Supplementary Tables 1-3, Figure 1 from Targeting the Lymphotoxin-β Receptor with Agonist Antibodies as a Potential Cancer Therapy

Improvements in transfusion medicine, blood typing, and donor screening have reduced the likelihood of a reaction from administration of blood products. In an effort to prevent transfusion reactions, our current standard is to administer acetaminophen 650mg and diphenhydramine 25mg, 30 minutes before transfusions. This practice may unnecessarily place patients at increased risk for complications including falls, cognitive impairment, and hepatic toxicity. P=Adult BMT/oncology patient; I=Does the prophylactic use of acetaminophen and diphenhydramine; C=versus no premedication; O=affect the incidence of blood transfusion reactions? The evidence based practice committee (EBPC) at a comprehensive cancer center in the Midwest conducted a literature review to determine if evidence supported the efficacy for reducing transfusion reactions with routine premedication utilizing acetaminophen and diphenhydramine. The search strategy included electronic searches of numerous resources including PubMed (MEDLINE), CINAHL (Cumulative Index to Nursing and Allied Health Literature), Medline, Elton B. Stephens Company (EBSCO), Ovid, and Scopus to gather established information. The results of one prospective, randomized, double-blind, placebo-controlled and two retrospective reviews were analyzed and failed to show any benefit to premedication prior to transfusion of blood products. Based on the evidence, the EBPC sought to obtain a baseline rate of transfusion reaction at their facility. Between January and April 2014, a retrospective review was conducted on 137 transfusion patients to determine the frequency of transfusion-related reactions. Four transfusion reactions were reported; all four patients had been premedicated prior to transfusion. Six patients did not receive any premedication; none of which had a transfusion-related reaction. The chair of the EBPC presented these findings at BMTQI. Committee concerns included the possible increase in confounding febrile reactions that might lead to unnecessary admissions and febrile transfusion reaction work-ups. A compromise was reached with the BMTQI committee approving the elimination of Benadryl as a routine premedication. Next steps will include implementation of this new protocol with additional data collection on rate of transfusion reactions for the next three months. If results concur with reviewed evidence, this protocol change will be expanded across the facility and can be a platform for other centers who are still using premedication.

After performing thousands of bone marrow biopsies (BMX), our nurse lead biopsy staff started identifying what is likely intuitively known about specific diseases and their BMX characteristics. We began collecting data both on core lengths and hemodilution rates to see if our data validates our suspicions that patients with plasma cell dyscrasias (PCD) have significantly smaller core lengths and myeloproliferative disorders (MPD) have significantly higher hemodilution rates. This was a chart review from pathology reports on 3199 patients with BMX's performed at Siteman Cancer Center between January 1st 2006 and December 30th 2011. A convenience sample of patients having their BMX at our center was utilized. We reviewed patients BMX results a week following the BMX procedure to monitor job performance of the BMX staff. Data collected included diagnosis, gross bone marrow core length, evaluable bone marrow core, and pathology reported bone marrow aspirate hemodilution. For this review PCD included MM, Monoclonal gammopathy of undetermined significance (MGUS), and Amyloidosis. MPD included Chronic Myeloid Leukemia (CML), Polycythemia Vera (PV), and Myelofibrosis (MF). Our data showed that the mean bone marrow core length among all samples was 1.9 centimeters (cm) with the average evaluable bone marrow core length being 1.3 cm. Mean PCD bone marrow core length was found to be much lower at 1.6 cm with evaluable core length of 1.1 cm. Our mean hemodiluted bone marrow aspirates rate was 12.2%. The percent of CLL hemodiluted bone marrow aspirate was 15.4% which included 2 dry taps and MPD was 17.3% and included 1 dry tap. Of interest, the percent of hemodiluted samples seen in patients with Mastocytosis was 55.6%. In reviewing our data, we noted that only 19 of 3199 patients had a dry tap which seems quite low. It is our thought that this is a reflection of the patient population we treat. Most of the newly diagnosed AML's have their BMX in the hospital. See chart for full data compilation. In our review, it appears that plasma dyscrasias have a bone marrow core less than the average reviewed and CLL, MPD, and Mastocytosis have a higher hemodilution rate in comparison. We are going to review the data further to see if differences can be seen in older vs. younger patients.Tabled 1TOTALALLAAAMLCLLMPDHDMDSMastoPCDNHLOtherTotal Bone Marrow Biopsies3199170549651252157831732598508137Average Core in Centimeters1.91.91.91.91.91.92.01.82.01.62.01.9Average Evaluable Core in CM.1.31.41.41.31.41.31.61.31.51.11.61.3% per Captured41.1%5.3%1.7%30.2%3.9%6.7%2.4%9.9%1.0%18.7%15.9%4.3%% Hemodiluted per Aspirates12.2%9.4%11.9%12.3%15.4%17.3%7.8%10.2%55.6%9.1%6.7%15.7%Drytab1900321121531 Open table in a new tab

