Eapen K. Jacob

Carboxypeptidase M: a biomarker for the discrimination of well-differentiated liposarcoma from lipoma

The discrimination between well-differentiated liposarcomas/atypical lipomatous tumors and lipomas can be diagnostically challenging at the histological level. However, cytogenetic identification of ring and giant rod chromosomes supports the diagnosis of well-differentiated liposarcoma/atypical lipomatous tumor. These abnormal chromosomes are mainly composed of amplified genomic sequences derived from chromosome 12q13-15, and contain several genes, including MDM2, CDK4 (SAS), TSPAN31, HMGA2, and others. MDM2 is consistently amplified in well-differentiated liposarcomas/atypical lipomatous tumors, and up to 25% in other sarcomas. As part of a large genomic study of lipomatous neoplasms, we initially found CPM to be consistently amplified in well-differentiated liposarcomas/atypical lipomatous tumors. To further explore this initial finding, we investigated the copy number status of MDM2 and CPM by fluorescent in situ hybridization (FISH) on a series of 138 tumors and 17 normal tissues, including 32 well-differentiated liposarcoma/atypical lipomatous tumors, 63 lipomas, 11 pleomorphic lipomas, 2 lipoblastomas, 30 other tumors and 17 normal fat samples. All 32 well-differentiated liposarcoma/atypical lipomatous tumors showed amplification of MDM2 and CPM, usually >20 copies per cell. The other tumors lacked MDM2 and/or CPM amplification. Chromogenic in situ hybridization confirmed the above results on a subset of these tumors (n=27). These findings suggest that identification of CPM amplification could be used as an alternative diagnostic tool for the diagnosis of well-differentiated liposarcoma/atypical lipomatous tumors.

Positive virtual crossmatch with negative flow crossmatch results in two cases

Pre-transplant (Tx) presence of HLA antibodies (HLA-Ab) especially donor specific antibodies (DSA) has been correlated with post-Tx rejection. While crossmatch (XM) is the specific method to identify DSA, logistical reasons prevent performing a prospective XM in all transplants. In such cases DSA as identified by solid-phase assay (SPA) are being used to perform a virtual crossmatch (VXM). We present two cases, a heart-lung transplant and a kidney transplant, for which testing detected a presumptive DSA with discordant results: a negative flow cytometric crossmatch (FXM) and a positive VXM using SPA. The subsequent investigation determined the antibody, in both cases, was presumably directed against an epitope of a HLA-B*44 antigen found on the single antigen beads (SAB) used in the SPA but not against the native form on the donor lymphocytes used in the FXM. Manufacturing of SAB beads results in denaturation of epitopes, majority of which are removed from the final product, but residual amount is present on the final product. Denaturation of majority of antigen epitopes on single antigen beads did not remove the activity of the recipients antibodies but it did diminish the activity of positive control serum. This indicates denaturation of some of the HLA-B*44 antigen during manufacturing of the SAB may have lead to the reactivity. Antibody mediated rejection does not appear to be associated with the titer of this antibody to denatured antigen in the first case and so clinical relevance of such antibodies is unclear. Subsequently a second case of discordant FXM and VXM was identified in a potential kidney transplant patient who went on to an uneventful transplant. In this case, lymphocytes from the donor were positively shown to express HLA-B*44:02 using known anti- HLA-B*44:02 control serum. Platelets identified as HLA-B*44:02 could adsorb the anti-HLA-B*44:02 from the control serum activity but not from that of the recipients anti- HLA-B 44 antibody adding evidence that this antibody should best be classified as a false positive finding. The presence of such an antibody if misidentified may result in unnecessary therapy being instituted or the inappropriate denial of an organ for transplantation.

Combined corticosteroid and catecholamine stimulation of aqueous humor flow

PURPOSE Epinephrine is known to stimulate aqueous humor flow in humans. Corticosteroids are known to augment the effect of beta-adrenergic agonists on the ciliary body. This experiment was carried out to determine whether a corticosteroid can increase the stimulatory effect of epinephrine on aqueous humor flow. METHODS Twenty human volunteers were studied for 24 hours. Hydrocortisone was given orally and epinephrine was given intravenously during sleep while aqueous flow was monitored. Flow was compared with a second 24-hour study when oral and intravenous placebos were given. The sequence of administration was randomized. Subjects and investigators were masked. The flows also were compared with a previously published study in which the same dose of epinephrine was administered without steroid. RESULTS Epinephrine plus hydrocortisone compared with placebos increased aqueous flow 42% in subjects during sleep. The combination of epinephrine and hydrocortisone was a more potent stimulus to aqueous flow than epinephrine alone, which increased aqueous flow by only 27% (P = 0.045). CONCLUSION The two major hormones of the adrenal gland work in concert to increase the rate of aqueous humor flow in humans.

Infusion technique of hematopoietic progenitor cells and related adverse events (CME)

The use of hematopoietic progenitor cell (HPC) transplant has risen over the past two decades. A variety of adverse events (AEs) of varying severity have been noted during HPC infusions. These AEs have been associated with several factors such as the amount of dimethyl sulfoxide and white blood cells in the HPC product. We performed a single‐institution retrospective analysis to determine the effect of two different HPC infusion techniques, manual push with syringes versus infusion from bags with the aid of gravity, on the occurrence of infusion‐related AEs.

