Identification of Tumor-Associated Antigens from Medullary Breast Carcinoma by a Modified SEREX Approach

Abstract

Medullary breast carcinoma (MBC) is a relatively rare malignancy with heavy lymphocytic infiltration that despite cytologically anaplastic features and high mitotic index has more favorable prognosis than other types of breast cancer. Lymphocytic infiltration of tumors reflects ongoing immune response against tumor antigens which could represent a great interest as potential targets for cancer immunotherapy. The search for MBC antigens by SEREX methodology has not been successful due to a very high titer of false positive clones, representing immunoglobulin genes. Here, we describe a novel approach for generating cDNA expression libraries from MBC tumor samples which are depleted of IgG cDNA clones and, therefore, are suitable for the identification of novel tumor-associated antigens (TAA) by SEREX approach. Modified methodology allowed us to isolate a panel of known and novel TAA which are currently under further investigation.

Autoantibody Response to ZRF1 and KRR1 SEREX Antigens in Patients with Breast Tumors of Different Histological Types and Grades

“…In this investigation, our attention was focused on two potential tumor-associated antigens, namely, Zuotin-related factor 1 (ZRF1) and R motif-containing protein 1 (KRR1), previously identified by SEREX analysis of medullary breast carcinoma cDNA library [12, 15]. These antigens had tumor-associated serological profile of autoantibodies according to the data of preliminary phage based allogeneic screening with sera of medullary breast carcinoma patients [12].…”

“…These antigens had tumor-associated serological profile of autoantibodies according to the data of preliminary phage based allogeneic screening with sera of medullary breast carcinoma patients [12]. Recombinant plasmids ZRF1/pBK-CMV and KRR1/pBK-CMV were obtained by “ in vivo excision” procedure of cDNA of phage λ positive clones of MBC cDNA library during previous investigation [12, 15]. …”

“…Today more than 2000 TAAs have been identified by different approaches including SEREX, SEPRA, phage display, microarray, and other high throughput technologies [14, 23]. Previously, we have described 81 autoantigens identified from breast, colon, and thyroid cancers by SEREX approach [10, 12, 13, 15, 16]. Original modification of SEREX technique for the generation of medullary breast carcinoma cDNA expression library depleted from IgG genes has been developed [12].…”

“…Previously, we have described 81 autoantigens identified from breast, colon, and thyroid cancers by SEREX approach [10, 12, 13, 15, 16]. Original modification of SEREX technique for the generation of medullary breast carcinoma cDNA expression library depleted from IgG genes has been developed [12]. As a result, MBC cDNA library suitable for serological screening was created and 41 MBC-derived autoantigens have been identified [12].…”

“…Original modification of SEREX technique for the generation of medullary breast carcinoma cDNA expression library depleted from IgG genes has been developed [12]. As a result, MBC cDNA library suitable for serological screening was created and 41 MBC-derived autoantigens have been identified [12]. Preliminary phage based allogenic screening revealed that 18 of them had exhibited cancer-related autoantibody profile and reacted only with sera of medullary breast carcinoma patients but not with sera of healthy donors [12, 15].…”