B cell activation factor of the TNF family (BAFF) is a potent B cell survival factor. BAFF overexpressing transgenic mice (BAFF-Tg mice) exhibit features of autoimmune disease, including B cell hyperplasia and hypergammaglobulinemia, and develop fatal nephritis with age. However, basal serum IgA levels are also elevated, suggesting that the pathology in these mice may be more complex than initially appreciated. Consistent with this, we demonstrate here that BAFF-Tg mice have mesangial deposits of IgA along with high circulating levels of polymeric IgA that is aberrantly glycosylated. Renal disease in BAFF-Tg mice was associated with IgA, because serum IgA was highly elevated in nephritic mice and BAFF-Tg mice with genetic deletion of IgA exhibited less renal pathology. The presence of commensal flora was essential for the elevated serum IgA phenotype, and, unexpectedly, commensal bacteria-reactive IgA antibodies were found in the blood. These data illustrate how excess B cell survival signaling perturbs the normal balance with the microbiota, leading to a breach in the normal mucosal-peripheral compartmentalization. Such breaches may predispose the nonmucosal system to certain immune diseases. Indeed, we found that a subset of patients with IgA nephropathy had elevated serum levels of a proliferation inducing ligand (APRIL), a cytokine related to BAFF. These parallels between BAFF-Tg mice and human IgA nephropathy may provide a new framework to explore connections between mucosal environments and renal pathology.

Abstract The lymphotoxin-β receptor (LTβR) is a tumor necrosis factor receptor family member critical for the development and maintenance of various lymphoid microenvironments. Herein, we show that agonistic anti-LTβR monoclonal antibody (mAb) CBE11 inhibited tumor growth in xenograft models and potentiated tumor responses to chemotherapeutic agents. In a syngeneic colon carcinoma tumor model, treatment of the tumor-bearing mice with an agonistic antibody against murine LTβR caused increased lymphocyte infiltration and necrosis of the tumor. A pattern of differential gene expression predictive of cellular and xenograft response to LTβR activation was identified in a panel of colon carcinoma cell lines and when applied to a panel of clinical colorectal tumor samples indicated 35% likelihood a tumor response to CBE11. Consistent with this estimate, CBE11 decreased tumor size and/or improved long-term animal survival with two of six independent orthotopic xenografts prepared from surgical colorectal carcinoma samples. Targeting of LTβR with agonistic mAbs offers a novel approach to the treatment of colorectal and potentially other types of cancers. (Cancer Res 2006; 66(19): 9617-24)

The cell surface co-stimulatory protein CD154 (CD40L) is a target for monoclonal antibody (mAb) inhibitors of T-cell mediated immune diseases. This protein, like most other members of the TNF ligand family, forms homotrimeric complexes on the cell surface and in solution, with a three-fold axis of symmetry. We find that several different anti-CD154 monoclonal antibodies form distinctive complexes with soluble CD154. These soluble complexes have been analyzed using size exclusion chromatography, static and dynamic light scattering, and electron microscopy and shown to consist of caged structures of various geometries. The cell surface complexes have been analyzed by confocal microscopy and, depending on the mAb, remain as small, separate complexes or form large aggregates. The formation of these complexes in solution is likely to have an impact on measures of affinity, while the cell surface complexes could affect binding potency and provoke other biological effects.