Rheotherapy for vascular disorders.

Patients with vascular disorders are seldom offered apheresis in he management of their symptoms. In this article we review the different apheretical techniques used in these situations.

Hypertriglyceridemia: apheretic treatment.

Patients with extremely high triglyceride levels and associated lipemia are at high risk for acute pancreatitis. Two factors can increase triglyceride-rich lipoproteins; one is overproduction and other is a defect in clearance. Either mechanism can cause hypertriglyceridemia and both may exist simultaneously. Causes can be either primary or secondary. Plasmapheresis is efficacious for severe Hypertriglyceridemia in patients who have not responded to previous therapies. We have treated 15 cases of hypertriglyceridemia complicating the course of patients receiving Cyclosporin A after bone marrow transplantation. Five patients were treated with plasmapheresis, the other ten with cascade filtration. The removal rate for triglycerides was 58.0% for patients treated by cascade filtration and 63.5% for patients treated by plasmapheresis. The removal rates for triglycerides were low possibly as a consequence of early saturation of the filter.

Multicomponent collection as of 2005.

Multicomponent apheresis (MCA) begun in Genoa in 1985 in autologous terms. Named "sequestration" it was the preoperative collection of autologous components (RBC-plasma-platelets) using the same apparatus and harness ready for intraoperative blood salvage. In 1986 the technique was applied to donor platelet apheresis with the goal of reducing the costs of platelet collection and concurrently reducing the risks of viral disease transmission to haematological patients who did receive, in the same transfusion event, the necessary blood components obtained from the same donor. The results of this application were maximized by the so called aggressive MCA by which in selected donors, it is possible to collect 2 units of platelets along with 1 or 2 units of PRBCs. These applications were made possible by the development of the concept of dry-platelet collection according to which platelets are collected in only 20-25 mL of plasma and subsequently resuspended in non-plasma solutions such as T-Sol. A last development of MCC is for RBC apheresis, with the collection of 1-2 units of RBC independently of platelet collection. This is going to be the first step of apheresis as the unique modality of collecting blood leaving the bags to history of blood transfusion. Interestingly it took 15 years to MCC to be rediscovered and appreciated worldwide both for its intrinsic cost saving capabilities offered along with an increasing safety for patients. In terms of donor acceptance it is our experience that, since 1989 no donor has refused MCC, consisting at least in the concurrent collection of plasma along with platelets, but also RBC and or a second unit of platelets.

Rheopheresis for sudden hearing loss (SHL).

Rheopheresis has met an increasing interest and application in different disease conditions affecting microcirculation. Its last applications are for the management of Macular and Cochlear disorders. The sudden hearing loss syndrome is a condition that affects 20 out of 100,000 persons per year, reduces the patient's social interaction and quality of life. It is associated with vascular and coagulation risk factors and it is considered as a result of local hypoperfusion secondary both to inflammatory and dysimmune conditions determining inadequate NO release and endothelial dysfunction. Rheopheresis treatment is a new approach which brings to satisfactory clinical results. Cascade filtration, heparin induced lipid precipitation and conventional plasma exchange are equally effective, and only 2 sessions are required for producing long lasting benefits. Our experience is with 60 patients, 90% of which getting from partial to complete recovery after treatment as measured by pure tone audiometry.

Nine years of cascade filtration for thrombotic thrombocytopenic purpura.

