ALBERT

Description: Abstract 4860 Our flow cytometry laboratory processed 38 bone marrow (BM) samples (24 males and 14 females) from patients with Myelodisplasic Syndrome diagnosis, from January 2008 to May 2009. Upon examination, most BM smears corresponded with an hypercellular bone marrow and increased intermediate and immature cells. All patients classified according to OMS for Myelodisplasic Syndromes. Twenty-four (n=24) patients (65%) presented RA, five (n=5) patients (13%) RAEB-1, one (n=1) 1% presented RARS, and eight (n=8) patients (21%) classified as RAEB-II. Cytogenetic result analyses for all patients classified according with the Index Prognostic Standard Score (IPSS). Twenty-five (n=25) patients or 66% corresponded with a good cytogenetic risk; seven (n=7) or 20% corresponded with an intermediate risk and six (n=6) or 14% with poor risk. Cell cytometry results corresponded with increased immature cells. In 20 patients, the DR/CD34 average 27,29% (range 0-67); in 24 patients, the DR/CD33 average 31,80% (range 6-100); in 29 patients, the DR/CD117 average 18,89% (range 0-43); in 7 patients, the CD33/CD34 average 22,14% (range 0-50); in 5 patients, the CD38 average 47,60% (range 32-61); in 4 patients, the BCL2 average 21,75% (14-26 and in 3 patients, the DR/CD64 average 44,33% (range 30-60). All patients with high average of DR/CD64 classified as CMML. Patients with RAEB-II diagnosis classified as MDS in transformation. Most patients displayed 10% CD34+ values higher than normal human average. We conclude the cytometry is an important tool in the differential diagnosis of MDS. Disclosures No relevant conflicts of interest to declare.

Description: Classic Hodgkin Lymphoma (cHL) is a germinal center derived lymphoma with 8,500-9,000 new cases/year diagnosed in the US. Despite 90% stage I cHL patients can respond to current systemic therapy, this drops to 60%, when diagnosed in advanced stages. Furthermore, 20-30% of diagnosed patients, would be refractory or would relapse and have a poor prognosis. Refractory and relapsed disease (RRD) is currently the challenge when treating cHL patients. There is no specific therapy to offer rather than rescue chemotherapy schemes, which fails in 50% of the cases and associates with high risk severe toxicity. This highlights the need to deeper understand the cHL molecular biology, the screening for molecular markers suitable to identify the risk of refractory and relapse disease and specific therapeutic directed-targets. We have previously reported that the alternative NFkB pathway, mediated by Rel-B and NIK (NFkB Inducing Kinase), plays an important role in cHL survival. Its constitutive activation sustains high BCL2 expression levels and seems to be involved in the RRD. BCL2 was found as a specific Rel-B target gene in cHL cells by ChIP-Seq (Chromatin Immunoprecipitation sequencing) and expression arrays. BCL2 exogenous expression was enough to partially rescue the death induce in cHL cells, which highlight the relevance of this alternative NFkB pathway target gene. Since the BCL2 data was obtained in human cHL cell lines established from patients with refractory and relapsed disease, we decided to analyze whether mediators of this pathway and BCL2 could be useful as prognosis markers and would represent potential targetable factors in both refractory and relapsed disease. We analyzed NIK and BCL2 citoplasm expression in Hodgkin Reed-Sternberg cells (HRS) in the lymph node biopsies of 113 cHL naïve of therapy patients by inmunohistochemistry [52 female Md age and (range) 36 (6-88), 61 male 40.7 (9-78)]. The follow-up period range from 6 to 136 months. The univariate analysis showed no correlation between NIK or BCL2 expression and the prognosis clinical and pathological parameters, including the PET Scan indicated at the end of the first line treatment, neither the molecular markers routinely assayed. The statistical significance was maintained in multivariate analysis (Logistic and Cox Regression p=0.01). NIK expression did not associate with prognosis but the BCL2 expression level correlated with lack of response to conventional therapy and both early and late disease progression. The survival analysis, using the Kaplan-Meir curves, showed that patients with ≥60% positive HRS cells had a shorter disease-free survival (DFS) [Log Rank Test (Mantel Cox) p=0.002] and a reduced overall survival (OS) [Log Rank Test (Mantel Cox) p=0.02]. L1236, U-H01, KM-H2, SUPDH1 and L540, human cHL cell lines that express BCL2 protein, were sensitive to venetoclax, a specific BCL2 inhibitor. The drug induced a cell cycle arrest in S-Phase when treated with 1uM each 24 hours during 10 days, as compared to wild type cells and cells treated with the vehicle. In summary, we found that the alternative NFkB pathway plays a role in the refractory and relapsed classic Hodgkin Lymphoma disease, being BCL2 one of its key downstream target genes. BCL2 can be used as a prognosis marker determined by routine immunohistochemistry at diagnosis of the primary disease. BCL2 expression correlated with refractory disease to first line conventional therapy and disease progression. Based on the venetoclax effect in cHL cell lines we believe BCL2 directed-therapy in cHL should be considered in the subgroup of cHL patients that express this protein in ≥60% HRS cells in the lymph node biopsy performed at diagnosis. Disclosures No relevant conflicts of interest to declare. OffLabel Disclosure: venetoclax used to specifically block BCL2.

