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The galactose oxidase-Schiff (GOS) reaction detects D-galactose-β-[1,3]-N-acetyl-D-galactosamine. This is a T-antigen expressed in mucus from malignant cells and colonic mucosa adjacent to cancer but not in normal mucosa. Previous studies using a qualitative GOS assay proved to be of limited value for the detection of colorectal neoplasia. We used a newly developed quantitative GOS assay to determine its potential as an early detection biomarker for colorectal cancer. We completed a multi-center, prospective, cross-sectional cohort validation study consisting of 70 normal controls, 23 high-risk normal patients (polyp history or family history of colorectal cancer (CRC) with currently normal colonoscopy), 137 patients with adenomatous polyps, and 69 with colorectal cancers. Prior to colonoscopy, two samples of stool were collected via a rectal exam: one for FOBT, and one for GOS. The area under the ROC curve (AUC) for detecting colonic adenomas and cancer for normal colons, computed with logistic regression was 0.69 for GOS, 0.62 for FOBT, and 0.73 for GOS combined with FOBT. Adding GOS to FOBT did not significantly change the ROC of FOBT alone. GOS does not appear to be a suitable marker of colorectal neoplasia.

Specimen collection is an integral component of clinical research. Specimens from subjects with various stages of cancers or other conditions, as well as those without disease, are critical tools in the hunt for biomarkers, predictors, or tests that will detect serious diseases earlier or more readily than currently possible. Analytic methodologies evolve quickly. Access to high-quality specimens, collected and handled in standardized ways that minimize potential bias or confounding factors, is key to the “bench to bedside” aim of translational research. It is essential that standard operating procedures, “the how” of creating the repositories, be defined prospectively when designing clinical trials. Small differences in the processing or handling of a specimen can have dramatic effects in analytical reliability and reproducibility, especially when multiplex methods are used. A representative working group, Standard Operating Procedures Internal Working Group (SOPIWG), comprised of members from across Early Detection Research Network (EDRN) was formed to develop standard operating procedures (SOPs) for various types of specimens collected and managed for our biomarker discovery and validation work. This report presents our consensus on SOPs for the collection, processing, handling, and storage of serum and plasma for biomarker discovery and validation.

Advanced-stage follicular B-cell lymphoma is considered incurable. Anti-CD20 radioimmunotherapy is effective in patients who have had a relapse after chemotherapy or who have refractory follicular lymphoma, but it has not been tested in previously untreated patients.Seventy-six patients with stage III or IV follicular lymphoma received as initial therapy a single course of treatment with 131I-tositumomab therapy (registered as Tositumomab and Iodine I 131 Tositumomab [the Bexxar therapeutic regimen]). This consisted of a dosimetric dose of tositumomab and 131I-labeled tositumomab followed one week later by a therapeutic dose, delivering 75 cGy of radiation to the total body.Ninety-five percent of the patients had any response, and 75 percent had a complete response. The use of polymerase chain reaction (PCR) to detect rearrangement of the BCL2 gene showed molecular responses in 80 percent of assessable patients who had a clinical complete response. After a median follow-up of 5.1 years, the actuarial 5-year progression-free survival for all patients was 59 percent, with a median progression-free survival of 6.1 years. The annualized rate of relapse progressively decreased over time: 25 percent, 13 percent, and 12 percent during the first, second, and third years, respectively, and 4.4 percent per year after three years. Of 57 patients who had a complete response, 40 remained in remission for 4.3 to 7.7 years. Hematologic toxicity was moderate, with no patient requiring transfusions or hematopoietic growth factors. No cases of myelodysplastic syndrome have been observed.A single one-week course of 131I-tositumomab therapy as initial treatment can induce prolonged clinical and molecular remissions in patients with advanced follicular lymphoma.

