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The Impact of Thermotherapy on Immune Response

October 5th 2010Evelyn LakeMBP 1028

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Presentation Outline

What constitutes an immune response?

Range of thermotherapy temperatures: Cryotherapy Mild hyperthermia Extreme hyperthermia

Summary and conclusions Questions?www.physicscentral.

com

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The Immune Response

“The body's response to defend against attacks from disease-causing agents”

Ways to elicit a response = any form of stress1. Cold shock (mild or extreme)2. Oxidative shock3. Heat shock (mild to extreme)4. Heavy metals5. Pathological condition (ischemia or reperfusion)6. Inflammation7. Tissue damage8. Infection9. Mutated proteins (associated with disease)

www.bcm.edu.htm

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The Immune Response

“The body's response to defend against attacks from disease-causing agents”

Ways to elicit a response = any form of stress1. Cold shock (mild or extreme)2. Oxidative shock3. Heat shock (mild to extreme)4. Heavy metals5. Pathological condition (ischemia or reperfusion)6. Inflammation7. Tissue damage8. Infection9. Mutated proteins (associated with disease)

www.bcm.edu.htm

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The Immune Response How can you measure an immune

response?1. Chances for survival2. Number of metastases and/or size of tumor

(cancer)3. Levels of Heat Shock Protein (HSP) expressed4. Peptide antigen expression5. T-cell response

Can be a positive or negative response1. Suppression (+/-)2. Enhancement (+/-)

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Extreme Cryotherapy

Treatment: (-40⁰C)Cryoablation

The Immune Response: Can be beneficial or detrimental

Both immune suppression and immune stimulation have been observed

www.ctsnet.org

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Extreme Cryotherapy: Example Many factors may effect the immune response to

cryoablation:1. Type of tumor2. Volume of tumor3. Freezing method (rate, temperature, equipment)

Example: Rate of Freeze Alters the Immunologic Response After Cryoablation of Breast Cancer

Measured immune response: 1. T-cells2. Survival3. Tumor metastases

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Extreme Cryotherapy: Example Many factors may effect the immune response to

cryoablation:1. Type of tumor2. Volume of tumor3. Freezing method (rate, temperature, equipment)

Example: Rate of Freeze Alters the Immunologic Response After Cryoablation of Breast Cancer

Measured immune response: 1. Survival2. Tumor metastases3. T Cell expression

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T Cells Thymus Cells VERY large family of cells

with diverse functions w.r.t. cell-mediated immunity

Type of white blood cell called lymphocytes

Effector T cells have been shown to mediate the regression of pulmonary metastases

Play a critical role in mediating tumor regression [Winter, 2007]

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Extreme Cryotherapy: ExamplePurpose: Examine the mechanism by

which cryoablation may stimulate immune response w.r.t. different freeze rates

Low Freeze Rate: 10% argon duty cycle (on for 1 of every 10 seconds) for a total a few minutes (until tumor is frozen)

High Freeze Rate: 100% argon duty cycle for 30 seconds (until tumor is frozen)

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Extreme Cryotherapy: Example

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Extreme Cryotherapy: Example

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Extreme Cryotherapy: ExampleConclusion:

High freeze rate cryoablation caused a significant increase in tumor-specific T cells, a reduction in pulmonary metastases, and improved survival

Low freeze rate cryoablation caused an increase in regulatory T cells, an increase in pulmonary metastases, and was detrimental to survival

The techniques employed to freeze tissue can alter the immune response from stimulatory to suppressive

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Mild HyperthermiaThe Treatment:

Fever (39.5-43 ⁰C for 30 mins. - 8 hrs.)The Immune Response: Can manipulate

treatment to be Inhibitory of autoimmune or inflammatory, or Stimulatory of malignant or infectious disease

Immune Markers: Heat shock proteins (HSP) and glucose-regulated proteins (grp)

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Heat Shock Proteins (HSP)

Classified by size Two main types:

1. Molecular chaperones2. Proteases

Have dual function (intra.vs. extracellular)

VERY large family of proteins which are over expressed when an organism is under stress

alfa.di.uminho.pt/~pedrogabriel/research.htm

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Mild Hyperthermia: Examples Heat shock proteins as a vaccine:

1. Delivered via necrotic tumor cell death 2. Delivered via injection

Two examples:1. Heat Shock Protein 70 Gene Therapy Combined

with Hyperthermia using Magnetic Nanoparticles [Ito, 2003]

2. Characterization of Heat Shock Protein 110 and Glucose-Regulated Protein 170 as Cancer Vaccines and the Effect of Fever-Range Hyperthermia on Vaccine Activity [Wang, 2010]

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Mild Hyperthermia: Example 1 Mice infected with melanoma cells were

treated with gene (HSP70) and hyperthermia therapy

Tumor tissue was heated to 43 ⁰C for a period of 30 minutes 24hrs. after HSP70 gene injection

Heating was insufficient to kill tumor tissue, purpose was to observe the effects of combined heating and gene therapy

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Mild Hyperthermia: Example 1

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Mild Hyperthermia: Example 1

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Mild Hyperthermia: Example 2Example: Characterization of Heat Shock

Protein 110 and Glucose-Regulated Protein 170 as Cancer Vaccines and the Effect of Fever-Range Hyperthermia on Vaccine Activity [Wang, 2010]

Mice inflicted with colon cancer were treated with heat shock protein injections, a subset of which were also treated with mild hyperthermia

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Mild Hyperthermia: Example 2 Awake mice were heated to 39.5 ⁰C

for 6hrs. following protein injection

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Mild Hyperthermia: Conclusions

Example 1 Example 2Tumor was used as HSP

sourceHSP was injected into

animalTumor was targeted for

heatingEntire animal was heated

Heated to 43 ⁰C for 30 minutes

Heated to 39.5 ⁰C for 6hr.