Description: Thrombosis and/or disseminated intravascular coagulation (DIC) are complications specifically associated with the use of factor IX complex in some patients. Assuming that these complications might result from zymogen overload, we have produced, using diethylaminoethyl (DEAE)- Sephadex (Pharmacia, Piscataway, NJ) and sulfated dextran chromatography, a factor IX concentrate (coagulation factor IX) that is essentially free of prothrombin, factor VII, and factor X. Factor IX specific activity is at least 5 U/mg protein, a 250-fold purification compared to plasma. Amounts of factors II, VII, and X are less than 5 units each per 100 units of factor IX. The concentrate is essentially free of activated clotting factors and contains no added heparin. In the rabbit stasis model, a dose of 200 factor IX U/kg was less thrombogenic than 100 factor IX U/kg of the DEAE-Sephadex eluate from which the concentrate was derived. Infusion of 200 factor IX U/kg did not induce DIC in the nonstasis rabbit model, whereas 100 factor IX U/kg of the DEAE-Sephadex eluate resulted in DIC in this model. Several factor IX lots were found to have shortened nonactivated partial thromboplastin times (PTTs), but were nonthrombogenic in both animal models. These data indicate that coagulation factor IX concentrate is less thrombogenic than factor IX complex.

Description: INTRODUCTION: A sizable proportion of elderly acute myeloid leukemia (AML) patients receive frontline hypomethylating agents (HMAs), namely azacitidine (AZA) and decitabine (DAC), as they are deemed unfit for intensive chemotherapy (ICT) by their treating physicians. A foreseeable high early death (ED) rate and lack of overall survival (OS) benefit under ICT are the main drivers for this decision. Several groups have published different predictive tools for ED or OS in elderly patients receiving ICT but, since ED in patients treated by HMAs is lower, the research activity has been restricted to OS in this population. METHODS: 415 elderly AML patients (264 M, 152 F) aged 61-90 receiving frontline HMAs (AZA 297, DAC 118), either in daily practice or within clinical trials (AZA 27, DAC 17), with complete relevant clinical information (see Table I) were available from the PETHEMA epidemiologic AML registry (NCT02006004). We analyzed the predictive value for ED (8wk) of the prognostic factors for OS/ED in AML included in the Walter, MRC/LRF, ALFA and ALMA scoring systems, namely age, WBC count, performance status (PS), MRC 2010 cytogenetics, platelet count and secondary disease, as well as the type of HMA. The potential predictors were categorized following previous published models (Walter, MRC/LRF, ALFA, ALMA). Cumulative early death rate at 8 weeks was calculated by the life-time method and the relevant strata were tested for univariate significance by the Wilcoxon test. All significant covariates were included in a Cox multivariate regression model and those significant for death at 8wk were included in a new predictive tool (HMA-EDS). Patients were assigned randomly in a 1:1 ratio to a training cohort (TC) and a validation cohort (VC). The different scoring systems (Walter, MRC/LRF, ALFA, ALMA, HMA-EDS) were checked for their prognostic impact on ED. Finally the 95% CI for the expected death rate at 8wk for the different strata of the new model was calculated for the full patient series. RESULTS: 51 patients out of 415 died and 13 were lost to follow-up before day 56 (cumulative ED rate at 8wk 13%, 95%CI 9-17%). Age, cytogenetics, secondary AML, platelet count and type of HMA were not significantly associated to ED. PS and WBC count strata confirmed their prognostic utility both in univariate and multivariate analysis (Table II). We developed the HMA-EDS by adding WBC (cutoffs 10 and 50, scores 1/2/3) and PS (0-1/2-4, scores 0/1) that classified patients in low-risk (score 1-2/ 84.6% of patients) and high risk (3-4/ 15.4% of patients) strata. When the prognostic utility for ED in the TC and the VC for the different scoring AML systems were checked, only HMA-EDS predicted ED in both cohorts (see Table III). The new EDS discriminates 2 different strata for ED at 8wk in unfit AML patients treated by HMA (see Figure 1 & Table III), namely a lower-risk group (ED rate 10%, 95% CI 6-14) and a high-risk group (ED rate 26%, 95% CI 14-38). CONCLUSIONS: WBC count and PS are the main predictors for ED in unfit AML patients treated by HMAs. A new tool (HMA-EDS) discriminates two different risk groups and supersedes other previously published prognostic systems (Walter's, Wheatley's MRC/LRF, ALFA and ALMA) for this purpose. This score could be useful to select patients for front-line HMA or even HMAs-based combination therapies, given that several cycles are usually needed to achieve a clinical response. We suggest that other patient-related covariates such as geriatric assessment be checked in future studies. Disclosures