Outcome of Extracorporeal Photopheresis as an Add-On Therapy for Antibody-Mediated Rejection in Lung Transplant Recipients.

INTRODUCTION: The diagnosis and treatment of antibody-mediated rejection (AMR) after lung transplantation has recently gained recognition within the transplant community. Extracorporeal photopheresis (ECP), currently used to treat chronic lung allograft dysfunction, modulates various pathways of the immune system known to be involved in AMR. We hypothesize that adding ECP to established AMR treatments could prevent the rebound of donor-specific antibodies (DSA). OBJECTIVES: This study aimed to analyze the role of ECP as an add-on therapy to prevent the rebound of DSA. METHODS: Lung transplant recipients who received ECP as an add-on therapy for pulmonary AMR between January 2010 and January 2019 were included in this single-center retrospective analysis. Baseline demographics of the patients, as well as their immunological characteristics and long-term transplant outcomes, were analyzed. RESULTS: A total of 41 patients developed clinical AMR during the study period. Sixteen patients received ECP as an add-on therapy after first-line AMR treatment. Among the 16 patients, 2 (13%) had pretransplant DSA, both against human leukocyte antigen (HLA) class I (B38, B13, and C06). Fifteen patients (94%) developed de novo DSA (dnDSA), i.e., 10 (63%) against class I and 14 (88%) against class II. The median time to dnDSA after lung transplantation was 361 days (range 25-2,548). According to the most recent International Society of Heart and Lung Transplantation (ISHLT) consensus report, 2 (13%) patients had definite clinical AMR, 6 (38%) had probable AMR, and 7 (44%) had possible AMR. The median mean fluorescence intensity (MFI) of dnDSA at the time of clinical diagnosis was 4,220 (range 1,319-10,552) for anti-HLA class I and 10,953 (range 1,969-27,501) for anti-HLA class II antibodies. ECP was performed for a median of 14 cycles (range 1-64). MFI values of dnDSA against HLA classes I and II were significantly reduced over the treatment period (for anti-class I: 752; range 70-2,066; for anti-class II: 5,612; range 1,689-21,858). The 1-year survival rate was 55%. No adverse events related to ECP were reported in any of the patients. CONCLUSIONS: ECP is associated with a reduction of dnDSA in lung transplant recipients affected by AMR. Prospective studies are warranted to confirm the beneficial effects of ECP in the setting of AMR.

Health care providers' perceptions of family caregivers' involvement in consultations within a geriatric hospital setting.

This study explored health professionals' (HPs') experiences of interacting with family care-givers (FCs), and the strategies they employ during these interactions. Qualitative methods involved audio-taped and transcribed in-depth semi-structured interviews with 21 HPs (doctors, nurses) from the geriatric wards of two tertiary hospitals. Framework methods were used to analyze data. Seven main themes emerged: Variation in family behaviours; FCs face many challenges; Psychosocial factors influence FCs' behaviours; Attitudes, competing responsibilities and lack of resources influencing HPs' strategies and behaviours; Strategies employed by HPs to improve communication; HPs' self-care strategies; Impact of interactions on HPs and the healthcare system. Inter-relationships between the themes were then integrated into a three-part model. This study offered insights into HPs' perspectives, experiences, and behaviours in geriatric consultations where FCs are present. Findings emphasised the need for HP training programs to improve communication and collaboration between HPs, patients, and FCs.

Function of platelets in apheresis platelet concentrates and in patient blood after transfusion as assessed by Impact-R.

BACKGROUND: Platelet (PLT) transfusion is a mainstream therapy for preventing or treating bleeding episodes in patients with thrombocytopenia. The efficacy is usually estimated from the corrected count increment of PLTs after transfusion, which does not assess PLT function. We therefore evaluated PLT function in blood samples of patients with thrombocytopenia before and after transfusion. STUDY DESIGN AND METHODS: PLT function was assessed in 24 chemotherapy-treated patients and in the PLT concentrates (PCs) by the Impact-R (DiaMed). This device evaluates PLT adhesion and aggregation recorded as surface coverage (%) and size of aggregates (AS microm(2)). P-selectin expression was determined by flow cytometry. RESULTS: The PCs were stored for a median of 70 hours before transfusion. An analysis stratified by the median storage of PCs (<70 hr or >70 hr) showed no differences in the SC, the AS, and P-selectin expression between these concentrates' groups. Transfusion resulted in an increase of adhering PLTs in the patients after transfusion. There were no differences in the AS and in P-selectin expression before and after transfusion, but the AS increased after transfusion upon ex vivo exposure to adenosine 5'-diphosphate. P-selectin expression was significantly lower in the patient group receiving PCs stored for more than 70 hours. CONCLUSION: The current trial shows the feasibility of using the Impact-R to assess the function of transfused PLTs in the patient's blood stream.

Platelet function under high-shear conditions from platelet concentrates.