Treatment was effective w.r.t. survival

Treatment was effective w.r.t. survival

Tumor size was observed to decrease

Tumor size was observed to decrease

Treatments were each very effective Both forms of mild hyperthermia, in conjunction

with artificially stimulated immune responses

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Extreme Hyperthermia The Treatment: High-intensity focused

ultrasound (56-80⁰C for ~1 second) The Immune Response: Changes in T-

cell, HSP, and protein antigen expression Observed:

1. Spontaneous regression of tumor metastases2. Better survival after treatment3. Changes in all immune response agents listed

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Conclusions Thermotherapy offers an advantage over

traditional surgical treatments:1. Recuperation and hospital time is minimized by less

invasive procedure2. A beneficial immune response makes treatment

more effective Future Work:

1. Further study into the mechanism of immune response

2. Optimize immunological stimulation▪ Technique optimization (Ex. Freeze rate)▪ Response supplementation (Ex. HSP vaccination)

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References1. Antitumor effects of combined therapy of recombinant heat shock protein 70 and hyperthermia using

magnetic nanoparticles in an experimental subcutaneous murine melanoma (2004) Akira Ito, Fumiko Matsuoka, Hiroyuki Honda, Takeshi Kobayashi. Cancer Immunol Immunother 53: 26-32

2. Heat shock protein 70 egen therapy combined with hyperthermia using magnetic nanoparticles (2003) Akira Ito, Fumiko Matsuoka, Hiroyuki Honda, and Takeshi Kobayahsi. Cancer Gene Therapy 10: 918-925

3. Intracellular and extracellular functions of heat shock proteins: repercussions in cancer therapy (2007) E. Schmill, M. Gehrmann, M. Brunet, G. Multhoff, and C. Garrido. Journal of Leukocyte Biology 81: 15-27

4. Characterization of heat shock protein 110 and glucose-regulated protein 170 as cancer vaccines and the effect of fever-range hyperthermia on vaccine activity (2001) Xiang-Yang Wang, Latif Kazim, Elizabeth A. Repasky, and John R. Subjeck. J. Immunol. 166: 490-497

5. Regulatory potential of fever-range whole body hyperthermia on lagerhans cells and lymphocytes in an antigen-dependent cellular immune response (2001) Julie R. Ostberg, Caren Gellin, Rahul Patel and Elizabeth A. Repasky. J. Immunol. 167: 2666-2670

6. Role of the heat shock response and molecular chaperones in oncogenesis and cell death (2000) Caroline Jolly, Richard I. Morimoto. Journal of the National Cancer Institute. Vol. 92, No. 19, 1564-1572

7. Regulatory effects of fever-range while-body hyperthermia on the LPS-induced acute inflammatory response (2000) Julie R. Ostberg, Shannon L. Taylor, Heinz Baumann, and Elizabeth A. Repasky. Journal of Leukocyte Biology. Vol. 68 815-820

8. Effects of mild perioperative hypothermia on cellular immune response (1998) Benzion Beilin, M.D., Yehuda Shavit, Ph.D, Jacob Razumovsky, M.D., Yaacov Wolloch, M.D, Alexander Zeidel, M.D., Hanna Bessler, Ph.D. Anesthesiology V.89 No. 5 1133-1140

9. Investigation of the mechanism and the effect of cryoimmunology in the copenhagen rat (2000) Nathan E. Hoffmann, James E. Coad, Christopher S. Huot, David J. Swanlund and John C. Bischof. Cryobiology, Vo. 42, Issue 1 59-68

10. Enhanced tumor metastases in rats following cryosurgery of primary tumor (1982) Yamashita T., Hayakawa K, Hosokawa M, Kodama T, Inoue N, Tomita K, Kobayahsi H. PMID 73(2) 222-8

11. The cryobiology of cryosurgical injury (2002) Nathan E. Hoffmann, and John C. Bischof. Urology Elsevier Science Inc. (Suppl 2A) 40-49

12. Rate of freeze alters the immunologic response after cryoablation (2010) Michael S. Sabel, MD, Gang Su, Kent A. Griffith, MPH, MS, and Alfred E. Chang, MD. Surgical Oncology 17: 1187-1193

13. Host antitumour immune responses to HIFU ablation (2007) F. Wu, L. Zhou, W.R. Chen. Int. J. Hyperthermia 23(2): 165-171

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Questions?Thank you!