Ramos: Daiichi Sankyo: Honoraria; Celgene: Consultancy, Honoraria, Membership on an entity's Board of Directors or advisory committees, Research Funding; Janssen: Consultancy, Honoraria; Abbvie: Honoraria. Fernandez:Incyte: Membership on an entity's Board of Directors or advisory committees, Speakers Bureau; Teva: Membership on an entity's Board of Directors or advisory committees, Research Funding, Speakers Bureau; Daiichi Sankyo: Consultancy, Membership on an entity's Board of Directors or advisory committees, Research Funding, Speakers Bureau; Janssen: Membership on an entity's Board of Directors or advisory committees, Research Funding, Speakers Bureau; Celgene: Consultancy, Membership on an entity's Board of Directors or advisory committees, Research Funding, Speakers Bureau; Karyopharm: Membership on an entity's Board of Directors or advisory committees, Research Funding; Abbvie: Membership on an entity's Board of Directors or advisory committees; Pfizer: Membership on an entity's Board of Directors or advisory committees, Research Funding, Speakers Bureau; Novartis: Membership on an entity's Board of Directors or advisory committees, Research Funding, Speakers Bureau.

Description: The recent success of immunotherapy using chimeric antigen receptor modified T cells (CAR T) in B-cell malignancies highlights the potential of these cytotoxic "drugs" for cancer therapy. CAR T therapies generally rely upon manufacturing approaches that include prior T cell activation through engagement of the TCR and costimulatory receptors followed by ex vivo expansion of patient-derived T cells over days to weeks. We previously reported that CD3/CD28 stimulation prior to transduction promotes progressive T cell effector differentiation over time in culture with loss of CAR T cell potency (Ghassemi et al. 2018 PMID: 30030295). Since cell division is not a prerequisite for lentiviral vector-mediated gene delivery, we hypothesized that lentiviral transduction of quiescent T cells without prior activation will enhance engraftment and persistence of CART cells that is associated with long-term leukemia control. Here, we show that functional CD19-specific CAR T cells (CART19) can be generated in as little as 24 hours using lentiviral vectors without the need for prior T cell activation. We showed using a non-optimized process that a mean of 6.5% (range 2%-10%) of freshly isolated quiescent T cells can be transduced using an infrared red fluorescent protein (iRFP)-expressing lentiviral vector with slower kinetics of expression compared with activated T cell transduction (peak at 96 hrs vs. 48 hrs for quiescent and activated T cells, respectively). Although substantially less efficient compared to activated T cells, transduction was detected across all T cells subsets with central memory T cells showing the greatest transduction efficiency with a mean of 4-fold greater transduction compared with naïve T cells. Somewhat unexpectedly, CART19 cells generated from quiescent T cells using a CD19-specific CAR vector showed a 3-5 fold greater transgene expression compared with iRFP vectors transduced at similar MOI. However, we show that CAR expression can occur in quiescent T cells even without reverse transcription or integrase function, so called "pseudotransduction". Importantly, we show that this CAR expression produces T cells with cytolytic activity and effector cytokine production in response to antigen that is similar to activated and transduced CAR T cells. Using the well-characterized Nalm6 model of acute lymphoblastic leukemia, we show that CART19 cells generated by transduction of quiescent T cells for 16 hours followed by washing to remove vector exhibit dose-dependent anti-leukemic activity that is durable with injection of as little as 2x105 total T cells. We estimate the latter to contain ~2x104 T cells with integrated lentiviral vector based upon transduction efficiency determined in studies using non-tumor bearing mice. (Fig 1). In summary, our results support the need for further investigation of CAR T cells that are generated using engineering of quiescent T cells. Taking advantage of the ability of lentiviral vectors to transfer genes to quiescent T cells, the highly abbreviated and simplified manufacturing approach described here has the potential to enhance therapeutic potency while also substantially reducing the materials and labor costs associated with current manufacturing approaches that use activated and expanded T cells. In addition, the rapid nature of this manufacturing has the potential to extend the population of patients that may be treated with these therapies by shortening the interval between aphaeresis collection and re-infusion of CAR T cells, which prevents the treatment of some patients with rapidly progressive disease. Disclosures Ghassemi: Novartis: Patents & Royalties. Milone:Novartis: Patents & Royalties, Research Funding.