BACKGROUND: Platelet (PLT) collection and storage affect the functional capacity of PLTs in PLT concentrates (PCs). Therefore, PLTs' functional quality should be studied before transfusion. STUDY DESIGN AND METHODS: PCs (n = 15) were collected by a standard apheresis procedure (Trima, Gambro BCT) and were stored for 7 days. Samples were taken to assess PLT adhesion and aggregate formation by a cone and plate analyzer (Impact-R, DiaMed) on Days 1, 3, 5, and 7 after harvesting. This device allows testing PLT function under high-shear stress close to physiologic conditions. Concomitantly, P-selectin expression and the residual responsiveness to TRAP-6 were determined by flow cytometry. RESULTS: PLT adhesion, as measured by surface coverage, decreased during the entire observation period; likewise, the size of aggregates was significantly lower on Days 5 and 7 compared to Day 1 (p < 0.02). P-selectin expression increased from Day 5 to Day 7 (p < 0.04), whereas TRAP-6-inducible expression remained stable until Day 5 of storage and decreased significantly on Day 7 (p = 0.04). CONCLUSIONS: Our results show that high-shear-induced PLT adhesion and aggregation on the polystyrene surface deteriorate upon storage, suggesting decreased PLT function in vivo. Thus, the Impact-R may be a useful tool to assess the functional capacity of PLTs under various PLT harvesting and storage procedures.

Monitoring survival and function of transfused platelets in Bernard-Soulier syndrome by flow cytometry and a cone and plate(let) analyzer (Impact-R).

BACKGROUND: Bernard-Soulier syndrome (BSS) patients may repeatedly require transfusion of platelets (PLTs). The hemostatic competence of transfused PLTs requires monitoring. STUDY DESIGN AND METHODS: Flow cytometry and a cone and plate(let) analyzer (Impact-R, DiaMed) were used to monitor survival and function of transfused PLTs in a 7-year-old girl with BSS undergoing surgery. Flow cytometry was applied to differentiate autologous PLTs from transfused PLTs by staining for CD42b. The Impact, which measures PLT adhesion and aggregation in response to high shear stress, was used to evaluate PLT function. RESULTS: Transfused PLTs were detectable by flow cytometry for 1 week after transfusion. While the patient's PLTs did not respond to high shear stress before transfusion, a normal response was documented by the Impact on the day after transfusion and 1 week thereafter. CONCLUSION: Transfused PLTs were detectable by flow cytometry, and their functional activity was demonstrated by the Impact.

Agonist-inducible platelet activation in chronic idiopathic autoimmune thrombocytopenia.

OBJECTIVE: There are only few studies on agonist-inducible platelet activation in chronic idiopathic autoimmune thrombocytopenia (cAITP). MATERIALS AND METHODS: We compared agonist (TRAP-6, ADP, Arachidonic acid, Epinephrine, and Ristocetin) -inducible P-selectin expression and PAC-1 binding in 40 patients with cAITP (f/m ratio 23/17) with those in 20 healthy controls. Results were correlated with platelet counts, detectable platelet antibodies, and reticulated platelets. RESULTS: The in vivo activation of platelets determined the in vitro inducible response to agonists. The stronger the in vivo activation the less the number of platelets responding to agonists, as illustrated by the inverse correlation of P-selectin expression ex vivo and the relative increase after the exogenous addition of agonists. The agonist-inducible platelet activation was not associated with the presence of detectable platelet antibodies to GPIb/IX or GPIIb/IIIa. Agonist-inducible platelet activation was also not correlated with counts of reticulated platelets. CONCLUSION: Agonist-inducible activation of platelets in cAITP is affected mainly by their in vivo activation.

Agonists-induced platelet activation varies considerably in healthy male individuals: studies by flow cytometry.

Flow cytometric evaluation of platelet function extends our understanding of platelets' role in various clinical conditions associated with either bleeding disorders, thrombosis, or monitoring of antiplatelet therapy. The use of suboptimal concentrations of various agonists may allow assessing the "activatability" of platelets. We determined platelet responsiveness to thrombin-receptor-activating peptide-6, arachidonic acid, adenosine 5c-diphosphate (ADP), epinephrine, collagen, and ristocetin at suboptimal concentrations by determination of P-selectin expression and binding of PAC-1 in 26 healthy male individuals. The response varied considerably from one individual to the next. However, within individuals, responses to all agonists except collagen correlated strongly (p<0.05), suggesting a global variability of platelet responses. Moreover, P-selectin expression and PAC-1 binding were strongly correlated (p<0.05). Interestingly, with epinephrine, PAC-1 positive events outnumbered P-selectin positive events, while this was not seen with the other agonists. Thus, epinephrine may specifically affect the conformational switch mechanism and receptor clustering. Our data indicate that the in vitro response to suboptimal concentrations of agonists varies, but individuals with selective platelet defects may still be identified based on data obtained with the various agonists.

Flow cytometric evaluation of platelet activation in chronic autoimmune thrombocytopenia.

Platelets from 42 patients (platelet counts median 42x10(9)/L, range 3-223x10(9)/L) with chronic idiopathic autoimmune thrombocytopenia (cAITP) were investigated for P-Selectin expression and PAC-1 binding. The results showed that the levels of P-Selectin positive platelets (n=20) were higher in cAITP than in controls (P<0.0001), and correlated with platelet counts (P=0.04). PAC-1 binding was increased in only six patients, and not correlated with platelet counts. There was no correlation of P-Selectin or PAC-1 with detectable platelet antibodies. Thus, platelets are activated in cAITP, but platelets, characterized by PAC-1 binding, are rare. These are either needed to maintain vascular integrity, or underwent premature sequestration.