CD20-targeted radioimmunotherapy is a promising new treatment for B-cell non-Hodgkin lymphoma (NHL). We now provide updated and long-term data on 59 chemotherapy-relapsed/refractory patients treated with iodine 131I tositumomab in a phase I/II single-center study. Fifty-three patients received individualized therapeutic doses, delivering a specified total-body radiation dose (TBD) based on the clearance rate of a preceding dosimetric dose. Six patients received dosimetric doses only. Dose-escalations of TBD were conducted separately in patients who had or had not undergone a prior autologous stem cell transplant (ASCT) until a nonmyeloablative maximally tolerated TBD was established (non-ASCT = 75 cGy, post-ASCT = 45 cGy). Fourteen additional non-ASCT patients were treated with 75 cGy. Unlabeled antibody was given prior to labeled dosimetric and therapeutic doses to improve biodistribution. Forty-two (71%) of 59 patients responded; 20 (34%) had complete responses (CR). Thirty-five (83%) of 42 with low-grade or transformed NHL responded versus 7 (41%) of 17 with de novo intermediate-grade NHL (P = .005). For all 42 responders, the median progression-free survival was 12 months, 20.3 for those with CR. Seven patients remain in CR 3 to 5.7 years. Sixteen patients were re-treated after progression; 9 responded and 5 had a CR. Reversible hematologic toxicity was dose limiting. Only 10 patients (17%) had human anti-mouse antibodies detected. Long-term, 5 patients developed elevated thyroid-stimulating hormone levels, 5 were diagnosed with myelodysplasia and 3 with solid tumors. A single, well-tolerated treatment with iodine 131I tositumomab can, therefore, produce frequent and durable responses in NHL, especially low-grade or transformed NHL.

Starting with properly collected, processed, annotated, and stored human specimens is critical for successful research using unsupervised, high-throughput technologies for discovery, validation, and translation from such samples. Samples that are not reproducibly collected and managed undermine data generated in platforms, such as mass spectroscopy–based proteomic technologies. The key factors that determine quality of human biosamples are use of consistent methodologies for collection, handling, processing, and storing specimens; understanding the potential sources of bias; and thoroughly annotating the life-cycle of the specimen. These factors apply when a new biospecimen resource is being created or when previously collected or banked samples are being used. Standard operating procedures permit control of bias and ensure that high-quality biosamples result in reproducible data. Compliance with increasingly complex human subject protection regulations governing the collection, management, storage, sharing, and use of human biosamples is essential to ensure that data will be published in high-quality journals, that intellectual property is protected, and that data generated using biosamples can be used as part of submissions to governmental regulatory bodies for product approvals.

Adoptive immunotherapy using in vitro activated T-cells can mediate the destruction of metastatic tumor deposits in animal models. These antitumor effector cells can be generated from mice with subcutaneous tumor by the sequential in vitro activation of tumor-draining lymph node (TDLN) cells with monoclonal antibody to the T-cell receptor complex (anti-CD3) and expansion in low concentrations of interleukin-2 (IL-2). In this animal model, the concomitant presence of visceral tumor can suppress the sensitization of tumor-reactive TDLN cells. We investigated whether IL-1 alpha added during in vitro activation and/or expansion of TDLNs could augment their antitumor activity in adoptive therapy. Mice were inoculated subcutaneously with MCA 205 tumor. TDLN cells were harvested and activated in vitro with 1 microgram/ml anti-CD3 for 2 days (anti-CD3 phase), followed by expansion in 10 U/ml IL-2 for 3 days (IL-2 phase). Experimental cultures had IL-1 (10-10,000 U/ml) added in either or both phases. After the 5-day culture period, cells were counted to determine in vitro cellular proliferation and then adoptively transferred to mice bearing 3-day established lung metastases to assess in vivo antitumor efficacy. IL-1 added during the anti-CD3 or IL-2 phase did not alter in vitro cellular proliferation. The presence of IL-1 during the anti-CD3 phase led to the generation of cells that were significantly more therapeutically efficacious than cells generated in the absence of IL-1. The effect of IL-1 during anti-CD3 activation appeared to be dose dependent in the concentration range 10-1,000 U/ml. The addition of IL-1 during the IL-2 phase only did not enhance the antitumor reactivity of the activated cells. The beneficial effect of IL-1 during the anti-CD3 activation phase was specific for the tumor against which the TDLN had been initially sensitized; also, there was no evidence that non-tumor bearer lymphocytes developed significant antitumor reactivity when "activated" with IL-1 and anti-CD3. Furthermore, addition of IL-1 during the anti-CD3 activation phase abrogated suppression induced by the concomitant presence of lung metastases during subcutaneous tumor growth. IL-1 appears to up-regulate the in vitro activation of tumor-reactive lymphocytes derived from TDLN, in both the presence and the absence of visceral metastases.

