1. Evolution of the Functional Profile of HIV-Specific CD8+ T cells in a Cohort of Long Term Nonprogressors M López, N Rallón, A Peris, M Salgado, B Rodés, V Soriano, J González-Lahoz, and JM Benito Hospital Carlos III, Madrid, Spain

2. A small proportion of HIV-subjects is able to control virus replication and halt disease progression A small proportion of HIV-subjects is able to control virus replication and halt disease progression A higher and/or better HIV-specific immune response in these patients is believed to contribute to their favourable outcome A higher and/or better HIV-specific immune response in these patients is believed to contribute to their favourable outcome However, it is not well understood what characteristics of HIV-specific immune response better correlate with protection However, it is not well understood what characteristics of HIV-specific immune response better correlate with protection

3. Transversal studies have suggested that a polyfunctional HIV-specific immune response and directed against Gag may be involved in the control of HIV viral replication Transversal studies have suggested that a polyfunctional HIV-specific immune response and directed against Gag may be involved in the control of HIV viral replication

4. Nat Med. 2007 Jan;13(1):46-53Blood. 2006 Jun 15;107(12):4781-9

5. The stability or the potential evolution over time of cellular immune response in the absence of antiretroviral therapy, especially in long term non-progressors (LTNP) has been scarcely analysed

6. Herein, we have studied the evolution of HIV specific CD8+ T cell response in LTNP followed for four years OBJECTIVE

7. 10 HIV+ patients belonging to a cohort of well- characterized LTNP followed for four years were included in this study 10 HIV+ patients belonging to a cohort of well- characterized LTNP followed for four years were included in this study LTNP : LTNP : Serologically proven HIV-1 infection lasting for over 10 years, Serologically proven HIV-1 infection lasting for over 10 years, CD4 counts always above 500 cells/  l and CD4 counts always above 500 cells/  l and lack of HIV-related symptoms in the absence of any antiretroviral therapy lack of HIV-related symptoms in the absence of any antiretroviral therapy METHODOLOGY

8. Three functions of CD8+ T-cells (production of MIP1 , IL2 and TNF  ) were simultaneously examined in response to HIV-Gag and Nef peptide-pools using multiparameter flow cytometry Three functions of CD8+ T-cells (production of MIP1 , IL2 and TNF  ) were simultaneously examined in response to HIV-Gag and Nef peptide-pools using multiparameter flow cytometry

9. LTNP P A Gag-specific CD8+ T cell responses MIP1β CD8- ECD TNF α - PECY7 IL2-PE SS FS MIP1β LymphocytesCD8+ cells

10. All variables were expressed as median [interquartile range] All variables were expressed as median [interquartile range] Differences between values at baseline and after four years were evaluated using non- parametric tests Differences between values at baseline and after four years were evaluated using non- parametric tests

11. RESULTS

12. n CD4 count (cell/  l) Viral Load (Log ARN copies / ml) At baseline 10 After 4 years of follow up n.s 619[300]3.2[1.6]10 p n.s n.s: no significant difference, p>0,05 754[400] 2.6[0.8] Characteristics of Patients

13. HIV- GAG SPECIFIC CD8+ T CELL RESPONSE

14. After 4 years After 4 years of follow up of follow up % CD8+ T cells 4 2 1 0 At baseline At baseline * p<0.05 3 After 4 years After 4 years of follow up of follow up % CD8+ T cells 4 2 1 0 At baseline At baseline * p<0.05 3 Global HIV-GAG specific CD8+ T cell response in LTNPs at baseline and after 4 years of follow up

15. 0 50 100 MIPβ TNFα IL2 At baseline At baseline After 4 years After 4 years of follow up of follow up Percentage of patients Prevalence of HIV-Gag CD8+ T cell response in LTNPs at baseline and after 4 years of follow up

16. MIPβ TNFα IL2 % CD8+ T cells 3 2 1 0 At baseline At baseline After 4 years After 4 years of follow up of follow up * * * p<0.05 Levels of seven different functional subsets of CD8+ T cells in response to HIV-GAG in LTNPs at baseline and after 4 years of follow up

17. Contribution of seven different functional subsets of CD8+ T cells to the global HIV-GAG response in LTNPs at baseline and after 4 years of follow up

18. Contribution of seven different functional subsets of CD8+ T cells to the global HIV-GAG response in each LTNP at baseline and after 4 years of follow up

19. HIV- NEF SPECIFIC CD8+ T CELL RESPONSE

20. % CD8+ T cells 2 1 0 At baseline At baseline After 4 years After 4 years of follow up of follow up * p<0.05 Global HIV-NEF specific CD8+ T cell response in LTNPs at baseline and after 4 years of follow up

21. Prevalence of HIV-Nef CD8+ T cell response in LTNPs at baseline and after 4 years of follow up

22. Levels of seven different functional subsets of CD8+ T cells in response to HIV-Nef in LTNPs at baseline and after 4 years of follow up

23. Contribution of seven different functional subsets of CD8+ T cells to the global HIV-NEF response in LTNPs at baseline and after 4 years of follow up

24. Contribution of seven different functional subsets of CD8+ T cells to the global HIV-NEF response in each LTNP at baseline and after 4 years of follow up

25. HIV-specific CD8+ responses are maintained over time in LTNP. The functional profile of this response may evolve in a different manner depending on the targeted HIV protein. Functionality of Gag specific CD8+ T cells tends to increase over time, highlighting its importance in controlling HIV replication. CONCLUSIONS

26. Patients ACKNOWLEDGMENTS Laboratory Immunology Group Mariola López Sara Lozano Norma Ibon Rallón Alejandra Peris Clara Restrepo Jose Miguel Benito To all staff at the Molecular Biology Lab Clinic Juan González-Lahoz Pablo Labarga Pablo Barreiro Francisco Blanco Luz Martín-Carbonero Pablo Rivas Eugenia Vispo Jose Medrano Vicente Soriano Funding