HIV-1 envelope glycoproteins isolated from Viremic non-progressors HIV-infected individuals are fully functional and cytophatic

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1 HIV-1 envelope glycoproteins isolated from Viremic non-progressors HIV-infected individuals are fully functional and cytophatic Silvia Marfil 1, Veronique Hebman 2, Sara Marrero-Hernandez 3, Daniel Márquez-Arce 3, Romina Cabrera-Rodríguez 3, Maria Soledad Varela 3, Concepción Casado 4, Cecilia Cabrera 1, Víctor Urrea 1, María Pernas 4, Bonaventura Clotet 1, Cecilio López-Galíndez 4, Martine Biard- Piechazyck 2, Agustín Valenzuela 3, Julià Blanco 1,5 1 Inst. de Recerca de la Sida IrsiCaixa, Inst. de Recerca Germans Trias i Pujol (IGTP), Badalona, Spain. 2 CNRS, Centre d études d agents Pathogènes et Biotechnologies pour la Santé, Montpellier, France 3 Lab. Inmunología Celular y Viral, Fac. Ciencias de la Salud, Univ. de La Laguna (ULL), Tenerife, Spain 4 Instituto de Salud Carlos III, Madrid Spain. 5 Universitat de ViC-UCC, Barcelona, Spain 217/11/3

2 VIREMIC NON PROGRESSION HIV-1 disease progression, CD4<35 // <3 year SP LTNP, VL>2x1 3 LTNP-EC, VL<5, VL>1 4 // CD4>5 //>2 year

3 HYPOTHESIS AND OBJECTIVES Viruses isolated from show low replicative fitness. Weber et al. Impaired human immunodeficiency virus type 1 replicative fitness in atypical viremic non-progressor individuals. AIDS Res Ther 14, Viruses isolated from show preserved nef function. Heigele et al. Viremic long-term nonprogressive HIV-1 infection is not associated with abnormalities in known Nef functions. RETROVIROLOGY 11, Viral fitness is determined by Env function. Rangel et al. Role of the human immunodeficiency virus type 1 envelope gene in viral fitness. J Virol 77: To analyze the role of Env in the phenotype To compare Env functions between and

4 SAMPLES (n=4) (n=5) P-value CD4 Abs (cells/ul) * VL (copies/ml) ns Time from seroconversion (years) 5.7 HAART ns Gender (% of male) 1 1 ns * Curriu et al. Viremic HIV Infected Individuals with High CD4 T Cells and Functional Envelope Proteins Show Anti-gp41 Antibodies with Unique Specificity and Function. PLoS One 7: e

5 Relative Fusion (% of BaL) Relative Fusion (% of BaL) Relative Fusion (% of BaL) ENV CLONING AND SELECTION 5 p= p=.65 p=.3 M-W test K-W test (n=44) (n=56) ns (n=15) (n=17) Selected clones show similar activity and heterogeneity than full set.

6 FUSOGENICITY (FUSION/EXPRESSION) EXPRESSION (% of Env+ve cells) EXPRESSION AND FUSION (Cell-to-cell fusion) tat 4 env T cells 1 + Env Expression 2 TZM-bl cells 6 h Light Env clones from and show similar expression and specific fusogenicity.

7 RELATIVE FUSION (% of BaL) Env +ve 293T cells (%) INFECTIVITY (Log TTCID5/mL) INFECTIVITY (Cell-free virus infection) env psg T cells Light 2 TZM-bl cells titer 48 h 5 r =.361 p= r =,4728 p=, INFECTIVITY (Log TCID 5 /ml) INFECTIVITY (Log TCID 5 /ml) Env clones from and show similar infectivity.

8 MULTIVARIATE ANALYSIS OF INFECTIVITY INFECTIVITY vs ESTIMATE SD Z-score P-value Expression ** Fusogenicity Group ( or ) Infectivity of Env clones is related to expression but not to fusogenicity or their origin ( or ).

9 EXPRESSION (% of Env+ve cells) HIV TRANSFER (p24+ cells, %) CD4 BINDING AND SIGNALING CAPACITY Blanco et al. High level of coreceptor-independent HIV transfer induced by contacts between primary CD4 T cells.. J Biol Chem 279: , env T cells 1 psg3 + Primary CD4 T cells 2 h Valenzuela et al. HDAC 6 regulates HIV-1 infection. Mol Biol Cell 16:11, h WB CEM.NKR-CCR5 cells CD4 binding and signaling are similar in and.

10 CYTOPATHICITY (Autophagy induction) Autophagosomes identified by puncta using anti LC3 antibody. No information on primary isolates or CCR5-using viruses

11 LC3 puncta/cell AUTOPHAGY (LC3 puncta/cell) CYTOPATHICITY (Autophagy induction) env 293T cells + Analysis of autophagy by endogenous LC3 labelling HUT CCR5+ T cells NK RIS h r =.731 p = FUSOGENICITY (%) Env isolated from are cytophatic Cytopathicity is associated with fusogenic activity.

12 CONCUSIONS Small differences in fusogenicity hardly justify the phenotype Infectivity, CD4 binding and signaling are similar in both groups. Cytopathicity as measured by autophagy induction is similar in both groups. (First description of autophagy induction by primary Env) IMMUNOLOGICAL FACTORS??

13 THANKS Silvia Marfil Victor Urrea Elisabet García Jorge Carrillo Marta Curriu Bonaventura Clotet Julià Blanco Sara Marrero-Hernandez Daniel Márquez-Arce Romina Cabrera-Rodríguez Maria Soledad Varela Agustín Valenzuela Concha Casado María Pernas Cecilio López-G. Veronique Hebman Martine Biard-P. Elisabet Gómez Cecilia Cabrera

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