experiments. *P < .05. The Journal of Thoracic and Cardiovascular Surgery 2000 119, 39-45DOI: (10.1016/S0022-5223(00)70215-7) Copyright © 2000 Mosby, Inc. Terms and Conditions

Fig. 4 Stimulation of AEC proliferation by the pooled anti-HLA serum. A549 cells were starved for 48 hours and then incubated in the presence of the pooled anti-HLA serum, NHS (negative control), or complete medium (positive control) for 24, 48, and 72 hours. Cell proliferation was determined by tritiated thymidine incorporation. The results of quadruplicate cultures (n = 4) are expressed as the mean values ± SD and are representative of 3 different experiments. *P < .05. The Journal of Thoracic and Cardiovascular Surgery 2000 119, 39-45DOI: (10.1016/S0022-5223(00)70215-7) Copyright © 2000 Mosby, Inc. Terms and Conditions

Fig. 5 Stimulation of AEC proliferation by anti-HLA+ sera from lung transplant recipients with BOS at the time of the study. A549 cells were starved for 48 hours and then incubated in the presence of anti-HLA+ sera from lung transplant recipients with BOS at the time of the study, pooled anti-HLA serum (positive control), or NHS (negative control) for 24, 48, and 72 hours. Cell proliferation was determined by tritiated thymidine incorporation. The results of quadruplicate cultures (n = 4) are expressed as the mean values ± SD and are representative of 3 different experiments. The Journal of Thoracic and Cardiovascular Surgery 2000 119, 39-45DOI: (10.1016/S0022-5223(00)70215-7) Copyright © 2000 Mosby, Inc. Terms and Conditions

Fig. 6 Lack of stimulation of AEC proliferation by anti-HLA– sera from lung transplant recipients without BOS at the time of the study. A549 cells were starved for 48 hours and then incubated in the presence of anti-HLA– sera from lung transplant recipients without BOS at the time of the study, pooled anti-HLA serum, or NHS for 24, 48, and 72 hours. Cell proliferation was determined by tritiated thymidine incorporation. The results of quadruplicate cultures (n = 4) are expressed as the mean values ± SD and are representative of 3 different experiments. The Journal of Thoracic and Cardiovascular Surgery 2000 119, 39-45DOI: (10.1016/S0022-5223(00)70215-7) Copyright © 2000 Mosby, Inc. Terms and Conditions