Production and of monoclonal antibodies to simian immunodeficiency virus envelope glycoproteins

Molecular and biological characterization of simian immunodeficiency virus macaque strain 32H proviral clones containing nef size variants

“…The denatured proteins were separated on 12.5 % polyacrylamide gels and blotted onto nitrocellulose membranes. After blocking of non-specific binding with 5 % skimmed milk (Marvel), the membranes were incubated with a 1 : 100 dilution of a SIVmac251 transmembrane glycoprotein-specific mouse monoclonal antibody, KK7 (Kent et al, 1991), or SIVmac32H-positive macaque serum. 125I-labelled protein A was used to detect bound antibody.…”

“…As simian immunodeficiency virus (SIV) is the closest relative of HIV-1 and HIV-2 (Hirsch et al, 1987;Franchini et al, 1987;Chakrabarti et al, 1987;Guyader et al, 1987), the rhesus macaque animal model has been used in many of the vaccine trials to date (Carlson et at., 1990;Desrosiers et at., I989;Johnson et aL, 1992;Marthas et al, 1990;Murphey-Corb et al, t989;Cranage IP: 54.202.233.140 On: Sun, 13 May 2018 06:24:58 530 E. W. Rud and others et al, 1992). Assessment of the protection induced in these studies was complicated because both the inactivated whole virus vaccines and challenge virus were produced on human cells and that even formalin-fixed uninfected human cells can protect a proportion of vaccines under these conditions (Stott et al, 1991). The anti-human cell antibodies from the protected monkeys seem to correlate with protection (Langlois et al, 1992).…”

Stimulation of Specific Immune Responses to Simian Immunodeficiency Virus Using Chimeric Hepatitis B Core Antigen Particles

“…After a further 1 h incubation at 37 °C, the plates were washed and developed with an enzyme substrate as previously described . Reactivity of the mouse anti-SIV V2 monoclonal antibody (MAb) KK13 (Kent et al, 1991(Kent et al, , 1992 with V2 peptides expressed in CFPs was assayed by a modified double-antibody sandwich ELISA (Voller et al, 1976). Guinea-pig polyclonal antiHBcAg serum (100-fold dilution) was used to capture serial dilutions (starting from 100 Ixg/ml) of CFPs in PBS/2% dried milk powder.…”

Comparison of serum antibody reactivities to a conformational and to linear antigenic sites in the external envelope glycoprotein of simian immunodeficiency virus (SIVmac) induced by infection and vaccination

“…Plates were blocked by incubation with 50 lal/well PBS containing 0.5 % (v/v) Tween 20 (PBST) and 10 % (v/v) fetal calf serum (blocking buffer A) for 90 min. The plates were washed five times with PBST and each well was incubated for 90 min with 50 gl of a solution consisting of biotinylated monoclonal antibody KK17 (Kent et al, 1991) and a 1/200 dilution of monkey serum in blocking buffer A. In initial experiments, serial dilutions of monkey sera were mixed with labelled KK17 and endpoint titres were determined.…”

“…This report describes antibody responses to the SIVma e external glycoprotein measured in rhesus macaques infected with either the uncloned SIVma c 32H isolate or the J5 molecular clone of SIVma c 32H (Rud et al, 1992) and in animals vaccinated with formalin-inactivated SIV .... 32H using alum or syntex adjuvant formulation (saf-1) adjuvants. Reactivity to a non-continuous virus neutralization epitope was measured by competitive binding assay using a monoclonal antibody with specificity for the SIVma e envelope (Kent et al, 1991. The response to linear antigenic determinants of the SIV .... external glycoprotein was measured by binding to a series of 20-mer overlapping peptides.…”