title: Structural characteristics correlate with immune responses induced by HIV envelope glycoprotein vaccines.
abstract: HIV envelope glycoprotein (Env) is the target for inducing neutralizing antibodies. Env is present on the virus surface as a trimer, and, upon binding to CD4, a cascade of events leads to structural rearrangement exposing the co-receptor binding site and entry into the CD4+ host target cells. We have designed monomeric and trimeric Env constructs with and without deletion of the variable loop 2 (ΔV2) from SF162, a subtype B primary isolate, and performed biophysical, biochemical and immunological studies to establish a potential structure–functional relationship. We expressed these Envs in CHO cells, purified the proteins to homogeneity and performed biophysical studies to define the binding properties to CD4, structural characteristics and exposure of epitopes recognized by b12 and CD4i mAb (17B) on both full-length and mutant HIV Env proteins. Parameters evaluated include oligomerization state, number and affinity of CD4 binding sites, enthalpy and entropy of the Env–CD4 interaction and affinity for b12 and 17b mAbs. We observed one CD4 binding site per monomer and three active CD4 binding sites per trimer. A40-fold difference in affinity of the gp120 monomer vs. the o-gp140 trimer towards CD4 was observed (Kd = 58 nM and 1.5 nM, respectively),whereas only a 2-fold difference was observed for the V2 deleted Envs (Kd of gp120ΔV2 = 19 nM, Kd of o-gp140DV2 = 9.3 nM). Monomers had 3-fold higher affinity to the mAb 17b and at least 3-fold weaker affinity to b12 compared to trimers, with gp120DV2 having the weakest affinity for b12 (Kd = 446 nM). Affinity of CD4 binding correlated with proportion of the antibodies induced against the conformational epitopes by the corresponding Envs, and changes in mAb binding correlated with the induction of antibodies directed against linear epitopes. Furthermore,biophysical analysis reveals that the V2 deletion has broad structural implications in the monomer not shared by the trimer, and these changes are reflected in the quality of the immune responses induced in rabbits. These data suggest that biophysical characteristics of HIV Env, such as affinity for CD4, and exposure of important neutralizing epitopes, such as those recognized by b12 mAb, may be important predictors of its in vivo efficacy and may serve as important surrogate markers for screening Env structures as potential vaccine candidates.
authors: Chiron Vaccines, Immunology and Cell Biology, 4560 Horton Street, Emeryville, CA 94608, USA. victoria_sharma@chiron.com
authors: Victoria A
authors: VA
authors: Sharma
authors: Elaine
authors: E
authors: Kan
authors: Yide
authors: Y
authors: Sun
authors: Ying
authors: Y
authors: Lian
authors: Jimna
authors: J
authors: Cisto
authors: Verna
authors: V
authors: Frasca
authors: Susan
authors: S
authors: Hilt
authors: Leonidas
authors: L
authors: Stamatatos
authors: John J
authors: JJ
authors: Donnelly
authors: Jeffrey B
authors: JB
authors: Ulmer
authors: Susan W
authors: SW
authors: Barnett
authors: Indresh K
authors: IK
authors: Srivastava
mesh_terms: D016915:AIDS Vaccines / Q000737:chemistry / Q000235:genetics / Q000276:immunology; D000818:Animals; D057134:Antibodies, Neutralizing / Q000276:immunology; D000915:Antibody Affinity; D001665:Binding Sites; D015704:CD4 Antigens / Q000737:chemistry / Q000235:genetics; D016466:CHO Cells; D006224:Cricetinae; D015483:HIV Antibodies / Q000276:immunology; D015658:HIV Infections / Q000276:immunology / Q000517:prevention & control; D006801:Humans; D016056:Immunodominant Epitopes / Q000737:chemistry / Q000235:genetics / Q000276:immunology; D011817:Rabbits; D013329:Structure-Activity Relationship; D054299:env Gene Products, Human Immunodeficiency Virus / Q000737:chemistry* / Q000235:genetics / Q000276:immunology*
chemical_list: D016915:AIDS Vaccines; D057134:Antibodies, Neutralizing; D015704:CD4 Antigens; D015483:HIV Antibodies; D016056:Immunodominant Epitopes; D054299:env Gene Products, Human Immunodeficiency Virus