The binding affinity of double‐stranded RNA motifs to HIV‐1 Tat protein affects transactivation and the neutralizing capacity of anti‐Tat antibodies elicited after intranasal immunization

• Published in: European journal of immunology 2005-05, Vol.35 (5), p.1521-1529
• Main Authors: Partidos, Charalambos D., Hoebeke, Johan, Moreau, Emmanuel, Chaloin, Olivier, Tunis, Mélanie, Belliard, Guillaume, Briand, Jean‐Paul, Desgranges, Claude, Muller, Sylviane
• Format: Article
• Language: English
• Subjects: Adjuvants; Administration, Intranasal; Animals; Antibody Specificity - immunology; dsRNA motifs; Enzyme-Linked Immunosorbent Assay; Female; Gene Products, tat - immunology; Gene Products, tat - metabolism; HIV-1 - immunology; HIV-1 - metabolism; HIV‐1 Tat; Human immunodeficiency virus 1; Interferon-gamma - immunology; Interleukin-2 - immunology; Mice; Mice, Inbred BALB C; Mucosal immunization; RNA, Double-Stranded - administration & dosage; RNA, Double-Stranded - immunology; RNA, Double-Stranded - metabolism; RNA, Viral - administration & dosage; RNA, Viral - immunology; RNA, Viral - metabolism; Surface Plasmon Resonance; T-Lymphocytes - immunology; tat Gene Products, Human Immunodeficiency Virus; Tat‐TAR interaction; Transcriptional Activation
• ISSN: 0014-2980; 1521-4141
• DOI: 10.1002/eji.200425676

In this study we examined the hypothesis that the binding affinity of two double‐stranded (ds) RNA motifs to HIV‐1 Tat protein might affect transactivation and the type of anti‐Tat immune responses. Using surface plasmon resonance technology we demonstrated the capacity of the poly(A):poly(U) (pA:pU) motif to bind with high affinity to a totally synthetic Tat protein and to inhibit more efficiently the Tat/transactivation response element (TAR) RNA interaction as compared to the poly(I):poly(C) (pI:pC) motif. Furthermore, the pA:pU motif was tenfold more effective in inhibiting Tat‐driven transactivation than the pI:pC motif. Following intranasal immunization of mice, both dsRNA motifs enhanced the antibody (serum and mucosal) and cellular responses to Tat. However, only the serum samples of mice immunized with Tat + pI:pC inhibited Tat‐driven transactivation. The profile of serum antibody subclasses together with the secreted cytokines by Tat‐stimulated splenocyte cultures indicated that both dsRNA motifs favored the induction of a balanced Th1 and Th2 immune response. The demonstration in this study that two dsRNA motifs had a marked effect on Tat/TAR RNA interaction and on the neutralizing capacity of anti‐Tat specific antibody responses highlights their potential for biological applications and the importance of selecting the appropriate motif as an adjuvant for vaccine design.