1. NK cells kill targets that do not express HLA class I

2. Methods for addressing the NK cell dysregulation PBMCs from XMRV infected patients with low NK cell function were activated with the mitogen PHA and treated with Ampligen The effects on NK cell (CD56+) phenotype were determined by flow cytomentry Signaling changes due to the treatment were detected via cytokine analysis The change in XMRV copy number was detected with qRT-PCR

3. Ampligen significantly amplifies the degranulation of NK cells – Ampligen may increase the ability of CD56+ cells to degranulate when encountering a target Ampligen significantly increases the production of Perforin and GranzymeB – Ampligen may promote synthesis of cytotoxic proteins- perforin and GranzymeB Ampligen may regulate NK and inflammatory signaling molecules – Anti-viral as well as immune-stimulating cytokines are upregulated Preliminary results from 8 study subjects  Ampligen significantly amplifies the degranulation of NK cells  Ampligen may increase the ability of CD56+ cells to degranulate when encountering a target  Ampligen significantly increases the production of Perforin and GranzymeB  Ampligen may promote synthesis of cytotoxic proteins- perforin and GranzymeB  Ampligen may regulate NK and inflammatory signaling molecules  Anti-viral as well as immune-stimulating cytokines are upregulated

4. Preliminary Results Cont. XMRV copy number is modulated by Ampligen When treated with Ampligen, qRT-PCR indicates a decrease in some patients and an increase in others. This may suggest why Ampligen worsens some CFS patients and not others. Stratification for Ampligen treatment must consider XMRV status.

5. Infectious XMRV was detected in lymphocytes and plasma from >75% of CFS patients CFS-XMRV can be transmitted cell-free from patient plasma to human prostate and T cell lines and to primary T cells An immune response to the virus was detected in a majority of CFS subjects XMRV in CFS and prostate cancer are closely related and form a distinct phylogenetic branch There is a highly significant association between the XMRV retrovirus and Chronic Fatigue Syndrome Infectious XMRV was detected in lymphocytes and plasma from >75% of CFS patients XMRV in CFS and prostate cancer are closely related and form a distinct phylogenetic branch An immune response to the virus was detected in a majority of CFS subjects CFS-XMRV can be transmitted cell-free from patient plasma to human prostate and T cell lines and to primary T cells

6. From WPI Judy A Mikovits Vincent Lombardi Max Pfost Kathryn Hagen From Cleveland Clinic Robert Silverman Jaydip Das Gupta Robert Silverman Jaydip Das Gupta Cancer and Inflammation Program: Frank Ruscetti Mike Dean Bert Gold Dan Bertolette Ying Huang Laboratory of Cancer Prevention: Sandra Ruscetti Charlotte Hanson Jami Troxler Cari Petrow-Sadowski Rachel Bagni Kunio Nagashima Judy A Mikovits Vincent Lombardi Max Pfost Kathryn Hagen Cancer and Inflammation Program: Frank Ruscetti Mike Dean Bert Gold Dan Bertolette Ying Huang Laboratory of Cancer Prevention: Sandra Ruscetti Charlotte Hanson Jami Troxler Cari Petrow-Sadowski Rachel Bagni Kunio Nagashima Robert Silverman Jaydip Das Gupta Robert Silverman Jaydip Das Gupta The CFS patients in the US Acknowledgements: