Local intragraft humoral immune responses in chronic lung allograft dysfunction

Abstract

Background: Donor human leukocyte antigen (HLA)-specific antibodies (DSA) and non-HLA antibodies can cause allograft injury, possibly leading to chronic lung allograft dysfunction (CLAD) after lung transplantation. It remains unclear whether these antibodies are produced locally in the graft or derived solely from circulation. We hypothesized that DSA and non-HLA antibodies are produced in CLAD lungs.

Methods: Lung tissue was prospectively collected from 15 CLAD patients undergoing retransplantation or autopsy. 0.3 g of fresh lung tissue was cultured for 4 days without or with lipopolysaccharide or CD40L: lung culture supernatant (LCS) was sampled. Protein eluate was obtained from 0.3 g of frozen lung tissue. The mean fluorescence intensity (MFI) of DSA and non-HLA antibodies was measured by Luminex and antigen microarray, respectively.

Results: LCS from all 4 patients who had serum DSA at lung isolation were positive for DSA, with higher levels measured after CD40L stimulation (CD40L⁺LCS). Of these, only 2 had detectable DSA in lung eluate. MFI of non-HLA antibodies from CD40L⁺LCS correlated with those from lung eluate but not with those from sera. Flow cytometry showed higher frequencies of activated lung B cells in patients whose CD40L⁺LCS was positive for DSA (n = 4) or high non-HLA antibodies (n = 6) compared to those with low local antibodies (n = 5). Immunofluorescence staining showed CLAD lung lymphoid aggregates with local antibodies contained larger numbers of IgG⁺ plasma cells and greater IL-21 expression.

Conclusions: We show that DSA and non-HLA antibodies can be produced within activated B cell-rich lung allografts.

Keywords: antibody-mediated rejection; chronic lung allograft dysfunction; donor-specific antibodies; lung transplant; lymphoid aggregates; non-HLA antibodies; tertiary lymphoid organs.