In vitro characterisation of anti-major histocompatibility complex class I-mediated transfusion-related acute lung injury (TRALI) — ASN Events
  1. Schedule
  2. Poster Session Three
  3. In vitro characterisation of anti-major histocompatibility complex class I-mediated transfusion-related acute lung injury (TRALI)

In vitro characterisation of anti-major histocompatibility complex class I-mediated transfusion-related acute lung injury (TRALI) (#423)

Annette Sultana 1 2 3 , Melinda M Dean 1 2 4 , Michael C Reade 2 5 , Robert L Flower 1 2 4 , John-Paul Tung 1 2 3 4
  1. Australian Red Cross Blood Service, Kelvin Grove, QLD, Australia
  2. Faculty of Medicine, University of Queensland, Brisbane, QLD, Australia
  3. Critical Care Research Group, The Prince Charles Hospital , Chermside, QLD, Australia
  4. Faculty of Health, Queensland University of Technology, Brisbane, QLD, Australia
  5. Joint Health Command, Australian Defence Force, Canberra , ACT, Australia

Introduction

Transfusion-related acute lung injury (TRALI) is a leading cause of transfusion-related mortality. In vivo mouse models of anti-major histocompatibility complex (MHC) class I antibody mediated-TRALI have provided insights into mechanisms causing TRALI; however, the clinical relevance of these insights remains unclear. To facilitate translation to the clinical setting a human in vitro TRALI model was developed.

Methods

Human lung microvascular endothelial cells (HLMVECs) were co-cultured with buffer ± lipopolysaccharide (LPS; E. coli O55:B5; 0.2, 1 or 2µg/ml) ± freshly isolated neutrophils. In subsequent experiments either buffer, Tumour necrosis factor (TNF)-α (0.002, 0.01 or 0.02µg/ml) or an anti-MHC-ABC monoclonal antibody (MoAb; clone W6/32, G46-2.6, and/or 246-B8.E7; 0.6, 3.33 or 9.33µg/ml) were added after the neutrophils. HLMVEC activation (intracellular adhesion molecule (ICAM)-1 expression) was assessed using flow cytometry.

Results

Co-culture with LPS resulted in a concentration-dependent increase in HLMVEC ICAM-1 expression, with or without neutrophils. Addition of TNF-α to LPS (0.2µg/ml)-treated HLMVECs further increased HLMVEC ICAM-1 expression. This demonstrated that the LPS concentration used in the model still enabled detection of further increases in ICAM-1 expression. In the absence of LPS, anti-MHC class I MoAb-mediated-HLMVEC activation was not evident regardless of MoAb clone or concentration. However, in the presence of LPS alone or co-culture with MoAbs (W6/32, 246-B8.E7 or W6/32+246-B8.E7+G46-2.6) ICAM-1 expression increased.

Discussion and Conclusions

Mouse in vivo models of TRALI tested a large panel of anti-MHC class I MoAb, of which only the 34-1-2s clone induced TRALI. Human cells are likely to react with different anti-MHC class I MoAbs, so testing of a wider panel of anti-human MHC class I MoAbs was required. Having now identified several MoAbs that activate HLMVECs, this in vitro model will provide a useful tool in characterising mechanisms that contribute to anti-MHC class I-mediated TRALI in humans and identify potential risk reduction strategies.

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