Description: Background: The trombopoietin receptor agonists (TRAs) romiplostim and eltrombopag are effective and safe in the treatment of chronic immune thrombocytopenia (ITP). However, when no response is achieved or when adverse events occur with one TRA the value of the sequential use of romiplostim and eltrombopag has not been clearly established. Here we have evaluated the efficacy and tolerance of using eltrombopag after romiplostim in ITP. Methods: Fifty-one primary ITP patients (aged 18 years or more) who had been sequentially treated first with romiplostim and then with eltrombopag in the Spanish Eltrombopag Registry were retrospectively evaluated. In accordance with the usual standards, complete response was defined as a platelet count of 100x109/L and a response as a platelet count of 30x109/L or a count of at least twice the initial (pre-treatment) value. This study was performed in accordance with the standards of the Helsinki declaration and approved by the Hospital Universitario de Burgos Ethics Committee. Results: The median age of our cohort was 49 [range, 18–83] years. There were 32 women and 19 men. According to the standard definition, patients were allocated to newly diagnosed (n=2), persistent (n=5) and chronic (n=44) ITP groups. The median number of therapies prior to administration of eltrombopag was 4 [range, 2–9], including splenectomy (39%), rituximab (33%) and romiplostim (100%). The median duration of romiplostim use before switching to eltrombopag was 12 (IQR 5–21) months. The reasons for switching from the romiplostim to eltrombopag were: lack of efficacy of romiplostim (n=25), patient's preference (n=16), platelet-count fluctuation (n=6), and side-effects (n=4). The initial response rate to eltrombopag was 41/51 (80.5%), including 67% (n=34) of cases with complete remission. After a median follow-up of 13 months with eltrombopag, 39 patients maintained their response. When eltrombopag was used for patients who were refractory to the maximum romiplostim dose the initial response rate of eltrombopag was 25%. However, 83% of patients who relapsed after their initial response to romiplostim responded to eltrombopag. Sixteen romiplostim responders requested their physicians to switch them to eltrombopag because they preferred an oral drug. The efficacy was maintained after switching in all 16 patients. In the platelet-count fluctuation group, the initial response rate was also 100%. All 4 patients who were switched to eltrombopag because they experienced side-effects of romiplostim achieved complete remission with eltrombopag and their adverse events were resolved. 16 / 51 (33%) patients experienced one or more adverse event during treatment with eltrombopag. The frequency of grade 3–4 adverse events during treatment with eltrombopag was 9.8%. Conclusion: The use of eltrombopag after romiplostim for treating ITP is effective and safe. The reason for discontinuing romiplostim was associated with the response to eltrombopag. Disclosures No relevant conflicts of interest to declare.

