Acute cellular rejection (ACR) frequently occurs following lung transplantation (LuTx) and represents a risk factor for the development of chronic lung allograft dysfunction (CLAD) as well as long-term survival. The histopathological diagnosis of ACR carries a burden of interobserver variability. The widespread utilization and cost-effectiveness of immunohistochemistry (IHC) was proven beneficial in diagnosing rejection in human kidney transplantations and LuTx rat models. However, its potential for ACR detection in patients remains unexplored. We analyzed surface markers (CD3, CD4, CD8, CD20, CD68, CD47, PD-1, PD-L1, and CD31/PECAM-1) on lung tissue cryobiopsy samples collected within 6 months post-LuTx from 60 LuTx recipients, 48 of whom were diagnosed with ACR. Additionally, serum samples from 51 patients were analyzed using a multiplex bead-based Luminex assay. The cytokines and markers included PD-L1, IL2, TNFα, IFNγ, and Granzyme B. We observed a significant increase in PD-L1 tissue expression within the rejection group, suggesting a concerted effort to suppress immune responses, especially those mediated by T-cells. Furthermore, we noted significant differences in PECAM-1 levels between ACR/non-ACR. Additionally, peripheral blood C-reactive-protein levels tended to be higher in the ACR group, while Luminex serum analyses did not reveal any significant differences between groups. In conclusion, our findings suggest the potential value of PECAM-1 and PD-L1 markers in diagnosing ACR.
- Klíčová slova
- acute cellular rejection, checkpoint inhibitors, immunohistochemistry, luminex, lung transplantation,
- MeSH
- akutní nemoc MeSH
- antigeny CD274 * metabolismus krev MeSH
- antigeny CD31 * metabolismus MeSH
- biologické markery * krev metabolismus MeSH
- dospělí MeSH
- imunohistochemie MeSH
- lidé středního věku MeSH
- lidé MeSH
- plíce patologie MeSH
- rejekce štěpu * diagnóza krev MeSH
- senioři MeSH
- transplantace plic * škodlivé účinky MeSH
- Check Tag
- dospělí MeSH
- lidé středního věku MeSH
- lidé MeSH
- mužské pohlaví MeSH
- senioři MeSH
- ženské pohlaví MeSH
- Publikační typ
- časopisecké články MeSH
- Názvy látek
- antigeny CD274 * MeSH
- antigeny CD31 * MeSH
- biologické markery * MeSH
- CD274 protein, human MeSH Prohlížeč
- PECAM1 protein, human MeSH Prohlížeč
Compared to solid tumors, the role of PD-L1 in hematological malignancies is less explored, and the knowledge in this area is mostly limited to lymphomas. However, several studies indicated that PD-L1 is also overexpressed in myeloid malignancies. Successful treatment of the acute myeloid leukemia (AML) is likely associated with elimination of the residual disease by the immune system, and possible involvement of PD-L1 in this process remains to be elucidated. We analyzed PD-L1 expression on AML primary cells by flow cytometry and, in parallel, transcript levels were determined for the transcription variants v1 and v2. The ratio of v1/v2 cDNA correlated with the surface protein amount, and high v1/v2 levels were associated with worse overall survival (p = 0.0045). The prognostic impact of PD-L1 was limited to AML with mutated nucleophosmin and concomitant internal tandem duplications in the FLT3 gene (p less than 0.0001 for this particular AML subgroup).
- Klíčová slova
- AML, CD34, FLT3-ITD, NPM1, PD-1, PD-L1 transcript, leukemia,
- MeSH
- akutní myeloidní leukemie krev genetika MeSH
- antigeny CD274 krev genetika metabolismus MeSH
- jaderné proteiny genetika MeSH
- lidé MeSH
- messenger RNA genetika metabolismus MeSH
- mutace MeSH
- nádorové biomarkery krev genetika metabolismus MeSH
- nukleofosmin MeSH
- tyrosinkinasa 3 podobná fms genetika MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- Názvy látek
- antigeny CD274 MeSH
- CD274 protein, human MeSH Prohlížeč
- FLT3 protein, human MeSH Prohlížeč
- jaderné proteiny MeSH
- messenger RNA MeSH
- nádorové biomarkery MeSH
- NPM1 protein, human MeSH Prohlížeč
- nukleofosmin MeSH
- tyrosinkinasa 3 podobná fms MeSH
The expression on the surface of tumor cells of ligands for the PD-1 inhibitory receptor prevents the antitumor immune response and is considered to be a negative prognostic factor in a variety of solid tumors as well as in hematologic malignancies. To determine if it were possible to analyze PD-L1 with PCR-based methods, we assessed the expression of PD-L1 in primary samples from patients with acute myeloid leukemia, in healthy donors, and in a panel of cell lines, by means of flow cytometry, RT-PCR, and Western blotting. Although the surface density of the protein was not correlated with the amount of expressed full-length mRNA, we found a statistically significant positive correlation between PD-L1 surface density and the ratio of two transcript variants (variant 1/variant 2). Our PCR-based method allows for retrospective examination of PD-L1 surface expression from frozen cDNA samples, without the need for a reference gene. Our results also suggest that variant 2, which is produced by alternative splicing, negatively regulates PD-L1 protein expression on the cell surface. In addition, PD-L1 exposition on the cell surface is clearly associated with a shift of electrophoretic mobility, observed on Western blots. This finding can explain the relatively large variability in PD-L1 apparent molecular weight reported in the literature and offers an alternate means for the assessment of PD-L1 surface expression. Cancer Immunol Res; 4(10); 815-9. ©2016 AACR.
- MeSH
- akutní myeloidní leukemie genetika imunologie MeSH
- alternativní sestřih MeSH
- antigeny CD274 biosyntéza krev genetika MeSH
- genetická variace MeSH
- lidé MeSH
- messenger RNA genetika MeSH
- nádorové biomarkery biosyntéza krev genetika MeSH
- nádorové buňky kultivované MeSH
- nádorové proteiny biosyntéza krev genetika MeSH
- regulace genové exprese u nádorů imunologie MeSH
- RNA nádorová genetika MeSH
- western blotting metody MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- Názvy látek
- antigeny CD274 MeSH
- CD274 protein, human MeSH Prohlížeč
- messenger RNA MeSH
- nádorové biomarkery MeSH
- nádorové proteiny MeSH
- RNA nádorová MeSH
