1. Immunohistochemical Differentiation between
Invasive Squamous Cell Carcinoma and
Carcinoma-In-Situ of the Uterine Cervix
Ruben Flam-Shepherd
Dr. Golnar Rasty
ANA498Y
University of Toronto

2. Introduction
 Cervical Cancer
 2nd most common cancer in women15
 Squamous Cell Carcinoma (SQCC) most common kind15
 Arises from CIS
 Differentiation between CIS and SQCC important
 Previous Study
 SMA and CD34
 Hypothesis: Immune reaction to CIS and Invasive SQCC is different

3. Materials and Methods
 Pathology archive at UHN searched
 2001-2009
 37 cases retrieved
 Immunohistochemistry performed
 For CD3, CD4, CD8, CD20, CD138
 Slides were scored
 Data sets compared using Pearson’s Chi-Squared Test

4. Materials and Methods
Scoring
Figure 1. Sample CD3 staining
receiving a score of 1/0
Figure 2. Sample CD4 staining
receiving a score of 1/0
Figure 3. Sample CD8 staining
receiving a score of 1/0
Figure 4. Sample CD20 staining
receiving a score of 0/0
Figure 5. Sample CD138 staining
receiving a score of 0/0
Figure 6. Sample CIS staining
receiving a score of 2

5. CD3
 Involved in T-Cell Receptor (TCR) Complex17
 2 heterodimers present
 T-Cells are triggered by peptide
 Presented by Major Histocompatibility Complex (MHC)
 CD3 involved in activating signal cascade
 Immunoreceptor tyrosine-based activation motifs (ITAMEs)21
 Phosphorylated by lymphocyte cell-specific protein tyrosine kinase (p56lck)
 Play role in neuronal morphogenesis2

6. CD4
 Interacts with MHC II16
 Forms complex with p56lck 16
 Targeted by HIV (gp120)3
 Induces conformational change in gp120
 gp120 subsequently binds to coreceptors CCR5 or CXCR4
 CD4 counts are used to determine progression of HIV6
 1450 x 106 CD4 T-Cells/L (normal)
 400 x 106 CD4 T-Cells/L (HIV)
 200 x 106CD4 T-Cells/L (AIDS)

7. CD8
 Present as either a homodimer or heterodimer10
 αα/αβ
 Interacts with MHC I10
 Forms complex with p56lck
 Different effects
 αα – forms coreceptor with TCR complex
 αβ – regulatory role inhibiting αα form
 CD8 counts increase during HIV infection8
 Counts correlated to HIV RNA levels

8. CD20
 Integral membrane protein14
 Mainly intracellular component
 2 loops (18 amino acids) exposed extracellularly
 First expressed during early pre-B Cell developmen 19
 Negligible blood serum levels.
 Not shed from cell surface9
 Negligible blood serum levels.

9. CD138
 Also known as Syndecan-118
 Family of cell surface proteoglycans
 Expressed in surface epithelial cells and plasma cells20
 Some functions:
 Catalyze bind of ligands to proper receptors
 Adhesion at cell-to-cell interphases
 Mice homozygous null for CD1381
 Normal development
 Normal fertility

10. Results
Immunohistochemistry – CD3 Scoring
0
27%
1
6%
1A
7%
2
60%
2A
0%
Invasion
0
14%
1
13%
1A
13%
2
60%
2A
0%
CIS w/Invasion Present
0
9%
1
18%
1A
14%
2
59%
2A
0%
CIS w/o Invasion
No significant differences or correlations

11. Results
Immunohistochemistry – CD4 Scoring
0
20%
1
20%
1A
0%
2
60%
2A
0%
Invasion
0
13%
1
20%
1A
7%
2
60%
2A
0%
CIS w/Invasion Present
0
4%
1
14%
1A
18%
2
64%
2A
0%
CIS w/o Invasion
No significant differences or correlations

12. Results
Immunohistochemistry – CD8 Scoring
0
27%
1
40%
1A
0%
2
33%
2A
0%
Invasion
0
27%
1
20%
1A
13%
2
40%
2A
0%
CIS w/Invasion Present 0
4%
1
41%
1A
18%
2
32%
2A
5%
CIS w/o Invasion
Significant differences between 1 and 3 (p < 0.005) and 2 and 3 (p < 0.01)

13. Results
Immunohistochemistry – CD20 Scoring
0
67%
1
27%
1A
0%
2
6%
2A
0%
Invasion
0
47%
1
27%
1A
13%
2
13%
2A
0%
CIS w/Invasion Present
0
41%
1
9%
1A
23%
2
0%
2A
27%
CIS w/o Invasion
Significant differences between 1 and 3 (p < 0.05) and 2 and 3 (p<0.05)

14. Results
Immunohistochemistry – CD138 Scoring
0
67%
1
13%
1A
7%
2
13%
2A
0%
Invasion
0
60%
1
27%
1A
6% 2
7%
2A
0%
CIS w/Invasion Present
0
14%
1
18%
1A
23%
2
45%
2A
0%
CIS w/o Invasion
Significant differences between 1 and 3 (p < 0.0005) and 2 and 3 (p<0.001)

15. Conclusion
 Decreased expression of certain markers in invasive conditions
 CD8
 CD20
 CD138
 Future directions
 Repeat trials with larger sample size
 Investigate underlying mechanism

16. Acknowledgments
 I’d like to thank Dr. Rasty for her time, support and knowledge during this and past research
opportunities

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