Thirty-one previously untreated patients with follicular low-grade B-cell non-Hodgkin's lymphoma expressing the CD20 antigen were treated with iodine-131 tositumomab therapy between 1996 and 1998. The therapy led to a temporary depletion of peripheral blood B-lymphocytes. Recovery of B-cells occurred in most cases by 3 to 6 months and in all patients by 12 months posttherapy. A temporary decline in T-cell subpopulations, but no reduction in serum immunoglobulin levels, could be observed. ELISA techniques were used to detect specific antibodies against rubella, mumps, varicella zoster, measles, and tetanus. Almost all patients remained seropositive against the different antigens during the 1- to 2-year follow-up. No significant reduction in antibody concentrations to tetanus or measles could be detected. The data show that acquired humoral immunity against common antigens appears to be preserved despite a temporary loss of B-lymphocytes.

Background: Curcumin, a dietary polyphenol derivative of turmeric, has potent cancer reductive activity in in vivo colon carcinogenesis models. We administered curcumin at doses of 2 g or 4 g to separate groups of 20 healthy human smokers with > 8 aberrant crypt foci (ACF). The 2 g dose showed no ACF reduction while the 4 g dose showed a 46% reduction (p Methods: Curcuminoid products were extracted from homogenized biopsy tissue and ultacentrifuged plasma and evaporated under argon. The separation was performed on a Symmetry® C18 column. The mobile phase used under gradient conditions was 25% acetonitrile with 74.9% containing 0.1% acetic acid (25:74.9:0.1; v/v/v; A) and 100% acetonitrile containing 0.1% acetic acid (99.9:0.1; v/v; B) The flow rate was 0.3 mL/min and injection volume for all samples was 10 µL. The lower limit of detection for curcumin from human plasma was 5 ng/mL and from human colonic mucosa was 50 ng/5mg tissue. Results: See Table below (summarized by pre and post treatment dose for plasma and mucosal concentrations) Conclusions: Native curcumin is not detectable in the majority of biopsy and plasma samples. Conjugated curcumin is detectable in all plasma samples and the majority of mucosal biopsies with a significant increase in plasma concentrations at 4g. We postulate the conjugates detected in the rectal mucosa are delivered systemically from absorption and conjugation higher in the gastrointestinal tract and may have biologic activity. Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 1286. doi:10.1158/1538-7445.AM2011-1286

Curcumin, a dietary polyphenol derivative of turmeric, has potent cancer preventive activity in in vivo colon carcinogenesis models. To determine whether curcumin has pharmacodynamic effects on the human colon, we administered curcumin at doses of 2 g or 4 g to separate groups of 20 healthy human smokers who had 8 or more aberrant crypt foci on screening colonoscopy. We removed 3 aberrant crypt foci and 4 flat mucosal biopsies under magnifying endoscopy prior to and after 28 days of daily curcumin dosing. We enrolled 41 subjects (66% African Americans, 27% Caucasian, mean age 55+/−6.5 years). PGE2 and 5-HETE concentrations on tissue biopsies (assayed with liquid chromatography tandem mass spectroscopy) are shown in the Table. Curcumin concentrations assayed by HPLC in the colonic mucosa were undetectable ( (Support: NCI N01-35610, F. Meyskens PI Univ California Irvine) Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 2906.