Description: Leukemia relapse occurring in donor cells, so called donor cell leukemia (DCL) after allogeneic hematopoietic stem cell transplantation has been previously reported in the literature. Some authors have suggested that the development of DCL is perhaps a more common occurrence than traditionally thought. Donor cell myeloma (DCM) seems to be less frequent than DCL. This 46-year old male when first seen in 2000 was diagnosed with stage IIIa multiple myeloma. A monoclonal IgA kappa spike was recorded at diagnosis. Treatment with melphalan and prednisone was delivered every four to six weeks for a total of 22 courses. Fourty months after the initial diagnosis, an M2 acute myelogenous leukemia was identified. Treatment with chemotherapy resulted in complete remission. Matched UCB cells were localized at the London Cord Blood Bank. The UCB belonged to a male product of a white western European mother and a black Nigerian father who was a carrier of hemoglobin S. Hemoglobins A, F and S were detected in the UCB, consonant with sickle cell trait. The patient was allografted employing the "Mexican" NST conditioning regimen, granulocyte count recovered to more than 0.5 x 109/L on day 14, with the platelet count never dropping below 20 x 109/L. On day +40, the polymorphic microsatellite markers revealed mixed chimerism. The hemoglobin S gene was identified on day +20 and on day +60, full chimerism was shown. Cyclosporine A was stopped on day +350. The patient returned 170 months after the transplant with low back pain and the bone marrow aspiration disclosed 80% abnormal plasma cells, an IgA kappa monoclonal spike of 3.1 gr/dl, and complete chimerism. Malignant plasma cells were sorted by means of flow cytometry before genetic fingerprinting; cells were stained with an admixture of fluorescent monoclonal antibodies and cells co-expressing dim CD45, bright CD38 and CD56 were sorted out to ≥99% purity. Sorted cells were shown to have donor origin (Figure 1). The patient was treated with thalidomide, dexamethasone and bortezomib and the monoclonal spike disappeared; an autologous stem cell transplant is planned. Most people consider that the development of a malignancy in the cells of the donor is a rare event and very few prospective studies have analyzed the real prevalence of this phenomenon. Prospectively, we have found that 7% (95% CI 2.9 to 13.6%) of patients with leukemic activity after an allogeneic graft do have a donor cell-derived leukemia; this figure contrasts with those described elsewhere in non-prospective studies. A major problem in the analysis of donor cell derived malignancies is that demonstration of the donor cell origin of malignant activity. In this case, the demonstration of DNA of the donor in the fluorescence-activated sorted malignant plasma cells is indicative of the origin of the myeloma cells. Interestingly, the immunoglobulin type produced by the initial myeloma cells is the same as that of the donor-cell myeloma; Despite being two myelomas producing the same immunoglobulin subtype, both should be considered as de novomalignancies and as such, treated; we have previously shown that donor cell leukemias do have a response when treated as de novo, non-secondary leukemias. To our best knowledge, this is the second report of DCM following allogeneic HSCT. Prior to this case, Kim et al reported a DCM after an allogeneic transplant in a patient with refractory anemia with ringed sideroblasts. Previously, two cases have been reported of donor-origin MM, but they occurred in patients who underwent solid organ transplantation of the kidney and heart-lung. Kumar et alreported a case of DCM developing after unrelated allogeneic HSCT in the both donor and recipient but they did not conducted a comprehensive molecular cytogenetic study. In the case published by Maestas et al, an abnormal proliferation of plasma cells was identified in the donor, thus making possible that a malignant plasma cell clone was already present in the donor stem cells. In summary, we have clearly shown that this patient has had three different malignancies: 1) De novomultiple myeloma, 2) Secondary acute myelogenous leukemia and 3) De novodonor cell-derived multiple myeloma. The mechanisms involved in these episodes could be useful to better understand tumorigenesis. Disclosures No relevant conflicts of interest to declare.

