Date Approved

5-12-2026

Graduate Degree Type

Thesis

Degree Name

Biomedical Sciences (M.H.S.)

Degree Program

Biomedical Sciences

First Advisor

Kristin Renkema PhD

Second Advisor

Elena Tonc PhD

Third Advisor

Kathryn Haley PhD

Academic Year

2025/2026

Abstract

The immune system is a complex, highly coordinated network of cells and signaling molecules that protects the body from infection and disease. Immunology is therefore a fundamental component of undergraduate education for students pursuing careers in medicine and biomedical research. Despite its importance, many undergraduate immunology courses lack a laboratory component. To address this gap, we developed an open-access immunology laboratory manual and evaluated its ability to generate meaningful data using primary murine cells and commercially available cell lines. Given the cost and logistical constraints of using mouse models in an undergraduate laboratory setting, we evaluated whether immortalized cell lines could serve as accessible alternatives. Female C57BL/6 mice and the murine-derived cell lines RAW 264.7 and CTLL-2 were used to model innate and adaptive immune cell activation, respectively. Macrophages were stimulated with lipopolysaccharide (LPS) and IFN-α, followed by assessment of MHC-II and CD86 expression via flow cytometry and TNF-α production by ELISA. T cells were stimulated with anti-CD3, anti-CD28, and IL-2, with activation assessed by CD44 and CD69 expression. RAW 264.7 macrophages exhibited increased MHC-II and CD86 expression and elevated TNF-α production following stimulation, consistent with responses observed in primary macrophages. In contrast, CTLL-2 T cells demonstrated little to no upregulation of CD44 and CD69 and did not recapitulate the activation observed in primary T cells. These findings indicate that RAW 264.7 cells represent a cost-effective alternative to primary macrophages for modeling innate immune activation in an undergraduate immunology laboratory course, whereas CTLL-2 cells require continued optimization or the use of alternative models.

Download

Share

COinS