jurkovic.ppt
TRANSCRIPT
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Immune Response and Splenomegaly in B16 Melanoma
Dominika A JurkovicAdvisor: Dr. SpilatroApril 2, 2003
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Overview of Presentation
Background Information Hypothesis Procedures Data Conclusions Discussion Future Experiments
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Background Information
What is melanoma? B16 Melanoma Splenomegaly
www.melanoma.com
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Hypothesis
Splenomegaly in mice implanted with B16 melanoma is an immunological response to the developing tumor
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Spleen Size in B16 Melanoma
Purpose is to examine Spleen enlargement in response to a developing tumor
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Procedure
Establish Tumor Line
Implantation Allow to grow Excise tissue Weigh tissues Preserve tissues Appearance of B16 Melanoma in Mice
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Spleen Size Over Time
0
0.02
0.04
0.06
0.08
0.1
0.12
0.14
0.16
0.18
0 3 6 9 12 15 18
Day of Experiment
Sp
lee
n S
ize
(g
)
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Spleen Enlargement
Size Comparison of Day 0 and Day 18 Spleen
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Spleen Weight Compared to Tumor Size
0
0.02
0.04
0.06
0.08
0.1
0.12
0.14
0.16
0.18
0.2
0 0.5 1 1.5 2
Average Tumor Size (g)
Avera
ge S
ple
en
Siz
e (
g)
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Procedure for Cell-Free Homogenate Homogenize B16
Tissue Centrifuge Filter Sterilization Inject 8µg Protein
Inject PBS Buffer
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Spleen Enlargement in Control Groups
0%
25%
50%
75%
100%
125%
150%
175%
200%
225%
250%
PBS Buffer Cell-free Extract B16 Melanoma
Antigen
Inc
rea
se
in S
ize
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Summary
Spleen enlarges as the tumor grows PBS Control negative Cell free extract less effect indicating
response to soluble tumor antigens
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Histological Changes in the Spleen
Examine histology of spleen in normal, melanoma implanted, and cell-free extract injected mice
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Procedures
Fixation Embedding Staining
Microtome
Nikon camera
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Anatomy and Function of the Spleen
Sears, D. 1997 Gude, W. 1982
Primary Follicles
Secondary Follicle
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Structural Changes
Day 0 Day 18
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Histology Results
*, #, ^ =Significant Difference
Normal B16 Melanoma Cell-Free ExtractAverage Diameter of Follicles 279.21um +/- .509* 249.49 +/- .394* 311.11 +/- .564
Follices per mm2 61.55/mm2 +/- 9.71#^ 55.02/mm2 +/- 5.03# 38.2/mm2 +/- 8.79^Total Follicles of Cross-section 72.66 +/- 17.01* 126.93 +/- 20.49* 74 +/- 12.16
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Summary
Structural changes indicate a humoral response is occurring in the B16 Melanoma
Humoral response is seen in Cell-free extract injected mice, but not to the same degree
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Overall Conclusions
Splenomegaly is an immune response to B16 Melanoma
Histology of spleen indicates a humoral immune response to B16 melanoma suggesting production of B16 specific antigens
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Discussion The immune system can recognize
cancer cells as antigens, or foreign invaders
Researchers have identified that some cancer cells do carry antigens on their surfaces.
By training the immune system to attack these antigens, or "build up an immunity," doctors may be able to halt the disease
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Future Experiments
Confirm type of immune response Identify specific antigen associated with
the melanoma tissue Examine splenomegaly in other mice
strains implanted with B16 melanoma
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Acknowledgements
Dr. Spilatro-Advisor Dr. Brown and the
capstone class Dr. Kramer
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Questions?