Recommendations for breast cancer education for African American women below screening age.

Although statistically unlikely, early-onset breast cancer tends to be more aggressive and leads to greater mortality than breast cancer among women of screening age. Young African American women are disproportionately impacted by early-onset breast cancer compared to women of other races. Given the racial disparities and because young women are typically not the primary audience for breast cancer educational messaging, there is a need to identify recommendations for age-appropriate breast cancer education for African American women below mammogram eligibility. Through N = 30 key informant interviews with young African American breast cancer survivors, family members of young survivors, community organization leaders and healthcare providers, we identified breast cancer educational message content and communication channels relevant for these women. Participants recommended that message content should emphasize the need to address family cancer history and self-advocacy in healthcare encounters in addition to concerns about loss of womanhood, financial costs and opportunity costs associated with preventive healthcare visits. Breast cancer messages for this audience should consider the influences of earlier life stage, culture and race. Recommended communication channels highlighted use of social media and videos. Findings will inform future age-appropriate educational messaging aimed at eliminating early-onset breast cancer disparities disproportionately impacting young African American women.

Human IL-6 enhances human lymphocyte engraftment and activation but not human antibody production in SCIDhu PBL mice.

The SCIDhu PBL model of human Ig production was modified by using human interleukin-6 (hIL-6) secreting tumors for continuous hIL-6 production, in vivo. On day one, SCID mice were injected subcutaneously with 200 microliters PBS (control mice), 10(4) SP2/0-Ag14 cells (IL-6+ mice) or 10(4) hIL-6 secreting SP2/0-hIL6.17 cells (IL-6- mice). The mice were reconstituted with human PBMC on day two and immunized with 100 micrograms of tetanus toxoid (TT) on days two and fifteen. Serum hIL-6 concentrations in IL-6+ mice ranged between 2.9 and 38.1 ng/ml by days 26-33. IL-6+ mice had enlarged spleens and lymph nodes (LN). Flow cytometry and histology showed that SCIDhu PBL mouse spleen, LN and peritoneal exudate cells (PEC) contained mostly murine myeloid lineage cells. In addition, many more human B cells, T cells and IL-2R(+)-activated lymphocytes were present in spleen, LN and PEC of IL-6+ mice. Despite enhanced lymphocyte engraftment and activation, by day 14 IL-6+ mice produced up to 6-fold less TT-specific IgG relative to total IgG than either control group. TT-specific and total Ig sera concentrations were equivalent in all three groups on days 26-33. Our results suggest that sustained circulating hIL-6 enhanced human delayed-type hypersensitivity (DTH)-like inflammatory responses with consequential inhibition of TT-specific IgG production in SCIDhu PBL mice.

Evaluation of Bcl-2/B cell transgenic mice (B6) for hybridoma production.

Bcl-2 is an oncogene associated with prevention of apoptosis in a variety of cell types. Bcl-2 expression in B lymphoid cells prolongs antibody production, in vitro and in vivo. A line of transgenic mice (B6) has been developed that expresses human Bcl-2 in the B cells of SWR/SJL mice. B6 transgenic, nontransgenic littermates, and BALB/c mice were immunized with beta-galactosidase (B-gal) or sheep red blood cells (SRBC). The number of spleen cells recovered from immunized B6 mice was 3-4 times greater than syngeneic, nontransgenic littermates or BALB/c mice. Spleen cells from B-gal or SRBC immune B6, SWR/SJL, and BALB/c mice were fused with P3 myeloma cells to produce hybridomas. Forty-eight percent of the wells plated with fused B6 spleen cells produced B-gal-specific antibodies compared to 14% from BALB/c and 12% from SWR/SJL. Antibody-specific wells were subcloned, resulting in enhanced recovery of antigen-specific subclones with B6-derived fusions compared to controls. In the SRBC fusions, 17% of the wells plated with fused B6 spleen cells produced SRBC-specific antibodies compared to 6% for BALB/c and SWR/SJL spleens. After subcloning, B6-derived clones produced 8% positive subclones compared to 9.5% from SWR/SJL and 3.5% from BALB/c. Comparison of the isotype distribution of subclones showed a higher ratio of IgG antibodies compared to IgM from B6 mice in the B-gal fusions. IgA antibodies were recovered only from B6 mice. These data indicate that B6 transgenic mice that overexpress Bcl-2 in their B cells may be superior to other mouse strains for production of antigen-specific hybridomas.

Production and in vivo characterization of a bifunctional antibody (IVA039.1) with specificity for the mouse interleukin-2 receptor and vinca alkaloids.

The autoreactive T cell plays a pivotal role in the pathogenesis of type I diabetes in humans and in rodent animal models. Elimination or attenuation of these cells may provide a means to treat the disease. The use of antibodies directed to T cells has shown varying degrees of effectiveness in the treatment of autoimmune disease. The use of a bifunctional antibody directed to T cells with a cytolytic agent may provide an additional level of therapeutic efficacy compared to anti-T-cell antibodies alone. To test this hypothesis, we prepared a bifunctional antibody (IVA039.1) with specificity for the mouse interleukin-2 (IL-2) receptor and vinca alkaloids. The antibody was derived from the fusion of vinca immune spleen cells with PC61 5.3, a hybridoma that produces rat anti-mouse IL-2 receptor antibody. IVA039.1 was purified by affinity chromatography through Protein A and anti-vinca affinity columns followed by TSK-DEAE high-pressure liquid chromatography (HPLC). Bifunctionality of the antibody was confirmed by fluorescence-activated cell sorting (FACS) analysis, enzyme-linked immunoadsorbent assay (ELISA) and a cell assay designed to measure simultaneously both IL-2 receptor and vinca reactivities. The biodistribution of IVA039.1 was determined in normal and streptozotocin-complete Freund's adjuvant (CFA) induced diabetic mice. Enhanced uptake of IVA039.1 was observed in the pancreata, spleens, and lymph nodes of diabetic compared to normal mice. These data suggest that bifunctional antibodies that can deliver cytolytic agents to T cells may be appropriate candidates for the treatment of diabetes and other autoimmune diseases.

Production of IgG monoclonal antibodies to the tumor-associated antigen, CA-195.

The isotype of a monoclonal antibody is closely associated with its biologic activity. Certain immunoglobulin subclasses are more effective than others regarding their ability to execute complement-mediated lysis of cells, antibody-dependent cell-mediated cytotoxicity, and tumor localization. Many potential targets for cancer immunotherapy are tumor-associated antigens with high percentages of carbohydrate. Immunizations of mice with carbohydrate antigens usually produce IgM and IgG3 antibodies. The use of different adjuvants in immunization protocols has been associated with the induction of isotype-specific antibody responses. In experiments reported here, we compare the use of four different adjuvants on the generation of an IgG immune response to the carbohydrate-rich, tumor-associated antigen, CA-195. We report the production of IgG1 and IgG2a monoclonal antibodies (mAbs) to CA-195.