Thrombotic thrombocytopenic purpura (TTP) continues to attract physicians' attention because of its unpredictable course and underlying pathogenetic mechanisms. It is also attractive because of its optimal response to therapeutic plasma exchange (PE) even in the most severe cases. The usual approach to its treatment is conventional PE with plasma substitution and only recently fresh frozen plasma (FFP) has been substituted by cryodepleted or virally inactivated plasma with clinical results that are, if not better, at least comparable with the ones attained when FFP is employed. Nonetheless, no consensus exists regarding the optimal PE fluid and this is not of marginal interest as even after PE treatment mortality is still from 15 to 20%. On the contrary, some consensus exists on the pathogenetic relevance of the exceedingly large Von Willebrand (vWF) multimers whose presence parallels the clinical course and appears to be determined by the abnormalities in the production or function and survival of vWF-cleaving proteases which is auto-antibody mediated. In both cases plasmapheresis without plasma infusion is relatively ineffective, perhaps because it does not increase protease activity. Paradoxically, cascade filtration (CF) can produce the same favorable results without replacing any protease activity. As with CF, the replacement is the autologous plasma with approximately 20% levels of vWF, fibrinogen, fibronectin, IgM and circulating immune complexes and 75% of albumin, IgGs, AT III and proteins whose molecular weight exceeds 250-300000 Da. Our experience with CF for TTP began in 1994. Since then, 16 patients have undergone CF combined with decreasing amounts of FFP supplementation and since 1998 without any allogeneic FFP supplementation in 9 cases. Twenty-four patients (96%) treated with no or minimal amounts of FFP survived but four (16%) experienced from one to four recurrences associated with cutaneous, paradental, cholecystic and vaginal infections. Only one patient died (5%) after the second CF procedure. There were no untoward effects related to the procedure itself and up to 18 procedures in one patient were carried out over 16 months, 10 with the patient in her sixth month of pregnancy and four in the post partum period in preparation for a splenectomy. Remission was achieved after an average number of treatments (10.7 +/- 6.8); a result that compares favorably with those of our historical control group of 47 cases (14 +/- 13). The patient's exposure to allogenic plasma which was 10.8 +/- 4.6 plasma U/session was reduced to 0 in 10 patients, to 1.4 +/- 1.2 and 4.4 +/- 2.3 plasma U/session, respectively, for seven and nine patients receiving PE + CF and CF with same plasma supplementation as described in our previous article. Based on our experience, we believe CF is presently the optimal treatment for patients with classic, sporadic TTP.

Cord blood (CB) stem cells for wound repair. Preliminary report of 2 cases.

In 2 patients, to promote skin wound/lesion repair we used fibrin-platelet glue combined with HLA compatible (2 mismatches accepted) buffy coats containing CD 34+ cord blood cells. The fibrin platelet glue was prepared with autologous apheresis platelets and cryoprecipitate. The original product was divided into 3 and 4 aliquots respectively for a correspondent number of applications. At each application, the margins of the lesion were infiltrated with 3 ml of cord blood buffy coat, containing 30 x 10(3) CD 34+ cells. No graft versus tissue reaction was seen in our patients in a follow-up of 3-7 months. The level of improvement, scored arbitrarily from 0 to 4, was 3 and 4, respectively. Our conclusion is that the use of cord blood cells along with fibrin platelet glue is of clinical interest.

Factors predicting response and graft-versus-host disease after donor lymphocyte infusions: a study on 593 infusions.

In the present study, we analyze factors predicting graft-versus-host disease (GvHD) and response after donor lymphocyte infusions (DLI). A total of 100 patients received 593 DLI between June 1990 and December 2000 in a bulk dose (n=14) or in escalating dose infusions (n=86). Patients were analyzed after stratification for type of relapse: (1). molecular relapse (n=6), (2). cytogenetic relapse (n=20), (3). chronic phase of chronic myeloid leukemia (CML) or complete remission of other disease post chemotherapy (n=24), (4). CML in accelerated/blastic phase (n=14), (5). resistant disease not responding to chemotherapy (n=36). The proportion of responders to DLI in these five groups was 100, 90, 75, 36 and 0% (P<0.0001). Factors predicting response by multivariate analysis were type of relapse (P<0.0001), post-DLI GvHD (P=0.005), pancytopenia (P=0.008), and a diagnosis of CML (P=0.04). Acute GvHD (grades II-IV) occurred in 21 patients (21%), and correlated in multivariate analysis with pancytopenia and less than four DLI. Other predictors of GvHD were the number of CD3+cells/infusion and serum levels of gamma-glutamyl transferase (gammaGT). The actuarial probability of treatment-related mortality was 9% for HLA identical siblings and 44% for alternative donor transplants (P=0.006). Response to DLI is predicted by tumor burden and is associated with GvHD and pancytopenia.