Abstract 3759 Poster Board III-695 Purpose Tositumomab and iodine I 131 tositumomab (Bexxar® Therapeutic Regimen) has been found effective in relapsed/refractory follicular lymphoma. We now report updated results of a single center, single-arm, Phase II trial of a single one-week course of this treatment for 76 previously untreated, stage III and IV, follicular lymphoma patients (reporting period June 1996 to May 2009). Patients and Methods Patients had a median age of 49 years (range 23 to 69) and received a dosimetric dose followed by a single total body dose of 75 cGy iodine I-131 tositumomab one week later. Seventy percent had stage IV disease and 70% of patients had histological grade 1 follicular lymphoma, 29% had grade 2, and 1 patient had mantle-cell lymphoma. Bone marrow involvement was present in 64% of patients; 43% of patients had at least one tumor 3 5 cm in diameter; LDH was elevated in 30%. Overall, 35% of patients had high risk Follicular Lymphoma International Prognostic Index (FLIPI) scores and 50% had intermediate risk scores. Patients entered long-term follow-up after disease progression or after 2 years on study. Response, second malignancy occurrence and thyroid medication use were assessed every 6 months for 5 years and yearly thereafter up to 12 years post treatment. Results As previously reported (NEJM 352:441, 2005), the overall response rate was 97% with 57 patients (75%) achieving a complete remission. Hematologic toxicity, although common, was modest to moderate (grade 4 neutropenia in 5% of patients and no grade 4 thrombocytopenia). After a median of 10 years follow-up (range 0.7 to 12.3 years), the median duration of response was 6 years (95% CI: 2.5, 10.8), with approximately 40% remaining progression-free at 10 years. For the 57 complete responders, median progression-free survival was 10.9 years (95% CI: 8.3, NR). Ten-year overall survival was approximately 82%. Five cases (7%) of hypothyroidism occurred 0.5 to 2.9 years after treatment and were managed with thyroid hormone replacement. Eleven patients (14%) were diagnosed with second malignancies including 4 skin neoplasms (2 basal cell and 2 squamous cell) and 7 visceral neoplasms (3 breast, 2 prostate, 1 endometrial cancer, 1 glioblastoma). One case of myelodsyplastic syndrome was diagnosed about 8 years after treatment. Conclusion A single course of treatment with Bexxar therapeutic regimen can commonly produce durable responses, especially durable complete responses lasting over a decade in patients with untreated follicular lymphoma. Further studies comparing this monotherapy to other regimens, including combination therapies, are warranted. Disclosures: Kaminski: GlaxoSmithKline: Honoraria, Patents & Royalties, Research Funding, Speakers Bureau. Off Label Use: Radioimmunotherapeutic for treatment of frontline treatment of follicular lymphoma. On-label use is for relapsed/refractory patients. Horner:GlaxoSmithKline: Employment. Williams:GlaxoSmithKline: Employment. Vleisides:GlaxoSmithKline: Employment. Wahl:Nordion: Honoraria; GlaxoSmithKline: Patents & Royalties.

• Antitumor effector T cells can be generated from tumordraining lymph node cells by activation with anti-CD3 monoclonal antibody and interleukin 2. Fresh tumordraining lymph node cells do not have antitumor activity, but after anti-CD3 monoclonal antibody–interleukin 2 activation, these cells mediate immunologically specific tumor regression in vivo. We examined the effect of interleukin 1 α administration on the development of effector T cells derived from tumor-draining lymph nodes. Mice were inoculated with tumor cells, and interleukin 1α (180 to 720 ng) was administered on days 0, 2, and 4. Tumordraining lymph nodes harvested 10 to 14 days later were activated in anti-CD3 monoclonal antibody for 2 days followed by expansion in interleukin 2 for 3 days. Antitumor efficacy of the activated cells was assessed in the adoptive immunotherapy of lung micrometastases. Administration of interleukin 1α enhanced the antitumor reactivity of effector cells derived from tumor-draining lymph nodes without changing the immunologic specificity of the cells. Administration of interleukin 1 α did not alter the phenotype of fresh or activated tumor-draining lymph node cells. Interleukin 1α deserves further investigation as an immune adjuvant in adoptive immunotherapy. ( Arch Surg . 1991;126:1503-1509)