Description: Abstract 3011 Poster Board II-987 Somatic mutation of the X- linked gene PIG-A results in partial or absolute deficiency of cell surface expression of glycosylphosphatidylinositol-anchored proteins (GPI-APs). In humans, this genetic event in a hematopoietic stem cell leads to the blood disease paroxysmal nocturnal hemoglobinuria (PNH). PNH has two major clinical presentations: predominantly hemolytic without overt marrow failure, referred to as classic PNH, and PNH clonal expansion in the setting of marrow failure, or the aplastic anemia/PNH syndrome (AA/PNH). The diagnosis of PNH is based on the presence of GPI-AP-deficient clones mainly within erythrocytes and granulocytes. Since PNH is a stem cell disorder, all hematopoietic lineages are involved and GPI-AP-deficient clones are found among platelets, monocytes and lymphocytes. Except for their different clinical presentations, classic PNH and AA-PNH have never been shown to have specific discriminating biological features. Indeed, we previously reported no difference in the expression profile of CD34 cells in hemolytic and aplastic forms of PNH (Chen G et al, Leukemia 2005; 19:862). As marrow failure in PNH, as in aplastic anemia, is immune-mediated, and T cells are affected by the PIG-A mutation, we hypothesized that the mutant subset of lymphocytes might differ in PNH. We compared by microarray the T cell compartment in both forms of the disease. We sorted peripheral blood GPI-AP-normal and -deficient pooled CD4 and CD8 T cells using CD59 and CD55 as GPI-AP markers, and subjected each subset to array hybridization and analysis. Samples were obtained from 1 patient with classic PNH, 3 patients with AA-PNH syndrome and 3 age-matched normal donors as controls. In preliminary control experiments, we determined using microarray of total RNA derived from healthy donors' T cells, with and without GPI-AP-specific antibodies, the absence of altered gene expression pattern or activation of specific genes due to selective antibody binding to normal cells. By principal component analysis (PCA), the phenotype of the PNH clone in classic PNH was divergent from the AA-PNH clone and from that of normal donors. By consistency test, only 15 probesets/genes resulted common in the comparison between GPI-AP-deficient and GPI-AP-normal cells between classic PNH and AA-PNH syndrome. Using Ingenuity Software, we found that the represented probesets/genes were involved in some major canonical pathways such as cell death, immune and lymphatic system development, immune response, and cell-to-cell signaling interaction. In order to investigate the different gene expression profile derived from specific subset of T cells, we sorted GPI-AP-deficient and GPI-AP-normal CD8 T cells using CD48 and CD52 as markers. When we compared GPI-AP-deficient and GPI-AP-normal cells between classic PNH and AA-PNH syndrome, we found that only 23 probesets/genes were up-regulated and 132 down-regulated confirming the divergent gene expression profile between the two diseases (Figure 1). We conclude that the restricted similarity in the gene expression profile in the GPI-AP-deficient T cells between classic PNH and AA-PNH syndrome leads to a specific “signature” of genes that might support the different clinical presentation and evolution of the two diseases.Figure 1.Divergent gene expression profile between GPI-AP-deficient and GPI-AP-normal cells in classic PNH and AA-PNH syndromeFigure 1. Divergent gene expression profile between GPI-AP-deficient and GPI-AP-normal cells in classic PNH and AA-PNH syndrome Disclosures: No relevant conflicts of interest to declare.

Description: Three patients with thrombotic thrombocytopenic purpura (TTP) were treated by infusion of normal plasma with dramatic responses. The plasmas collected from these patients during relapse induced in vitro aggregation of washed platelets from both normal donors and the patients during remission. The platelet aggregating factor was not dialyzable or adsorbable by Al(OH)3 and was not inactivated by diisopropylfluorophosphate, hirudin, or heparin in the presence of normal amounts of antithrombin. In contrast to the platelet aggregation induced by platelet isoantibody, the platelet aggregating activity of TTP plasma diminished as a function of time when it was incubated with normal plasma at 37 degrees C. These observations suggest that at least some instances of TTP appear to be due to deficiency of a plasma inhibitor to counteract a platelet aggregating factor demonstrated to be present in the plasma of these patients.

Description: Three 59Fe-labeled nonheme components of the cytosol were identified when rabbit reticuloyctes were incubated with 59Fe-labeled plasma under conditions in which the iron supply was not limiting. Two of these components were identified as ferritin and transferrin. The latter was characterized by gel filtration as having apparent molecular weight higher than transferrin, indicating that the transferrin may be complexed to another moiety. The third component, referred to as iron- binding protein-I (IBP-I), is as yet uncharacterized. When the reticulocytes were incubated with unlabeled plasma after pulse-labeling with 59Fe-labeled plasma, 59Fe radioactivity in these cytosol components decreased; after 15 min of chase, the 59Fe in ferritin, transferrin, and IBP-I fell to 64.6%, 26.5%, and 65.8% of the initial values, respectively. A good correlation existed between the decrease of 59Fe in these three nonheme compartments and the associated increase in 59Fe-heme. The data presented suggest that cytosol ferritin, transferrin, and IBP-I are intermediates in the transport of 59Fe from the plasma membrane to the mitochondria.