The role of autologous fibrin-platelet glue in plastic surgery: a preliminary report.

To promote wound healing, we used autologous fibrin-platelet glue in 14 patients with skin and soft tissue losses caused by recent trauma or chronic pathology. The level of improvement was scored, arbitrarily, from 0 to 4. Very favourable results (score 3-4) were seen in 11 out of 14 patients. The glue preparation is very easy, inexpensive and creates excellent and stable hemostasis. From a general point of view, we have confirmed the utility of fibrin-platelet glue in terms of reduced infections and length of hospital stay.

Evaluation of dosed red blood cell units collected with Gambro BCT--TRIMA.

The increasing need of collecting high quality blood components and of improving the overall productivity of a blood centre requires the utilisation of a new innovative process that combines high speed collection with an automated process and blood component tailoring to fit individual patient requirements. We collected dosed Red Blood Cell (dRBC) units on 64 donors, eligible as regular donors on the Gambro BCT TRIMA using the dRBC collection protocol. The collection target was set to 180 ml packed Red Blood Cells (pRBCs) in 225 ml total collection volume (n = 7), or 300 ml pRBCs in 375 ml total collection volume (n = 33) or 360 ml in 450 ml (n = 24), depending on donor's hematological profile and blood volemia. Saline was infused as the replacement fluid at a 120%) collection:infusion ratio. Donor per cent hematocrit was (mean +/- S.D.) 43.7 +/- 4.0% and TBV = 4.99 +/- 0.69 1. The procedures yielded 100 +/- 6% of predicted yield, with a hematocrit of 78.2 +/- 6.6% in 29 +/- 3 min. Hb content was 99.9 +/- 21.8 in all procedures, or 61.5-94.4-118.6 g in the three groups, respectively. After the addition of the SAG-M storage solution, the hematocrit was 56.3 +/- 6.2%. No adverse reactions have been reported by the donors and all pPRBC units were transfused to patients without any transfusion reaction being reported by clinicians. The dRBC protocol is well tolerated by donors without any side effects, other than normal effects of regular blood donation. Higher pRBC productivity can be reached with a safe and automated process in conjunction with a high and consistent product quality easily matching the donor collection criteria and pRBC unit standards. Tailoring of pRBC units can result in an improved patient transfusion support.

Double plateletpheresis (DPA) and tailored RBC collection with the Excel-Pro: preliminary results.

The Dideco Excel-Pro is frequently used for double plateletpheresis (DPA) when the platelet precount exceeds 280 x 10(3)/l. Platelets are collected as "dry platelets" and the resuspension solution is added when the procedure is over. Even when DPA is carried out the product volume prior to resuspension may be as low as 60 ml. As a result, a third product may be collected along with platelets. Our priority is to collect RBCs and, depending on the donor's BW, tailored RBC collections are carried out. This means that from 400 to 480 ml of PRBC (70% hct) are collected from donors whose BW exceeds 75 kg. The results of the last 27 DPA/tailored PRBC collections are: Donors gender and BW (kg), 19M/8F: 88.4 +/- 7.3 Hemoglobin (g/dl): 15.4 +/- 1.3 Platelet precount (x 10(3)/microl): 308 +/- 45 Volume of blood processed (1): 5.5 Procedure time (min): 81 +/- 3 Platelet yield (x 10(11)): 6.8 +/- 0.6 Avg Hemoglobin content of PBRC (g): 102.6 +/- 12.3 WBC contamination of the platelets: 6.8 +/ -10(5).

Cellular contamination of plasma collected with various apheresis systems.

