Abstract A074: Early detection of breast implant associated anaplastic large cell lymphoma by multiplex lateral flow assay of CD30 and IL-10
Authors: Xu Peng , Kourentzi Katerina , Willson Richard , Kadin Marshall 
Published: November 2024

Abstract
Introduction: More than 1300 women have developed a unique breast implant associated anaplastic large cell lymphoma (BIA-ALCL) around breast implants. BIA-ALCL presents as an accumulation of fluid (seroma) a median of 9 years after implant placement. When detected early, survival with surgery alone is nearly 100%. Remaining patients present with masses and/or lymph node metastases requiring radiation, chemotherapy and/or immunotherapy and fatalities have occurred. Methods: Preliminary studies identified CD30 and IL-10 as the most sensitive and specific biomarkers for BIA-ALCL in seroma fluids. Therefore, we constructed a multiplex LFA to detect both CD30 and IL-10 in 30 mL seroma fluid within 20 minutes. LFA is a point of care immunoassay using capture and detection antibodies that detect separate epitopes of the target molecule. The detection antibody is linked to a colored particle; in this multiplex assay CD30 antibody was linked to blue and IL-10 to red particles. Separate positive test lines (blue and red) indicate presence of CD30 or IL-10. A control line confirms migration of the seroma fluid past the test lines. To determine the dynamic range of sensitivity we spiked recombinant CD30 and IL-10 into benign seromas. Test and control lines were quantitated with NIH Image J software and ratios of test line to control line calculate for each sample. Forty-eight hour supernatants of supernatants of BIA-ALCL lines TLBR1 and TLBR2 were tested for IL-10 and CD30. Fifteen patients, 8 with BIA-ALCL and 7 with benign seromas provided consent for testing of their seroma fluids. Results: Multiplex CD30/IL10 LFA produced positive test lines for neat and 1:10 dilutions of 48 hour supernatnats of BIA-ALCL lines TLBR1 and TLBR2. Both IL-10 and CD-30 could be detected on the multiplex LFA strip using a 1:3 dilution of seroma fluids. The multiplex LFA shows both IL-10 and CD30 are significantly higher in malignant samples (IL-10 TL/CL: malignant, 0.2±0.16 n=8 vs benign, 0.004±0.007 n=7, p&lt;0.05; CD30 TL/CL: malignant, 0.38±0.2 vs benign, 0.02±0.02, p&lt;0.001, two-way ANOVA). Importantly, when either IL-10 or CD30 yields a faint positive test line, the other is not detected reducing the possibility of false positives. Conclusions: The CD30/IL-10 multiplex LFA had 100% positive and negative predictive value in this pilot study. The assay was able to distinguish lymphoma from recurrent breast cancer and more common benign seromas. The use of this multiplex assay mitigates positive false positive results from more sensitive CD30 LFA which has been detected in some benign seromas. This LFA will potentially allow early pre-operative detection of ALCL allowing surgeons to plan their surgical approach. The LFA can replace more costly time-consuming flow cytometry, immunohistochemistry, cyto- and histopathologic studies which require at least 10 ml of fluid. The LFA can be performed outside of medical centers in rural and underserved areas. A larger prospective multicenter study including intra-operative sampling is planned to validate these results.
Citation Format: Peng Xu, Katerina Kourentzi, Richard Willson, Marshall Kadin. Early detection of breast implant associated anaplastic large cell lymphoma by multiplex lateral flow assay of CD30 and IL-10 [abstract]. In: Proceedings of the AACR Special Conference: Liquid Biopsy: From Discovery to Clinical Implementation; 2024 Nov 13-16; San Diego, CA. Philadelphia (PA): AACR; Clin Cancer Res 2024;30(21_Suppl):Abstract nr A074.