ABO blood group incompatibility as an adverse risk factor for outcomes in patients with myelodysplastic syndromes and acute myeloid leukemia undergoing HLA‐matched peripheral blood hematopoietic cell transplantation after reduced‐intensity conditioning

ABO incompatibility is not a contraindication to hematopoietic cell transplantation (HCT), but it has been associated with additional risks including delayed engraftment, pure red cell aplasia (PRCA), and higher transfusion needs. Data on these events and on patient survival after reduced‐intensity conditioning (RIC) HCT are limited.

Defining a reference range for cold agglutinin titers

The cold agglutinin (CAGG) titer is offered at our institution to aid in diagnosing cold agglutinin disease (CAD). Our goal was to create a seasonally adjusted reference range using prospective samples and compare it to a reference range generated retrospectively.

Clinical Outcomes of HLA-DPB1 Mismatches in 10/10 HLA-Matched Unrelated Donor-Recipient Pairs Undergoing Allogeneic Stem Cell Transplant

HLA‐DPB1 matching may impact allogeneic hematopoietic stem cell transplantation (ASCT) outcomes; however, this locus is not in linkage disequilibrium with the remainder of the HLA genes. After classifying HLA‐DPB1 mismatches based on T‐cell epitope, avoiding non‐permissive mismatches may impact survival. We tested this hypothesis at a single academic institution.

Clinically significant anti‐A1 in a presumed ABO‐identical hematopoietic stem cell transplant recipient: a case report

BACKGROUND: Subgroups of the blood group A (ABO) are generally not considered ABO incompatible for hematopoietic progenitor cell (HPC) transplant.

Current Practices and Supply Logistics Do Not Support HLA Matching for Red Cell Transfusions

In some sense the first suggestion becomes moot if the second is practical, as it would also address the accepted cause of alloimmunization, the presence of mismatched white blood cells. Such matching would allow for decreased nonself HLA exposure be it from red cells or white cells. However, is this concept practical or even advisable? Simple maneuvers such as leukoreduction (LR) likely add 30% to the blood management and logistical costs of blood components currently (2). Extensive HLA matching would incur enormous costs including HLA typing of donors and recipients and incur huge logistical costs in the attempt to transport units to the appropriate locations. Vassallo estimated in platelet refractoriness, in which HLA class I A and B loci are only considered, that most blood banks could only provide HLA identical units in 10–20% of cases (3). To be effective in regards to transplants, some of which are affected by HLA class I and II, the authors’ concept would need to match for 6–10 loci thereby increasing the difficulty of obtaining a match by many orders of magnitude. The National Marrow Donor Program (NMDP) is designed to provide such matching for hematopoietic stem cell transplants yet even with it’s 9 million potential donors approximately 35% of searches yield partial matches or mismatches (4). One must remember that in HPC transplants only one dose is needed. In blood component therapy, it is the rare patient that needs only one transfusion of red blood cells or platelets. Such needs would require a program with a donor pool many times the size of the NMDP. (Additionally the assertion that the NMDP registry could be used for this purpose is inappropriate as NMDP donors have registered to donate hematopoietic stem cells for transplants not blood donation for elective surgeries.)

Extracorporeal Photopheresis Improves Survival in Hematopoietic Cell Transplant Patients with Bronchiolitis Obliterans Syndrome Without Significantly Impacting Measured Pulmonary Functions

We carried out the first matched retrospective cohort study aimed at studying the safety and efficacy of extracorporeal photopheresis (ECP) for bronchiolitis obliterans syndrome (BOS) after allogeneic hematopoietic cell transplantation (HCT). Medical records of 1325 consecutive adult patients who underwent HCT between 2005 and 2015 were reviewed. Seventy-four patients (median age, 51 years) with a diagnosis of BOS were included in the study. After propensity-score matching for BOS severity, 26 patients who underwent ≥3 months of ECP were matched to 26 non-ECP-treated patients, who were assigned an index date corresponding to the ECP start date for their matched pairs. The rate of decline in FEV1 percentage predicted (FEV1PP) decreased after ECP initiation (and after index date in the non-ECP group), with no significant difference between the 2 groups (P = .33). On a multivariable analysis that included baseline transplant and pulmonary function test variables, matched related donor HCT (HR, .1; 95% CI, .03 to .5; P = .002), ECP (HR, .1; 95% CI, .01 to .3; P = .001), and slower rate of decline in FEV1PP before the ECP/index date (HR, .7; 95% CI, .6 to .8; P = .001) were associated with a better overall survival. At last follow-up, non-ECP-treated patients were more likely to be on >5 mg daily dose of prednisone (54% versus 23%; P = .04) and had a greater decline in their Karnofsky performance score (mean difference, -9.5 versus -1.6; P = .06) compared with ECP-treated-patients. In conclusion, compared with other BOS-directed therapies, ECP was found to improve survival in HCT patients with BOS, without significantly impacting measured pulmonary functions. These findings need prospective validation in a larger patient cohort.