Efforts to improve the purity of blood products have mainly focused on reducing white blood cell (WBC) levels in cellular blood products. Relatively little attention has been given to the cellular purity of plasma. We evaluated plasma units collected on six apheresis systems: Dideco Excel, Haemonetics-MCS+, Fresenius-AS-Tech 204, Baxter-Amicus, Gambro BCT COBE Spectra and Gambro BCT-Trima. Collected plasma volumes averaged 300-350 ml for the various systems. Plasma samples were analyzed for platelet (PLT) content (Technicon H3, Bayer) and residual WBC (Imagn 2000). Results are given below. Platelet levels were consistently low for MCS+, COBE Spectra and Trima (all <50 x 10(3) microl(-1)), and were highest with AS204. Residual WBC levels were relatively low in all systems except MCS+. Extremely low levels were observed in Trima plasma. All of the Trima and Spectra units contained <1 x 10(6) WBC per product. With Excel, AS-Tech 204 and Amicus, 1 to 2 units were found to have >1 x 10(6) WBC, while almost all units from MCS+ exceeded this limit. Different levels of plasma purity were obtained with different apheresis systems. The Gambro BCT COBE Spectra and Trima systems were found to achieve consistently low levels of both platelets and WBC.

Cascade filtration (CF) with the Haemonetics MCS+: a new technical adaptation.

CF was introduced in clinical medicine in 1980. Up to now, exclusively two-vein procedures have been carried out with some limitations to expansion of this technique. In this report we describe the very first application of single-needle CF carried out with Haemonetics MCS + apparatus. Twenty procedures were completed without any untoward effect in patients suffering from TTP, post-hepatitic cryoblobulinemia, familial hypercholesterolemia and acute Guillan-Barrè Syndrome. From 1 to 4 sessions were carried out per patient with the expected laboratory and clinical results. The only limit is the procedure time that averages 231 +/- 48 min., approximately 40% longer than two needle procedures.

Lepromatous vasculitis successfully treated by plasma exchange (PE).

Vascular involvement is presently considered a "common pathway" in a number of diseases that is mediated by circulating immune complexes (CIC). CIC are found in the circulation when the disease is active and in single patients their level may parallel disease activity. Lepromatous leprosis is characterized by the presence of CIC and deposits of immunoglobulins and complement in vascular lesions of the different organs and an Arthus-like mechanism is considered as the basis for the clinical picture. The same mechanism is considered to play an essential pathophysiologic role in Lucio's phenomenon, which is characterized by lymphohistiocytic vascular infiltrates with or without thrombosis and secondary cutaneous infarction. Lepromatous vascular involvement is mediated by CIC whose antigen composition is known, the same as it is with HCV mediated cryoglobulinemia, HBV positive panarteritis nodosa, rheumatoid vasculitis, or Wagner's granulomatosis, which are usually treated by PE [1-3]. PE has been employed for lepromatous vasculitis since 1979 [4] and other cases have been successfully treated afterwards [5,6]. We report on another patient successfully treated by plasma exchange.

The MCS + revision A2 for highly efficient PBSC collection.

Autologous PBSC transplantation is an integral component of the management of hemato-oncology patients. In order to reduce the number of sessions needed to collect the desired number of repopulating cells there has been significant research activity in developing progressively more and more effective technologies and techniques. Recently our group has been involved in the rejuvenation of the MCS + apparatus for both platelet and PBSC collection. The so called "version A2 protocol" is aimed at collecting PBSC in a very efficient way. This protocol is characterized by high blood flow rates both in the collection and reinfusion (80 ml/min) recirculation (56 ml/min) and collection phases (30 ml/min). Only one recirculation is carried out every 5 cycles and only from 5 to 7 are carried out for a single procedure. Twenty-seven collections were carried out of which 25 were evaluable in terms of PBSC efficiency. These averaged 68.8% in an average procedure time of 3.5-5 h for processing an average of 7,052 ml of blood. The RBC contamination was reduced to approximately 5.02 g of hemoglobin and the average volume of the product to 177 ml. If these results are confirmed, the gap between CFC and DFC PBSC is progressively closing.